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Sample records for babesia equi trophozoites

  1. Real-time PCR for detection of Theileria equi and Babesia caballi ...

    African Journals Online (AJOL)

    Real-time PCR for detection of Theileria equi and Babesia caballi parasites in ticks. ... This study aimed to develop a real-time PCR screening test for Babesia caballi and Theileria equi in ticks. Adult D. reticulatus were ... This test is suitable for application in epidemiological surveillance of equine babesiosis and theileriosis.

  2. Differential expression of three members of the multidomain adhesion CCp family in Babesia bigemina, Babesia bovis and Theileria equi.

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    Reginaldo G Bastos

    Full Text Available Members of the CCp protein family have been previously described to be expressed on gametocytes of apicomplexan Plasmodium parasites. Knocking out Plasmodium CCp genes blocks the development of the parasite in the mosquito vector, making the CCp proteins potential targets for the development of a transmission-blocking vaccine. Apicomplexans Babesia bovis and Babesia bigemina are the causative agents of bovine babesiosis, and apicomplexan Theileria equi causes equine piroplasmosis. Bovine babesiosis and equine piroplasmosis are the most economically important parasite diseases that affect worldwide cattle and equine industries, respectively. The recent sequencing of the B. bovis and T. equi genomes has provided the opportunity to identify novel genes involved in parasite biology. Here we characterize three members of the CCp family, named CCp1, CCp2 and CCp3, in B. bigemina, B. bovis and T. equi. Using B. bigemina as an in vitro model, expression of all three CCp genes and proteins was demonstrated in temperature-induced sexual stages. Transcripts for all three CCp genes were found in vivo in blood stages of T. equi, and transcripts for CCp3 were detected in vivo in blood stages of B. bovis. However, no protein expression was detected in T. equi blood stages or B. bovis blood stages or B. bovis tick stages. Collectively, the data demonstrated a differential pattern of expression of three orthologous genes of the multidomain adhesion CCp family by B. bigemina, B. bovis and T. equi. The novel CCp members represent potential targets for innovative approaches to control bovine babesiosis and equine piroplasmosis.

  3. Assessment of theileria equi and babesia caballi infections in equine populations in Egypt by molecular, serological and hematological approaches

    Science.gov (United States)

    Background: Equine piroplasmosis caused by Theileria equi, Babesia caballi, or both, cause significant economic losses in the equine industry and remains uncontrolled in Egypt. Methods: T. equi and B. caballi infections were assessed in blood from 88 horses and 51 donkeys from different localities ...

  4. Seroprevalence of Theileria equi and Babesia caballi in horses in Spain

    OpenAIRE

    Montes Cort?s, Maria Guadalupe; Fern?ndez-Garc?a, Jos? Luis; Habela Mart?nez-Est?llez, Miguel ?ngel

    2017-01-01

    Equine piroplasmoses are enzootic parasitic diseases distributed worldwide with high incidence in tropical and subtropical regions. In Spain, there is insufficient epidemiological data about equine piroplasmoses. The main aim of the present study was therefore to estimate the prevalence of Theileria equi and Babesia caballi in five regions and obtain information about the risk factors. This study was conducted in the central and south-western regions of Spain, using indirect fluorescence anti...

  5. [A Duplex PCR Method for Detection of Babesia caballi and Theileria equi].

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    Zhang, Yang; Zhang, Yu-ting; Wang, Zhen-bao; Bolati; Li, Hai; Bayinchahan

    2015-04-01

    To develop a duplex PCR assay for detection of Babesia caballi and Theileria equi. Two pairs of primers were designed according to the BC48 gene of B. caballi and 18 s rRNA gene of T. equi, and a duplex PCR assay was developed by the optimization of reaction conditions. The specificity, sensitivity and reliability of the method were tested. The horse blood samples of suspected cases were collected from Yili region, and detected by the duplex PCR, microspopy, conventional PCR, and fluorescence quantitative PCR, and the results were compared. Using the duplex PCR assay, the specific fragments of 155 bp and 280 bp were amplified from DNA samples of B. caballi and T. equi, respectively. No specific fragment was amplified from DNA samples of B. bigemina, Theilerdia annulata, Theilerdia sergenti, Toxoplasma gondii, Neospora caninum, and Trypanosoma evansi. The limit of detection was 4.85 x 10(5) copies/L for B. caballi DNA and 4.85 x 10(4) copies/µl for T. equi DNA, respectively. Among the 24 blood samples, 11 were found B. caballi-positive by the duplex PCR assay, and 18 were T. equi-positive. The coincidence rate of microscopy, conventional PCR, and fluorescence quantitative PCR with duplex PCR was 91.7% (22/24), 95.8% (23/24), and 95.8% (23/24), respectively. A duplex PCR assay for simultaneous detection of B. caballi and T. equi is established.

  6. Prevalance of Theileria equi and Babesia caballi in Donkeys from Eastern Turkey in Winter Season

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    I. Balkaya*, A. E. Utuk1 and F. C. Piskin1

    2010-10-01

    Full Text Available The aim of this study was to determine the prevalence of Theileria equi and Babesia caballi by the examination of Giemsa-stained blood smears and c-ELISA method in donkeys, selected randomly from Erzurum, the largest province of Eastern Turkey. The specimens were consisted of 92 thin blood-smears and 75 sera during winter season. As result of microscopic examination no parasite was detected. Of the 75 sera, 3 (4% and 1 (1.33% samples were positive for the presence of T. equi and B. caballi antibodies, respectively. We couldn’t detect mix infection with both parasites. This study indicated the prevalence of T. equi and B. caballi in donkeys for the first time from Eastern Turkey in winter season.

  7. Microscopic and Molecular Detection of Theileria (Babesia) Equi Infection in Equids of Kurdistan Province, Iran.

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    Habibi, Gholamreza; Esmaeilnia, Kasra; Hablolvarid, Mohammad Hasan; Afshari, Asghar; Zamen, Mohsen; Bozorgi, Soghra

    2016-01-01

    Equine piroplasmosis (EP) is the cause of persistent tick-borne infection with no symptoms, but the most important problem of EP is due to the persistent carrier state. Carrier animals to Babesia (Theileria) equi (Laveran 1901) and B. caballi (Nuttall, 1910) infestation could be identified by extremely sensitive PCR-based method. The purpose of this study was to identify the causative agents of equine piroplasmosis based on molecular and microscopic assays in equids from Kurdistan Province, Iran. Thirty one horse and mule blood samples were used with history of living in Kurdistan Province of Iran. The blood specimens were utilized for T. equi and B. caballi DNA identification by PCR and Giemsa stained smears for microscopic observation. The results clearly showed the presence of B. (Theileria) equi DNA in 30 of 31 blood samples (96.77%), but the microscopic examination revealed the 3 of 31 positive Babesia like organisms in the red blood cells (9.67%). The obtained results demonstrated the presence of hidden B. (Theileria) equi infection in horses with previous habitance in Kurdistan Province of Iran. The carrier animals became a main source of infection and can transmit the disease. Therefore, hidden infection might be considered as a health threatening and limiting factor in animals used in therapeutic antisera research and production centers.

  8. Seroprevalence of Babesia caballi and Theileria equi in five draught equine populated metropolises of Punjab, Pakistan.

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    Hussain, Muhammad Hammad; Saqib, Muhammad; Raza, Fahad; Muhammad, Ghulam; Asi, Muhammad Nadeem; Mansoor, Muhammad Khalid; Saleem, Muhammad; Jabbar, Abdul

    2014-05-28

    Equine piroplasmosis (EP) caused by intraerythrocytic parasites (Theileria equi and Babesia caballi) is an emerging equine disease of world-wide distribution. In Pakistan, the prevalence and incidence of EP are unknown. In order to obtain the first insights into the prevalence of the disease, a total of 430 equids, including 33 mules, 65 horses and 332 donkeys, aging from ≤ 5 to ≥ 10 years of either sex, from five metropolises of Punjab, Pakistan, were serologically tested for the presence of antibodies directed against B. caballi and T. equi, using a competitive enzyme-linked immunosorbent assay (cELISA). Out of 430 equid serum samples tested, 226 (52.6%, 95% CI 47.7-57.4) were found cELISA positive for EP (T. equi and/or B. caballi infections). The overall seroprevalence of EP was 41.2% (95% CI 36.5-46.0) for T. equi and 21.6% (95% CI 17.8-25.8) for B. caballi. A small proportion of equids (10.2%, 95% CI 7.5-13.5) was seropositive for both T. equi and B. caballi. Seroprevalence of T. equi was significantly higher (P<0.01) in equines from the metropolis of Lahore (66.7%, 95% CI 54.3-77.6) and in horses (56.9%, 95% CI 44.0-69.2). Multivariable logistic regression model analysis indicated that factors associated with prevalence of EP were being an equine species kept in metropolis Lahore (OR=4.24, 95% CI 2.28-7.90), horse (OR=2.82, 95% CI 1.53-5.20) and male equids (OR=1.81, 95% CI 1.15-2.86). Copyright © 2014 Elsevier B.V. All rights reserved.

  9. Seroprevalence of Theileria equi and Babesia caballi in horses in Spain

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    Montes Cortés Maria Guadalupe

    2017-01-01

    Full Text Available Equine piroplasmoses are enzootic parasitic diseases distributed worldwide with high incidence in tropical and subtropical regions. In Spain, there is insufficient epidemiological data about equine piroplasmoses. The main aim of the present study was therefore to estimate the prevalence of Theileria equi and Babesia caballi in five regions and obtain information about the risk factors. This study was conducted in the central and south-western regions of Spain, using indirect fluorescence antibody testing (IFAT in 3,100 sera samples from apparently healthy horses of different ages, breeds, coat colours, genders and geographical locations. The overall seroprevalence was 52%, consisting of 44% seropositive for T. equi and 21% for B. caballi. There was a significant association between age (p < 0.0001, breed (p < 0.004, geographical location (p < 0.0001 and the seroprevalence, but neither the coat colour nor the gender was significantly associated with prevalence. In addition, it was proved that most of the geographic areas showed a moderate to high prevalence. The statistical κ value was used to compare the results obtained by the IFAT and the competitive enzyme-linked immunosorbent assay (cELISA utilised to test some samples (n = 108 and showed a higher concordance for T. equi (κ = 0.68 than for B. caballi (κ = 0.22. Consequently, this revealed the importance of developing an appropriate technique to detect each haemoparasite.

  10. Clinical investigation on Theileria equi and Babesia caballi infections in Italian donkeys.

    Science.gov (United States)

    Laus, Fulvio; Spaterna, Andrea; Faillace, Vanessa; Veronesi, Fabrizia; Ravagnan, Silvia; Beribé, Francesca; Cerquetella, Matteo; Meligrana, Marina; Tesei, Beniamino

    2015-04-28

    Interest in the welfare and diseases of donkeys is constantly increasing in several countries. Despite this, clinical research into donkeys needs to be in continual development since they show different reactions compared to horses in many conditions, including infectious diseases, and need specific clinical and therapeutic approaches. No reports are currently available on clinical and clinical pathology data regarding donkeys with natural piroplasms infection. Venous blood samples were taken from one hundred and thirty eight donkeys and underwent indirect fluorescent antibody test (IFAT) to detect IgG antibodies against Theileria equi and Babesia caballi and real-time polimerase chain reaction (PCR) to detect Babesia spp. and Theileria spp. Clinical examinations, haematological analyses and serum bilirubin evaluation were also performed and compared with positive or negative status. A seroprevalence of 40.6% and 47.8% was found for T. equi and B. caballi, respectively; double positivity was detected in 19.6% of the animals. PCR results showed that 17.4% of the animals tested positive for T.equi and 3.6% for B. caballi with no double positivity. Twelve donkeys (8.7%) had clinical signs consistent with chronic forms of the disease and no acute forms were detected. Fifty-eight donkeys had haematological and serum bilirubin alterations and 56 (96.6%) of them were IFAT and/or PCR positive. Changes in erythrocyte number, packed cell volume, hemoglobin concentration, mean corpuscular hemoglobin, platelets number and total bilirubin were significantly associated with positive and symptomatic animals. Nonspecific clinical presentation seems to be very common in donkeys and several clinical pathology alterations persist after natural infection. Therefore, apparently healthy donkeys can have masked but severe clinical pathology alterations. Acute forms are very seldom observed in donkeys. Clinical monitoring of chronically infected donkeys is recommended since such animals

  11. Detection of Theileria equi and Babesia caballi using microscopic and molecular methods in horses in suburb of Urmia, Iran

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    Farnaz Malekifard

    2014-06-01

    Full Text Available Equine piroplasmosis is a severe disease of horses caused by the intra-erythrocyte protozoan, Theileria equi and Babesia caballi. The aim of this study was to identify equine piroplasmosis based on molecular and morphometrical features in horses in suburb of Urmia, West Azerbaijan province, Iran. From April to September 2011, a total number of 240 blood samples were collected randomly from horses of 25 villages. The specimens were transferred to the laboratory and the blood smears stained with Geimsa, and the morphological and biometrical data of parasite in any infected erythrocyte were considered. Extracted DNA from each blood sample was used in multiplex PCR in order to confirm the presence of B. caballi and T. equi. Microscopic observation on 240 blood smears determined that 15 (6.25% and 5 (2.80% samples were infected by T. equi and B. caballi, respectively. The mixed infections occurred in 2 (0.83% samples. The results of the PCR assays showed 26 (10.83%, 14 (5.83% and 4 (1.66% were distinguished as T. equi, B. caballi and mixed infection, respectively. Differences in infection rates were statistically nonsignificant between male and female horses and among different age groups. Our findings indicated that T. equi and B. caballi were prevalent in horse population.

  12. Comparative Bioinformatics Analysis of Transcription Factor Genes Indicates Conservation of Key Regulatory Domains among Babesia bovis, Babesia microti, and Theileria equi.

    Science.gov (United States)

    Alzan, Heba F; Knowles, Donald P; Suarez, Carlos E

    2016-11-01

    Apicomplexa tick-borne hemoparasites, including Babesia bovis, Babesia microti, and Theileria equi are responsible for bovine and human babesiosis and equine theileriosis, respectively. These parasites of vast medical, epidemiological, and economic impact have complex life cycles in their vertebrate and tick hosts. Large gaps in knowledge concerning the mechanisms used by these parasites for gene regulation remain. Regulatory genes coding for DNA binding proteins such as members of the Api-AP2, HMG, and Myb families are known to play crucial roles as transcription factors. Although the repertoire of Api-AP2 has been defined and a HMG gene was previously identified in the B. bovis genome, these regulatory genes have not been described in detail in B. microti and T. equi. In this study, comparative bioinformatics was used to: (i) identify and map genes encoding for these transcription factors among three parasites' genomes; (ii) identify a previously unreported HMG gene in B. microti; (iii) define a repertoire of eight conserved Myb genes; and (iv) identify AP2 correlates among B. bovis and the better-studied Plasmodium parasites. Searching the available transcriptome of B. bovis defined patterns of transcription of these three gene families in B. bovis erythrocyte stage parasites. Sequence comparisons show conservation of functional domains and general architecture in the AP2, Myb, and HMG proteins, which may be significant for the regulation of common critical parasite life cycle transitions in B. bovis, B. microti, and T. equi. A detailed understanding of the role of gene families encoding DNA binding proteins will provide new tools for unraveling regulatory mechanisms involved in B. bovis, B. microti, and T. equi life cycles and environmental adaptive responses and potentially contributes to the development of novel convergent strategies for improved control of babesiosis and equine piroplasmosis.

  13. Comparative Bioinformatics Analysis of Transcription Factor Genes Indicates Conservation of Key Regulatory Domains among Babesia bovis, Babesia microti, and Theileria equi.

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    Heba F Alzan

    2016-11-01

    Full Text Available Apicomplexa tick-borne hemoparasites, including Babesia bovis, Babesia microti, and Theileria equi are responsible for bovine and human babesiosis and equine theileriosis, respectively. These parasites of vast medical, epidemiological, and economic impact have complex life cycles in their vertebrate and tick hosts. Large gaps in knowledge concerning the mechanisms used by these parasites for gene regulation remain. Regulatory genes coding for DNA binding proteins such as members of the Api-AP2, HMG, and Myb families are known to play crucial roles as transcription factors. Although the repertoire of Api-AP2 has been defined and a HMG gene was previously identified in the B. bovis genome, these regulatory genes have not been described in detail in B. microti and T. equi. In this study, comparative bioinformatics was used to: (i identify and map genes encoding for these transcription factors among three parasites' genomes; (ii identify a previously unreported HMG gene in B. microti; (iii define a repertoire of eight conserved Myb genes; and (iv identify AP2 correlates among B. bovis and the better-studied Plasmodium parasites. Searching the available transcriptome of B. bovis defined patterns of transcription of these three gene families in B. bovis erythrocyte stage parasites. Sequence comparisons show conservation of functional domains and general architecture in the AP2, Myb, and HMG proteins, which may be significant for the regulation of common critical parasite life cycle transitions in B. bovis, B. microti, and T. equi. A detailed understanding of the role of gene families encoding DNA binding proteins will provide new tools for unraveling regulatory mechanisms involved in B. bovis, B. microti, and T. equi life cycles and environmental adaptive responses and potentially contributes to the development of novel convergent strategies for improved control of babesiosis and equine piroplasmosis.

  14. Detection of Babesia caballi and Theileria equi in Blood from Equines from Four Indigenous Communities in Costa Rica

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    María Fernanda Posada-Guzmán

    2015-01-01

    Full Text Available A cross-sectional study was carried out in four indigenous communities of Costa Rica to detect presence and prevalence of Babesia caballi and Theileria equi and to investigate factors associated with presence of these hemoparasites. General condition of horses (n=285 was evaluated, and hematocrits and hemoglobin were determined from blood samples of 130 horses, which were also analyzed using blood smears, nested polymerase chain reaction (PCR, and immunosorbent assay (c-ELISA. The general condition of the horses (n=285 in terms of their body and coat was between regular and poor, and hematocrit and hemoglobin average values were low (19% and 10.65 g/dL, resp.. Erythrocyte inclusions were observed in 32 (24.6% of the samples. Twenty-six samples (20.0% gave positive results for B. caballi and 60 (46.2% for T. equi; 10 horses (7.7% showed mixed infection, when analyzed by PCR. Using c-ELISA, it was found that 90 (69.2% horses had antibodies against B. caballi and 115 (88.5% against T. equi, while 81 (62.3% showed mixed reactions. There were no factors associated with the presence of B. caballi and T. equi. These results contrast with results previously obtained in equines in the Central Valley of Costa Rica.

  15. Detection of Babesia caballi and Theileria equi in Blood from Equines from Four Indigenous Communities in Costa Rica.

    Science.gov (United States)

    Posada-Guzmán, María Fernanda; Dolz, Gaby; Romero-Zúñiga, Juan José; Jiménez-Rocha, Ana Eugenia

    2015-01-01

    A cross-sectional study was carried out in four indigenous communities of Costa Rica to detect presence and prevalence of Babesia caballi and Theileria equi and to investigate factors associated with presence of these hemoparasites. General condition of horses (n = 285) was evaluated, and hematocrits and hemoglobin were determined from blood samples of 130 horses, which were also analyzed using blood smears, nested polymerase chain reaction (PCR), and immunosorbent assay (c-ELISA). The general condition of the horses (n = 285) in terms of their body and coat was between regular and poor, and hematocrit and hemoglobin average values were low (19% and 10.65 g/dL, resp.). Erythrocyte inclusions were observed in 32 (24.6%) of the samples. Twenty-six samples (20.0%) gave positive results for B. caballi and 60 (46.2%) for T. equi; 10 horses (7.7%) showed mixed infection, when analyzed by PCR. Using c-ELISA, it was found that 90 (69.2%) horses had antibodies against B. caballi and 115 (88.5%) against T. equi, while 81 (62.3%) showed mixed reactions. There were no factors associated with the presence of B. caballi and T. equi. These results contrast with results previously obtained in equines in the Central Valley of Costa Rica.

  16. Seroprevalence and risk factors associated with Babesia caballi and Theileria equi infections in donkeys from Southern Italy.

    Science.gov (United States)

    Piantedosi, D; D'Alessio, N; Di Loria, A; Di Prisco, F; Mariani, U; Neola, B; Santoro, M; Montagnaro, S; Capelli, G; Veneziano, V

    2014-12-01

    Equine piroplasmosis (EP) has been frequently described in donkeys in subtropical and tropical regions, but published data reflecting large scale surveys are very limited in Europe. The seroprevalence of Babesia caballi and Theileria equi was determined in a donkey population from Campania Region in Southern Italy using a commercial indirect fluorescent antibody test (IFAT), and the risk factors associated with the occurrence of the infection were assessed. Of 203 samples, the overall seroprevalence for EP was 57.1% (116/203), with 35.5% (72/203) for B. caballi and 44.3% (90/203) for T. equi. Co-infection was detected in 46 donkeys (22.6%). The distribution of IFAT antibody titres to B. caballi was: 1:80 (n= 67), 1:160 (n= 2), 1:320 (n= 3); while the distribution of IFAT antibody titres to T. equi was: 1:80 (n= 25), 1:160 (n= 42), 1:320 (n= 12), 1:640 (n= 8), 1:1280 (n= 3). All examined donkeys were asymptomatic, except one adult male (with a titre of 1:640 against T. equi) that showed clinical signs corresponding to the acute stage of EP, reported for the first time in Italy. The unique risk factor associated with a higher B. caballi seroprevalence was the presence of horses in the farms, while risk factors associated with a higher T. equi seroprevalence were poor body condition, presence of ruminants in the farms and milk production. The results indicate a high level of exposure in donkeys living in Southern Italy and suggest that donkeys may be an important reservoir of EP. Copyright © 2014 Elsevier Ltd. All rights reserved.

  17. A field survey for the seroprevalence of Theileria equi and Babesia caballi in donkeys from Nuu Division, Kenya.

    Science.gov (United States)

    Oduori, David O; Onyango, Solomon C; Kimari, Joseph N; MacLeod, Ewan T

    2015-07-01

    Equine piroplasmosis is one of the most significant tick-borne disease of equids. The prevalence of this disease in donkeys of semi-arid Kenya remains largely unexplored. The primary objective of this study was to demonstrate the extent to which donkeys in Nuu division, Kenya have been exposed to the haemoprotozoans Babesia caballi and Theileria equi, the causative agents of equine piroplasmosis. The study also assessed the effect of age and sex on seroprevalence. A stratified sampling approach was used and three hundred and fourteen donkeys were sampled across nine sub-locations in Nuu division, Mwingi district. Serodiagnosis was via competitive inhibition enzyme linked immunosorbent assays (cELISA). The seroprevalence of T. equi was 81.2% (95% CI: 76.4-85.4). There was no significant difference in sub-location seropositivity, gender seropositivity or age related seropositivity. Antibodies against B. caballi were not detected (95% CI: 0-1.2). Findings from this study suggest that T. equi infection is endemic in Nuu division, Mwingi where it exists in a state of endemic stability. Existence of the infection should be communicated to animal health practitioners and donkey owning communities in the area. Copyright © 2015 Elsevier GmbH. All rights reserved.

  18. Molecular and serological detection of Theileria equi, Babesia caballi and Anaplasma phagocytophilum in horses and ticks in Maranhão, Brazil

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    Rita de Maria Seabra Nogueira

    Full Text Available ABSTRACT: Equine piroplasmosis is a tick-borne disease caused by the intraeytrhocytic protozoans Babesia caballi and Theileria equi. It has been reported as a main equine parasitic disease. In addition, Anaplasma phagocytophilum, the causative agent of granulocytic ehrlichiosis, causes a seasonal disease in horses. Both diseases, can be detrimental to animal health. In this sense, blood samples and ticks were collected from 97 horses raised in the microregion of Baixada Maranhense, Maranhão State, Brazil. Serum samples were subjected to Indirect Fluorescence Antibody Test (IFAT and blood samples and ticks to Polymerase Chain Reaction (PCR to evaluate the infection by Theileria equi, Babesia caballi and Anaplasma phagocytophilum. The overall seroprevalence was 38.14%, 18.55% and 11.34% for T. equi, B. caballi and A. phagocytophilum, respectively. The results of PCR from blood samples showed 13.40% and 3.09% positive samples to T. equi and B. caballi, respectively. A total of 170 tick specimens were collected and identified as Dermacentor nitens, Amblyomma cajennense sensu lato and Rhipicephalus (Boophilus microplus. It was detected 2.35% (4/170 and 0.59% (1/170 positive tick samples by PCR for T. equi and B. caballi, respectively. All samples were negative to A. phagocytophilum. No statically difference (p>0.05 was observed when gender, age, use of ectoparasiticide and tick presence were analyzed. A BLASTn analysis of the sequenced samples indicated 97 to 100% similarity with T. equi 18S rRNA gene sequences in GenBank and 98 to 100% with B. caballi. Genetic analysis classified the obtained sequences as T. equi and B. caballi cluster, respectively. It can be concluded that these pathogens occur and are circulating in the studied area.

  19. An enzyme-linked immunosorbent assay for the detection of IgG antibodies against Babesia equi in horses Um ensaio imunoenzimático para a detecção de anticorpos IgG contra Babesia equi em eqüinos

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    Cristiane Divan Baldani

    2004-10-01

    Full Text Available A crude antigenic preparation of Babesia equi was used to develop and establish the suitability of an enzyme-linked immunosorbent assay (ELISA for the detection of parasite carriers. Optimal dilutions of the antigen, using positive and negative reference sera, were determined by checkboard titrations. The specificity and sensitivity of the ELISA were 100 %. A total of 90 serum samples were taken from horses from the Northeast region of São Paulo State and examined for diagnosis of equine B. equi infection by ELISA. Approximately 75% (n=67 of all the horses tested were found serologically positive for B. equi. These results suggest that the ELISA described may prove to be an appropriate serological test for epidemiological studies on B. equi infections in the field and that equine piroplasmosis is a cause for serious concern in the State of São Paulo, Brazil.Um ensaio de imunoadsorção enzimática (ELISA baseado em antígeno bruto de Babesia equi foi desenvolvido para a detecção de portadores crônicos de babesiose eqüina. As diluições ótimas do antígeno e dos soros controles positivo e negativo foram determinadas através de titulação em bloco. A sensibilidade e especificidade do teste foram de 100%. O ELISA foi empregado em 90 soros de eqüinos da região Nordeste do Estado de São Paulo, de modo que aproximadamente 75% (n=67 dos eqüinos foram positivos para B. equi. Estes resultados sugerem que o ELISA descrito pode ser utilizado no diagnóstico sorológico de B. equi e que a piroplasmose eqüina é um problema sério no Estado de São Paulo, Brasil.

  20. Comparative bioinformatics analysis of transcription factor genes indicates conservation of key regulatory domains among babesia bovis, babesia microti and theileria equi.

    Science.gov (United States)

    Apicomplexa tick borne hemoparasites including B. bovis, B. microti, and Theileria equi are responsible for bovine and human babesiosis and equine theileriosis respectively. These neglected parasites of vast medical, epidemiological, and economic impact have complex life cycles in their vertebrate a...

  1. Quest for the piroplasms in camels: Identification of Theileria equi and Babesia caballi in Jordanian dromedaries by PCR

    Czech Academy of Sciences Publication Activity Database

    Qablan, M.; Sloboda, M.; Jirků, Milan; Oborník, Miroslav; Dwairi, S.; Amr, Z. S.; Hořín, P.; Lukeš, Julius; Modrý, David

    2012-01-01

    Roč. 186, 3/4 (2012), s. 456-460 ISSN 0304-4017 R&D Projects: GA ČR GA206/09/0927 Institutional research plan: CEZ:AV0Z60220518 Keywords : Babesia * Theileria * Camelus * Jordan * Host specificity * Diagnosis Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 2.381, year: 2012

  2. Prevalence of Theileria equi and Babesia caballi as well as the identification of associated ticks in sympatric Grevy's zebras (Equus grevyi) and donkeys (Equus africanus asinus) in northern Kenya.

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    Hawkins, Elaine; Kock, Richard; McKeever, Declan; Gakuya, Francis; Musyoki, Charles; Chege, Stephen M; Mutinda, Mathew; Kariuki, Edward; Davidson, Zeke; Low, Belinda; Skilton, Robert A; Njahira, Moses N; Wamalwa, Mark; Maina, Elsie

    2015-01-01

    The role of equine piroplasmosis as a factor in the population decline of the Grevy's zebra is not known. We determined the prevalence of Babesia caballi and Theileria equi in cograzing Grevy's zebras (Equus grevyi) and donkeys (Equus africanus asinus) in northern Kenya and identified the associated tick vectors. Blood samples were taken from 71 donkeys and 16 Grevy's zebras from March to May 2011. A nested PCR reaction using 18s ribosomal (r)RNA primers on 87 blood spots showed 72% (51/71; 95% confidence interval [CI] 60.4-81.0%) of donkeys and 100% (16/16; 95% CI, 77.3-100%) of Grevy's zebras were T. equi positive. No samples were positive for B. caballi. Sequence comparison using the National Center for Biotechnology Information's basic local alignment search tool identified homologous 18s rRNA sequences with a global geographic spread. The T. equi-derived sequences were evaluated using Bayesian approaches with independent Metropolis-coupled Markov chain Monte Carlo runs. The sequences clustered with those found in Sudan, Croatia, Mongolia, and the US, with statistical support greater than 80% for the two main clades. Hyalomma tick species were found on both donkeys and Grevy's zebras, whereas Rhipicephalus pulchellus was found exclusively on Grevy's zebras and Hyalomma marginatum rupfipes on donkeys. The prevalence of T. equi was 100% in Grevy's zebras and 72% in donkeys with common tick vectors identified. Our results suggest that donkeys and Grevy's zebras can be asymptomatic carriers and that piroplasmosis is endemic in the study area.

  3. Detection of Babesia hongkongensis sp. nov. in a Free-Roaming Felis catus Cat in Hong Kong

    OpenAIRE

    Wong, Samson S. Y.; Poon, Rosana W. S.; Hui, Janet J. Y.; Yuen, Kwok-Yung

    2012-01-01

    Intraerythrocytic Babesia-like trophozoites were seen in postmortem kidney sections of a free-roaming cat in Hong Kong. DNA sequences of the 18S rRNA and mitochondrial cytochrome b genes had only 96.7% and 90.4% nucleotide identity with known Babesia sequences. We propose that this new species be named Babesia hongkongensis.

  4. Detection of Babesia hongkongensis sp. nov. in a free-roaming Felis catus cat in Hong Kong.

    Science.gov (United States)

    Wong, Samson S Y; Poon, Rosana W S; Hui, Janet J Y; Yuen, Kwok-Yung

    2012-08-01

    Intraerythrocytic Babesia-like trophozoites were seen in postmortem kidney sections of a free-roaming cat in Hong Kong. DNA sequences of the 18S rRNA and mitochondrial cytochrome b genes had only 96.7% and 90.4% nucleotide identity with known Babesia sequences. We propose that this new species be named Babesia hongkongensis.

  5. Babesia sp

    Directory of Open Access Journals (Sweden)

    Endang Setiyani

    2012-11-01

    Full Text Available Babesia merupakan parasit yang dapat menimbulkan infeksi pada manusia melalui gigitan sengkenit. Penyakitnya disebut babesiosis atau piroplasmosis yaitu suatu penyakit hewan yang bisa menular ke manusia (zoonosis yang disebabkan oleh protozoa parasit spesies Babesia seperi parasit malaria, ia juga menginfeksi set darah merah binatang liar maupun binatang peliharaan dengan gejala mirip demam malaria, yaitu demam disertai anemi hemolitik.

  6. A case of human infection with a novel Babesia species in China.

    Science.gov (United States)

    Man, Su-Qin; Qiao, Ke; Cui, Jie; Feng, Meng; Fu, Yong-Feng; Cheng, Xun-Jia

    2016-03-29

    Babesiosis is an uncommon but emerging tick-borne disease caused by the genus Babesia. In this case study, we report a case of human infection with a novel Babesia sp. in China. The patient in question had been suffering from repetitive occurrences of mild fever of unknown origin and fatigue for 10 years. Ring forms, tetrads, and one or two dots of chromatin or trophozoite-like organisms were observed in the patient's thin blood smears and bone marrow smears. Using a confocal laser-scanning microscope, it was observed that the patient's serum had reactivity with the surface proteins of the B. microti strain. Electron microscopy revealed oval red blood cells with 1 ~ 2 μm of knob protrusions in the cellular membrane. The results of the Babesia-specific nested PCR assay for 18S rRNA confirmed the presence of Babesia infection. The construction of a phylogenetic relationship showed clustering with B. microti and B. duncani, which was identified as a novel Babesia species and named as Babesia sp. XXB/HangZhou. Azithromycin, doxycycline, and moxifloxacin hydrochloride were shown to relieve symptoms but were not as effective after continuous usage. After atovaquone (Mepron®) administration, the patient recovered from fever and tested negative for detection of Babesia-specific genes. Babesia sp. XXB/HangZhou is a novel Babesia species, which causes mild babesiosis in an immunocompetent patient.

  7. Axenic isolation of viable Giardia muris trophozoites.

    Science.gov (United States)

    Tillotson, K D; Buret, A; Olson, M E

    1991-06-01

    Large numbers of viable Giardia muris trophozoites were isolated from the duodenum of experimentally infected mice 6 days after inoculation with 1,000 G. muris cysts. A series of shaking, incubation, and washing steps in the presence of the broad-spectrum antibiotic piperacillin readily provided 4.9 +/- 1.5 x 10(5) G. muris trophozoites per mouse, free of detectable contaminant organisms. Anaerobic and microaerophilic culturing and scanning electron microscopy demonstrated axenic status and high purity of the isolates. The viability of trophozoites was 98 +/- 2%. Application of this technique should permit novel immunological and epidemiological analyses of G. muris infection and biochemical investigations of this protozoan parasite.

  8. Babesia behnkei sp. nov., a novel Babesia species infecting isolated populations of Wagner's gerbil, Dipodillus dasyurus, from the Sinai Mountains, Egypt.

    Science.gov (United States)

    Bajer, Anna; Alsarraf, Mohammed; Bednarska, Małgorzata; Mohallal, Eman M E; Mierzejewska, Ewa J; Behnke-Borowczyk, Jolanta; Zalat, Sammy; Gilbert, Francis; Welc-Falęciak, Renata

    2014-12-09

    Although a number of new species of Babesia/Theileria have been described recently, there are still relatively few reports of species from Africa. In this study based on the evaluation of morphology and phylogenetic relationships, we describe a novel species from Wagner's gerbil, Babesia behnkei n. sp. Rodents (n = 1021) were sampled in four montane valleys (wadies) in 2000, 2004, 2008 and 2012 in the Sinai Mountains, Egypt. The overall prevalence of Babesia spp. was highest in the Wagner's gerbil (Dipodillus dasyurus; 38.7%) in comparison to the prevalence in the spiny mice species, Acomys dimidiatus and A. russatus. Morphological investigations were conducted for the comparison of trophozoites of the novel species of Babesia with the B. microti King's 67 reference strain. Thirty-two isolates derived from D. dasyurus over a 9 year period (2004-2012) from two wadies (29 isolates from Wadi Gebel and 3 from Wadi El-Arbaein) were investigated by microscopic, molecular and phylogenetic analysis. A near-full-length sequence of the 18S rRNA gene and the second internal transcribed spacer (ITS2) region were amplified, sequenced and used for the construction of phylogenetic trees. A novel species of Babesia was identified in two isolated populations of D. dasyurus. Phylogenetic analysis of 18S rDNA and ITS2 sequences revealed that B. behnkei n. sp. is most closely related to B. lengau from cheetahs from South Africa and to Nearctic species found only in North America (the pathogenic B. duncani and B. conradae) and that it is more distant to the cosmopolitan rodent parasite B. microti. Trophozoites of B. behnkei were smaller and less polymorphic than trophozoites of B. microti. Babesia behnkei n. sp. is a novel species of the 'Duncani group' maintained in isolated populations of Dipodillus dasyurus occurring in the Sinai Mountains of Egypt.

  9. Development of a pan-Babesia FRET-qPCR and a survey of livestock from five Caribbean islands.

    Science.gov (United States)

    Li, Jing; Kelly, Patrick; Zhang, Jilei; Xu, Chuanling; Wang, Chengming

    2015-09-30

    Babesia spp. are tick-borne protozoan hemoparasites and the second most common blood-borne parasites of mammals, in particular domestic animals. We used the Clustal Multiple Alignment program and 18S rRNA gene sequences of 22 Babesia species from GenBank to develop a PCR that could detect a wide variety of Babesia spp. in a single reaction. The pan-Babesia FRET-qPCR we developed reliably detected B. gibsoni, B. canis, B. vogeli, B. microti, B. bovis, and B. divergens under controlled conditions but did not react with closely related species, mainly Hepatozoon americanum, Theileria equi, and Toxoplasma gondii. When we tested the pan-Babesia FRET-qPCR on DNA of whole blood from 752 cattle, sheep, goats, donkeys and horses from five Caribbean islands, we detected Babesia spp. expected to be present in the animals, mainly B. bovis and B. bigemina in cattle and B. caballi in horses and donkeys. Further, we found that animals were not uncommonly infected with species of Babesia usually associated with other hosts, mainly B. vogeli and B. gibsoni in cattle, sheep and goats, B. rossi in goats, and B. caballi in goats and sheep. Finally, the pan-Babesia FRET-qPCR enabled us to identify unknown species of Babesia in cattle, goats, sheep and donkeys. Overall, 70 % (525/752) of the animals we tested were positive confirming earlier limited studies that infections with Babesia spp. are common in livestock in the Caribbean.

  10. EquiMar

    DEFF Research Database (Denmark)

    Johnstone, C. M.; McCombes, T.; Bahaj, A. S.

    2011-01-01

    the performance evaluation of such systems in order to address this deficiency. This paper reports the development of a set of ‘Best Practices’ within the ECFPVII EquiMar project to be adopted for the performance quantification of wave and tidal energy converters as they evolve from an engineering concept......At the present time there are no approved standards or recognised best practices being implemented for the performance appraisal and benchmarking of wave and tidal energy converters. As such, this develops considerable misunderstanding between device developers, testing centres, investors....../ financiers etc when attempting to quantify the performance of a device since it makes it very difficult to reference and benchmark the performance of a marine energy converter. The EC Framework Programme VII EquiMar project has set out to develop a suite of Best Practices to be adopted when undertaking...

  11. [Sensibility of Entamoeba histolytica trophozoites to ivermectin].

    Science.gov (United States)

    González-Salazar, Francisco; Mata-Cárdenas, Benito D; Vargas-Villareal, Javier

    2009-01-01

    Amebiasis caused by Entamoeba histolytica is a problem of public world health. The most frequent clinical presentation are the dysentery and the amebic liver abscess. Fifty millions of cases and more than 100.000 deaths for this disease are reported annually worldwide. The life cycle of E. histolytica has two phases: trophozoite and cyst. Trophozoites are the causal agent of disease. The effective treatment for the amebiasis includes drugs with serious collateral effects. Ivermectin is a macrolid with activity against endoparasites and ectoparasites causing strongiloidosis, filariasis, oncocercosis, scabiasis and pediculosis. The use of ivermectin has been extended almost worldwide; it is recognized as a safe drug. The main objective of this study was to determine in vitro sensibility of trophozoites of E. histolytica was to the treatment with ivermectin. To determine the sensibility of the parasites to the drug, E. histolytica was cultivated in PEHPS medium. During its logarithmic growth phase the trophozoites were exposed to different concentrations of ivermectin. As controls other antiparasitic drugs were used. For each drug, serial dilutions were prepared, and mixed in culture tubes with parasites (2 x 104 cells/ml). They were incubated for 72 h and then the percentage of growth inhibition was calculated by Probit analysis. Ivermectin showed activity against trophozoites of E. histolytica. The 50% of growth inhibition of ivermectin was 6.40 mg/ml. This dose was higher than for other anti parasitic drugs. Its activity in vivo in animal models remains to be demonstrated.

  12. Modification of host erythrocyte membranes by trypsin and chymotrypsin treatments and effects on the in vitro growth of bovine and equine Babesia parasites.

    Science.gov (United States)

    Okamura, Masashi; Yokoyama, Naoaki; Takabatake, Noriyuki; Okubo, Kazuhiro; Ikehara, Yuzuru; Igarashi, Ikuo

    2007-02-01

    In the present study, we investigated the effects of protease pretreatments of host erythrocytes (RBC) on the in vitro growth of bovine Babesia parasites (Babesia bovis and B. bigemina) and equine Babesia parasites (B. equi and B. caballi). The selected proteases, trypsin and chymotrypsin, clearly modified several membrane proteins of both bovine and equine RBC, as demonstrated by SDS-PAGE analysis; however, the protease treatments also modified the sialic acid content exclusively in bovine RBC, as demonstrated by lectin blot analysis. An in vitro growth assay using the protease-treated RBC showed that the trypsin-treated bovine RBC, but not the chymotrypsin-treated ones, significantly reduced the growth of B. bovis and B. bigemina as compared to the control. In contrast, the growth of B. equi and B. caballi was not affected by any of these proteases. Thus, the bovine, but not the equine, Babesia parasites require the trypsin-sensitive membrane (sialoglyco) proteins to infect the RBC.

  13. Evaluation of PCR and multiplex PCR in relation to nested PCR for diagnosing Theileria equi

    Directory of Open Access Journals (Sweden)

    Danielle C. Leal

    2011-07-01

    Full Text Available Conventional PCR (PCRTeq for diagnosing Theileria equi and multiplex PCR (M/PCRTeq-Bc for diagnosing T. equi and Babesia caballi were comparatively evaluated with nested PCR (N/PCR-Teq for diagnosing equine piroplasmosis. In DNA sensitivity determinations, in multiple dilutions of equine blood that had tested positive for T. equi, PCR-Teq and N/PCR-Teq detected hemoparasite DNA in the larger dilutions (1:128, but did not differ significantly from the M/PCRTeq-Bc (1:64. In analyses on equine serum tested by ELISA, there was high agreement between this serological test and PCR-Teq (k = 0.780 and moderate agreement with N/PCR-Teq (k = 0.562 and M/PCRTeq-Bc (k = 0.488. PCR-Teq found a higher frequency of T. equi both in extensively and intensively reared horses, but this was not significant in relation to N/PCR-Teq (P>0.05, and both PCRs indicated that there was an endemic situation regarding T. equi in the population of horses of this sample. PCR-Teq was only significantly different from M/PCR-Teq-Bc (P<0.05. PCR-Teq presented high sensitivity and specificity, comparable to N/PCR-Teq, but with the advantage of higher speed in obtaining results and lower costs and risks of laboratory contamination. This accredits PCR-Teq for epidemiological studies and for determinations on affected horses.

  14. Generation and characterization of monoclonal antibodies against Giardia muris trophozoites.

    Science.gov (United States)

    Heyworth, M F; Ho, K E; Pappo, J

    1989-11-01

    Mouse monoclonal antibodies (mAb) were produced against Giardia muris trophozoite surface antigens. To generate B-cell hybridomas, P3/NS1/1-Ag4-1 myeloma cells were fused with splenic lymphocytes from BALB/c mice that had been immunized parenterally with G. muris trophozoites. Hybridoma culture supernatants were screened for mAb by flow cytometry of G. muris trophozoites incubated with culture supernatant followed by fluorescein-conjugated anti-mouse IgG and IgM. Flow cytometry showed three types of trophozoite staining by mAb: (i) bright staining of greater than 90% of trophozoites, with aggregation of the organisms; (ii) bright staining of approximately 90% of trophozoites, with little or no aggregation; (iii) dull staining of approximately 20% of trophozoites, without aggregation. Western blotting of mAb on G. muris trophozoite antigens separated by polyacrylamide gel electrophoresis showed that a mAb exhibiting the third of these flow cytometry staining patterns recognized trophozoite antigens of MW approximately 31,000 and 35,000. Immunoprecipitation studies indicated that the same mAb specifically precipitated two 125I-labelled trophozoite surface antigens of MW approximately 30,000. Monoclonal antibodies generated in this study may facilitate the purification and biochemical characterization of trophozoite antigens that are targets for protective intestinal antibody in G. muris-infected mice.

  15. Intravascular detection of Giardia trophozoites in naturally infected mice

    Directory of Open Access Journals (Sweden)

    El-Shewy K.A.

    2003-06-01

    Full Text Available During routine transmission electron microscopic (TEM examination of mice naturally infected with Giardia muris, an intense infection with Giardia trophozoites was demonstrated within intestinal and renal tissues. Examination of randomly taken sections from these heavily infected tissues revealed marked deep affection with mixed pathology. Duodenal sections were found loaded with Giardia trophozoites in intimate contact with necrotic gut cells. Some of these trophozoites were detected within central lacteal of damaged villi and nearby blood vessels. Interestingly, and for the first time to be demonstrated, morphologically identical G. muris trophozoite was detected in a renal blood vessel. An intense cellular immune reaction was obviously demonstrated with remarkable interaction between giant macrophages and the trophozoites particulates. Involvement of deep tissues by Giardia trophozoites and their presence within vascular channels could open up questions about the possible invasive and disseminative behavior of G. muris, particularly in heavily and naturally infected hosts.

  16. A molecular and haematological study of Theileria equi in Balkan donkeys.

    Science.gov (United States)

    Davitkov, Dajana; Davitkov, Darko; Vucicevic, Milos; Stanisic, Ljubodrag; Radakovic, Milena; Glavinic, Uros; Stanimirovic, Zoran

    2017-06-01

    Equine piroplasmosis in donkeys has been recognised as a serious problem of major economic importance. The present molecular study is the first investigation of the presence of Theileria equi and Babesia caballi in Balkan donkeys and of the possible haematological alterations related to it. A total of 70 apparently healthy donkeys from Serbia were included in this study. The overall prevalence of T. equi infection in donkeys tested with multiplex PCR was 50%. There was no B. caballi-positive sample. Infections in donkeys included in this study seem to be associated with decreased red blood cell count, haemoglobin concentration, haematocrit and platelet count, and with increased white blood cell count, mean corpuscular haemoglobin and mean corpuscular haemoglobin concentration. Altered haematological parameters in donkeys can lead to a decrease in working capacity and production performance. Further molecular research and long-term monitoring of equine piroplasmosis is needed in Serbia and throughout Europe.

  17. Babesial vector tick defensin against Babesia sp. parasites.

    Science.gov (United States)

    Tsuji, Naotoshi; Battsetseg, Badgar; Boldbaatar, Damdinsuren; Miyoshi, Takeharu; Xuan, Xuenan; Oliver, James H; Fujisaki, Kozo

    2007-07-01

    Antimicrobial peptides are major components of host innate immunity, a well-conserved, evolutionarily ancient defensive mechanism. Infectious disease-bearing vector ticks are thought to possess specific defense molecules against the transmitted pathogens that have been acquired during their evolution. We found in the tick Haemaphysalis longicornis a novel parasiticidal peptide named longicin that may have evolved from a common ancestral peptide resembling spider and scorpion toxins. H. longicornis is the primary vector for Babesia sp. parasites in Japan. Longicin also displayed bactericidal and fungicidal properties that resemble those of defensin homologues from invertebrates and vertebrates. Longicin showed a remarkable ability to inhibit the proliferation of merozoites, an erythrocyte blood stage of equine Babesia equi, by killing the parasites. Longicin was localized at the surface of the Babesia sp. parasites, as demonstrated by confocal microscopic analysis. In an in vivo experiment, longicin induced significant reduction of parasitemia in animals infected with the zoonotic and murine B. microti. Moreover, RNA interference data demonstrated that endogenous longicin is able to directly kill the canine B. gibsoni, thus indicating that it may play a role in regulating the vectorial capacity in the vector tick H. longicornis. Theoretically, longicin may serve as a model for the development of chemotherapeutic compounds against tick-borne disease organisms.

  18. Novel type of linear mitochondrial genomes with dual flip-flop inversion system in apicomplexan parasites, Babesia microti and Babesia rodhaini

    Directory of Open Access Journals (Sweden)

    Hikosaka Kenji

    2012-11-01

    Full Text Available Abstract Background Mitochondrial (mt genomes vary considerably in size, structure and gene content. The mt genomes of the phylum Apicomplexa, which includes important human pathogens such as the malaria parasite Plasmodium, also show marked diversity of structure. Plasmodium has a concatenated linear mt genome of the smallest size (6-kb; Babesia and Theileria have a linear monomeric mt genome (6.5-kb to 8.2-kb with terminal inverted repeats; Eimeria, which is distantly related to Plasmodium and Babesia/Theileria, possesses a mt genome (6.2-kb with a concatemeric form similar to that of Plasmodium; Cryptosporidium, the earliest branching lineage within the phylum Apicomplexa, has no mt genome. We are interested in the evolutionary origin of linear mt genomes of Babesia/Theileria, and have investigated mt genome structures in members of archaeopiroplasmid, a lineage branched off earlier from Babesia/Theileria. Results The complete mt genomes of archaeopiroplasmid parasites, Babesia microti and Babesia rodhaini, were sequenced. The mt genomes of B. microti (11.1-kb and B. rodhaini (6.9-kb possess two pairs of unique inverted repeats, IR-A and IR-B. Flip-flop inversions between two IR-As and between two IR-Bs appear to generate four distinct genome structures that are present at an equi-molar ratio. An individual parasite contained multiple mt genome structures, with 20 copies and 2 – 3 copies per haploid nuclear genome in B. microti and B. rodhaini, respectively. Conclusion We found a novel linear monomeric mt genome structure of B. microti and B. rhodhaini equipped with dual flip-flop inversion system, by which four distinct genome structures are readily generated. To our knowledge, this study is the first to report the presence of two pairs of distinct IR sequences within a monomeric linear mt genome. The present finding provides insight into further understanding of evolution of mt genome structure.

  19. Description of Babesia duncani n.sp. (Apicomplexa: Babesiidae) from humans and its differentiation from other piroplasms.

    Science.gov (United States)

    Conrad, Patricia A; Kjemtrup, Anne M; Carreno, Ramon A; Thomford, John; Wainwright, Katlyn; Eberhard, Mark; Quick, Rob; Telford, Sam R; Herwaldt, Barbara L

    2006-06-01

    The morphologic, ultrastructural and genotypic characteristics of Babesia duncani n.sp. are described based on the characterization of two isolates (WA1, CA5) obtained from infected human patients in Washington and California. The intraerythrocytic stages of the parasite are morphologically indistinguishable from Babesia microti, which is the most commonly identified cause of human babesiosis in the USA. Intraerythrocytic trophozoites of B. duncani n.sp. are round to oval, with some piriform, ring and ameboid forms. Division occurs by intraerythrocytic schizogony, which results in the formation of merozoites in tetrads (syn. Maltese cross or quadruplet forms). The ultrastructural features of trophozoites and merozoites are similar to those described for B. microti and Theileria spp. However, intralymphocytic schizont stages characteristic of Theileria spp. have not been observed in infected humans. In phylogenetic analyses based on sequence data for the complete18S ribosomal RNA gene, B. duncani n.sp. lies in a distinct clade that includes isolates from humans, dogs and wildlife in the western United States but separate from Babesia sensu stricto, Theileria spp. and B. microti. ITS2 sequence analysis of the B. duncani n.sp. isolates (WA1, CA5) show that they are phylogenetically indistinguishable from each other and from two other human B. duncani-type parasites (CA6, WA2 clone1) but distinct from other Babesia and Theileria species sequenced. This analysis provides robust molecular support that the B. duncani n.sp. isolates are monophyletic and the same species. The morphologic characteristics together with the phylogenetic analysis of two genetic loci support the assertion that B. duncani n.sp. is a distinct species from other known Babesia spp. for which morphologic and sequence information are available.

  20. Antibody response to Giardia muris trophozoites in mouse intestine.

    Science.gov (United States)

    Heyworth, M F

    1986-05-01

    The protozoan parasite Giardia muris colonizes the mouse small intestinal lumen. This parasite is cleared immunologically from the intestine of normal mice. In contrast, T-lymphocyte-deficient (nude) mice have an impaired immunological response to G. muris and become chronically infected. In the present study, trophozoites were harvested from the intestinal lumen of immunocompetent BALB/c mice and nude mice and examined for surface-bound mouse immunoglobulins by immunofluorescence microscopy. Immunoglobulin A (IgA) and IgG, but not IgM, were detected on trophozoites obtained from BALB/c mice, from day 10 of the infection onwards. Trophozoites from nude mice showed very little evidence of surface-bound mouse immunoglobulin at any time during the 5-week period immediately following infection of these animals with G. muris cysts. Intestinal G. muris infection was cleared by the BALB/c mice but not by the nude animals. The data suggest that parasite-specific IgA and IgG bind to G. muris trophozoites in the intestinal lumen of immunocompetent BALB/c mice. Intestinal antibodies that bind to trophozoite surfaces are likely to play an important part in the clearance of G. muris infection by immunocompetent mice. The inability of nude mice to clear this infection at a normal rate is likely to be due to impairment of Giardia-specific intestinal antibody production.

  1. Aortitis with bacteraemia by Streptococcus equi Zooepidemicus

    International Nuclear Information System (INIS)

    Betancur, Carlos Alberto; Giraldo, Juan David; Saldarriaga Eugenia Lucia

    2005-01-01

    Infections by Streptococcus equi subspecies zooepidemicus occur in animals. In human beings these infections are generally accidental, and few cases have been reported. We present the case of a 56-year-old male, a butcher, who presented with abdominal pain. Aneurismatic dilatation of the aorta below the renal arteries was documented by CT-scanning. A purulent collection and arterial ulceration were found during surgery; Streptococcus equi zooepidemicus was isolated from the collection and from blood cultures

  2. Bacteremia due to Rhodococcus equi in an immunocompetent infant

    OpenAIRE

    P Devi; S Malhotra; A Chadha

    2011-01-01

    Rhodococcus equi , previously known as Corynebacterium equi, is one of the most important causes of zoonotic infection in grazing animals. Increased cases of human infection with R. equi have been reported especially in immunocompromised patients. Infection in immunocompetent patients is extremely rare. We report a case of R. equi bacteremia in a 26-day-old immunocompetent infant with recurrent swellings on different parts of the body. To the best of our knowledge, this is the first ever repo...

  3. Bacteremia due to Rhodococcus equi in an immunocompetent infant

    Directory of Open Access Journals (Sweden)

    P Devi

    2011-01-01

    Full Text Available Rhodococcus equi , previously known as Corynebacterium equi, is one of the most important causes of zoonotic infection in grazing animals. Increased cases of human infection with R. equi have been reported especially in immunocompromised patients. Infection in immunocompetent patients is extremely rare. We report a case of R. equi bacteremia in a 26-day-old immunocompetent infant with recurrent swellings on different parts of the body. To the best of our knowledge, this is the first ever report of R. equi bacteremia from an immunocompetent patient from Northern India.

  4. Sequence heterogeneity in the 18S rRNA gene in Theileria equi from horses presented in Switzerland.

    Science.gov (United States)

    Liu, Qin; Meli, Marina L; Zhang, Yi; Meili, Theres; Stirn, Martina; Riond, Barbara; Weibel, Beatrice; Hofmann-Lehmann, Regina

    2016-05-15

    A reverse line blot (RLB) hybridization assay was adapted and applied for equine blood samples collected at the animal hospital of the University of Zurich to determine the presence of piroplasms in horses in Switzerland. A total of 100 equine blood samples were included in the study. The V4 hypervariable region of the 18S rRNA gene was amplified by polymerase chain reaction and analyzed using the RLB assay. Samples from seven horses hybridized to a Theileria/Babesia genus-specific and a Theileria genus-specific probe. Of these, two hybridized also to the Theileria equi-specific probe. The other five positive samples did not hybridize to any of the species-specific probes, suggesting the presence of unrecognized Theileria variants or genotypes. The 18S rRNA gene of the latter five samples were sequenced and found to be closely related to T. equi isolated from horses in Spain (AY534822) and China (KF559357) (≥98.4% identity). Four of the seven horses that tested positive had a documented travel history (France, Italy, and Spain) or lived abroad (Hungary). The present study adds new insight into the presence and sequence heterogeneity of T. equi in Switzerland. The results prompt that species-specific probes must be designed in regions of the gene unique to T. equi. Of note, none of the seven positive horses were suspected of having Theileria infection at the time of presentation to the clinic. Clinicians should be aware of the possibility of equine piroplasma infections outside of endemic areas and in horses without signs of piroplasmosis. Copyright © 2016 Elsevier B.V. All rights reserved.

  5. On reflectionless equi-transmitting matrices

    Directory of Open Access Journals (Sweden)

    Pavel Kurasov

    2014-01-01

    Full Text Available Reflectionless equi-transmitting unitary matrices are studied in connection to matching conditions in quantum graphs. All possible such matrices of size 6 are described explicitly. It is shown that such matrices form 30 six-parameter families intersected along 12 five-parameter families closely connected to conference matrices.

  6. Production of recombinant EMA-1 protein and its application for the diagnosis of Theileria equi using an enzyme immuno assay in horses from São Paulo State, Brazil Produção da proteína recombinante EMA-1 e sua aplicação para o diagnóstico baseadono imuno ensaio enzimático de Theileria equi em equinos do Estado de São Paulo, Brasil

    Directory of Open Access Journals (Sweden)

    Cristiane Divan Baldani

    2011-03-01

    Full Text Available The erythrocytic-stage surface protein, Equi Merozoite Antigen 1 (EMA-1, is a major candidate for the development of a diagnostic antigen for equine piroplasmosis. In order to establish an effective diagnostic method for practical use, the gene encoding the entire EMA-1 of Theileria equi Jaboticabal strain was cloned and expressed in Escherichia coli as a histidine-tagged protein (His6-EMA1. The expressed EMA-1 reacted with specific antibodies in Western blot and had an apparent molecular mass of 34 kDa which was largely consistent with its theoretical value. The nucleotide sequence of the EMA-1 gene of Jaboticabal strain was comparatively analyzed with other published sequences. The results indicated a high degree of homology with EMA-1 genes of all other strains isolated from various countries. The recombinant purified His6-EMA1 protein was tested in an enzyme-linked immunosorbent assay (ELISA for the detection of antibodies anti-T. equi in horses. The ELISA clearly differentiated T. equi-infected from Babesia caballi-infected horse sera or normal horse sera. Field serum samples collected from horses in the State of São Paulo, Southeastern Brazil, were examined for the diagnosis of T. equi infection by ELISA. Of 170 samples analyzed, 95.88% (163/170 were positive for T. equi infection. These results suggest that the His6-EMA1 protein expressed in E. coli could be a reliable immunodiagnostic antigen for ELISA test and that T. equi infection is a serious concern in the State of São Paulo, Brazil.A proteína de superfície eritrocitária, Antígeno 1 do Merozoíta de Theileria equi (EMA-1, é um potencial candidato para o desenvolvimento de antígenos de valor diagnóstico para a piroplasmose equina. Com o objetivo de estabelecer um método de diagnóstico efetivo e prático, o gene EMA-1 da amostra Jaboticabal - SP de T. equi foi clonado e expresso em Escherichia coli contendo uma cauda de poli-histidina (His6-EMA1. O EMA-1 expresso reagiu com

  7. Intravascular detection of Giardia trophozoites in naturally infected mice. An electron microscopic study.

    Science.gov (United States)

    el-Shewy, K A; Eid, R A

    2003-06-01

    During routine transmission electron microscopic (TEM) examination of mice naturally infected with Giardia muris, an intense infection with Giardia trophozoites was demonstrated within intestinal and renal tissues. Examination of randomly taken sections from these heavily infected tissues revealed marked deep affection with mixed pathology. Duodenal sections were found loaded with Giardia trophozoites in intimate contact with necrotic gut cells. Some of these trophozoites were detected within central lacteal of damaged villi and nearby blood vessels. Interestingly, and for the first time to be demonstrated, morphologically identical G. muris trophozoite was detected in a renal blood vessel. An intense cellular immune reaction was obviously demonstrated with remarkable interaction between giant macrophages and the trophozoites particulates. Involvement of deep tissues by Giardia trophozoites and their presence within vascular channels could open up questions about the possible invasive and disseminative behavior of G. muris, particularly in heavily and naturally infected hosts.

  8. Diversity of seM in Streptococcus equi subsp. equi isolated from strangles outbreaks.

    Science.gov (United States)

    Libardoni, Felipe; Vielmo, Andréia; Farias, Luana; Matter, Letícia Beatriz; Pötter, Luciana; Spilki, Fernando Rosado; de Vargas, Agueda Castagna

    2013-03-23

    Strangles is the main upper respiratory tract disease of horses. There are currently no studies on the changes in alleles of the M protein gene (seM) in Brazilian isolates of Streptococcus equi ssp. equi (S. equi). This study aimed to analyze and differentiate molecularly S. equi isolates from equine clinical specimens from southern Brazil, between 1994 and 2010. seM alleles were analyzed in 47 isolates of S. equi obtained from clinical cases of strangles (15 Thoroughbred horses, 29 Crioulo breed horses and three Brasileiro de Hipismo--BH). seM alleles characterization was performed by comparing variable region sequences of the seM gene. The alleles were also phylogenetically grouped by Neighbor-joining analysis, which demonstrated the geographic distribution of those in properties from southern Brazil. Fifteen alleles of the gene seM were found among the 47 S. equi isolates analyzed. Among these, only one allele (seM-61), which was identified in seven isolates (14.9%), was found in the database PubMLST-seM. Within the new alleles, allele seM-115 was the most prevalent, having been found in 13 isolates (27.7%), followed by allele seM-117 in 10 isolates (21.3%). In the Brazilian horse population studied, there is greater diversity of M protein alleles in S. equi isolates compared to worldwide data deposited in PubMLST-seM. Among the 15 seM alleles identified, only one allele sequence was previously published. The alleles identification is important to control the disease by guiding selection of strains for the manufacture of commercial and autogenous vaccines. Copyright © 2012 Elsevier B.V. All rights reserved.

  9. Giardia muris and Giardia duodenalis groups: ultrastructural differences between the trophozoites

    OpenAIRE

    Sogayar,Maria Inês L.; Gregório,Elisa Aparecida

    1989-01-01

    Trophozoites of the Giardia muris group from hamsters, domestic rats and mice and of the Giardia duodenalis group from hamsters and domestic rats were examined by transmission electron microscopy. The basic ultrastructure of the trophozoites was similar. Differences were shown in the morphology of the ventrolateral flange of the trophozoites of Giardia muris and Giardia duodenalis groups. Marginal plates are less developed in the species of the Giardia duodenalis group. In this group, the dis...

  10. Babesia peircei sp. nov. from the jackass penguin

    African Journals Online (AJOL)

    1992-01-09

    Jan 9, 1992 ... An avian piroplasm, Babesia peircei sp. nov. is described from the jackass penguin Spheniscus demersus. Morphological differences between Babesia peircei sp. nov. and the other valid Babesia spp. are discussed together with the possible vectors. 'n Voal-piroplasma, Babesia peircei sp. nov. afkomstig ...

  11. Relative susceptibility of Giardia muris trophozoites to killing by mouse antibodies of different isotypes.

    Science.gov (United States)

    Heyworth, M F

    1992-02-01

    The aim of this work was to examine the ability of mouse IgA, IgG, and IgM anti-Giardia antibodies to kill Giardia muris trophozoites in the presence and absence of complement. Using a 2-color flow cytometry assay, binding of antibody to trophozoites was assessed with fluorescein-conjugated anti-mouse immunoglobulin, and percentages of killed trophozoites were quantified by staining with propidium iodide. Trophozoites were killed in the presence of complement by IgG3 and IgM anti-trophozoite monoclonal antibodies. Anti-trophozoite IgA, obtained from the intestinal lumen of G. muris-infected BALB/c mice, became bound to trophozoites in vitro but did not kill these organisms in the presence or absence of complement. The results suggest that clearance of G. muris infection by intestinal IgA directed against G. muris trophozoites does not involve antibody-dependent killing of trophozoites in the intestinal lumen.

  12. Lineages of Streptococcus equi ssp. equi in the Irish equine industry.

    Science.gov (United States)

    Moloney, Emma; Kavanagh, Kerrie S; Buckley, Tom C; Cooney, Jakki C

    2013-01-01

    Streptococcus equi ssp. equi is the causative agent of 'Strangles' in horses. This is a debilitating condition leading to economic loss, yard closures and cancellation of equestrian events. There are multiple genotypes of S. equi ssp. equi which can cause disease, but to date there has been no systematic study of strains which are prevalent in Ireland. This study identified and classified Streptococcus equi ssp. equi strains isolated from within the Irish equine industry. Two hundred veterinary isolates were subjected to SLST (single locus sequence typing) based on an internal sequence from the seM gene of Streptococcus equi ssp equi. Of the 171 samples which successfully gave an amplicon, 162 samples (137 Irish and 24 UK strains) gave robust DNA sequence information. Analysis of the sequences allowed division of the isolates into 19 groups, 13 of which contain at least 2 isolates and 6 groups containing single isolates. There were 19 positions where a DNA SNP (single nucleotide polymorphism) occurs, and one 3 bp insertion. All groups had multiple (2-8) SNPs. Of the SNPs 17 would result in an amino acid change in the encoded protein. Interestingly, the single isolate EI8, which has 6 SNPs, has the three base pair insertion which is not seen in any other isolate, this would result in the insertion of an Ile residue at position 62 in that protein sequence. Comparison of the relevant region in the determined sequences with the UK Streptococcus equi seM MLST database showed that Group B (15 isolates) and Group I (2 isolates), as well as the individual isolates EI3 and EI8, are unique to Ireland, and some groups are most likely of UK origin (Groups F and M), but many more probably passed back and forth between the two countries. The strains occurring in Ireland are not clonal and there is a considerable degree of sequence variation seen in the seM gene. There are two major clades causing infection in Ireland and these strains are also common in the UK.

  13. The Effectiveness of Anti-R. equi Hyperimmune Plasma against R. equi Challenge in Thoroughbred Arabian Foals of Mares Vaccinated with R. equi Vaccine

    Directory of Open Access Journals (Sweden)

    Osman Erganis

    2014-01-01

    Full Text Available This study aimed to determine the effectiveness of a pregnant mare immunization of a Rhodococcus equi (R. equi vaccine candidate containing a water-based nanoparticle mineral oil adjuvanted (Montanide IMS 3012 inactive bacterin and virulence-associated protein A (VapA, as well as the administration of anti-R. equi hyperimmune (HI plasma against R. equi challenge in the mares’ foals. The efficacy of passive immunizations (colostral passive immunity by mare vaccination and artificial passive immunity by HI plasma administration was evaluated based on clinical signs, complete blood count, blood gas analysis, serological response (ELISA, interleukin-4 (IL-4 and interferon gamma (IFN-γ, total cell count of the bronchoalveolar lavage fluids (BALF samples, reisolation rate of R. equi from BALF samples (CFU/mL, lung samples (CFU/gr, and lesion scores of the organs and tissue according to pathological findings after necropsy in the foals. The vaccination of pregnant mares and HI plasma administration in the foals reduced the severity of R. equi pneumonia and lesion scores of the organs and tissue by 3.54-fold compared to the control foals. This study thus indicates that immunization of pregnant mares with R. equi vaccine candidate and administration of HI plasma in mares’ foals effectively protect foals against R. equi challenge.

  14. Giardia agilis: Ultrastructure of the Trophozoites in the Frog Intestine

    Directory of Open Access Journals (Sweden)

    Maria Inês L Sogayar

    1998-05-01

    Full Text Available Intestine samples of Bufo sp. tadpoles with parasitism confirmed for Giardia agilis were studied by transmission electron microscopy. The G. agilis trophozoites were long and thin. The plasma membrane was sometimes undulated and the cytoplasm, adjacent to the dorsal and ventral regions, showed numerous vacuoles. The two nuclei presented prominent nucleoli. The cytoplasm was electron-dense with free ribosomes, glycogen and rough endoplasmic reticulum-like structures. Polyhedral inclusions were observed in the cytoplasm and outside the protozoan; some of these inclusions exhibited membrane disruption. The flagella ultrastructure is typical, with the caudal pair accompanied by the funis. Next to the anterior pair, osmiophilic material was noticed. The ventro-lateral flange was short and thick, supported by the marginal plates that penetrated into its distal extremity; only its distal portion had adjacent osmiophilic filament. The G. agilis trophozoites showed the general subcellular feature of the genus. However, the ventro-lateral flange ultrastructure was an intermediate type between G. muris and G. duodenalis.

  15. COMPARABLE CYTOLOGICAL DIAGNOSTIC OF BLOOD SMEARS ON BABESIA INFECTION

    Directory of Open Access Journals (Sweden)

    Pokhyl S.І.

    2015-05-01

    warmed (t = 36.0 ± 2.0°С commercial matrix solutions of eosin, azure and methylene blue were applied one by one. The smears were rinsed (1-2 seconds in distilled water and dehydrated. The procedure ended with short-term drying in a diffused stream of warm dry air (Samsung house fan, power 220 W. The results were compared with intact control. Smears were contrasted and analysed under a microscope LOMU (LOMO, Russia: x 300; x400; x1000; x1350 and photographed with a digital camera “Canon EOS-3000”. Results. Blood samples infected with Babesia species were collected (may-october from naturally (promenade in forest-park tick-borne infected dogs (Canis familiaris in all Kharkov region and sity. All (experimental animals were monitored twice daily by veterinary doctors for clinical signs and had rectal temperatures taken (authors have a greate thankness for the cooperation and consolidation Chif -Mr. Yu. V. Al’okhin and veterinary personal of Kharkov Center of Clinical Veterinary. Blood was drawn daily for hematocrit determination and peripheral blood smear were made from ear vien blood to determine parasitemia status. As result of the analysis of blood smears it was found out that against a background of orange erythrocyte cytoplasm the preparation area easily revealed crimson- and red-lilac pyriform (n = 8-12 in the field of vision of the preparation, annular (n = 9-16 in the field of vision, amoebiform haemoparasites and those with other shapes (Σ=13, thereby indicating a high level of infection (81.8 %. Owing to their own chromatophilic feature, protozoan cells looked geometrically marked and clearly contrasted against a background of the saturated red-violet colour of nuclei. The developed technique of staining facilitated: a more qualitative analysis of ontogenetic staging (III of Babesia (trophozoites, merozoites, sporozoites; improvement of differential diagnosis of the haemoparasites with blood platelets (the latter were distinguished from cells of the

  16. Desenvolvimento de vacina recombinante de proteína M de Streptococcus equi subsp. equi

    OpenAIRE

    MACIEL, Liana Flores

    2012-01-01

    A equinocultura no Brasil ganha espaço em setores ligados ao lazer, cultura e turismo, sendo responsável por milhões de empregos. A Adenite Equina causada pelo Streptococcus equi subsp. equi é uma doença do aparelho respiratório de elevado impacto econômico, gerando gastos com mão-de-obra e perda de desempenho dos animais. Para amenizar este problema, medidas profiláticas são importantes, como por exemplo, a vacinação. Porém, as vacinas disponíveis no mercado protegem apenas 50% dos animai...

  17. Immune mechanisms in Babesia-infected animals

    International Nuclear Information System (INIS)

    Phillips, R.S.

    1980-01-01

    The course of a Babesia infection depends on the species of host and parasite involved. Animals infected with virulent babesias may need chemotherapy before acquired immunity develops. Maintenance of immunity is not dependent on the presence of the parasite. Babesia infections are characteristically of long duration. The immune response to babesias includes both humoral and cellular components. Antibody levels detected in serodiagnostic tests do not relate to levels of resistance to the parasite. Some antibodies, however, appear to be protective. Antiparasitic antibodies may damage parasites in or outside the red cell and act as opsonins. T-cell-deficient and anti-lymphocyte-serum-treated rodents are more susceptible to rodent piroplasms indicating a role for T-cells as either helper cells and/or as mediators of cell-mediated immunity (CMI). There is indirect evidence of CMI, but the cell-mediated mechanisms involved in parasite killing are not known. In domestic animals immunity is largely species- and strain-specific. Antigenic variation by babesias occurs. In rodents, however, there is cross-immunity between different species of rodent piroplasms and between rodent piroplasms and some malaria parasites. Prior infection with agents such as BCG, and Corynebacterium parvum, gives mice non-specific resistance to rodent piroplasms possibly mediated through a soluble non-antibody factor. This factor may also be liberated during piroplasm infections and by being toxic to malaria parasites account for heterologous immunity. Active immunization against babesias has been achieved with avirulent strains, irradiated parasites and dead parasites in adjuvant. During Babesia infections the primary and, to a lesser degree, the secondary immune response to heterologous antigens can be depressed. Maximum depression coincides with peak parasitaemia when antigen priming may be abolished completely. Immunosuppression during Babesia infections can prolong or exacerbate concurrent

  18. EquiX-A Search and Query Language for XML.

    Science.gov (United States)

    Cohen, Sara; Kanza, Yaron; Kogan, Yakov; Sagiv, Yehoshua; Nutt, Werner; Serebrenik, Alexander

    2002-01-01

    Describes EquiX, a search language for XML that combines querying with searching to query the data and the meta-data content of Web pages. Topics include search engines; a data model for XML documents; search query syntax; search query semantics; an algorithm for evaluating a query on a document; and indexing EquiX queries. (LRW)

  19. Management of Rhodococcus equi pneumonia in foals

    Directory of Open Access Journals (Sweden)

    Johns I

    2013-11-01

    Full Text Available Imogen Johns Department of Clinical Sciences and Services, Royal Veterinary College, North Mymms, UK Abstract: Rhodococcus equi, a gram-positive facultative intracellular bacterial pathogen, is the most important cause of pneumonia in foals aged 3 weeks to 5 months. The disease occurs worldwide, resulting in significant morbidity and mortality on endemically affected farms. Foals appear to become infected early in life, but clinical signs are typically delayed until 1–3 months of age because of the insidious nature of the disease. Although pneumonia is the most common clinical manifestation, up to 74% of foals may concurrently have extrapulmonary disorders, including both extrapulmonary infections (abdominal abscessation, colitis, osteomyelitis and immune-mediated disorders (nonseptic synovitis, uveitis. Diagnosis is based on the combination of clinical signs and abnormalities on hematologic screening and thoracic imaging in an appropriately aged foal and is confirmed by bacteriologic culture of the organism. Management of R. equi infections, in particular on farms with endemic disease, combines appropriate treatment of affected foals with preventative measures targeted at preventing infection and identifying foals before the development of severe disease. The combination of rifampin and a macrolide antimicrobial is recommended for treatment, as the combination is synergistic, reaches high intracellular concentrations, and should minimize the development of antimicrobial resistance. The prognosis for survival for foals with R. equi pneumonia is good, especially in foals mildly or subclinically affected, as is the prognosis for future athletic performance. Screening for early identification before the development of clinical signs has been advocated on endemically affected farms, although the most appropriate method, the timing of screening, and the selection of foals requiring treatment have yet to be determined. Recent evidence suggests that

  20. A comparison of the Giardia lamblia trophozoite and cyst transcriptome using microarrays

    Directory of Open Access Journals (Sweden)

    Widmer Giovanni

    2011-05-01

    Full Text Available Abstract Background Compared with many protists, Giardia lamblia has a simple life cycle alternating between cyst and trophozoite. Most research on the molecular biology of Giardia parasites has focused on trophozoites and the processes of excystation and encystation, whereas cysts have attracted less interest. The striking morphological differences between the dormant cyst and the rapidly dividing and motile trophozoite implies profound changes in the metabolism as the parasite encysts in the host's intestine and excysts upon ingestion by a new host. Results To investigate the magnitude of the transcriptional changes occurring during the G. lamblia life cycle we compared the transcriptome of G. lamblia trophozoites and cysts using single-color oligonucleotide microarrays. Cysts were found to possess a much smaller transcriptome, both in terms of mRNA diversity and abundance. Genes encoding proteins related to ribosomal functions are highly over-represented. The comparison of the transcriptome of cysts generated in culture or extracted from feces revealed little overlap, raising the possibility of significant biological differences between the two types of cysts. Conclusions The comparison of the G. lamblia cyst and trophozoite transcriptome showed that transcripts of most genes are present at a lower level in cysts. This global view of the cyst and trophozoite transcriptome complements studies focused on the expression of selected genes during trophozoite multiplication, encystation and excystation.

  1. Seroprevalence of Rhodococcus equi in horses in Israel

    Directory of Open Access Journals (Sweden)

    Sharon Tirosh-Levy

    2017-06-01

    Full Text Available Rhodococcus equi is a common cause of pneumonia in foals and has extensive clinical, economic and possibly zoonotic consequences. This bacterium survives well in the environment and may be considered as normal flora of adult horses. Certain strains of this bacterium are extremely virulent in foals, and early identification and intervention is crucial for prognosis. Rhodococcus equi is endemic in many parts of the world and occasionally isolated in Israel. This study was designed to evaluate R. equi seroprevalence in adult horses in Israel to indirectly indicate the potential level of exposure of susceptible foals. Sera were collected from 144 horses during spring 2011 and from 293 horses during fall 2014, and the presence of antibodies against virulent R. equi was detected by enzyme-linked immunosorbent assay. Equine seroprevalence of R. equi was found to be 7.6% in 2011 and 5.1% in 2014. Only one farm had seropositive horses in 2011, whereas several farms had seropositive horses in 2014. No significant risk factors for seropositivity were found. Rhodococcus equi appears to be endemic in Israel. This is the first survey of R. equi in Israel that provides information on the epidemiology of this important bacterium.

  2. Seroprevalence of Rhodococcus equi in horses in Israel.

    Science.gov (United States)

    Tirosh-Levy, Sharon; Gürbilek, Sevil E; Tel, Osman Y; Keskin, Oktay; Steinman, Amir

    2017-06-26

    Rhodococcus equi is a common cause of pneumonia in foals and has extensive clinical, economic and possibly zoonotic consequences. This bacterium survives well in the environment and may be considered as normal flora of adult horses. Certain strains of this bacterium are extremely virulent in foals, and early identification and intervention is crucial for prognosis. Rhodococcus equi is endemic in many parts of the world and occasionally isolated in Israel. This study was designed to evaluate R. equi seroprevalence in adult horses in Israel to indirectly indicate the potential level of exposure of susceptible foals. Sera were collected from 144 horses during spring 2011 and from 293 horses during fall 2014, and the presence of antibodies against virulent R. equi was detected by enzyme-linked immunosorbent assay. Equine seroprevalence of R. equi was found to be 7.6% in 2011 and 5.1% in 2014. Only one farm had seropositive horses in 2011, whereas several farms had seropositive horses in 2014. No significant risk factors for seropositivity were found. Rhodococcus equi appears to be endemic in Israel. This is the first survey of R. equi in Israel that provides information on the epidemiology of this important bacterium.

  3. Infection of dogs with Babesia canis in Gwagwalada metropolis of ...

    African Journals Online (AJOL)

    ADEYEYE

    2014-10-30

    Oct 30, 2014 ... determine the prevalence of Babesia canis and the correlation of infection with age, sex, breed, types of management and presence .... Table 3: Breed distribution of Babesia infection in dogs in Gwagwalada Area Council, FCT. Breed ... the management style of the dogs and infection with. Babesia. Table 5 ...

  4. Rhodococcus equi Parte 2: imunologia e profilaxia Rhodococcus equi Part 2: immunology and profilaxy

    Directory of Open Access Journals (Sweden)

    Ana Carolina Rusca Correa Porto

    2011-12-01

    Full Text Available Rhodococcus equi é um patógeno ubíquo e habitante da flora intestinal dos equinos de importância na neonatologia equina. Todos os potros são expostos à doença ao nascimento, porém alguns a desenvolvem e outros não. Este artigo revisa características da resposta imune, tanto em adultos competentes quanto em potros suscetíveis ao patógeno. A resposta imune humoral é abordada, incluindo uma discussão sobre o uso do plasma hiperimune como ferramenta profilática. O papel dos mecanismos de imunidade inata na suscetibilidade de alguns potros ao R. equi é também abordado. Da mesma maneira, os componentes envolvidos na resposta cito-mediada são revisados, com atenção particular às pesquisas direcionadas ao desenvolvimento de uma vacina efetiva para ser utilizada em potros.Rhodococcus equi is an important pathogen in equine neonatology that is ubiquitous and a normal intestinal inhabitant of equines. All foals are exposed at birth, however, some foals develop disease and others do not. This article reviews concepts of the equine immune response, both in the immune adult and susceptible neonate, with respect to this pathogen. Humoral immune responses are addressed, with a discussion on the use of hyperimmune plasma as a prophylactic tool. The role that innate immune mechanisms play in the susceptibility of some foals to R. equi infection is also highlighted. Likewise, cell-mediated immune components are reviewed, with particular attention directed towards research to develop an effective vaccine for foals.

  5. First evidence of Babesia venatorum and Babesia capreoli in questing Ixodes ricinus ticks in the Czech Republic

    Czech Academy of Sciences Publication Activity Database

    Venclíková, Kristýna; Mendel, Jan; Betášová, Lenka; Hubálek, Zdeněk; Rudolf, Ivo

    2015-01-01

    Roč. 22, č. 2 (2015), s. 212-214 ISSN 1232-1966 EU Projects: European Commission(XE) 261504 - EDENEXT Institutional support: RVO:68081766 Keywords : Babesia sp. EU1 * Babesia venatorum * Babesia capreoli * Ixodes ricinus * ixodid ticks Subject RIV: EE - Microbiology, Virology Impact factor: 0.895, year: 2015

  6. Giardia muris trophozoite antigenic targets for mouse intestinal IgA antibody.

    Science.gov (United States)

    Heyworth, M F; Vergara, J A

    1994-02-01

    The aim of this work was to characterize Giardia muris trophozoite proteins that are targets for intestinal anti-trophozoite IgA in G. muris-infected mice. Intestinal secretions were obtained from immunocompetent BALB/c mice that had been infected with G. muris cysts 4-5 weeks previously and from control uninfected BALB/c mice. Flow cytometry of G. muris trophozoites that had been incubated with intestinal secretions and with fluorescein isothiocyanate-conjugated anti-mouse IgA showed that anti-trophozoite IgA was present in intestinal secretions obtained from infected BALB/c mice. By immunoblotting on G. muris trophozoite proteins separated by one-dimensional gel electrophoresis, this IgA recognized at least one trophozoite protein of molecular mass of approximately 80 kDa. The 80-kDa G. muris protein(s) has a molecular mass similar to that described for cysteine-rich surface proteins of the human parasite Giardia lamblia.

  7. An Adenoviral Vector Based Vaccine for Rhodococcus equi.

    Directory of Open Access Journals (Sweden)

    Carla Giles

    Full Text Available Rhodococcus equi is a respiratory pathogen which primarily infects foals and is endemic on farms around the world with 50% mortality and 80% morbidity in affected foals. Unless detected early and treated appropriately the disease can be fatal. Currently, there is no vaccine available to prevent this disease. For decades researchers have endeavoured to develop an effective vaccine to no avail. In this study a novel human adenoviral vector vaccine for R. equi was developed and tested in the mouse model. This vaccine generated a strong antibody and cytokine response and clearance of R. equi was demonstrated following challenge. These results show that this vaccine could potentially be developed further for use as a vaccine to prevent R. equi disease in foals.

  8. Virulence of Rhodococcus equi Isolated from Cats and Dogs

    OpenAIRE

    Takai, Shinji; Martens, Ronald J.; Julian, Alan; Garcia Ribeiro, Márcio; Rodrigues de Farias, Marconi; Sasaki, Yukako; Inuzuka, Kazuho; Kakuda, Tsutomu; Tsubaki, Shiro; Prescott, John F.

    2003-01-01

    Nine cat isolates and nine dog isolates of Rhodococcus equi from clinical material were investigated for the presence of the virulence-associated antigens (VapA and VapB) and virulence plasmids. Five of the cat isolates and one dog isolate were VapA positive and contained an 85-kb type I or an 87-kb type I plasmid. The remaining 12 isolates were avirulent R. equi strains and contained no virulence plasmids.

  9. The life cycle of Babesia bovis and Babesia bigemina in ticks and cattle in South Africa

    OpenAIRE

    2015-01-01

    Ph.D. Bovine babesiosis, or redwater, is at present known to be caused by Babesia bigemina and Babesia bovis in the Republic of South Africa. Until recently, however, the only information on the natural transmission of these parasites in the country was based on observations made during the early part of this century and information on the developmental cycle of the parasites in their vectors was superficial or nonexistent ...

  10. Babesia peircei sp. nov. from the jackass penguin | Earlé | African ...

    African Journals Online (AJOL)

    An avian piroplasm, Babesia peircei sp. nov. is described from the jackass penguin Spheniscus demersus. Morphological differences between Babesia peircei sp. nov. and the other valid Babesia spp. are discussed together with the possible vectors.

  11. Morphological Findings in Trophozoites during Amoebic Abscess Development in Misoprostol-Treated BALB/c Mice

    Directory of Open Access Journals (Sweden)

    Andrés Aceves-Cano

    2015-01-01

    Full Text Available During amoebic liver abscess (ALA formation in susceptible animals, immune response is regulated by prostaglandin E2 (PGE2 dependent mechanisms. The aim of this study was to analyze the effect of misoprostol (MPL, a PGE1 analogue, on ALA formation in BALB/c mice. Male mice from BALB/c strain were intrahepatically infected with 7.5×105 trophozoites of E. histolytica strain HM1:IMSS and treated with 10−4 M of MPL daily until sacrifice at 2, 4, and 7 days postinfection (p.i.. ALA formation was evaluated at 2, 4, and 7 days postinfection; trophozoite morphology was analyzed using immunohistochemistry and image analysis. Results showed an increase in frequency of ALA formation in infected and MPL-treated mice only at 2 days p.i. (P=0.03. A significant diminution in the size of trophozoites was detected in abscesses from mice independently of MPL treatment (from 5.8±1.1 µm at 2 days p.i. to 2.7±1.9 µm at 7 days p.i. compared with trophozoites dimensions observed in susceptible hamsters (9.6±2.7 µm (P<0.01. These results suggest that MPL treatment may modify the adequate control of inflammatory process to allow the persistence of trophozoites in the liver; however, natural resistance mechanisms cannot be discarded.

  12. Detection of beta-tubulin in the cytoplasm of the interphasic Entamoeba histolytica trophozoites.

    Science.gov (United States)

    Gómez-Conde, Eduardo; Vargas-Mejía, Miguel Ángel; Díaz-Orea, María Alicia; Hernández-Rivas, Rosaura; Cárdenas-Perea, María Elena; Guerrero-González, Tayde; González-Barrios, Juan Antonio; Montiel-Jarquín, Álvaro José

    2016-08-01

    It is known that the microtubules (MT) of Entamoeba histolytica trophozoites form an intranuclear mitotic spindle. However, electron microscopy studies and the employment of anti-beta-tubulin (β-tubulin) antibodies have not exhibited these cytoskeletal structures in the cytoplasm of these parasites. The purpose of this work was to detect β-tubulin in the cytoplasm of interphasic E. histolytica trophozoites. Activated or non-activated HMI-IMSS-strain E. histolytica trophozoites were used and cultured for 72 h at 37 °C in TYI-S-33 medium, and then these were incubated with the anti-β-tubulin antibody of E. histolytica. The anti-β-tubulin antibody reacted with the intranuclear mitotic spindle of E. histolytica-activated trophozoites as control. In contrast, in non-activated interphasic parasites, anti-β-tubulin antibody reacted with diverse puntiform structures in the cytoplasm and with ring-shaped structures localized in the cytoplasm, cellular membrane and endocytic stomas. In this work, for the first time, the presence of β-tubulin is shown in the cytoplasm of E. histolytica trophozoites. Copyright © 2016 Elsevier Inc. All rights reserved.

  13. Invasion of erythrocytes by Babesia bovis

    NARCIS (Netherlands)

    Gaffar, Fasila Razzia

    2004-01-01

    In this thesis we investigated the invasion of erythrocytes taking place during the asexual erythrocytic blood stage of the apicomplexan parasites Babesia bovis parasite. Host cell invasion by apicomplexan parasites is a complex process requiring multiple receptor-ligand interactions, involving

  14. Babesia microti infection, eastern Pennsylvania, USA.

    Science.gov (United States)

    Acosta, Marcela E Perez; Ender, Peter T; Smith, Erin M; Jahre, Jeffrey A

    2013-07-01

    Infection with Babesia microti has not been well-described in eastern Pennsylvania, USA, despite the vector of this organism being prevalent. We report 3 cases of babesiosis in eastern Pennsylvania in persons without recent travel outside the region or history of blood transfusions, suggesting emergence of this infection.

  15. Rhodococcus equi: the many facets of a pathogenic actinomycete.

    Science.gov (United States)

    Vázquez-Boland, José A; Giguère, Steeve; Hapeshi, Alexia; MacArthur, Iain; Anastasi, Elisa; Valero-Rello, Ana

    2013-11-29

    Rhodococcus equi is a soil-dwelling pathogenic actinomycete that causes pulmonary and extrapulmonary pyogranulomatous infections in a variety of animal species and people. Young foals are particularly susceptible and develop a life-threatening pneumonic disease that is endemic at many horse-breeding farms worldwide. R. equi is a facultative intracellular parasite of macrophages that replicates within a modified phagocytic vacuole. Its pathogenicity depends on a virulence plasmid that promotes intracellular survival by preventing phagosome-lysosome fusion. Species-specific tropism of R. equi for horses, pigs and cattle appears to be determined by host-adapted virulence plasmid types. Molecular epidemiological studies of these plasmids suggest that human R. equi infection is zoonotic. Analysis of the recently determined R. equi genome sequence has identified additional virulence determinants on the bacterial chromosome. This review summarizes our current understanding of the clinical aspects, biology, pathogenesis and immunity of this fascinating microbe with plasmid-governed infectivity. Copyright © 2013 Elsevier B.V. All rights reserved.

  16. Giardia muris and Giardia duodenalis groups: ultrastructural differences between the trophozoites.

    Science.gov (United States)

    Sogayar, M I; Gregório, E A

    1989-01-01

    Trophozoites of the Giardia muris group from hamsters, domestic rats and mice and of the Giardia duodenalis group from hamsters and domestic rats were examined by transmission electron microscopy. The basic ultrastructure of the trophozoites was similar. Differences were shown in the morphology of the ventrolateral flange of the trophozoites of Giardia muris and Giardia duodenalis groups. Marginal plates are less developed in the species of the Giardia duodenalis group. In this group, the distal extremity of the lateral flange is short and thick and the marginal plate does not penetrate into the distal extremity of the flange. In the Giardia muris group, the ventro-lateral flange is well developed and narrow and the marginal plate penetrates the distal extremity of the flange. The osmiophilic lamella, which accompanies the dorsal surface of the marginal plate is seen only in the Giardia muris group.

  17. Giardia muris and Giardia duodenalis groups: ultrastructural differences between the trophozoites

    Directory of Open Access Journals (Sweden)

    Maria Inês L. Sogayar

    1989-08-01

    Full Text Available Trophozoites of the Giardia muris group from hamsters, domestic rats and mice and of the Giardia duodenalis group from hamsters and domestic rats were examined by transmission electron microscopy. The basic ultrastructure of the trophozoites was similar. Differences were shown in the morphology of the ventrolateral flange of the trophozoites of Giardia muris and Giardia duodenalis groups. Marginal plates are less developed in the species of the Giardia duodenalis group. In this group, the distal extremity of the lateral flange is short and thick and the marginal plate does not penetrate into the distal extremity of the flange. In the Giardia muris group, the ventro-lateral flange is well developed and narrow and the marginal plate penetrates the distal extremity of the flange. The osmiophilic lamella, which accompanies the dorsal surface of the marginal plate is seen only in the Giardia muris group.

  18. Development and application of loop-mediated isothermal amplification methods targeting the seM gene for detection of Streptococcus equi subsp. equi.

    Science.gov (United States)

    Hobo, Seiji; Niwa, Hidekazu; Oku, Kazuomi

    2012-03-01

    Loop-mediated isothermal amplification (LAMP) constitutes a potentially valuable diagnostic tool for rapid diagnosis of contagious diseases. In this study, we developed a novel LAMP method (seM-LAMP) to detect the seM gene of Streptococcus equi subsp. equi (S. equi), the causative agent of strangles in equids. The seM-LAMP successfully amplified the target sequence of the seM gene at 63°C within 60 min. The sensitivity of the seM-LAMP was slightly lower than the 2nd reaction of the seM semi-nested PCR. To evaluate the species specificity of the seM-LAMP, we tested 100 S. equi and 189 non-S. equi strains. Significant amplification of the DNA originating from S. equi was observed within 60 min incubation, but no amplification of non-S. equi DNA occurred. The results were identical to those of seM semi-nested PCR. To investigate the clinical usefulness of the methods, the seM-LAMP and the seM semi-nested PCR were used to screen 590 nasal swabs obtained during an outbreak of strangles. Both methods showed that 79 and 511 swabs were S. equi positive and negative, respectively, and the results were identical to those of the culture examination. These results indicate that the seM-LAMP is potentially useful for the reliable routine diagnosis of Streptococcus equi subsp. equi infections.

  19. COMPARABLE CYTOLOGICAL DIAGNOSTIC OF BLOOD SMEARS ON BABESIA INFECTION

    OpenAIRE

    Pokhyl S.І.; Torianik І.І.; Tymchenko О.М.; Chygyrynska N.А.; Kostyria І.А.

    2015-01-01

    In last time Babesiosis as a tick-borne hemoprotozoans human disease have a very important role in differentil diagnostics of modern infectious medicine. It caused by protozon of the genus Babesia, which invade and destory erythrocytes. Babesiosis olso has been called tick fever. So, Babesia has been known by other genus names, including Nuttallia, Microbabesia, Babesialla, and Gonderia. Because all Babesia species are piroplasms, a more inclusive term for anthropozoon...

  20. Current State in the Diagnosis of Blood Parasite Babesia SP

    OpenAIRE

    Suteky, Tatik

    2007-01-01

    Babesiosis merupakan penyakit parasit yang disebabkan oleh Babesia sp tidak hanya menyerang berbagai spesies hewan tetapi juga pada manusia. Kasus babesiosis pada orang yang disebabkan oleh Babesia microti telah dilaporkan diberbagai negara. Diagnosis untuk Babesia biasanya dilakukan secara mikroskopis dengan preparat apus darah, namun dengan metode ini sering tidak menunjukkan hasil akurat sehingga metode lain perlu dikembangkan. Metode seperti sub inokulasi pada hewan percobaan dan Quantita...

  1. Molecular evidence of a new Babesia sp. in goats.

    Science.gov (United States)

    Ozubek, Sezayi; Aktas, Munir

    2017-01-15

    In this study, a novel Babesia sp. infecting goats was detected and its phylogenetic relationship to related species was determined. A total of 200 blood samples collected from sheep (n=78) and goats (n=122) were examined in the study. The V4 hypervariable region of the 18S rRNA gene of the novel Babesia sp. was amplified by PCR and analysed using a reverse line blot hybridization assay adapted for small ruminants. Samples from seven goats hybridized to Theileria/Babesia catch-all and Babesia catch-all probes and did not hybridize to any species-specific probe tested, suggesting the presence of an unrecognized Babesia species or genotype. Sequencing results showed the isolate to clearly differ from ovine Babesia species and genotypes currently available in the GenBank database. The isolate showed 90.9%, 93.5%, and 93.4% identity to B. ovis, B. motasi, and B. crassa, respectively and 91-93% similarity to Babesia genotypes recently described in small ruminants. The highest homology (∼96-97%) observed was with Babesia odocoilei, Babesia sp. EU1, and Babesia divergens. The new isolate was provisionally designated Babesia sp. The study contributes to better insight into the distribution and phylogenetic diversity of piroplasms in small ruminants. The survey indicated a high prevalence of piroplasms in small ruminants (21.5%). Of those detected, T. ovis was the most prevalent (17%), followed by Babesia sp. (3.5%), and B. ovis (2%). Copyright © 2016 Elsevier B.V. All rights reserved.

  2. Detection of Babesia spp. in Dogs and Their Ticks From Peninsular Malaysia: Emphasis on Babesia gibsoni and Babesia vogeli Infections in Rhipicephalus sanguineus sensu lato (Acari: Ixodidae).

    Science.gov (United States)

    Prakash, Batah Kunalan; Low, Van Lun; Vinnie-Siow, Wei Yin; Tan, Tiong Kai; Lim, Yvonne Ai-Lian; Morvarid, Akhavan Rezaei; AbuBakar, Sazaly; Sofian-Azirun, Mohd

    2018-05-12

    Canine babesiosis is an emerging tick-borne disease with a worldwide distribution, including Malaysia. While the prevalence of Babesia has been documented from dogs in Malaysia, occurrence of Babesia has been relatively little studied in their tick vectors. Accordingly, a total of 240 dogs and 140 Rhipicephalus sanguineus sensu lato (s.l.) (Acari: Ixodidae) ticks from Malaysia were molecularly screened for the presence of Babesia protozoa in the present study. Babesia gibsoni was only detected in ticks (1.4%), whereas Babesia vogeli was detected in both ticks (1.4%) and dogs (2.1%). This study highlights the detection of B. gibsoni and B. vogeli for the first time, in both adult and nymphal stages of R. sanguineus s.l. in Malaysia, suggesting the potential role of this tick species in transmitting canine babesiosis.

  3. Streptococcus equi subsp zooepidemicus Invades and Survives in Epithelial Cells

    DEFF Research Database (Denmark)

    Skive, Bolette; Rohde, Manfred; Molinari, Gabriella

    2017-01-01

    Streptococcus equi subsp. zooepidemicus (S. zooepidemicus) is an opportunistic pathogen of several species including humans. S. zooepidemicus is found on mucus membranes of healthy horses, but can cause acute and chronic endometritis. Recently S. zooepidemicus was found able to reside in the endo......Streptococcus equi subsp. zooepidemicus (S. zooepidemicus) is an opportunistic pathogen of several species including humans. S. zooepidemicus is found on mucus membranes of healthy horses, but can cause acute and chronic endometritis. Recently S. zooepidemicus was found able to reside...

  4. First molecular evidence of potentially zoonotic Babesia microti and Babesia sp. EU1 in Ixodes ricinus ticks in Belgium

    OpenAIRE

    Lempereur, L.; De Cat, A.; Caron, Y.; Madder, M.; Claerebout, E.; Saegerman, C.; Losson, B.

    2011-01-01

    We report the first molecular evidence of the presence of Babesia sp. EU1 and Babesia microti in Ixodes ricinus ticks in Belgium. A 1-year national survey collected 1005 ticks from cats and dogs. A polymerase chain reaction technique amplifying a part of the 18S rRNA gene detected Babesia spp. in 11 out of 841 selected and validated tick extracts. Subsequent sequencing identified Ba. microti (n = 3) and Babesia sp. EU1 (n = 6). This study has demonstrated a low infection rate (1.31% with 95% ...

  5. Severe human Babesia divergens infection in Norway

    Directory of Open Access Journals (Sweden)

    K. Mørch

    2015-04-01

    Full Text Available Human babesiosis is a rare but potentially life-threatening parasitic disease transmitted by ixodid ticks, and has not previously been reported in Norway. We report a case of severe babesiosis that occurred in Norway in 2007. The patient had previously undergone a splenectomy. He was frequently exposed to tick bites in an area endemic for bovine babesiosis in the west of Norway. The patient presented with severe haemolysis and multiorgan failure. Giemsa-stained blood smears revealed 30% parasitaemia with Babesia spp. He was treated with quinine in combination with clindamycin, apheresis, and supportive treatment with ventilatory support and haemofiltration, and made a complete recovery. This is the first case reported in Norway; however Babesia divergens seroprevalence in cattle in Norway is high, as is the risk of Ixodes ricinus tick bite in the general population. Babesiosis should be considered in the differential diagnosis of unexplained febrile haemolytic disease.

  6. Severe human Babesia divergens infection in Norway.

    Science.gov (United States)

    Mørch, K; Holmaas, G; Frolander, P S; Kristoffersen, E K

    2015-04-01

    Human babesiosis is a rare but potentially life-threatening parasitic disease transmitted by ixodid ticks, and has not previously been reported in Norway. We report a case of severe babesiosis that occurred in Norway in 2007. The patient had previously undergone a splenectomy. He was frequently exposed to tick bites in an area endemic for bovine babesiosis in the west of Norway. The patient presented with severe haemolysis and multiorgan failure. Giemsa-stained blood smears revealed 30% parasitaemia with Babesia spp. He was treated with quinine in combination with clindamycin, apheresis, and supportive treatment with ventilatory support and haemofiltration, and made a complete recovery. This is the first case reported in Norway; however Babesia divergens seroprevalence in cattle in Norway is high, as is the risk of Ixodes ricinus tick bite in the general population. Babesiosis should be considered in the differential diagnosis of unexplained febrile haemolytic disease. Copyright © 2015. Published by Elsevier Ltd.

  7. Eugregarine trophozoite detachment from the host epithelium via epimerite retraction: Fiction or fact?

    Czech Academy of Sciences Publication Activity Database

    Valigurová, A.; Michalková, V.; Koudela, Břetislav

    2009-01-01

    Roč. 39, č. 11 (2009), s. 1235-1242 ISSN 0020-7519 R&D Projects: GA MŠk LC522 Institutional research plan: CEZ:AV0Z60220518 Keywords : Apicomplexa * epimerite retraction * trophozoite Subject RIV: GJ - Animal Vermins ; Diseases, Veterinary Medicine Impact factor: 3.819, year: 2009

  8. Molecular detection of Theileria and Babesia infections in cattle.

    Science.gov (United States)

    Altay, Kursat; Aydin, M Fatih; Dumanli, Nazir; Aktas, Munir

    2008-12-20

    This study was carried out to determine the presence and distribution of tick-borne haemoprotozoan parasites (Theileria and Babesia) in apparently healthy cattle in the East Black Sea Region of Turkey. A total of 389 blood samples were collected from the animals of various ages in six provinces in the region. Prevalence of infection was determined by reverse line blot (RLB) assay. The hypervariable V4 region of the 18S ribosomal RNA (rRNA) gene was amplified with a set of primers for members of the genera Theileria and Babesia. Amplified PCR products were hybridized onto a membrane to which generic- and species-specific oligonucleotide probes were covalently linked. RLB hybridization identified infection in 16.19% of the samples. Blood smears were also examined microscopically for Theileria and/or Babesia spp. and 5.14% were positive. All samples shown to be positive by microscopy also tested positive with RLB assay. Two Theileria (T. annulata and T. buffeli/orientalis) and three Babesia (B. bigemina, B. major and Babesia sp.) species or genotypes were identified in the region. Babesia sp. genotype shared 99% similarity with the previously reported sequences of Babesia sp. Kashi 1, Babesia sp. Kashi 2 and Babesia sp. Kayseri 1. The most frequently found species was T. buffeli/orientalis, present in 11.56% of the samples. T. annulata was identified in five samples (1.28%). Babesia infections were less frequently detected: B. bigemina was found in three samples (0.77%), B. major in two samples (0.51%) and Babesia sp. in five samples (1.28%). A single animal infected with T. buffeli/orientalis was also infected with B. bigemina.

  9. Stimulation and quantification of Babesia divergens gametocytogenesis

    Czech Academy of Sciences Publication Activity Database

    Jalovecká, Marie; Bonsergent, C.; Hajdušek, Ondřej; Kopáček, Petr; Malandrin, L.

    2016-01-01

    Roč. 9, č. 1 (2016), č. článku 439. ISSN 1756-3305 R&D Projects: GA ČR GA13-11043S; GA ČR GP13-27630P; GA ČR GJ15-12006Y Institutional support: RVO:60077344 Keywords : Babesia divergens * gametocytes * transmission * bdccp genes * qRT-PCR Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 3.080, year: 2016

  10. Peritonitis in a llama caused by Streptococcus equi subsp. zooepidemicus.

    Science.gov (United States)

    Hewson, J; Cebra, C K

    2001-01-01

    A 7-month-old, male llama was diagnosed with peritonitis caused by Streptococcus equi subsp. zooepidemicus. Clinical findings, medical treatment, and case outcome are described. Hematogenous dissemination from suspected pneumonia is proposed as the route of infection in this case. Possible transmission of the organism through contact with horses is discussed. PMID:11424579

  11. Severe Rhodococcus equi pneumonia: case report and literature review

    DEFF Research Database (Denmark)

    Vestbo, Jørgen; Lundgren, Jens Dilling; Gaub, J

    1991-01-01

    or sputum on standard media, but is frequently regarded as a contaminant. Mortality from Rhodococcus equi pneumonia is high (25%) and early surgical intervention has been recommended. Based on this review, the benefit of surgery seems dubious, whereas good results have been obtained using long......-term antibiotic treatment with erythromycin plus rifampicin, or vancomycin in combination with either of these antibiotics....

  12. Efficacy of a Parapoxvirus ovis-based immunomodulator against equine herpesvirus type 1 and Streptococcus equi equi infections in horses.

    Science.gov (United States)

    Ons, Ellen; Van Brussel, Leen; Lane, Stephen; King, Vickie; Cullinane, Ann; Kenna, Rachel; Lyons, Pamela; Hammond, Toni-Ann; Salt, Jeremy; Raue, Rudiger

    2014-10-10

    The efficacy of Zylexis®, an immunomodulator in horses based on inactivated Parapoxvirus ovis (iPPVO), was assessed using an equine herpesvirus type 1 (EHV-1) challenge model in the presence of a natural infection with Streptococcus equi equi (S. equi). Eleven horses were treated with iPPVO and twelve were kept as controls. Six horses were challenged with EHV-1 and commingled with the horses on study. Animals were dosed on Days -2, 0 (just before commingling) and Day 7. On Day 11 significantly less nasal discharge, enlarged lymph nodes, EHV-1 shedding and lower rectal temperatures were observed in the iPPVO-treated group. In addition, iPPVO-treated horses showed significantly fewer enlarged lymph nodes on Days 17 and 19, significantly less lower jaw swelling on Day 3 and significantly lower rectal temperatures on Days 12 and 13. Dyspnoea, depression and anorexia were only recorded for the control group. Following challenge seven out of 11 horses in the iPPVO treated group shed EHV-1 but on Days 11, 12, 13, 14, 15 and 16 quantitative virus detection in this group was significantly lower as compared to the controls. All animals shed S. equi but the percentage of animals with positive bacterial detection was lower in the iPPVO group than in the control group from Day 14 through Day 28. This difference was significant on Day 24. No injection site reactions or adverse events were observed. In conclusion, Zylexis administration is safe and reduced clinical signs and shedding related to both EHV-1 and S. equi infections. Copyright © 2014 Elsevier B.V. All rights reserved.

  13. First molecular evidence of potentially zoonotic Babesia microti and Babesia sp. EU1 in Ixodes ricinus ticks in Belgium.

    Science.gov (United States)

    Lempereur, Laetitia; De Cat, Ann; Caron, Yannick; Madder, Maxime; Claerebout, Edwin; Saegerman, Claude; Losson, Bertrand

    2011-02-01

    We report the first molecular evidence of the presence of Babesia sp. EU1 and Babesia microti in Ixodes ricinus ticks in Belgium. A 1-year national survey collected 1005 ticks from cats and dogs. A polymerase chain reaction technique amplifying a part of the 18S rRNA gene detected Babesia spp. in 11 out of 841 selected and validated tick extracts. Subsequent sequencing identified Ba. microti (n=3) and Babesia sp. EU1 (n=6). This study has demonstrated a low infection rate (1.31% with 95% CI: 0.65-2.33) of Babesia spp. carriage in I. ricinus ticks in Belgium but, for the first time, reports two potentially zoonotic species belonging to this genus. Coinfection with Ba. microti and Borrelia burgdorferi sensu stricto also was demonstrated. In addition, this study clearly demonstrates that inhibitors of polymerase chain reaction amplification are present in engorged ticks.

  14. Entamoeba histolytica and E. dispar trophozoites in the liver of hamsters: in vivo binding of antibodies and complement

    Directory of Open Access Journals (Sweden)

    Gomes Maria A

    2010-03-01

    Full Text Available Abstract Background Human amoebiasis is caused by the parasitic protozoan Entamoeba histolytica that lives in the large intestine of hosts, where can produce asymptomatic colonization until severe invasive infections with blood diarrhea and spreading to other organs. The amoebic abscesses in liver are the most frequent form of amoebiasis outside intestine and still there are doubts about the pathogenic mechanisms involved in their formation. In this study we evaluated the in situ binding of antibodies, C3 and C9 complement components on trophozoites, in livers of hamsters infected with E. histolytica or E. dispar. These parameters were correlated with the extension of the hepatic lesions observed in these animals and with trophozoites survivor. Methods Hamsters were inoculated intra-hepatically with 100,000 trophozoites of E. histolytica or E. dispar strain and necropsied 12, 24, 48, 72, 144 and 192 h after inoculation. Antibodies, C3 and C9 binding to trophozoites were detected by immunohistochemistry. The estimation of the necrosis area and the number of labeled trophozoites was performed using digital morphometry analysis. Results In the liver sections of animals inoculated with the amoebas, the binding of antibodies to E. histolytica trophozoites was significantly lower than to E. dispar trophozoites. Trophozoites of E. dispar were also more frequently vacuolated and high labeled cellular debris observed in the lesions. Positive diffuse reaction to C3 complement component was more intense in livers of animals inoculated with E. histolytica after 24 and 72 h of infection. C3(+ and C9(+ trophozoites were detected in the vascular lumen, granulomas and inside and in the border of necrotic areas of both infected group animals. C3(+ and C9(+ trophozoite debris immunostaining was higher in livers of E. dispar than in livers of E. histolytica. A positive correlation between necrotic areas and number of C9(+ trophozoites was observed in animals

  15. Amblyomma cajennense is an intrastadial biological vector of Theileria equi.

    Science.gov (United States)

    Scoles, Glen A; Ueti, Massaro W

    2013-10-23

    The apicomplexan hemoprotozoan parasite Theileria equi is one of the etiologic agents causing equine piroplasmosis, a disease of equines that is endemic throughout large parts of the world. Before 2009 the United States had been considered to be free of this parasite. Occasional cases had occurred but there was no evidence for endemic vector-borne transmission in the U.S. until a 2009 outbreak in Texas in which Dermacentor variabilis and Amblyomma cajennense were implicated as vectors. Although D. variabilis has previously been shown to be a competent laboratory vector, studies suggested A. cajennense was not a competent transstadial vector, even though the presence of this tick species on horses in South American is epidemiologicaly correlated with higher a prevalence of infection. In this study we tested the transstadial and intrastadial vector competence of D. variabilis and A. cajennense for T. equi. A tick passaged T. equi strain from the Texas outbreak and ticks colonized from engorged females collected off horses on the outbreak ranch in Texas were used for these studies. Nymph or adult ticks were fed on infected horses and transmission fed on naïve horses. Infections were tracked with PCR and serology, dissected tick tissues were tested with PCR. A. cajennense transmitted T. equi intrastadially when adult ticks acquired infection by feeding on an infected horse, and transmitted to a naïve host on subsequent reattachment and feeding. D. variabilis failed to transmit in the same experiment. Transstadial transmission was not successful for either tick species. PCR on DNA isolated from eggs of females that had fed on an infected horse suggests that there is no transovarial passage of this parasite by either tick species. This work confirms that ticks from the Texas population of A. cajennense are competent intrastadial vectors of T. equi. We propose that the most likely natural mode of transmission for this parasite/vector combination in the Texas outbreak

  16. Babesia spp. in European wild ruminant species: parasite diversity and risk factors for infection.

    Science.gov (United States)

    Michel, Adam O; Mathis, Alexander; Ryser-Degiorgis, Marie-Pierre

    2014-06-13

    Babesia are tick-borne parasites that are increasingly considered as a threat to animal and public health. We aimed to assess the role of European free-ranging wild ruminants as maintenance mammalian hosts for Babesia species and to determine risk factors for infection. EDTA blood was collected from 222 roe deer (Capreolus c. capreolus), 231 red deer (Cervus e. elaphus), 267 Alpine chamois (Rupicapra r. rupicapra) and 264 Alpine ibex (Capra i. ibex) from all over Switzerland and analysed by PCR with pan-Babesia primers targeting the 18S rRNA gene, primers specific for B. capreoli and Babesia sp. EU1, and by sequencing. Babesia species, including B. divergens, B. capreoli, Babesia sp. EU1, Babesia sp. CH1 and B. motasi, were detected in 10.7% of all samples. Five individuals were co-infected with two Babesia species. Infection with specific Babesia varied widely between host species. Cervidae were significantly more infected with Babesia spp. than Caprinae. Babesia capreoli and Babesia sp. EU1 were mostly found in roe deer (prevalences 17.1% and 7.7%, respectively) and B. divergens and Babesia sp. CH1 only in red deer. Factors significantly associated with infection were low altitude and young age. Identification of Babesia sp. CH1 in red deer, co-infection with multiple Babesia species and infection of wild Caprinae with B. motasi and Babesia sp. EU1 are novel findings. We propose wild Caprinae as spillover or accidental hosts for Babesia species but wild Cervidae as mammalian reservoir hosts for B. capreoli, possibly Babesia sp. EU1 and Babesia sp. CH1, whereas their role regarding B. divergens is more elusive.

  17. Interaction of Escherichia coli K1 and K5 with Acanthamoeba castellanii Trophozoites and Cysts

    OpenAIRE

    Matin, Abdul; Jung, Suk-Yul

    2011-01-01

    The existence of symbiotic relationships between Acanthamoeba and a variety of bacteria is well-documented. However, the ability of Acanthamoeba interacting with host bacterial pathogens has gained particular attention. Here, to understand the interactions of Escherichia coli K1 and E. coli K5 strains with Acanthamoeba castellanii trophozoites and cysts, association assay, invasion assay, survival assay, and the measurement of bacterial numbers from cysts were performed, and nonpathogenic E. ...

  18. In Vitro Toxicity Evaluation of Silver Nanoparticles on Entamoeba histolytica trophozoite

    Directory of Open Access Journals (Sweden)

    Zahra’a A. Ahmed

    2017-09-01

    Full Text Available The protozoan parasite Entamoeba histolytica is a causative agent of amoebiasis, where it causes millions of cases of dysentery and liver abscess each year. Metronidazole is a drug of choice against amoebiasis. The drug is a choice because of its efficacy and low cost, but at the same time it causes several adverse side effects; therefore, it is important to find effective medications to treat amoebiasis without any complications or any side effects. The aim of this study is to evaluate the effectiveness of different concentrations (50, 75 and 100 µg/ml of silver nanoparticle (AgNPs against trophozoites stages of E. histolytica in vitro. The results showed a significant decrease (p ≤ 0.05 in numbers of trophozoites stages after treated with AgNPs and metronidazole when it was compared with the control. Likewise, a significant difference (p ≤ 0.05 was also observed between AgNPs groups and metronidazole drug, while it did not significantly differ between different concentrations of AgNPs. The mortality rate values of the E. histolytica trophozoites after 48h incubation with AgNPs at a concentration of 50, 75 and 100 μg/ml, and metronidazole were 37.2%, 42.4%, 46.7% and 100%, respectively. The microscopic studies confirmed that AgNPs were effective enough to induce apoptosis. Based on our results, the anti-parasitic activity of AgNPs at different concentrations will reduce the mean number of E. histolytica trophozoites.

  19. Atomic force microscopic imaging of Acanthamoeba castellanii and Balamuthia mandrillaris trophozoites and cysts.

    Science.gov (United States)

    Aqeel, Yousuf; Siddiqui, Ruqaiyyah; Ateeq, Muhammad; Raza Shah, Muhammad; Kulsoom, Huma; Khan, Naveed Ahmed

    2015-01-01

    Light microscopy and electron microscopy have been successfully used in the study of microbes, as well as free-living protists. Unlike light microscopy, which enables us to observe living organisms or the electron microscope which provides a two-dimensional image, atomic force microscopy provides a three-dimensional surface profile. Here, we observed two free-living amoebae, Acanthamoeba castellanii and Balamuthia mandrillaris under the phase contrast inverted microscope, transmission electron microscope and atomic force microscope. Although light microscopy was of lower magnification, it revealed functional biology of live amoebae such as motility and osmoregulation using contractile vacuoles of the trophozoite stage, but it is of limited value in defining the cyst stage. In contrast, transmission electron microscopy showed significantly greater magnification and resolution to reveal the ultra-structural features of trophozoites and cysts including intracellular organelles and cyst wall characteristics but it only produced a snapshot in time of a dead amoeba cell. Atomic force microscopy produced three-dimensional images providing detailed topographic description of shape and surface, phase imaging measuring boundary stiffness, and amplitude measurements including width, height and length of A. castellanii and B. mandrillaris trophozoites and cysts. These results demonstrate the importance of the application of various microscopic methods in the biological and structural characterization of the whole cell, ultra-structural features, as well as surface components and cytoskeleton of protist pathogens. © 2014 The Author(s) Journal of Eukaryotic Microbiology © 2014 International Society of Protistologists.

  20. A new quantitative in vitro microculture method for Giardia duodenalis trophozoites.

    Science.gov (United States)

    Bénéré, Ely; da Luz, Raquel Andreia Inocêncio; Vermeersch, Marieke; Cos, Paul; Maes, Louis

    2007-11-01

    A reliable, rapid and low-cost method for drug sensitivity determination of Giardia duodenalis trophozoites (WB-strain) was developed in a 96-well plate. Using a standard inoculum of 5 x 10(4) trophozoites per well (300 microl), good growth was obtained after sealing the plate with an air-tight adhesive tape and incubation at 37 degrees C for 72 h in modified TYI-S-33 medium. Viable burdens were quantified using the formazan dyes MTT (100 microg/well) and XTT (20 microg/well) and the fluorescent substrate resazurin (2.5 microg/well). Prior removal of the culture medium is required since it causes spontaneous reduction of the substrate. Resazurin proved to be far superior to MTT and XTT with a level of sensitivity of about 3 x 10(4) trophozoites. Inhibitory concentrations (IC(50)) of several anti-giardial reference drugs were within the range of published values: metronidazole 2.25 microM, tinidazole 1.75 microM, albendazole 0.10 microM, furazolidone 2.00 microM and quinacrine 0.32 microM. The broad-spectrum antibiotics chloramphenicol, rifampicin, penicillin G+streptomycin and gentamycin were devoid of any inhibitory activity and are considered suitable for decontamination during excystation experiments.

  1. A new PCR assay for the detection and differentiation of Babesia canis and Babesia vogeli.

    Science.gov (United States)

    Annoscia, Giada; Latrofa, Maria Stefania; Cantacessi, Cinzia; Olivieri, Emanuela; Manfredi, Maria Teresa; Dantas-Torres, Filipe; Otranto, Domenico

    2017-10-01

    Babesia spp. are globally distributed tick-borne protozoan parasites that infect the red blood cells of a wide range of vertebrate hosts, including humans. Diagnosis of babesiosis is often impeded by the transient presence of the parasites in peripheral blood, as well as by their pleomorphic nature. Given the reports of an expanding and, in some cases, sympatric geographical distribution of Babesia canis and Babesia vogeli in dogs and associated vectors, in Europe, the development of time-efficient and cost-effective diagnostic tools to detect and differentiate these two species is warranted. In this study, we designed and developed a novel polymerase chain reaction (PCR) assay targeting the parasite cytochrome c oxidase subunit 1 (cox1) gene, for the simultaneous detection and differentiation of B. canis and B. vogeli. The analytical sensitivity of the PCR was evaluated using serial dilutions of genomic DNA extracted from individual and artificially mixed canine blood samples infected by B. canis (3×10 2 infected erythrocytes/ml, ie/ml) and B. vogeli (2.1×10 1 ie/ml). The analytical specificity of the assay was assessed using blood samples positive for Hepatozoon canis, Ehrlichia canis, Anaplasma platys, Babesia microti, Babesia rossi and Theileria annae (syn. Babesia vulpes). The clinical specificity of the PCR assay was evaluated on 147 blood samples from dogs and 128 tick specimens (Dermacentor reticulatus and Rhipicephalus sanguineus sensu lato). Species-specific bands of the expected sizes (i.e., 750bp for B. canis and 450bp for B. vogeli), and two bands in the mixed blood samples were obtained. The PCR assay developed herein detected a low number of infected erythrocytes (i.e., 3×10 -2 B. canis, 2.1×10 -2 B. vogeli ie/ml). Of the 147 blood samples, nine (6.1%) were positive for B. canis and six (4.1%) for B. vogeli, whereas only one tick (D. reticulatus) was positive for B. canis. This PCR assay represents a rapid and reliable tool for the diagnosis of B

  2. First evidence of Babesia venatorum and Babesia capreoli in questing Ixodes ricinus ticks in the Czech Republic.

    Science.gov (United States)

    Venclikova, Kristyna; Mendel, Jan; Betasova, Lenka; Hubalek, Zdenek; Rudolf, Ivo

    2015-01-01

    Ixodes ricinus is the most common tick species occurring in Central Europe and it serves as a principal vector of emerging human pathogens. The aim of this study was to determine the prevalence of Babesia spp. in host-seeking I. ricinus in urban and natural habitats. PCR was applied on samples to assess prevalence of Babesia spp. in questing ixodid ticks. Sequencing was used for Babesia species determination. 1,473 I. ricinus ticks (1,294 nymphs, 99 males and 80 females) were examined for the presence of Babesia spp. at the two study sites. Minimum infection rate for Babesia spp. was found to be 0.5% (infected I. ricinus nymphs were only detected in the natural ecosystem). Two Babesia species were identified by sequencing: B. venatorum (formerly called Babesia sp. EU1) and B. capreoli. The results obtained represent the first evidence of the occurrence of B. venatorum and B. capreoli in host-seeking I. ricinus ticks in the Czech Republic.

  3. Serological survey of Rhodococcus equi infection in horses in Hokkaido.

    Science.gov (United States)

    Sanada, Y; Noda, H; Nagahata, H

    1992-08-01

    Serological survey of Rhodococcus equi infection in horses in Hokkaido was performed using ELISA. Of 2,879 horse sera, 318 (11.0%) gave antibody-positive (OD greater than or equal to 0.3) reactions. The antibody-positive rate of female was significantly higher (p less than 0.01) than that of male, and no statistical difference between Anglo-Arab and thoroughbred was detected in the antibody-positive rate. The maximum antibody-positive rate (27.1%) was shown at 14 years of age. The antibody-positive rates on the 160 farms were found to vary widely from 0 to 78.9%. A significant difference (p less than 0.01) in the antibody-positive rate was detected among horse farms. It was elucidated that 100 (62.5%) out of 160 horse farms had an antibody-positive horse. These results indicate that R. equi was widespread on horse farms, and the level of environmental contamination with R. equi differed among horse farms.

  4. Immunogenicity of an electron beam inactivated Rhodococcus equi vaccine in neonatal foals.

    Directory of Open Access Journals (Sweden)

    Angela I Bordin

    Full Text Available Rhodococcus equi is an important pathogen of foals that causes severe pneumonia. To date, there is no licensed vaccine effective against R. equi pneumonia of foals. The objectives of our study were to develop an electron beam (eBeam inactivated vaccine against R. equi and evaluate its immunogenicity. A dose of eBeam irradiation that inactivated replication of R. equi while maintaining outer cell wall integrity was identified. Enteral administration of eBeam inactivated R. equi increased interferon-γ production by peripheral blood mononuclear cells in response to stimulation with virulent R. equi and generated naso-pharyngeal R. equi-specific IgA in newborn foals. Our results indicate that eBeam irradiated R. equi administered enterally produce cell-mediated and upper respiratory mucosal immune responses, in the face of passively transferred maternal antibodies, similar to those produced in response to enteral administration of live organisms (a strategy which previously has been documented to protect foals against intrabronchial infection with virulent R. equi. No evidence of adverse effects was noted among vaccinated foals.

  5. Babesia bigemina: in vitro cultivation and characterization

    International Nuclear Information System (INIS)

    Vega Y Murguia, C.A.

    1985-01-01

    An in vitro model for the continuous replication of Babesia bigemina was developed and this model was used to study the parasite's biology. Initially, infected erythrocytes from a calf inoculated with a strain of B. bigemina was suspended with normal bovine erythrocytes and the parasite propagated in vitro. The cultured organism was inoculated into another calf and reproduced the disease with typical signs. Babesia bigemina was reisolated in pure culture in vitro. The animal recovered after receiving specific treatment. A procedure was developed to cryopreserve infected erythrocytes and merozoites to initiate in vitro cultures. Homogeneous parasite populations were obtained by cloning by limiting dilution. Parasitic growth was detected between 16-28 days after dilutions were made. Three primary clones were selected for recloning. Infected erythrocytes from the original isolate nd the clones were concentrated by Percoll density gradients. Density values for paired and single infected cells were determined. Enzymatic content of concentrated infected cells was analyzed by starch gel electrophoresis. Enzymes LDH, GPI, and GDH were detected, but polymorphism among clones was not observed. The enzyme 6-PDG was not associated with the parasite. Separation of labelled proteins was done by SDS-PAGE. The separation patterns were similar for all samples. a 43 Kd polypeptide was detected in the B. bigemina culture supernatant

  6. Different Babesia canis isolates, different diseases.

    Science.gov (United States)

    Schetters, T P; Moubri, K; Précigout, E; Kleuskens, J; Scholtes, N C; Gorenflot, A

    1997-11-01

    Using surface immunofluorescence isolate-specific antigens were detected on the membrane of erythrocytes infected with Babesia parasites. In addition, the strains reacted differently with Plasmagel in that the European isolate (B.c. canis) could be purified on Plasmagel effectively, whereas infected erythrocytes of the South-African isolate (B.c. rossi) could not. Experimental infection of dogs with Babesia canis isolates from geographically different areas revealed different pathology. The European isolate obtained from France exhibited transient parasitaemia, usually below 1%, associated with low PCV values and congestion of internal organs. Clinical disease was correlated with an effect on the coagulation system, and not with peripheral parasitaemia. Infection of dogs with South-African-derived isolate induced high parasitaemia usually much higher than 1%, which required chemotherapeutic treatment. In these animals clinical disease was correlated with peripheral parasitaemia and not with parameters of the coagulation system. The results show that the etiology of disease caused by these isolates of B.c. canis and B.c. rossi is different. This might have implications for the development of vaccines against these infections.

  7. Characteristics, immunological events, and diagnostics of Babesia spp. infection, with emphasis on Babesia canis

    Directory of Open Access Journals (Sweden)

    Kostro Krzysztof

    2015-12-01

    Full Text Available Vector-borne infection constitutes a significant health issue in dogs worldwide. Recent reports point to an increasing number of canine vector-borne disease cases in European countries, including Poland. Canine babesiosis caused by various Babesia species is a protozoal tick-borne disease with worldwide distribution and significant veterinary importance. The development and application of molecular methods have increased our knowledge about canine babesiosis, its prevalence, and clinical and pathological aspects of the infection. Parasitologists and veterinary surgeons need an accurate description of the species responsible for canine babesiosis to improve diagnostic and therapeutic methods, as well as predictions for the course of the disease. Therefore, we decided to summarise recent knowledge concerning Babesia species and B. canis.

  8. Identification of Babesia canis genotypes in dogs from Lithuania

    OpenAIRE

    Tamoliūnaitė, Dovilė; Radzijevskaja, Jana; Paulauskas, Algimantas; Sabūnas, Vytautas; Karvelienė, Birutė; Zamokas, Gintaras

    2018-01-01

    Canine babesiosis is a widespread tick-borne disease caused by haematozoan parasites of the genus Babesia. The vast majority of clinical babesiosis cases in dogs in Europe is caused by Babesia canis. Canine babesiosis has become quite frequent in Lithuania during the past decade. Babesiosis caused by B. canis may range from mild to severe disease in dogs. Such difference in the virulence of B. canis strains is associated with genetic heterogeneity among B. canis strains. We aimed to investiga...

  9. Babesia microti, human babesiosis, and Borrelia burgdorferi in Connecticut.

    OpenAIRE

    Anderson, J F; Mintz, E D; Gadbaw, J J; Magnarelli, L A

    1991-01-01

    Babesia microti was isolated from a white-footed mouse (Peromyscus leucopus) that was captured in southeastern Connecticut in 1988, when the first human case of babesiosis acquired in Connecticut was recognized. To date, 13 cases of babesiosis have been reported in Connecticut, the largest number of human cases reported on the mainland United States. Two of nine patients quiried remembered a prior tick bite. Since Babesia parasites are known to be vectored only by ticks, we surmise that 12 of...

  10. Failure of pulmonary clearance of Rhodococcus equi infection in CD4+ T-lymphocyte-deficient transgenic mice.

    OpenAIRE

    Kanaly, S T; Hines, S A; Palmer, G H

    1993-01-01

    Pulmonary clearance of Rhodococcus equi requires functional T lymphocytes. In this study, CD8+ T-lymphocyte-deficient transgenic mice cleared virulent R. equi from the lungs while infection in CD4+ T-lymphocyte-deficient transgenic mice persisted. Although both CD4+ and CD8+ T cells function early in pulmonary defense against R. equi, clearance is dependent on CD4+ T lymphocytes.

  11. EQUINOS PORTADORES de Streptococcus equi subespécie equi: PREVALÊNCIA, FATORES DE RISCO E CARACTERIZAÇÃO DE ALELOS seM

    OpenAIRE

    Felipe Libardoni

    2015-01-01

    A adenite equina é uma doença infecto-contagiosa que acomete o trato respiratório superior, sendo uma das principais doenças respiratórias de equinos. O agente etiológico dessa enfermidade é o Streptococcus equi subespécie equi (S. equi), responsável por aproximadamente 30% das notificações em todo o mundo. Os principais sinais clínicos da adenite são febre, secreção nasal e enfartamento de linfonodos, que ocorre pela dificuldade de fagocitose do S. equi por células de defesa devido a presenç...

  12. Diversity of Babesia in Ixodes ricinus ticks in Poland.

    Science.gov (United States)

    Welc-Falęciak, R; Bajer, A; Paziewska-Harris, A; Baumann-Popczyk, A; Siński, E

    2012-01-01

    The aims of this study were: (1) to estimate Babesia prevalence in the most common species of tick in Poland, Ixodes ricinus, in two recreational areas (Urwitałt in the Mazury Lake District and Bielański Forest in Warsaw), and (2) to evaluate the molecular diversity of Babesia isolates in questing I. ricinus in Poland. Questing ticks were collected from vegetation in forest areas in Urwitałt near Mikołajki and in Bielański Forest (Warsaw). Purified genomic DNA was used with specific primers to amplify a fragment of the Babesia spp. 18S rRNA gene. Tick-drag indices for I. ricinus were high in both study areas, reaching somewhat higher values in Urwitałt than in Bielański Forest. The overall prevalence of Babesia spp. in examined ticks was 1.6%. In Urwitałt, two strains of B. microti were identified using rRNA sequences: the enzootic Munich strain and an isolate close to the zoonotic Jena strain. The proportion of infections due to these two strains in questing ticks reversed over a six-year period. During 3 years of study in Bielański Forest, all Babesia isolates obtained from I. ricinus were identical to Babesia sp. EU1 (B. venatorum), previously recognized as an agent of human babesiosis. This study has confirmed the presence of enzoonotic and zoonotic Babesia species/strains in the abundant human-biting tick I. ricinus in recreational areas in Poland. It has also shown that the distribution of different genotypes has changed over time, however the reasons for these fluctuations still remain to be investigated.

  13. Babesia spp. in ticks and wildlife in different habitat types of Slovakia.

    Science.gov (United States)

    Hamšíková, Zuzana; Kazimírová, Mária; Haruštiaková, Danka; Mahríková, Lenka; Slovák, Mirko; Berthová, Lenka; Kocianová, Elena; Schnittger, Leonhard

    2016-05-20

    Babesiosis is an emerging and potentially zoonotic disease caused by tick-borne piroplasmids of the Babesia genus. New genetic variants of piroplasmids with unknown associations to vectors and hosts are recognized. Data on the occurrence of Babesia spp. in ticks and wildlife widen the knowledge on the geographical distribution and circulation of piroplasmids in natural foci. Questing and rodent-attached ticks, rodents, and birds were screened for the presence of Babesia-specific DNA using molecular methods. Spatial and temporal differences of Babesia spp. prevalence in ticks and rodents from two contrasting habitats of Slovakia with sympatric occurrence of Ixodes ricinus and Haemaphysalis concinna ticks and co-infections of Candidatus N. mikurensis and Anaplasma phagocytophilum were investigated. Babesia spp. were detected in 1.5 % and 6.6 % of questing I. ricinus and H. concinna, respectively. Prevalence of Babesia-infected I. ricinus was higher in a natural than an urban/suburban habitat. Phylogenetic analysis showed that Babesia spp. from I. ricinus clustered with Babesia microti, Babesia venatorum, Babesia canis, Babesia capreoli/Babesia divergens, and Babesia odocoilei. Babesia spp. amplified from H. concinna segregated into two monophyletic clades, designated Babesia sp. 1 (Eurasia) and Babesia sp. 2 (Eurasia), each of which represents a yet undescribed novel species. The prevalence of infection in rodents (with Apodemus flavicollis and Myodes glareolus prevailing) with B. microti was 1.3 % in an urban/suburban and 4.2 % in a natural habitat. The majority of infected rodents (81.3 %) were positive for spleen and blood and the remaining for lungs and/or skin. Rodent-attached I. ricinus (accounting for 96.3 %) and H. concinna were infected with B. microti, B. venatorum, B. capreoli/B. divergens, Babesia sp. 1 (Eurasia), and Babesia sp. 2 (Eurasia). All B. microti and B. venatorum isolates were identical to known zoonotic strains from Europe. Less than 1

  14. In vitro induction of Entamoeba histolytica cyst-like structures from trophozoites.

    Directory of Open Access Journals (Sweden)

    Hugo Aguilar-Díaz

    Full Text Available Inhibition of encystment can be conceived as a potentially useful mechanism to block the transmission of Entamoeba histolytica under natural conditions. Unfortunately, amoeba encystment has not been achieved in vitro and drugs inhibiting the formation of cysts are not available. Luminal conditions inducing encystment in vivo are also unknown, but cellular stress such as exposure to reactive oxygen species from immune cells or intestinal microbiota could be involved. A role for certain divalent cations as cofactors of enzymes involved in excystment has also been described. In this study, we show that trophozoite cultures, treated with hydrogen peroxide in the presence of trace amounts of several cations, transform into small-sized spherical and refringent structures that exhibit resistance to different detergents. Ultrastructural analysis under scanning and transmission electron microscopy revealed multinucleated structures (some with four nuclei with smooth, thick membranes and multiple vacuoles. Staining with calcofluor white, as well as an ELISA binding assay using wheat germ agglutinin, demonstrated the presence of polymers of N-acetylglucosamine (chitin, which is the primary component of the natural cyst walls. Over-expression of glucosamine 6-phosphate isomerase, likely to be the rate-limiting enzyme in the chitin synthesis pathway, was also confirmed by RT-PCR. These results suggest that E. histolytica trophozoites activated encystment pathways when exposed to our treatment.

  15. Curcumin alters the cytoskeleton and microtubule organization on trophozoites of Giardia lamblia.

    Science.gov (United States)

    Gutiérrez-Gutiérrez, Filiberto; Palomo-Ligas, Lissethe; Hernández-Hernández, José Manuel; Pérez-Rangel, Armando; Aguayo-Ortiz, Rodrigo; Hernández-Campos, Alicia; Castillo, Rafael; González-Pozos, Sirenia; Cortés-Zárate, Rafael; Ramírez-Herrera, Mario Alberto; Mendoza-Magaña, María Luisa; Castillo-Romero, Araceli

    2017-08-01

    Giardia lamblia is a worldwide protozoan responsible for a significant number of intestinal infections. There are several drugs for the treatment of giardiasis, but they often cause side effects. Curcumin, a component of turmeric, has antigiardial activity; however, the molecular target and mechanism of antiproliferative activity are not clear. The effects of curcumin on cellular microtubules have been widely investigated. Since tubulin is the most abundant protein in the cytoskeleton of Giardia, to elucidate whether curcumin has activity against the microtubules of this parasite, we treated trophozoites with curcumin and the cells were analyzed by scanning electron microscopy and confocal microscopy. Curcumin inhibited Giardia proliferation and adhesion in a time-concentration-dependent mode. The higher inhibitory concentrations of curcumin (3 and 15μM) disrupted the cytoskeletal structures of trophozoites; the damage was evident on the ventral disk, flagella and in the caudal region, also the membrane was affected. The immunofluorescence images showed altered distribution of tubulin staining on ventral disk and flagella. Additionally, we found that curcumin caused a clear reduction of tubulin expression. By docking analysis and molecular dynamics we showed that curcumin has a high probability to bind at the interface of the tubulin dimer close to the vinblastine binding site. All the data presented indicate that curcumin may inhibit Giardia proliferation by perturbing microtubules. Copyright © 2017. Published by Elsevier B.V.

  16. Cell biological characterization of the malaria vaccine candidate trophozoite exported protein 1.

    Directory of Open Access Journals (Sweden)

    Caroline Kulangara

    Full Text Available In a genome-wide screen for alpha-helical coiled coil motifs aiming at structurally defined vaccine candidates we identified PFF0165c. This protein is exported in the trophozoite stage and was named accordingly Trophozoite exported protein 1 (Tex1. In an extensive preclinical evaluation of its coiled coil peptides Tex1 was identified as promising novel malaria vaccine candidate providing the rational for a comprehensive cell biological characterization of Tex1. Antibodies generated against an intrinsically unstructured N-terminal region of Tex1 and against a coiled coil domain were used to investigate cytological localization, solubility and expression profile. Co-localization experiments revealed that Tex1 is exported across the parasitophorous vacuole membrane and located to Maurer's clefts. Change in location is accompanied by a change in solubility: from a soluble state within the parasite to a membrane-associated state after export to Maurer's clefts. No classical export motifs such as PEXEL, signal sequence/anchor or transmembrane domain was identified for Tex1.

  17. Presence of potentially pathogenic Babesia sp. for human in Ixodes ricinus in Switzerland.

    Science.gov (United States)

    Casati, Simona; Sager, Heinz; Gern, Lise; Piffaretti, Jean-Claude

    2006-01-01

    We have designed and performed a new PCR method based on the 18S rRNA in order to individuate the presence and the identity of Babesia parasites. Out of 1159 Ixodes ricinus (Acari: Ixodidae) ticks collected in four areas of Switzerland, nine were found to contain Babesia DNA. Sequencing of the short amplicon obtained (411-452 bp) allowed the identification of three human pathogenic species: Babesia microti, B. divergens, for the first time in Switzerland, Babesia sp. EU1. We also report coinfections with B. sp. EU1-Borrelia burgdorferi sensu stricto and Babesia sp. EU1-B. afzelii.

  18. Platelet indices in dogs with Babesia rossi infection

    DEFF Research Database (Denmark)

    Goddard, Amelia; Leisewitz, Andrew L; Kristensen, Annemarie Thuri

    2015-01-01

    BACKGROUND: Thrombocytopenia without clinical bleeding is a consistent finding in virulent canine babesiosis. OBJECTIVES: The purpose of the study was to investigate the platelet index phenotype in Babesia rossi-infected dogs and the association with disease outcome. We hypothesized that an incre......BACKGROUND: Thrombocytopenia without clinical bleeding is a consistent finding in virulent canine babesiosis. OBJECTIVES: The purpose of the study was to investigate the platelet index phenotype in Babesia rossi-infected dogs and the association with disease outcome. We hypothesized...... that an increased proportion of large, activated platelets would be present. METHODS: Ninety-six infected and 15 control dogs were included. Babesia-infected dogs were further divided into survivors and nonsurvivors. Platelet count, mean platelet volume (MPV), platelet volume distribution width (PDW), plateletcrit...

  19. Radiological findings in three acquired immunodeficiency syndrome patients with Rhodococcus equi pneumonia

    International Nuclear Information System (INIS)

    Liu Jinxin; Tang Xiaoping; Zhang Lieguang

    2011-01-01

    Objective: To study the imaging appearances of Rhodococcus equi pneumonia in three patients with acquired immunodeficiency syndrome ( AIDS). Methods: Thoracic imaging appearances of' Rhodococcus equi pneumonia in three patients with AIDS were retrospectively analyzed. Results: The chest radiograph showed patchy consolidations and small nodules (n=3), large consolidations with multiple cavitations (n=2). CT showed large lobar or segmental consolidations with multiple cavitations (n=2), patchy consolidations (n=2), bronchiectasis (n=1), multiple small centrilobular nodules (n=2) and tree-in-bud patterns (n=2). Conclusion: The most common radiological findings in AIDS patients with Rhodococcus equi pulmonary infection are large consolidations with multiple cavitations and multiple centrilobular nodules. (authors)

  20. First record of Babesia sp. in Antarctic penguins.

    Science.gov (United States)

    Montero, Estrella; González, Luis Miguel; Chaparro, Alberto; Benzal, Jesús; Bertellotti, Marcelo; Masero, José A; Colominas-Ciuró, Roger; Vidal, Virginia; Barbosa, Andrés

    2016-04-01

    This is the first reported case of Babesia sp. in Antarctic penguins, specifically a population of Chinstrap penguins (Pygoscelis antarctica) in the Vapour Col penguin rookery in Deception Island, South Shetlands, Antarctica. We collected peripheral blood from 50 adult and 30 chick Chinstrap penguins. Examination of the samples by microscopy showed intraerythrocytic forms morphologically similar to other avian Babesia species in 12 Chinstrap penguin adults and seven chicks. The estimated parasitaemias ranged from 0.25×10(-2)% to 0.75×10(-2)%. Despite the low number of parasites found in blood smears, semi-nested PCR assays yielded a 274 bp fragment in 12 of the 19 positive blood samples found by microscopy. Sequencing revealed that the fragment was 97% similar to Babesia sp. 18S rRNA from Australian Little Penguins (Eudyptula minor) confirming presence of the parasite. Parasite prevalence estimated by microscopy in adults and chicks was higher (24% vs. 23.3%, respectively) than found by semi-nested PCR (16% vs. 13.3% respectively). Although sampled penguins were apparently healthy, the effect of Babesia infection in these penguins is unknown. The identification of Babesia sp. in Antarctic penguins is an important finding. Ixodes uriae, as the only tick species present in the Antarctic Peninsula, is the key to understanding the natural history of this parasite. Future work should address the transmission dynamics and pathogenicity of Babesia sp. in Chinstrap penguin as well as in other penguin species, such as Gentoo penguin (Pygoscelis papua) and Adélie penguin (Pygoscelis adeliae), present within the tick distribution range in the Antarctic Peninsula. Copyright © 2016 Elsevier GmbH. All rights reserved.

  1. Fatal Babesia canis canis infection in a splenectomized Estonian dog.

    Science.gov (United States)

    Tiškina, Valentina; Capligina, Valentina; Must, Külli; Berzina, Inese; Ranka, Renate; Jokelainen, Pikka

    2016-01-25

    A previously splenectomized dog from Estonia was presented with a sudden lack of appetite and discoloration of the urine. Despite supportive therapy, its condition deteriorated dramatically during 1 day. Severe thrombocytopenia and high numbers of protozoan hemoparasites were evident in blood smears, and the hematocrit dropped from 46 to 33 %. The dog was euthanized before specific antibabesial treatment was initiated. Blood samples from the dog and from two other dogs in the same household tested positive for Babesia using molecular methods, and the sequences of partial 18S rRNA gene confirmed the causative species as Babesia canis canis. The risk of severe, rapidly progressing babesiosis in splenectomized dogs merits awareness.

  2. In vitro activity of Arbutus unedo leaf extracts against Trichomonas vaginalis trophozoites.

    Science.gov (United States)

    Ertabaklar, Hatice; Kivçak, Bijen; Mert, Tuba; Ozensoy Töz, Seray

    2009-01-01

    Trichomonas vaginalis (T. vaginalis) is a flagellated protozoan commonly causing sexually transmitted disease. T. vaginalis infections are treated with a 5-nitroimidazole derivate. However, drug resistance has been known to occur for a long time and new alternatives are under investigation. Arbutus unedo is a wild plant mainly growing in maquis and rocky places of the seaboard in Southern Europe. In our study, ethanolic, water, hexane and ethyl acetate extracts of Arbutus unedo leaves were tested in vitro against T. vaginalis trophozoites and the ethyl acetate extract of Arbutus unedo leaves was found to be effective (Growth inhibition rate (GI): 100%, at the concentration of 500 microg/ml). It may be a promising anti-trichomonacidal agent in the future and further experiments are needed.

  3. Dietary lipids containing gangliosides reduce Giardia muris infection in vivo and survival of Giardia lamblia trophozoites in vitro.

    Science.gov (United States)

    Suh, M; Belosevic, M; Clandinin, M T

    2004-06-01

    We examined whether a ganglioside supplemented diet affected the course of Giardia muris infection in mice and survival of Giardia lamblia trophozoites in vitro. Female CD-1 mice were fed 1 of 5 experimental diets: standard lab chow as a control diet; semi-synthetic diets containing 20% (w/w) triglyceride based on the fat composition of a conventional infant formula; triglyceride diet; triglyceride diet containing a low level of ganglioside (0.1% w/w); and triglyceride diet containing a high level of ganglioside (1.0% w/w of diet). After 2 weeks of feeding, mice were inoculated with G. muris by gastric intubation and fed the experimental diets during the course of the infection. Cysts released in the faeces and trophozoites present in the small intestine were enumerated at various times post-infection. The average cyst output and the number of trophozoites during the course of the infection in mice fed ganglioside-containing diet were found to be significantly lower (3-log10 reduction) compared to animals fed control diets. The results of in vitro growth studies indicated that gangliosides may be directly toxic to the parasites. Thus, gangliosides have a protective effect against G. muris infection in vivo and affect the survival of G. lamblia trophozoites in vitro.

  4. Molecular characterization of Babesia peircei and Babesia ugwidiensis provides insight into the evolution and host specificity of avian piroplasmids

    Directory of Open Access Journals (Sweden)

    Michael J. Yabsley

    2017-12-01

    Full Text Available There are 16 recognized species of avian-infecting Babesia spp. (Piroplasmida: Babesiidae. While the classification of piroplasmids has been historically based on morphological differences, geographic isolation and presumed host and/or vector specificities, recent studies employing gene sequence analysis have provided insight into their phylogenetic relationships and host distribution and specificity. In this study, we analyzed the sequences of the 18S rRNA gene and ITS-1 and ITS-2 regions of two Babesia species from South African seabirds: Babesia peircei from African penguins (Spheniscus demersus and Babesia ugwidiensis from Bank and Cape cormorants (Phalacrocorax neglectus and P. capensis, respectively. Our results show that avian Babesia spp. are not monophyletic, with at least three distinct phylogenetic groups. B. peircei and B. ugwidiensis are closely related, and fall within the same phylogenetic group as B. ardeae (from herons Ardea cinerea, B. poelea (from boobies Sula spp. and B. uriae (from murres Uria aalge. The validity of B. peircei and B. ugwidiensis as separate species is corroborated by both morphological and genetic evidence. On the other hand, our results indicate that B. poelea might be a synonym of B. peircei, which in turn would be a host generalist that infects seabirds from multiple orders. Further studies combining morphological and molecular methods are warranted to clarify the taxonomy, phylogeny and host distribution of avian piroplasmids. Keywords: Africa, Babesia, Piroplasmida, Phalacrocoracidae, Spheniscidae, Tick-borne pathogen

  5. Subclinical Theileria Equi Infection and Rhabdomyolysis in Three Endurance Horses

    Directory of Open Access Journals (Sweden)

    A. Muñoz1,2*, R. G. M. Rodríguez2, C. Riber1,2, P. Trigo2, M. Gómez-Díez2 and F. Castejon2

    2013-04-01

    Full Text Available Three well-trained endurance horses, competing over different distances, developed sudden and unexpected rhabdomyolysis at the onset of exercise. They were treated and afterwards they did not have any other bout of muscle damage. All of them were positive to Theileria equi (polymerase reaction chain. The possible reasons of the rhabdomyolysis could have been the direct effect of the parasite on the muscle and/or the result of the anemia and a limited oxygen supply to the exercising muscles. It is suggested that the horses were carriers of Theileria and they developed clinical signs because of the immune-suppression caused by prolonged submaximal exercise and/or transportation.

  6. EquiFACS: The Equine Facial Action Coding System.

    Directory of Open Access Journals (Sweden)

    Jen Wathan

    Full Text Available Although previous studies of horses have investigated their facial expressions in specific contexts, e.g. pain, until now there has been no methodology available that documents all the possible facial movements of the horse and provides a way to record all potential facial configurations. This is essential for an objective description of horse facial expressions across a range of contexts that reflect different emotional states. Facial Action Coding Systems (FACS provide a systematic methodology of identifying and coding facial expressions on the basis of underlying facial musculature and muscle movement. FACS are anatomically based and document all possible facial movements rather than a configuration of movements associated with a particular situation. Consequently, FACS can be applied as a tool for a wide range of research questions. We developed FACS for the domestic horse (Equus caballus through anatomical investigation of the underlying musculature and subsequent analysis of naturally occurring behaviour captured on high quality video. Discrete facial movements were identified and described in terms of the underlying muscle contractions, in correspondence with previous FACS systems. The reliability of others to be able to learn this system (EquiFACS and consistently code behavioural sequences was high--and this included people with no previous experience of horses. A wide range of facial movements were identified, including many that are also seen in primates and other domestic animals (dogs and cats. EquiFACS provides a method that can now be used to document the facial movements associated with different social contexts and thus to address questions relevant to understanding social cognition and comparative psychology, as well as informing current veterinary and animal welfare practices.

  7. Restriction Fragment Length Polymorphisms of Virulence Plasmids in Rhodococcus equi

    Science.gov (United States)

    Takai, Shinji; Shoda, Masato; Sasaki, Yukako; Tsubaki, Shiro; Fortier, Guillaume; Pronost, Stephane; Rahal, Karim; Becu, Teotimo; Begg, Angela; Browning, Glenn; Nicholson, Vivian M.; Prescott, John F.

    1999-01-01

    Virulent Rhodococcus equi, which is a well-known cause of pyogranulomatous pneumonia in foals, possesses a large plasmid encoding virulence-associated 15- to 17-kDa antigens. Foal and soil isolates from five countries—Argentina, Australia, Canada, France, and Japan—were investigated for the presence of 15- to 17-kDa antigens by colony blotting, using the monoclonal antibody 10G5, and the gene coding for 15- to 17-kDa antigens by PCR. Plasmid DNAs extracted from positive isolates were digested with restriction endonucleases BamHI, EcoRI, EcoT22I, and HindIII, and the digestion patterns that resulted divided the plasmids of virulent isolates into five closely related types. Three of the five types had already been reported in Canadian and Japanese isolates, and the two new types had been found in French and Japanese isolates. Therefore, we tentatively designated these five types 85-kb type I (pREAT701), 85-kb type II (a new type), 87-kb type I (EcoRI and BamHI type 2 [V. M. Nicholson and J. F. Prescott, J. Clin. Microbiol. 35:738–740, 1997]), 87-kb type II (a new type), and 90-kb (pREL1) plasmids. The 85-kb type I plasmid was found in isolates from Argentina, Australia, Canada, and France. Plasmid 87-kb type I was isolated in specimens from Argentina, Canada, and France. The 85-kb type II plasmid appeared in isolates from France. On the other hand, plasmids 87-kb type II and 90-kb were found only in isolates from Japan. These results revealed geographic differences in the distribution of the virulence plasmids found in the five countries and suggested that the restriction fragment length polymorphism of virulence plasmids might be useful to elucidate the molecular epidemiology of virulent R. equi in the world. PMID:10488224

  8. Ocorrência de Babesia sp em pequenos roedores no Brasil Occurrence of Babesia sp in small rodents in Brazil

    OpenAIRE

    G.S. Gazeta; R.W. Carvalho; R.F. Avelar; M. Amorim; A.E. Aboud-Dutra

    2004-01-01

    Foi analisada a ocorrência de babesiose em pequenos roedores nos municípios de Silva Jardim e Nova lguaçu, Estado do Rio de Janeiro. Foram capturados 44 roedores de seis espécies diferentes e entre eles a prevalência da infecção foi de 27,3%. Rattus norvegicus foi considerado o principal reservatório (50,0%) e Oligoryzomys nigripes como novo hospedeiro para Babesia sp. Este foi o primeiro relato de Babesia sp. em roedores no Brasil. A freqüência de roedores positivos e o risco de infecção dos...

  9. Principales marcadores moleculares utilizados para la identificación de Babesia bovis y Babesia bigemina

    Directory of Open Access Journals (Sweden)

    Sandra Ríos T.

    2011-05-01

    Full Text Available Este artículo describe los principales marcadores moleculares utilizados para la identificación de B. bovis y B. bigemina reportados en la literatura científica. Para ello se diseñó una revisión sistemática a partir de la aplicación de la estrategia metodológica PICO modificada con el objetivo de definir las secuencias nucleotídicas detectadas en los diferentes sitios geográficos y su utilidad diagnóstica. Se realizó una búsqueda avanzada con los términos “Babesia bovis” y “DNA” y “Babesia bigemina” y “DNA” en las bases de datos ScienceDirect, SpringerLink y PubMed que después de ser filtradas permitieron obtener un resultado total de 68 artículos originales. Tanto los artículos incluidos como los excluidos fueron almacenados en tablas, en las cuales se presenta la justificación de su condición dentro del estudio. A los 68 artículos seleccionados se les aplicó una evaluación con criterios de inclusión y exclusión previamente definidos, de este modo, 21 artículos originales cumplieron con los criterios de inclusión y se incluyeron en el estudio. Se describe la utilidad de los marcadores moleculares referenciados en la literatura científica desde 1995 hasta el 2010: la subunidad pequeña RNAr, el gen citocromo b, gen msa-1 and msa-2c, el gen Bv, el factor de elongación alfa (EF-1α, el gen de la beta-Tubulina, SBP 1-2-3, y los RAP; su aplicación diagnóstica y su utilización en los diferentes sitios geográficos. Los marcadores moleculares utilizados para la detección de las babesias bovinas varían dependiendo de la región geográfica, grado de conservación genética y resultados de estudios previos que concluyen su utilidad diagnóstica.

  10. Evaluation of micro fatigue crack growth under equi-biaxial stress by membranous pressure fatigue test

    International Nuclear Information System (INIS)

    Iida, Satoshi; Abe, Shigeki; Nakamura, Takao; Kamaya, Masayuki

    2014-01-01

    For preventing nuclear power plant (NPP) accidents, NPPs are required to ensure system safety in long term safe operation under aging degradation. Now, fatigue accumulation is one of major ageing phenomena and are evaluated to ensure safety by design fatigue curve that are based on the results of uniaxial fatigue tests. On the other hand, thermal stress that occurs in piping of actual components is not uniaxial but equi-biaxial. For accurate evaluation, it is required to conform real circumstance. In this study, membranous pressure fatigue test was conducted to simulated equi-biaxial stress. Crack initiation and crack growth were examined by replica investigation. Calculation result of equivalent stress intensity factor shows crack growth under equi-biaxial stress is faster than under uniaxial stress. It is concluded that equi-biaxial fatigue behavior should be considered in the evaluation of fatigue crack initiation and crack growth. (author)

  11. Babesia bovis clones: biochemical and enzymatic characterization

    International Nuclear Information System (INIS)

    Rodriguez Camarillo, S.D.

    1985-01-01

    Studies were undertaken to generate additional knowledge of the biochemistry of Babesia bovis. A modified in vitro culture technique used for cloning B. bovis. This technique included a low oxygen concentration atmosphere (2%, O 2 , 5% CO 2 , 93% N 2 ) and 4 mm fluid level. Cultures initiated with one infected erythrocyte were maintained until parasitemias of positive wells reached 2% parasitemia. Primary clones were obtained and from these, nine clones were recloned twice and used for subsequent studies. A procedure was developed to concentrate and separate B. bovis merozoites and infected erythrocytes by Percoll density gradients. Merozoites separated at 1.087 g/ml specific density, whereas infected erythrocytes separated at 1.121 g/ml. Viability of purified parasites was not affected. Agarose gel electrophoresis was used to identify metabolic enzyme in B. bovis and B. bigemina. The enzymes LDH, GDH, GPI and HK were detected in both species. Molecular analysis by one and two-dimensional gel electrophoresis of proteins metabolically labeled with 35 S-methionine indicated that two clones, derived from the same field strain, were similar but not identical to the parent. Fewer proteins were observed in the parental strain. Growth of two 60-Co irradiated B. bovis clones indicated a dose-effect relationship. Growth of parasites exposed for the longest period was initially retarded but returned to normal growth after two or three subcultures. Cultures exposed for shorter periods were unaffected with respect to the rate of growth. Analysis of electrophoretic mobility of metabolic enzyme showed a change in migration pattern

  12. Detection of Theileria and Babesia species in ticks collected from cattle.

    Science.gov (United States)

    Ica, A; Vatansever, Z; Yildirim, A; Duzlu, O; Inci, A

    2007-09-01

    The present study was carried out to detect tick species that infest cattle, and Theileria and Babesia species transmitted by these ticks in Kayseri province (Turkey). A total of 300 cattle were examined for tick infestations. Of the 300 cattle, 117 (39%) were infested with ticks. A total of 1160 ticks belonging to 11 Ixodid genera were collected from the infested animals and their shelters. The most prevalent tick species was Boophilus annulatus 26.37% (306/1160) followed by Hyalomma marginatum marginatum 21.12% (245/1160) and Rhipicephalus turanicus 18.7% (217/1160). The collected ticks were separated into 43 tick pools, according to their species. These pools were examined for bovine Theileria and Babesia species (Theileria sp., Babesia sp., Theileria annulata, T. buffeli/orientalis, Babesia bigemina, B. bovis and B. divergens) by using the reverse line blotting method (RLB). Of the 43 tick pools examined, 6 (14%) were infected with B. bigemina, 4 (9.3%) with T. annulata, and 1 (2.3%) with Babesia sp., whereas 1 (2.3%) displayed mixed infection with T. annulata + B. bigemina. The sequence and phylogenetic analyses of Babesia sp., which could not be identified to the species level by RLB, were performed. In the phylogenetic tree, Babesia sp. (Kayseri 1) grouped with Babesia sp. (Kashi 2), Babesia sp. (Kashi 1), Babesia sp. (Xinjiang) and B. orientalis with 96.8-100% identity.

  13. Wild cervids are host for tick vectors of babesia species with zoonotic capability in Belgium.

    Science.gov (United States)

    Lempereur, Laetitia; Wirtgen, Marc; Nahayo, Adrien; Caron, Yannick; Shiels, Brian; Saegerman, Claude; Losson, Bertrand; Linden, Annick

    2012-04-01

    Babesiosis is a tick-borne disease caused by different species of intraerythrocytic protozoan parasites within the genus Babesia. Different species of Babesia are described as potentially zoonotic and cause a malaria-like disease mainly in immunocompromised humans. Interest in the zoonotic potential of Babesia is growing and babesiosis has been described by some authors as an emergent zoonotic disease. The role of cervids to maintain tick populations and act as a reservoir host for some Babesia spp. with zoonotic capability is suspected. To investigate the range and infection rate of Babesia species, ticks were collected from wild cervids in southern Belgium during 2008. DNA extraction was performed for individual ticks, and each sample was evaluated for the absence of PCR inhibition using a PCR test. A Babesia spp. genus-specific PCR based on the 18S rRNA gene was applied to validated tick DNA extracts. A total of 1044 Ixodes ricinus ticks were collected and 1023 validated samples were subsequently screened for the presence of Babesia spp. DNA. Twenty-eight tick samples were found to be positive and identified after sequencing as containing DNA representing: Babesia divergens (3), B. divergens-like (5), Babesia sp. EU1 (11), Babesia sp. EU1-like (3), B. capreoli (2), or unknown Babesia sp. (4). This study confirms the presence of potentially zoonotic species and Babesia capreoli in Belgium, with a tick infection rate of 2.7% (95% CI 1.8,3.9%). Knowledge of the most common reservoir source for transmission of zoonotic Babesia spp. will be useful for models assessing the risk potential of this infection to humans.

  14. Infection of dogs with Babesia canis in Gwagwalada metropolis of ...

    African Journals Online (AJOL)

    Epidemiological investigation was carried out to determine the prevalence of infection with Babesia canis in dogs in Gwagwalada metropolis of the Federal Capital Territory, Abuja Nigeria, from November 2013 to January 2014. Blood samples were collected from 101 dogs and examined for the parasite. Data obtained were ...

  15. Reclassification of Theileria annae as Babesia vulpes sp. nov.

    Science.gov (United States)

    Baneth, Gad; Florin-Christensen, Monica; Cardoso, Luís; Schnittger, Leonhard

    2015-04-08

    Theileria annae is a tick-transmitted small piroplasmid that infects dogs and foxes in North America and Europe. Due to disagreement on its placement in the Theileria or Babesia genera, several synonyms have been used for this parasite, including Babesia Spanish dog isolate, Babesia microti-like, Babesia (Theileria) annae, and Babesia cf. microti. Infections by this parasite cause anemia, thrombocytopenia, and azotemia in dogs but are mostly subclinical in red foxes (Vulpes vulpes). Furthermore, high infection rates have been detected among red fox populations in distant regions strongly suggesting that these canines act as the parasite's natural host. This study aims to reassess and harmonize the phylogenetic placement and binomen of T. annae within the order Piroplasmida. Four molecular phylogenetic trees were constructed using a maximum likelihood algorithm based on DNA alignments of: (i) near-complete 18S rRNA gene sequences (n = 76 and n = 93), (ii) near-complete and incomplete 18S rRNA gene sequences (n = 92), and (iii) tubulin-beta gene sequences (n = 32) from B. microti and B. microti-related parasites including those detected in dogs and foxes. All phylogenetic trees demonstrate that T. annae and its synonyms are not Theileria parasites but are most closely related with B. microti. The phylogenetic tree based on the 18S rRNA gene forms two separate branches with high bootstrap value, of which one branch corresponds to Babesia species infecting rodents, humans, and macaques, while the other corresponds to species exclusively infecting carnivores. Within the carnivore group, T. annae and its synonyms from distant regions segregate into a single clade with a highly significant bootstrap value corroborating their separate species identity. Phylogenetic analysis clearly shows that T. annae and its synonyms do not pertain to Theileria and can be clearly defined as a separate species. Based on the facts that T. annae and its synonyms have not been

  16. In vitro host erythrocyte specificity and differential morphology of Babesia divergens and a zoonotic Babesia sp. from eastern cottontail rabbits (Sylvilagus floridanus).

    Science.gov (United States)

    Spencer, Angela M; Goethert, Heidi K; Telford, Samuel R; Holman, Patricia J

    2006-04-01

    A Babesia sp. isolated from eastern cottontail rabbits (Sylvilagus floridanus) is morphologically similar and genetically identical, based on SSU rRNA gene comparisons, to 2 agents responsible for human babesiosis in the United States. This zoonotic agent is closely related to the European parasite, Babesia divergens. The 2 organisms were characterized by in vitro comparisons. In vitro growth of the rabbit Babesia sp. was supported in human and cottontail rabbit erythrocytes, but not in bovine cells. Babesia divergens was supported in vitro in bovine and human erythrocytes, but not in cottontail rabbit cells. Morphometric analysis classifies B. divergens as a small babesia in bovine erythrocytes, but the parasite exceeds this size in human erythrocytes. The rabbit Babesia sp. is large, the same size in both human or rabbit erythrocytes, and is significantly larger than B. divergens. Eight or more rabbit Babesia sp. parasites may occur within a single erythrocyte, sometimes in a floret array, unlike B. divergens. The erythrocyte specificity and morphological differences reported in this study agree with previous in vivo results and validate the use of in vitro methods for characterization of Babesia species.

  17. Bacteremia por Rhodococcus equi em paciente com síndrome da imunodeficiência adquirida: relato de caso Bacteremia due to Rhodococcus equi in a patient with acquired immunodeficiency syndrome: case report

    Directory of Open Access Journals (Sweden)

    Carina Secchi

    2006-12-01

    Full Text Available Rhodococcus equi é um importante agente de infecções zoonóticas, podendo causar sérias infecções em humanos, principalmente em pacientes imunocomprometidos. Neste estudo, nós relatamos o caso de uma bacteremia fatal devido a Rhodococcus equi em paciente com síndrome da imunodeficiência adquirida (HIV positivo.Rhodococcus equi is an important agent for zoonotic infections, and may cause serious infections in humans, especially immunocompromised patients. In this study, a case of fatal bacteremia due to Rhodococcus equi in a patient with acquired immunodeficiency syndrome (HIV positive is reported.

  18. Molecular cloning, characterization and antigenicity of Babesia sp. BQ1 (Lintan) (Babesia cf. motasi) apical membrane antigen-1 (AMA-1).

    Science.gov (United States)

    Niu, Qingli; Liu, Zhijie; Yang, Jifei; Guan, Guiquan; Pan, Yuping; Luo, Jianxun; Yin, Hong

    2017-04-01

    Apical membrane antigen-1 (AMA-1) has been described as a potential vaccine candidate in apicomplexan parasites. Here we characterize the ama-1 gene. The full-length ama-1 gene of Babesia sp. BQ1 (Lintan) (BLTAMA-1) is 1785 bp, which contains an open reading frame (ORF) encoding a 65-kDa protein of 594 amino acid residues; by definition, the 5' UTR precedes the first methionine of the ORF. Phylogenetic analysis based on AMA-1 amino acid sequences clearly separated Piroplasmida from other Apicomplexa parasites. The Babesia sp. BQ1 (Lintan) AMA-1 sequence is most closely associated with that of B. ovata and B. bigemina, with high bootstrap value. A recombinant protein encoding a conserved region and containing ectodomains I and II of BLTAMA-1 was constructed. BLTrAMA-1-DI/DII proteins were tested for reactivity with sera from sheep infected by Babesia sp. BQ1 (Lintan). In Western-blot analysis, native Babesia sp. BQ1 (Lintan) AMA-1 proteins were recognized by antibodies raised in rabbits against BLTrAMA-1 in vitro. The results of this study are discussed in terms of gene characterization, taxonomy and antigenicity.

  19. Ocorrência de Babesia sp em pequenos roedores no Brasil Occurrence of Babesia sp in small rodents in Brazil

    Directory of Open Access Journals (Sweden)

    G.S. Gazeta

    2004-12-01

    Full Text Available Foi analisada a ocorrência de babesiose em pequenos roedores nos municípios de Silva Jardim e Nova lguaçu, Estado do Rio de Janeiro. Foram capturados 44 roedores de seis espécies diferentes e entre eles a prevalência da infecção foi de 27,3%. Rattus norvegicus foi considerado o principal reservatório (50,0% e Oligoryzomys nigripes como novo hospedeiro para Babesia sp. Este foi o primeiro relato de Babesia sp. em roedores no Brasil. A freqüência de roedores positivos e o risco de infecção dos roedores não diferiram entre as áreas estudadas.The occurrence of babesiosis was studied in 44 small rodents of six species captured in Silva Jardim and Nova lguaçu counties, State of Rio de Janeiro, Brazil. The prevalence of injection was 27.3%. Rattus norvegicus was considered as the main reservoir and Oligoryzomys nigripes as a new host to Babesia sp. The frequency and the risk of rodent infection were considered equal among the studied areas. This is the first report of Babesia sp in small rodents in Brazil.

  20. Genomic evidence for the evolution of Streptococcus equi: host restriction, increased virulence, and genetic exchange with human pathogens.

    Directory of Open Access Journals (Sweden)

    Matthew T G Holden

    2009-03-01

    Full Text Available The continued evolution of bacterial pathogens has major implications for both human and animal disease, but the exchange of genetic material between host-restricted pathogens is rarely considered. Streptococcus equi subspecies equi (S. equi is a host-restricted pathogen of horses that has evolved from the zoonotic pathogen Streptococcus equi subspecies zooepidemicus (S. zooepidemicus. These pathogens share approximately 80% genome sequence identity with the important human pathogen Streptococcus pyogenes. We sequenced and compared the genomes of S. equi 4047 and S. zooepidemicus H70 and screened S. equi and S. zooepidemicus strains from around the world to uncover evidence of the genetic events that have shaped the evolution of the S. equi genome and led to its emergence as a host-restricted pathogen. Our analysis provides evidence of functional loss due to mutation and deletion, coupled with pathogenic specialization through the acquisition of bacteriophage encoding a phospholipase A(2 toxin, and four superantigens, and an integrative conjugative element carrying a novel iron acquisition system with similarity to the high pathogenicity island of Yersinia pestis. We also highlight that S. equi, S. zooepidemicus, and S. pyogenes share a common phage pool that enhances cross-species pathogen evolution. We conclude that the complex interplay of functional loss, pathogenic specialization, and genetic exchange between S. equi, S. zooepidemicus, and S. pyogenes continues to influence the evolution of these important streptococci.

  1. Wild Cervids Are Host for Tick Vectors of Babesia Species with Zoonotic Capability in Belgium

    OpenAIRE

    Lempereur, Laetitia; Wirtgen, Marc; Nahayo, Adrien; Caron, Yannick; Shiels, Brian; Saegerman, Claude; Losson, Bertrand; Linden, Annick

    2012-01-01

    Babesiosis is a tick-borne disease caused by different species of intraerythrocytic protozoan parasites within the genus Babesia. Different species of Babesia are described as potentially zoonotic and cause a malaria-like disease mainly in immunocompromised humans. Interest in the zoonotic potential of Babesia is growing and babesiosis has been described by some authors as an emergent zoonotic disease. The role of cervids to maintain tick populations and act as a reservoir host for some Babes...

  2. Babesia spp. in ticks and wildlife in different habitat types of Slovakia

    OpenAIRE

    Ham??kov?, Zuzana; Kazim?rov?, M?ria; Haru?tiakov?, Danka; Mahr?kov?, Lenka; Slov?k, Mirko; Berthov?, Lenka; Kocianov?, Elena; Schnittger, Leonhard

    2016-01-01

    Background Babesiosis is an emerging and potentially zoonotic disease caused by tick-borne piroplasmids of the Babesia genus. New genetic variants of piroplasmids with unknown associations to vectors and hosts are recognized. Data on the occurrence of Babesia spp. in ticks and wildlife widen the knowledge on the geographical distribution and circulation of piroplasmids in natural foci. Questing and rodent-attached ticks, rodents, and birds were screened for the presence of Babesia-specific DN...

  3. Characterization of Theileria equi genotypes in horses in Israel, the Palestinian Authority and Jordan.

    Science.gov (United States)

    Ketter-Ratzon, Dafna; Tirosh-Levy, Sharon; Nachum-Biala, Yaarit; Saar, Tal; Qura'n, Lara; Zivotofsky, Doni; Abdeen, Ziad; Baneth, Gad; Steinman, Amir

    2017-06-01

    Equine theileriosis caused by Theileria equi is endemic in the Middle East, where it causes a severe disease as well as widespread subclinical infection. The aim of this study was to evaluate the diversity of T. equi genotypes in Israel and the neighboring Palestinian Authority and Jordan. Blood samples from 355 horses from Israel, the Palestinian Authority and Jordan were tested for the prevalence of T. equi DNA. Two hundred and fourteen (60%) were found positive for T. equi infection by PCR. Of those, the 18S rRNA (1458bp) and the EMA-1 (745bp) genes of T. equi were sequenced from 15 horse samples that represent Israel's geographical distribution together with four samples from the Palestinian Authority and two from Jordan. The results were used for genotype characterization and phylogenetic analysis of T. equi in the equine population in Israel and its surroundings. Three 18S rRNA genotype clades were found in Israel (A, C and D) with clade D being the most prevalent and included all four isolates from the PA. In contrast, the EMA-1 gene showed little diversity with all sequences clustering in the same clade apart from one Jordanian sequence. Results suggest that although the Israeli horse population is small and relatively confined geographically, it is probable that the genetic variability, which was found among Israeli horses, is a result of introduction of horses from other countries. It also suggests that the EMA-1 gene is probably not a good target for the evaluation of variance in T. equi populations. Characterization of the different genotypes prevalent in a certain region is important in order to map out the intra-species sequence heterogeneity of the parasite, which is needed in order to develop new diagnostic tools and vaccines. Copyright © 2017 Elsevier GmbH. All rights reserved.

  4. Validation and use of an ELISA kit for the diagnosis of Babesia bovis in Cuba

    International Nuclear Information System (INIS)

    Blandino, T.; Alonso, M.; Barrera, M.; Mendoza, E.

    1998-01-01

    Babesia bovis, the most important etiological agent causing bovine babesiosis, is widely distributed in Cuba and affects mainly adult cattle. A survey of the prevalence of the disease in cattle using an ELISA kit (FAO/IAEA) revealed that 34.2% of the animals between 6 and 18 months of age were positive to Babesia bovis, whereas 69.9% on the cattle older than 18 months were positive. Antibodies to Babesia bovis were detected in 96.9% of calves vaccinated with an attenuated Babesia bovis vaccine. A good correlation was found between the results of ELISA kit with those from indirect immunofluorescence and immunoperoxidase tests developed in Cuba. (author)

  5. Detection of Babesia DNA in blood and spleen samples from Eurasian badgers (Meles meles) in Scotland.

    Science.gov (United States)

    Bartley, Paul M; Wilson, Cari; Innes, Elisabeth A; Katzer, Frank

    2017-08-01

    Babesia are intraerythrocytic parasites of importance worldwide within the fields of human and veterinary medicine, as some Babesia sp., including Babesia microti are potentially zoonotic and can cause fatal disease in both humans and animals. The aims of this study were to use a nested PCR (amplifying the 18S rRNA gene) to determine the presence and species of Babesia parasite DNA found in blood (n = 47) and spleen (n = 47) samples collected from Eurasian badgers (Meles meles) in Scotland. The results showed 28/47 (59·6%) blood and 14/47 (29·8%) spleen samples tested positive for the presence of Babesia DNA. Initial sequence analysis of the Babesia DNA identified three distinct sequence types (submitted to GenBank KX528553, KX528554 and KX528555), which demonstrated ⩾99% identity to Babesia sp. parasites previously identified in badgers in Spain (KT223484 and KT223485). Phylogenetic analysis showed that the three isolates are closely related to Babesia annae, B. microti and other Piroplasmida species found in wildlife. Further sequence analysis of the samples demonstrated that the badgers were routinely infected with more than one parasite isolate and there was also evidence of genetic recombination between the Babesia parasite isolates (submitted to GenBank KY250472 - KY250477).

  6. Detection and characterization of Babesia species in Ixodes ticks in Estonia.

    Science.gov (United States)

    Katargina, Olga; Geller, Julia; Vasilenko, Veera; Kuznetsova, Tatiana; Järvekülg, Lilian; Vene, Sirkka; Lundkvist, Åke; Golovljova, Irina

    2011-07-01

    The presence of Babesia spp. was studied in 2603 Ixodes ricinus and Ixodes persulcatus ticks collected at seven sites in Estonia. By reverse line blot screening, Babesia spp. was detected in 36 (1.4%) ticks, among them 18 (0.7%) were further recognized by a Babesia microti probe, 3 (0.1%) by a Babesia divergens probe, and the other 15 (0.6%) were recognized only by the universal Babesia spp. "catch all" probe. Sequence analyses of 6 of these 15 samples revealed that all of them belonged to Babesia sp. EU1. B. microti was detected in both tick species I. ricinus and I. persulcatus at the seven sites, whereas B. divergens-like and Babesia sp. EU1 were found only in I. persulcatus and I. ricinus, respectively. Genetic characterization based on partial 18S rRNA showed that the Estonian sequences of B. microti, B. divergens-like, and Babesia sp. EU1 share a high rate of similarity and are closely related to sequences from other European countries, Siberia, and United States. The present study demonstrated for the first time the existence and distribution of Babesia spp. in I. persulcatus and I. ricinus ticks in Estonia.

  7. Merozoite proteins from Babesia sp. BQ1 (Lintan) as potential antigens for serodiagnosis by ELISA.

    Science.gov (United States)

    Guan, G Q; Chauvin, A; Rogniaux, H; Luo, J X; Yin, H; Moreau, E

    2010-05-01

    Babesia sp. BQ1 (Lintan) is a Babesia isolated from sheep infested with Haemaphysalis qinghaiensis in China, and is closely related to B. motasi based on the 18S rRNA gene sequence. In the present study, an ELISA was developed with merozoite antigens of Babesia sp. BQ1 (Lintan) (BQMA) purified from in vitro culture. When the positive threshold was chosen as 30% of the antibodies rate, evaluated with 198 negative sera, the specificity was 95.5%. Except for Babesia sp. Tianzhu, there was no cross-reaction between BQMA and positive sera from Babesia sp. BQ1 (Ningxian)-, Babesia sp. Hebei-, Babesia sp. Xinjiang-, Theileria luwenshuni-, T. uilenbergi-, or Anaplasma ovis-infected sheep, which are the dominant haemoparasites of small ruminants in China. Specific antibodies against Babesia sp. BQ1 (Lintan) were produced 1 or 2 weeks post-infection and a high level of antibodies persisted for more than 8 months in experimentally infected sheep. This ELISA was tested on 974 sera collected from field-grazing sheep in 3 counties of Gansu province, northwestern China to evaluate the seroprevalence of Babesia sp. BQ1 (Lintan) infection and the average positive rate was 66.84%. The feasibility of increasing the specificity of this BQMA-based ELISA, by using some BQMA antigens for serodiagnosis is discussed.

  8. Interactions between the intestinal flagellates Giardia muris and Spironucleus muris and the blood parasites Babesia microti, Plasmodium yoelii and Plasmodium berghei in mice.

    Science.gov (United States)

    Brett, S J; Cox, F E

    1982-08-01

    In mice infected with the intestinal flagellates Giardia muris or Spironucleus muris, together with the blood parasites Babesia microti or Plasmodium yoelii, there is a temporary decrease of flagellate cyst output coincident with the peak of the blood parasite infections, followed by a rapid return to normal levels. This decrease in cyst output is correlated with decreased numbers of trophozoites in the small intestine. The effect on S. muris is more marked than that on G. muris. Neither blood parasites has any effect on the total duration of the flagellate infection and the flagellates do not affect the blood parasites. In mice infected with G. muris or S. muris and P. berghei there is also a decrease in cyst output but this is less apparent than in infections with B. microti or P. yoelii because of the fatal nature of the P. berghei infection. It is suggested that the decrease in cyst output is probably due to changes in the contents of the small intestine or to non-specific immunological factors rather than to specific immunological changes.

  9. Humoral immune responses of pregnant Guinea pigs Immunized with live attenuated Rhodococcus equi

    Directory of Open Access Journals (Sweden)

    Mawlood Abass Ali Al- Graibawi

    2018-02-01

    Full Text Available The potential to increase passive transfer of specific Rhodococcus equi (R.equi humoral immunity to newborn by preparturient vaccination of their dams was investigated in Pregnant Guinea pigs as a pilot study. Attenuated autogenous vaccine was prepared from a Congo red negative (CR- R.equi local isolate mixed with adjuvant (potassium alum sulphate, tested for sterility, safety and potency prior to vaccination .Two groups of pregnant G. pigs were used, the first group was vaccinated twice subcutaneously (S.C with the prepared vaccine at five and three weeks prior parturition, the second group was inoculated with adjuvant plus phosphate buffer saline (PBS twice s.c and kept as control. Offspring from the vaccinated dams had revealed high titers of specific R. equi antibody as detected by tube agglutination (TA and passive haemagglutination (PH test and showed protection against challenge dose. The results revealed that vaccination of pregnant G. pigs with the prepared attenuated vaccine was safe and efficient method to protect their offspring against experimental challenge with virulent R.equi. Vaccination was associated with increased humoral immune response in vaccinated group.

  10. The sensor kinase MprB is required for Rhodococcus equi virulence.

    Science.gov (United States)

    MacArthur, Iain; Parreira, Valeria R; Lepp, Dion; Mutharia, Lucy M; Vazquez-Boland, José A; Prescott, John F

    2011-01-10

    Rhodococcus equi is a soil bacterium and, like Mycobacterium tuberculosis, a member of the mycolata. Through possession of a virulence plasmid, it has the ability to infect the alveolar macrophages of foals, resulting in pyogranulomatous bronchopneumonia. The virulence plasmid has an orphan two-component system (TCS) regulatory gene, orf8, mutation of which completely attenuates virulence. This study attempted to find the cognate sensor kinase (SK) of orf8. Annotation of the R. equi strain 103 genome identified 23 TCSs encoded on the chromosome, which were used in a DNA microarray to compare TCS gene transcription in murine macrophage-like cells to growth in vitro. This identified six SKs as significantly up-regulated during growth in macrophages. Mutants of these SKs were constructed and their ability to persist in macrophages was determined with one SK, MprB, found to be required for intracellular survival. The attenuation of the mprB- mutant, and its complementation, was confirmed in a mouse virulence assay. In silico analysis of the R. equi genome sequence identified an MprA binding box motif homologous to that of M. tuberculosis, on mprA, pepD, sigB and sigE. The results of this study also show that R. equi responds to the macrophage environment differently from M. tuberculosis. MprB is the first SK identified as required for R. equi virulence and intracellular survival. Copyright © 2010 Elsevier B.V. All rights reserved.

  11. Efficacy of imidocarb dipropionate in eliminating Theileria equi from experimentally infected horses.

    Science.gov (United States)

    Grause, Juanita F; Ueti, Massaro W; Nelson, Jeffrey T; Knowles, Donald P; Kappmeyer, Lowell S; Bunn, Thomas O

    2013-06-01

    Theileria equi, one of the causative agents of equine piroplasmosis, is endemic in many regions of the world but is considered a 'foreign' animal disease in the USA. In an effort to prevent the importation of T. equi, stringent serological screening of horses is practiced prior to entry to the USA. Current regulatory options available where horses are found to be infected include permanent quarantine with or without chemotherapy, repatriation, or euthanasia. Chemotherapeutics that eliminate infection and subsequently transmission risk are critical in the management of infected horses. In this study, the efficacy of the drug imidocarb dipropionate against experimental T. equi infection was assessed. Of nine horses experimentally inoculated with T. equi isolated from an animal previously imported from Peru, six were treated with imidocarb dipropionate after the resolution of the acute phase of the disease. Elimination of the parasite was demonstrated in 5/6 by nested PCR, blood transfusions to naïve horses, and reversion to seronegative status. The findings support the use of this drug as a potential treatment option in controlling outbreaks of T. equi, and also suggest that 'combination testing' using both serological and PCR detection methods are necessary to demonstrate clearance of infection. Published by Elsevier Ltd.

  12. Rhodococcus equi venous catheter infection: a case report and review of the literature

    Directory of Open Access Journals (Sweden)

    Nahleh Zeina

    2011-08-01

    Full Text Available Abstract Introduction Rhodococcus equi is an animal pathogen that was initially isolated from horses and is being increasingly reported as a cause of infection in humans with impaired cellular immunity. However, this pathogen is underestimated as a challenging antagonist and is frequently considered to be a mere contaminant despite the potential for life-threatening infections. Most case reports have occurred in immunocompromised patients who have received organ transplants (for example kidney, heart, bone marrow or those with human immunodeficiency virus infection. Infections often manifest as pulmonary involvement or soft tissue abscesses. Bacteremia related to R. equi infections of tunneled central venous catheters has rarely been described. Case presentation We report the case of a 63-year-old non-transplant recipient, non-HIV infected Caucasian woman with endometrial carcinoma who developed recurrent bloodstream infections and septic shock due to R. equi and ultimately required the removal of her port catheter, a subcutaneous implantable central venous catheter. We also review the medical literature related to human infections with R. equi. Conclusion R. equi should be considered a serious pathogen, not a contaminant, particularly in an immunocompromised patient who presents with a central venous catheter-related bloodstream infection. Counseling patients with central venous catheters who participate in activities involving exposure to domesticated animals is recommended.

  13. First evidence of [i]Babesia venatorum[/i] and [i]Babesia capreoli[/i] in questing Ixodes ricinus ticks in the Czech Republic

    Directory of Open Access Journals (Sweden)

    Kristyna Venclikova

    2015-05-01

    Full Text Available Introduction and objective. [i]Ixodes ricinus[/i] is the most common tick species occurring in Central Europe and it serves as a principal vector of emerging human pathogens. The aim of this study was to determine the prevalence of [i]Babesia spp[/i]. in host-seeking [i]I. ricinus[/i] in urban and natural habitats. Materials and methods. PCR was applied on samples to assess prevalence of [i]Babesia spp.[/i] in questing ixodid ticks. Sequencing was used for [i]Babesia[/i] species determination. Results. 1,473 [i]I. ricinus[/i] ticks (1,294 nymphs, 99 males and 80 females were examined for the presence of [i]Babesia spp[/i]. at the two study sites. Minimum infection rate for [i]Babesia[/i] spp. was found to be 0.5% (infected I. ricinus nymphs were only detected in the natural ecosystem. Two[i] Babesia[/i] species were identified by sequencing: [i]B. venatorum[/i] (formerly called[i] Babesia[/i] sp. EU1 and [i]B. capreoli. [/i] Conclusions. The results obtained represent the first evidence of the occurrence of [i]B. venatorum[/i] and [i]B. capreoli[/i] in host-seeking[i] I. ricinus[/i] ticks in the Czech Republic.

  14. A survey of canine haemoprotozoan parasites from Turkey, including molecular evidence of an unnamed Babesia.

    Science.gov (United States)

    Aktas, Munir; Ozubek, Sezayi

    2017-06-01

    Canine tick-borne apicomplexan parasites have emerged in recent years, showing a wider geographic distribution and increased global prevalence. A reverse line blot assay was performed on 219 blood samples collected from domestic dogs for simultaneous detection of all named canine piroplasm species as well as Hepatozoon canis. Ten samples hybridized to the Theileria/Babesia and Babesia catch all probes but did not hybridize to any species-specific probe tested, suggesting the presence of an unrecognized Babesia species or genotype. Sequencing results showed 91.5%, 91.9%, 92.4%, 92.4%, and 89.2% similarity to B. canis, B. vogeli, B. rossi, B. gibsoni, and B. conradae, respectively. The highest homology (98.1-98.5%) observed was with unnamed Babesia sp. isolates (Ludhiana and Malbazar) described in dogs, Babesia sp. of buffalo origin, Babesia sp. Kashi 2, and Babesia orientalis, along with Babesia occultans of cattle origin. The partial cox1 sequence indicated that this isolate was most similar to Babesia sp. 1 HG-2012, with an identity of 86.5%. The survey revealed high prevalence of haemoprotozoans in domestic dogs (57.5%, CI 50.7-64.2), with Hepatozoon canis the most prevalent (54.3%, CI 47.5-61.117%), followed by Babesia sp. (4.6%, CI 2.2-8.2), B. vogeli (1.4%; CI 0.3-3.9), and B. canis (0.4%, CI 0-2.5). Combined infection of Hepatozoon canis and Babesia sp. was detected in five (2.3%, CI 0.7-5.2) samples and of H. canis and B. vogeli in two (0.9%, CI 0.1-3.2) dogs. The study contributes insight into the distribution and phylogenetic diversity of canine piroplasms in Turkey. Copyright © 2017 Elsevier Ltd. All rights reserved.

  15. Babesiosis caused by a large Babesia species in 7 immunocompromised dogs.

    Science.gov (United States)

    Sikorski, L E; Birkenheuer, A J; Holowaychuk, M K; McCleary-Wheeler, A L; Davis, J M; Littman, M P

    2010-01-01

    A large unnamed Babesia species was detected in a dog with lymphoma. It was unknown if this was an underrecognized pathogen. Report the historical and clinicopathologic findings in 7 dogs with babesiosis caused by a large unnamed Babesia species characterize the 18S ribosomal ribonucleic acid (rRNA) genes. Seven immunocompromised dogs from which the Babesia was isolated. Retrospective case review. Cases were identified by a diagnostic laboratory, the attending clinicians were contacted and the medical records were reviewed. The Babesia sp. 18S rRNA genes were amplified and sequenced. Six of 7 dogs had been splenectomized; the remaining dog was receiving oncolytic drugs. Lethargy, anorexia, fever, and pigmenturia were reported in 6/7, 6/7, 4/7, and 3/7 dogs. Laboratory findings included mild anemia (7/7) and severe thrombocytopenia (6/7). Polymerase chain reaction (PCR) assays used to detect Babesia sensu stricto species were all positive, but specific PCR assays for Babesia canis and Babesia gibsoni were negative in all dogs. The 18S rRNA gene sequences were determined to be identical to a large unnamed Babesia sp. previously isolated. Cross-reactive antibodies against other Babesia spp. were not always detectable. Five dogs were treated with imidocarb dipropionate and 1 dog with atovaquone/azithromycin; some favorable responses were noted. The remaining dog was untreated and remained a clinically stable carrier. Dogs with pigmenturia, anemia, and thrombocytopenia should be tested for Babesia sp. by PCR. Serology is not sufficient for diagnosis of this Babesia sp. Asplenia, chemotherapy, or both might represent risk factors for persistent infection, illness, or both.

  16. Peritoneal Effusion in a Dog due to Babesia gibsoni Infection

    OpenAIRE

    Gonde, Suresh; Chhabra, Sushma; Singla, L. D.; Bansal, B. K.

    2014-01-01

    A five-year-old male Labrador was presented to Teaching Veterinary Clinics of GADVASU with a primary complaint of distended abdomen, fever, and anorexia. The dog was found to be dull with elevated rectal temperature (104°F), heart rate (148 per minute), and respiration rate (58 per minute). Blood smear examination and PCR assay revealed that dog was positive for Babesia gibsoni. Elevated bilirubin, alanine amino transferase (ALT), alkaline phosphatase (ALP), creatinine, blood urea nitrogen (B...

  17. A unique case of Babesia gibsoni infected dog with paraplegia

    OpenAIRE

    Gonde, Suresh; Chhabra, S.; Uppal, S. K.; Singla, L. D.; Randhawa, S. S.

    2016-01-01

    A 4 year old male Rottweiler was presented to Teaching Veterinary Clinics of GADVASU with a primary complaint of paraplegia, fever and inappetence. The dog was found to be dull with elevated rectal temperature (105 °F), heart rate (158 per minute) and respiration rate (57 per minute). Blood smear examination and PCR assay revealed that dog was positive for Babesia gibsoni. Elevated bilirubin, alanine aminotransferase, alkaline phosphatase, creatinine, total leucocyte count, neutrophils and hy...

  18. Babesias of red deer (Cervus elaphus in Ireland

    Directory of Open Access Journals (Sweden)

    Zintl Annetta

    2011-01-01

    Full Text Available Abstract Blood samples were obtained from 38 wild red deer (Cervus elaphus at two sites in Ireland and subjected to PCR analysis of the 18S rRNA gene followed by sequencing. Two fragments of the 18S rRNA gene were generated by two different PCR protocols and subsequent sequencing suggested that at least six of the deer were infected by a babesia that, in those loci, is indistinguishable from Babesia divergens, an important tick-borne pathogen of cattle and of zoonotic significance. Additionally, a B. odocoilei-like parasite was detected in three samples and a babesia that did not match any sequences in the GenBank database was found in five samples. Neither B. capreoli nor B. venatorum (EU1 were found. There have been several reports of B. divergens occurring in deer species, including red deer, roe deer (Capreolus capreolus and reindeer (Rangifer tarandus. However, in view of recent re-sequencing of bovine-origin samples deposited previously in GenBank, it is unlikely that any of these sequences from deer are B. divergens. The present study describes the only deer piroplasm detected so far that shows complete identity with B. divergens, in just over half of the 18S rRNA gene. The entire gene of this deer parasite should be analysed and transmission experiments undertaken before the infectivity of B. divergens for red deer can be confirmed.

  19. First case of human babesiosis in Korea: detection and characterization of a novel type of Babesia sp. (KO1) similar to ovine babesia.

    Science.gov (United States)

    Kim, Jung-Yeon; Cho, Shin-Hyeong; Joo, Hyun-Na; Tsuji, Masayoshi; Cho, Sung-Ran; Park, Il-Joong; Chung, Gyung-Tae; Ju, Jung-Won; Cheun, Hyeng-Il; Lee, Hyeong-Woo; Lee, Young-Hee; Kim, Tong-Soo

    2007-06-01

    We report on the first case of human babesiosis in Korea. The intraerythrocytic parasite (KO1) in the patient's blood mainly appeared as paired pyriforms and ring forms; but Maltese cross forms were not seen, and the parasite showed morphological features consistent with those of the genus Babesia sensu stricto. The sequence of the 18S rRNA gene of KO1 was closely related to that of Babesia spp. isolated from sheep in China (similarity, 98%). The present study provides the first evidence of the presence of a hitherto unidentified, new type of Babesia parasite capable of infecting humans.

  20. Serological and molecular detection of Theileria equi in sport horses of northeastern Brazil.

    Science.gov (United States)

    Ferreira, Edlainne P; Vidotto, Odilon; Almeida, Jonatas C; Ribeiro, Luana P S; Borges, Marcos V; Pequeno, Walter H C; Stipp, Danilo T; de Oliveira, Celso J B; Biondo, Alexander W; Vieira, Thállitha S W J; Vieira, Rafael F C

    2016-08-01

    Theileriosis is a worldwide protozoal tick-borne disease caused by Theileria equi, which may produce a variety of clinical signs and turn infected horses into lifetime carriers. This study has aimed to perform a serological and molecular detection of T. equi and associated factors in sports horses from six areas of northeastern Brazil. In overall, 59.6% horses were positive by indirect immunofluorescence assay and 50.4% by polymerase chain reaction. No significant association was found when presence of ticks, age, gender, anemia or total plasma proteins was analyzed with seropositivity and molecular techniques. Although a significant association of infection was found in two cities. Thus, local risk factors other than presence of ticks, horse age, gender, anemia and total plasmatic proteins may dictate prevalence of T. equi infection in sports horses, even in highly endemic areas with no control of infection prior to horse competitions. Copyright © 2016 Elsevier Ltd. All rights reserved.

  1. Rapid and accurate identification of Streptococcus equi subspecies by MALDI-TOF MS

    DEFF Research Database (Denmark)

    Kudirkiene, Egle; Welker, Martin; Knudsen, Nanna Reumert

    2015-01-01

    phenotypic and sequence similarity between three subspecies their discrimination remains difficult. In this study, we aimed to design and validate a novel, Superspectra based, MALDI-TOF MS approach for reliable, rapid and cost-effective identification of SEE and SEZ, the most frequent S. equi subspecies.......3±7.5%). This result may be attributed to the highly clonal population structure of SEE, as opposed to the diversity of SEZ seen in horses. Importantly strains with atypical colony appearance both within SEE and SEZ did not affect correct identification of the strains by MALDI-TOF MS. Atypical colony variants...... are often associated with a higher persistence or virulence of S. equi, thus their correct identification using the current method strengthens its potential use in routine clinical diagnostics. In conclusion, reliable identification of S. equi subspecies was achieved by combining a MALDI-TOF MS method...

  2. The Antiphagocytic Activity of SeM of Streptococcus equi Requires Capsule.

    Science.gov (United States)

    Timoney, John F; Suther, Pranav; Velineni, Sridhar; Artiushin, Sergey C

    2014-01-01

    Resistance to phagocytosis is a crucial virulence property of Streptococcus equi (Streptococcus equi subsp. equi; Se), the cause of equine strangles. The contribution and interdependence of capsule and SeM to killing in equine blood and neutrophils were investigated in naturally occurring strains of Se. Strains CF32, SF463 were capsule and SeM positive, strains Lex90, Lex93 were capsule negative and SeM positive and strains Se19, Se1-8 were capsule positive and SeM deficient. Phagocytosis and killing of Se19, Se1-8, Lex90 and Lex93 in equine blood and by neutrophils suspended in serum were significantly (P ≤ 0.02) greater compared to CF32 and SF463. The results indicate capsule and SeM are both required for resistance to phagocytosis and killing and that the anti-phagocytic property of SeM is greatly reduced in the absence of capsule.

  3. Klossiella equi Infection in an Immunosuppressed Horse: Evidence of Long-Term Infection

    Directory of Open Access Journals (Sweden)

    Lora R. Ballweber

    2012-01-01

    Full Text Available A 13-year-old quarter horse gelding presented with a history of hematuria of approximately 1-year duration, anemia, weight loss over the previous six months, and bilateral nasal discharge of 2-week duration. It was determined that hematuria was most likely caused by the coccidian parasite Klossiella equi. Additional case workup suggested a diagnosis of pituitary pars intermedia dysfunction. Confirmatory testing was declined by the owners and the horse was discharged on medical therapy. Despite initial improvement after discharge, the horse developed unresolving sinusitis approximately 1 year later and was euthanized. Necropsy confirmed the presence of an adenoma of the pars intermedia of the pituitary gland, supporting the initial diagnosis. Additional findings included multiple developmental stages of K. equi present in the kidneys. This finding demonstrates infections with K. equi can be chronic in nature and supports the association of increased severity of klossiellosis and impaired immune function.

  4. Equi-axed and columnar grain growth in UO2

    International Nuclear Information System (INIS)

    White, R.J.

    1997-01-01

    The grain size of UO 2 is an important parameter in the actual performance and the modelling of the performance of reactor fuel elements. Many processes depend critically on the grain size, for example, the degree of initial densification, the evolution rate of stable fission gases, the release rates of radiologically hazardous fission products, the fission gas bubble swelling rates and the fuel creep. Many of these processes are thermally activated and further impact on the fuel thermal behavior thus creating complex feedback processes. In order to model the fuel performance accurately it is necessary to model the evolution of the fuel grain radius. When UO 2 is irradiated, the fission gases xenon and krypton are created from the fissioning uranium nucleus. At high temperatures these gases diffuse rapidly to the grain boundaries where they nucleate immobile lenticular shaped fission gas bubbles. In this paper the Hillert grain growth model is adapted to account for the inhibiting ''Zener'' effects of grain boundary fission gas porosity on grain boundary mobility and hence grain growth. It is shown that normal grain growth ceases at relatively low levels of irradiation. At high burnups, high temperatures and in regions of high temperature gradients, columnar grain growth is often observed, in some cases extending over more than fifty percent of the fuel radius. The model is further extended to account for the de-pinning of grains in the radial direction by the thermal gradient induced force on a fission gas grain boundary bubble. The observed columnar/equi-axed boundary is in fair agreement with the predictions of an evaporation/condensation model. The grain growth model described in this paper requires information concerning the scale of grain boundary porosity, the local fuel temperature and the local temperature gradient. The model is currently used in the Nuclear Electric version of the ENIGMA fuel modelling code. (author). 14 refs, 3 figs, 1 tab

  5. Identification of genomic loci associated with Rhodococcus equi susceptibility in foals.

    Directory of Open Access Journals (Sweden)

    Cole M McQueen

    Full Text Available Pneumonia caused by Rhodococcus equi is a common cause of disease and death in foals. Although agent and environmental factors contribute to the incidence of this disease, the genetic factors influencing the clinical outcomes of R. equi pneumonia are ill-defined. Here, we performed independent single nucleotide polymorphism (SNP- and copy number variant (CNV-based genome-wide association studies to identify genomic loci associated with R. equi pneumonia in foals. Foals at a large Quarter Horse breeding farm were categorized into 3 groups: 1 foals with R. equi pneumonia (clinical group [N = 43]; 2 foals with ultrasonographic evidence of pulmonary lesions that never developed clinical signs of pneumonia (subclinical group [N = 156]; and, 3 foals without clinical signs or ultrasonographic evidence of pneumonia (unaffected group [N = 49]. From each group, 24 foals were randomly selected and used for independent SNP- and CNV-based genome-wide association studies (GWAS. The SNP-based GWAS identified a region on chromosome 26 that had moderate evidence of association with R. equi pneumonia when comparing clinical and subclinical foals. A joint analysis including all study foals revealed a 3- to 4-fold increase in odds of disease for a homozygous SNP within the associated region when comparing the clinical group with either of the other 2 groups of foals or their combination. The region contains the transient receptor potential cation channel, subfamily M, member 2 (TRPM2 gene, which is involved in neutrophil function. No associations were identified in the CNV-based GWAS. Collectively, these data identify a region on chromosome 26 associated with R. equi pneumonia in foals, providing evidence that genetic factors may indeed contribute to this important disease of foals.

  6. Disseminated Rhodococcus equi infection in a patient with Hodgkin’s lymphoma

    Directory of Open Access Journals (Sweden)

    Mikić Dragan

    2014-01-01

    Full Text Available Introduction. Rhodococcus (R equi is an opportunistic, uncommon human pathogen that causes mainly infection in immunocompromised hosts. The disease is usually presented as subacute pneumonia that is mostly cavitary and sometimes bacteremic. Case report. We reported the extremly rare case of a 43-year-old woman with Hodgkin's lymphoma, who developed R. equi pulmonary infection after recieving multiple courses of chemotherapy. Secondary, the patient developed bacteremia, leading to sepsis and dissemination of R. equi infection in many extrapulmonary sites. At addmission the patient was febrile, tachypnoic, tachycardic, hypotensive, with facial edema, splenomegaly, positive meningeal signs, left hemiparesis and paraparesis. Laboratory data included erythrocyte sedimentation rate (ESR > 140 mm/h, C-reactive protein (CRP 143.0 mg/L, red blood cells (RBC 2.14 × 1012/L, whyite blood cells (WBC 2.8 × 109/L, lactate dehydrogenase (LDH 706 U/L, serum albumin 26 g/L, sodium 127 mmol/L and potassium 2.7 mmol/L. Blood culture and culture of sputum and empyema were positive for R. equi. Imaging studies demonstrated a large right cavitary pneumonia and abscess, empyema, pericarditis, mediastinal and intra-abdominal lymphadenopathy, brain and psoas abscesses, osteomyelitis and spondylodiscitis. The patient recovered completely after a 12-month treatment with combinations of parenteral and oral antibiotics (meropenem, vancomycin, teicoplanin, ciprofloxacin, rifampicin, macrolides etc, including drainage of abscesses and empyema. Eight years after completition of the treatment the patient was without recurrence of R. equi infection and lymphoma. Conclusion. Since the eradication od R. equi is very difficult, it is very important to make the diagnosis and initiate appropriate antibiotic therapy as soon as possible.

  7. Babesia ugwidiensis, a new species of Avian piroplasm from Phalacrocoracidae in South Africa

    Directory of Open Access Journals (Sweden)

    Peirce M.A.

    2012-11-01

    Full Text Available A new species of haematozoa, Babesia ugwidiensis sp. nov. from a cormorant is described. This is the first species of piroplasm to be recorded from the Phalacrocoracidae and the relationship of this parasite to other Babesia spp. from marine hosts is discussed.

  8. Babesia lengau sp. nov., a Novel Babesia Species in Cheetah (Acinonyx jubatus, Schreber, 1775) Populations in South Africa ▿

    Science.gov (United States)

    Bosman, Anna-Mari; Oosthuizen, Marinda C.; Peirce, Michael A.; Venter, Estelle H.; Penzhorn, Barend L.

    2010-01-01

    In a previous paper, we reported on a large number of cheetah blood specimens that gave positive signals only for Babesia and/or Theileria genus-specific probes on the reverse line blot (RLB) assay, indicating the presence of a novel species or variant of an existing species. Some of these specimens were investigated further by microscopic, serological, sequencing, and phylogenetic analyses. The near-full-length 18S rRNA genes of 13 samples, as well as the second internal transcribed spacer (ITS2) region, were amplified, cloned, and sequenced. A species-specific RLB probe, designed to target the hypervariable V4 region of the 18S rRNA gene for detection of the novel Babesia sp., was used to screen an additional 137 cheetah blood specimens for the presence of the species. The prevalence of infection was 28.5%. Here we describe the morphology and phylogenetic relationships of the novel species, which we have named Babesia lengau sp. nov. PMID:20519464

  9. Babesia lengau sp. nov., a novel Babesia species in cheetah (Acinonyx jubatus, Schreber, 1775) populations in South Africa.

    Science.gov (United States)

    Bosman, Anna-Mari; Oosthuizen, Marinda C; Peirce, Michael A; Venter, Estelle H; Penzhorn, Barend L

    2010-08-01

    In a previous paper, we reported on a large number of cheetah blood specimens that gave positive signals only for Babesia and/or Theileria genus-specific probes on the reverse line blot (RLB) assay, indicating the presence of a novel species or variant of an existing species. Some of these specimens were investigated further by microscopic, serological, sequencing, and phylogenetic analyses. The near-full-length 18S rRNA genes of 13 samples, as well as the second internal transcribed spacer (ITS2) region, were amplified, cloned, and sequenced. A species-specific RLB probe, designed to target the hypervariable V4 region of the 18S rRNA gene for detection of the novel Babesia sp., was used to screen an additional 137 cheetah blood specimens for the presence of the species. The prevalence of infection was 28.5%. Here we describe the morphology and phylogenetic relationships of the novel species, which we have named Babesia lengau sp. nov.

  10. SEROLOGICAL, PARASITOLOGICAL AND MOLECULAR ASSESSMENT OF Babesia bovis AND Babesia bigemina IN CATTLE FROM STATE OF MARANHÃO

    Directory of Open Access Journals (Sweden)

    FRANCISCO BORGES COSTA

    2015-01-01

    Full Text Available The aim of the present study was to determine the prevalence of Babesia bovis and Babesia bigemina in dairy cattle from São Luis Island in the state of Maranhão, Brazil. A total of 281 blood samples were collected. In total, 275 (97.9% animals were B. bovis-reactive and B. bigemina reactive in the Enzyme-Linked Immunosorbent Assay (ELISA. The microscopy examination detected 22 (7.8% animals that were positive for Babesia sp. and the Polymerase Chain Reaction (PCR analysis showed that 91 animals (32.38% and 23 animals (8.18% were positive for B. bovis and B. bigemina, respectively, while 17 animals (6.04% were co-infected. There is a high level of transmission of these protozoa in Maranhão, and the animals were naturally exposed. Therefore, it is possible to characterize the island as enzootic stability for babesiosis, indicat-ing a risk of financial losses when susceptible animals are introduced from areas of enzootic instability or free regions of B. bovis and B. bigemina.

  11. Use of Equipment Information System (EQUIS) to determine priority for purchasing safeguards equipment

    International Nuclear Information System (INIS)

    Silberberg, S.

    1988-01-01

    To manage its large world-wide inventory of safeguards equipment, the IAEA Safeguards department uses a computerized Equipment Information System (EQUIS). EQUIS data have been analyzed using Queueing Theory to determine if inventory is adequate to meet inspector demands and in those cases where it is inadequate, to indicate how many additional units should be procured. Results are tabulated for various types of non-destructive analysis (NDA) equipment. For equipment where there is a high turnover and hence a large amount of data, the analysis provides a powerful tool for assisting procurement decisions

  12. Pneumonia due to Rhodococcus equi in a non-Hodgkin's lymphoma patient: case report

    Directory of Open Access Journals (Sweden)

    Iuri de França Bonilha

    Full Text Available The authors reported a lung infection by Rhodococcus equi in a 25 years-old male patient admitted to hospital with cough, dyspnea, fever, and previous diagnosis of pleural effusion. R. equi was isolated from pleural fluid and the patient acquired nosocomial infection by Acinetobacter baumannii, isolated from chest drain. The patient was treated with antibiotics. During hospitalization, he was diagnosed with non-Hodgkin lymphoma of precursor T-cell lymphoblastic lymphoma subtype in biopsy of pleura. After undergoing surgery for pulmonary decortication for drain empyema, the patient died due to septicemia.

  13. Natural Infection of the South American Tapir ( Tapirus terrestris ) by Theileria equi.

    Science.gov (United States)

    Da Silveira, Alexandre Welzel; De Oliveira, Gustavo Gomes; Menezes Santos, Leandro; da Silva Azuaga, Lucas Bezerra; Macedo Coutinho, Claudia Regina; Echeverria, Jessica Teles; Antunes, Tamires Ramborger; do Nascimento Ramos, Carlos Alberto; Izabel de Souza, Alda

    2017-04-01

    Theileria equi is a tick-borne piroplasm considered endemic in equines in Brazil. The cohabitation of domestic and wild animals in areas of extensive cattle breeding favors the close contact between different species and the sharing of vectors and, consequently, pathogens. We report the natural infection of a young South American tapir ( Tapirus terrestris ) by T. equi in Mato Grosso do Sul, Brazil. Although it was not possible to associate the clinical and hematologic status of the animal with the infection by the protozoan parasite, our report represents an alert on the sharing of pathogens between domestic and wild animals.

  14. Identificação diferencial de Rhodococcus equi e Dietzia maris em bubalinos Differential identification of Rhodococcus equi and Dietzia maris in buffaloes

    Directory of Open Access Journals (Sweden)

    L.R. Viana

    2009-08-01

    Full Text Available Foram analisados 24 isolados bacterianos oriundos de leite e pele de búfalas (Bubalus bubalis, os quais foram previamente identificados como Rhodococcus equi com o auxílio de fenotipia concisa. Testes fenotípicos complementares e ferramentas moleculares foram utilizados com o objetivo de caracterizar esses isolados, bem como diferenciá-los de outros microrganismos intimamente relacionados. Observaram-se três fenótipos distintos, porém a identificação dos isolados foi inconclusiva. Apenas um dos isolados foi comprovado como sendo R. equi com a realização da PCR espécie-específica, sequenciamento e análise dos fragmentos de DNA. Os demais isolados só foram identificados pelo sequenciamento de fragmento do gene que codifica a região 16S do rRNA universal de bactérias, indicando tratar-se de Dietzia maris. O perfil de susceptibilidade aos antimicrobianos revelou maior resistência dos isolados de D. maris para oxacilina (96% e rifampicina (87%. O isolado de R. equi apresentou resistência à amicacina, oxacilina, penicilina, rifampicina e tetraciclina. Alerta-se para o risco da incorreta identificação dos isolados baseados em testes fenotípicos concisos e para a necessidade de utilização de testes complementares para diferenciação entre R. equi e D. maris.Twenty-four bacterial isolates from milk and skin of buffalo females (Bubalus bubalis, which previously had been identified as Rhodococcus equi by using a restricted number of phenotypical tests for bacterial characterization, were analyzed. The goal of this study was to perform the characterization of these isolates, as well as the differentiation of other microorganisms closely related by using additional phenotypical tests and molecular tools. Based on the phenotypical results, three different biotypes were obtained. However, the identification of the isolates was inconclusive. Only one of the isolates was confirmed as R. equi by the PCR specifically for this species, as

  15. Molecular diagnosis and phylogenetic analysis of Babesia bigemina and Babesia bovis hemoparasites from cattle in South Africa

    Science.gov (United States)

    2013-01-01

    Background Babesia parasites, mainly Babesia bovis and B. bigemina, are tick-borne hemoparasites inducing bovine babesiosis in cattle globally. The clinical signs of the disease include, among others, anemia, fever and hemoglobinuria. Babesiosis is known to occur in tropical and subtropical regions of the world. In this study, we aim to provide information about the occurrence and phylogenetic relationship of B. bigemina and B. bovis species in cattle from different locations in nine provinces of South Africa. A total of 430 blood samples were randomly collected from apparently healthy cattle. These samples were genetically tested for Babesia parasitic infections using nested PCR assays with species-specific primers. Results Nested PCR assays with Group I primer sets revealed that the overall prevalence of B. bigemina and B. bovis in all bovine samples tested was 64.7% (95% CI = 60.0-69.0) and 35.1% (95% CI = 30.6-39.8), respectively. Only 117/430 (27.2%) animals had a mixed infection. The highest prevalence of 87.5% (95% CI = 77.2-93.5) for B. bigemina was recorded in the Free State province collection sites (Ficksburg, Philippolis and Botshabelo), while North West collection sites had the highest number of animals infected with B. bovis (65.5%; 95% CI = 52.7-76.4). Phylograms were inferred based on B. bigemina-specific gp45 and B. bovis-specific rap-1 nucleotide sequences obtained with Group II nested PCR primers. Phylogenetic analysis of gp45 sequences revealed significant differences in the genotypes of B. bigemina isolates investigated, including those of strains published in GenBank. On the other hand, a phylogeny based on B. bovis rap-1 sequences indicated a similar trend of clustering among the sequences of B. bovis isolates investigated in this study. Conclusion This study demonstrates the occurrence of Babesia parasites in cattle from different provinces of South Africa. It was also noted that the situation of Babesia parasitic infection

  16. Molecular diagnosis and phylogenetic analysis of Babesia bigemina and Babesia bovis hemoparasites from cattle in South Africa.

    Science.gov (United States)

    Mtshali, Moses Sibusiso; Mtshali, Phillip Senzo

    2013-08-08

    Babesia parasites, mainly Babesia bovis and B. bigemina, are tick-borne hemoparasites inducing bovine babesiosis in cattle globally. The clinical signs of the disease include, among others, anemia, fever and hemoglobinuria. Babesiosis is known to occur in tropical and subtropical regions of the world. In this study, we aim to provide information about the occurrence and phylogenetic relationship of B. bigemina and B. bovis species in cattle from different locations in nine provinces of South Africa. A total of 430 blood samples were randomly collected from apparently healthy cattle. These samples were genetically tested for Babesia parasitic infections using nested PCR assays with species-specific primers. Nested PCR assays with Group I primer sets revealed that the overall prevalence of B. bigemina and B. bovis in all bovine samples tested was 64.7% (95% CI = 60.0-69.0) and 35.1% (95% CI = 30.6-39.8), respectively. Only 117/430 (27.2%) animals had a mixed infection. The highest prevalence of 87.5% (95% CI = 77.2-93.5) for B. bigemina was recorded in the Free State province collection sites (Ficksburg, Philippolis and Botshabelo), while North West collection sites had the highest number of animals infected with B. bovis (65.5%; 95% CI = 52.7-76.4). Phylograms were inferred based on B. bigemina-specific gp45 and B. bovis-specific rap-1 nucleotide sequences obtained with Group II nested PCR primers. Phylogenetic analysis of gp45 sequences revealed significant differences in the genotypes of B. bigemina isolates investigated, including those of strains published in GenBank. On the other hand, a phylogeny based on B. bovis rap-1 sequences indicated a similar trend of clustering among the sequences of B. bovis isolates investigated in this study. This study demonstrates the occurrence of Babesia parasites in cattle from different provinces of South Africa. It was also noted that the situation of Babesia parasitic infection in cattle from certain areas

  17. Isolation and characterization of Babesia pecorum sp. nov. from farmed red deer (Cervus elaphus).

    Science.gov (United States)

    Jouglin, Maggy; Fernández-de-Mera, Isabel G; de la Cotte, Nathalie; Ruiz-Fons, Francisco; Gortázar, Christian; Moreau, Emmanuelle; Bastian, Suzanne; de la Fuente, José; Malandrin, Laurence

    2014-08-26

    The diversity of Babesia species infecting cervids in parts of central and southern Spain was analyzed by collecting blood from farmed red deer (Cervus elaphus). Babesia sp. was isolated in vitro from two red deer herds in Cádiz and Ciudad Real. The number of Babesia sp. carriers differed between the two herds: 36/77 in Cádiz and 1/35 in Ciudad Real. Hyalomma lusitanicum was the most prevalent tick species identified on the Cádiz farm vegetation and on sampled animals, and is therefore a candidate vector. The molecular characteristics of 21 isolates were determined by complete (8 isolates) or partial (13 isolates) 18S rRNA gene sequencing. The sequences were highly similar (over 99.4% identity) and 6 sequence types were identified at the level of one herd only, demonstrating a rather high genetic diversity. They formed a monophyletic clade, and members of the three main sequence types shared a similar morphology and the same erythrocyte susceptibility pattern. This clade also included Babesia sp. Xinjiang isolated from sheep in China and Babesia sp. identified in giraffe in South Africa, with identities higher than 98.3% and statistically relevant phylogenetic support. None of the biological properties analyzed for both Babesia from red deer and Babesia sp. Xinjiang allowed their differentiation (ability to develop in vitro in erythrocytes from cattle and sheep, as well as in erythrocytes from different cervids, unsuccessful infection of calves). We propose the Babesia isolated from red deer as a new species named B. pecorum. Whether Babesia sp. Xinjiang and the Babesia characterized in South Africa belong to the same species is debated.

  18. Epidemiology, diagnosis and therapy of Theileria equi infection in Giza, Egypt

    Directory of Open Access Journals (Sweden)

    Fayez A. Salib

    2013-04-01

    Full Text Available Aim: To study the prevalence of Theileria equi among horses in different age groups, both sexes, months and seasons of the year, and regions of Giza governourate. Studying the changes in the blood picture, blood chemistries, liver enzymes associate with T.equi infections in horses. Evaluating IFA and CFT at different dilutions in the serodiagnosis of T.equi infections in horses. Evaluating four anti-Theileria medication regimens (diminazine aceturate, imidiocarb 7%, buparvaquone and a combination of imidiocarb 7% and buparvaquone in treatment of T.equi infections in horses. Materials and Methods: Total of 149 horses were examined by clinical signs and blood smears. Fortey whole blood samples from T.equi infected horses were examined to measure haemoglobin, total RBCs count and PCV. Fortey serum samples from T.equi infected horses were examined to measure total bilirubin, direct bilirubin, ALT and AST enzymes. Serum samples from T.equi infected (40 and non infected (14 horses were tested by indirect fluorescent antibody test (IFA and complement fixation test (CFT at different dilutions. Four groups of T.equiinfected horses (A,B,C,D, each group was represented by 10 horses and was separately treated with diminazine aceturate, imidiocarb 7%, buparvaquone and a combination of imidiocarb 7% and buparvaquone respectively. Results: the prevalence of T.equi was 41.61% in totally examined horses. The prevalence was higher in males than females. The highest prevalence was among age group ranged from 5-10 years as (22.81%. The highest prevalence was in July and was recorded as (25.81% and the disease was more prevalent in summer than winter. The highest prevalence was recorded in Nazlet-alsamman as (51.61%. Equine theileriosis was clinically characterized by fever, haemoglobinuria, oedema, anaemia and icterus. The best dilution for IFA was 1/160 where sensitivity, specificity and accuracy were the highest for this test as (98%, (92.86% and (97

  19. Strong conservation of rhoptry-associated-protein-1 (RAP-1) locus organization and sequence among Babesia isolates infecting sheep from China (Babesia motasi-like phylogenetic group).

    Science.gov (United States)

    Niu, Qingli; Valentin, Charlotte; Bonsergent, Claire; Malandrin, Laurence

    2014-12-01

    Rhoptry-associated-protein 1 (RAP-1) is considered as a potential vaccine candidate due to its involvement in red blood cell invasion by parasites in the genus Babesia. We examined its value as a vaccine candidate by studying RAP-1 conservation in isolates of Babesia sp. BQ1 Ningxian, Babesia sp. Tianzhu and Babesia sp. Hebei, responsible for ovine babesiosis in different regions of China. The rap-1 locus in these isolates has very similar features to those described for Babesia sp. BQ1 Lintan, another Chinese isolate also in the B. motasi-like phylogenetic group, namely the presence of three types of rap-1 genes (rap-1a, rap-1b and rap-1c), multiple conserved rap-1b copies (5) interspaced with more or less variable rap-1a copies (6), and the 3' localization of one rap-1c. The isolates Babesia sp. Tianzhu, Babesia sp. BQ1 Lintan and Ningxian were almost identical (average nucleotide identity of 99.9%) over a putative locus of about 31 Kb, including the intergenic regions. Babesia sp. Hebei showed a similar locus organization but differed in the rap-1 locus sequence, for each gene and intergenic region, with an average nucleotide identity of 78%. Our results are in agreement with 18S rDNA phylogenetic studies performed on these isolates. However, in extremely closely related isolates the rap-1 locus seems more conserved (99.9%) than the 18S rDNA (98.7%), whereas in still closely related isolates the identities are much lower (78%) compared with the 18S rDNA (97.7%). The particularities of the rap-1 locus in terms of evolution, phylogeny, diagnosis and vaccine development are discussed. Copyright © 2014 The Authors. Published by Elsevier B.V. All rights reserved.

  20. Disseminated Rhodococcus equi infection in a kidney transplant patient without initial pulmonary involvement

    NARCIS (Netherlands)

    Rahamat-Langendoen, Janette C.; van Meurs, Matijs; Zijlstra, Jan G.; Lo-Ten-Foe, Jerome R.

    2009-01-01

    Rhodococcus equi is increasingly recognized as an opportunistic pathogen in solid organ transplant recipients. Primary pulmonary involvement is the most common finding. We report a case of a 42-year-old female kidney transplant recipient who developed multiple disseminated abscesses caused by R.

  1. In vitro and intra-macrophage gene expression by Rhodococcus equi strain 103.

    Science.gov (United States)

    Rahman, Md Tanvir; Parreira, Valeria; Prescott, John F

    2005-09-30

    Rhodococcus equi is a facultative intracellular respiratory pathogen of foals that persists and multiplies within macrophages. In foals, virulence is associated with 80-90 kb plasmids, which include a pathogenicity island (PI) containing the virulence-associated protein (vap) gene family, but detailed understanding of the basis of virulence is still poor. A 60 spot-based DNA microarray was developed containing eight PI genes and 42 chromosomal putative virulence or virulence-associated genes selected from a recent partial genome sequence in order to study transcription of these genes by R. equi grown inside macrophages and under in vitro conditions thought to simulate those of macrophages. In addition to seven PI genes, nine chromosomal genes involved in fatty acid and lipid metabolism (choD, fadD13, fbpB), heme biosynthesis (hemE), iron utilization (mbtF), heat shock resistance and genes encoding chaperones (clpB, groEL), a sigma factor (sigK), and a transcriptional regulator (moxR) were significantly induced in R. equi growing inside macrophages. The pattern of R. equi chromosomal genes significantly transcribed inside macrophages largely differed from those transcribed under in vitro conditions (37 degrees C, pH 5.0 or 50mM H2O2 for 30 min). This study has identified genes, other than those of the virulence plasmid, the transcription of which is enhanced within equine macrophages. These genes should be investigated further to improve understanding of how this organism survives intracellularly.

  2. Diversion of phagosome trafficking by pathogenic Rhodococcus equi depends on mycolic acid chain length.

    Science.gov (United States)

    Sydor, Tobias; von Bargen, Kristine; Hsu, Fong-Fu; Huth, Gitta; Holst, Otto; Wohlmann, Jens; Becken, Ulrike; Dykstra, Tobias; Söhl, Kristina; Lindner, Buko; Prescott, John F; Schaible, Ulrich E; Utermöhlen, Olaf; Haas, Albert

    2013-03-01

    Rhodococcus equi is a close relative of Mycobacterium spp. and a facultative intracellular pathogen which arrests phagosome maturation in macrophages before the late endocytic stage. We have screened a transposon mutant library of R. equi for mutants with decreased capability to prevent phagolysosome formation. This screen yielded a mutant in the gene for β-ketoacyl-(acyl carrier protein)-synthase A (KasA), a key enzyme of the long-chain mycolic acid synthesizing FAS-II system. The longest kasA mutant mycolic acid chains were 10 carbon units shorter than those of wild-type bacteria. Coating of non-pathogenic E. coli with purified wild-type trehalose dimycolate reduced phagolysosome formation substantially which was not the case with shorter kasA mutant-derived trehalose dimycolate. The mutant was moderately attenuated in macrophages and in a mouse infection model, but was fully cytotoxic.Whereas loss of KasA is lethal in mycobacteria, R. equi kasA mutant multiplication in broth was normal proving that long-chain mycolic acid compounds are not necessarily required for cellular integrity and viability of the bacteria that typically produce them. This study demonstrates a central role of mycolic acid chain length in diversion of trafficking by R. equi. © 2012 Blackwell Publishing Ltd.

  3. MICs of Oxazolidinones for Rhodococcus equi Strains Isolated from Humans and Animals

    OpenAIRE

    Bowersock, Terry L.; Salmon, Sarah A.; Portis, Ellen S.; Prescott, John F.; Robison, Denise A.; Ford, Charles W.; Watts, Jeffrey L.

    2000-01-01

    Eperezolid and linezolid are representatives of a new class of orally active, synthetic antimicrobial agents. The in vitro activity values (MICs) of linezolid, eperezolid, and comparator antibiotics against 102 strains of Rhodococcus equi isolated from humans and animals were determined. Linezolid was more active than eperezolid against the strains tested; premafloxacin was the most active comparator antibiotic.

  4. Streptococcus equi subspecies zooepidemicus Infections in Humans by Zoonotic Transmission from Horses

    Centers for Disease Control (CDC) Podcasts

    2013-06-12

    Dr. Mike Miller reads an abridged version of the Emerging Infectious Diseases’ article, Streptococcus equi subspecies zooepidemicus Infections in Humans by Zoonotic Transmission from Horses.  Created: 6/12/2013 by National Center for Emerging and Zoonotic Infectious Diseases (NCEZID).   Date Released: 7/3/2013.

  5. Rhodococcus equi infection after alemtuzumab therapy for T-cell prolymphocytic leukemia

    NARCIS (Netherlands)

    Meeuse, Jan J.; Sprenger, Herman G.; Van Assen, Sander; Leduc, Dominique; Daenen, Simon M.G.J.; Arends, Jan P.; van der Werf, Tjipke

    2007-01-01

    Rhodococcus equi, mainly known from veterinary medicine as a pathogen in domestic animals, can also cause infections in immunocompromised humans, especially in those with defects in cellular immunity. Alemtuzumab, an anti-CD52 monoclonal antibody, causes lymphocytopenia by eliminating CD52-positive

  6. Recognition of a 30,000 MW antigen of Giardia muris trophozoites by intestinal IgA from Giardia-infected mice.

    Science.gov (United States)

    Heyworth, M F; Pappo, J

    1990-08-01

    The principal aims of this work were (i) to identify the molecular weight (MW) of Giardia muris trophozoite antigens that are recognized by IgA in small intestinal secretions from G. muris-infected mice, and (ii) to determine whether mouse intestinal Giardia-specific IgA is directed against trophozoite surfaces. BALB/c mice were infected with G. muris cysts, and intestinal secretions were harvested from these mice at various times after the start of Giardia infection, and from uninfected mice. Flow cytometry showed that intestinal IgA from G. muris-infected mice, but not from uninfected mice, became bound to trophozoite surfaces in vitro. Western blotting of trophozoite proteins with mouse intestinal secretions showed that IgA from Giardia-infected mice reacted specifically with a broad protein band of approximately 30,000 MW. This finding suggests that one or more trophozoite proteins of approximately 30,000 MW are targets for intestinal antibody in mice infected with G. muris.

  7. Peritoneal Effusion in a Dog due to Babesia gibsoni Infection

    Directory of Open Access Journals (Sweden)

    Suresh Gonde

    2014-01-01

    Full Text Available A five-year-old male Labrador was presented to Teaching Veterinary Clinics of GADVASU with a primary complaint of distended abdomen, fever, and anorexia. The dog was found to be dull with elevated rectal temperature (104°F, heart rate (148 per minute, and respiration rate (58 per minute. Blood smear examination and PCR assay revealed that dog was positive for Babesia gibsoni. Elevated bilirubin, alanine amino transferase (ALT, alkaline phosphatase (ALP, creatinine, blood urea nitrogen (BUN, total leucocyte count, hypoalbuminaemia, and hypoproteinaemia were haematobiochemical alterations. Radiography and ultrasonography showed ground glass appearance and anechoic area of abdomen, respectively.

  8. A recently identified ovine Babesia in China: serology and sero-epidemiology.

    Science.gov (United States)

    Guan, Guiquan; Ma, Miling; Liu, Aihong; Ren, Qiaoyun; Wang, Jinming; Yang, Jifei; Li, Anyan; Liu, Zhijie; Du, Pengfei; Li, Youquan; Liu, Qing; Zhu, Hai; Yin, Hong; Luo, Jianxun

    2012-12-01

    Babesia sp. in Xinjiang, transmitted by Hyalomma, is a large Babesia that is infective for small ruminants, but it has almost no pathogenicity in healthy sheep. On the basis of the sequences of the 18S rRNA and internal transcribed spacer (ITS) genes, morphological characteristics, vector tick species and pathogenicity it was identified recently as a novel Babesia species. In the present study, an enzyme-linked immunosorbent assay (ELISA) was developed using soluble merozoite antigens of Babesia sp. in Xinjiang (BXJMA) derived from in vitro culture. When the positive threshold was chosen as 24.65% of the specific mean antibody rate, the specificity and sensitivity were both 97.3%. There was no cross-reaction between BXJMA and positive sera from sheep infected with other Chinese ovine piroplasms or Anaplasma ovis in the ELISA and western blotting. Specific antibodies against Babesia sp. in Xinjiang could be detected 2 weeks post infection and a high level of antibodies persisted for more than 12 weeks in experimentally infected sheep. The ELISA was tested on 3857 sera collected from small ruminants in 50 prefectures of 22 provinces to evaluate the sero-epidemiology of Babesia sp. in Xinjiang infection, and the average positive rate was 31.66%. These data provide that the developed ELISA is a powerful tool for the sero-diagnosis of Babesia sp. in Xinjiang and confirm that it is a novel species. Copyright © 2012 Elsevier Ireland Ltd. All rights reserved.

  9. Structure of the virulence-associated protein VapD from the intracellular pathogen Rhodococcus equi

    International Nuclear Information System (INIS)

    Whittingham, Jean L.; Blagova, Elena V.; Finn, Ciaran E.; Luo, Haixia; Miranda-CasoLuengo, Raúl; Turkenburg, Johan P.; Leech, Andrew P.; Walton, Paul H.; Murzin, Alexey G.; Meijer, Wim G.; Wilkinson, Anthony J.

    2014-01-01

    VapD is one of a set of highly homologous virulence-associated proteins from the multi-host pathogen Rhodococcus equi. The crystal structure reveals an eight-stranded β-barrel with a novel fold and a glycine rich ‘bald’ surface. Rhodococcus equi is a multi-host pathogen that infects a range of animals as well as immune-compromised humans. Equine and porcine isolates harbour a virulence plasmid encoding a homologous family of virulence-associated proteins associated with the capacity of R. equi to divert the normal processes of endosomal maturation, enabling bacterial survival and proliferation in alveolar macrophages. To provide a basis for probing the function of the Vap proteins in virulence, the crystal structure of VapD was determined. VapD is a monomer as determined by multi-angle laser light scattering. The structure reveals an elliptical, compact eight-stranded β-barrel with a novel strand topology and pseudo-twofold symmetry, suggesting evolution from an ancestral dimer. Surface-associated octyl-β-d-glucoside molecules may provide clues to function. Circular-dichroism spectroscopic analysis suggests that the β-barrel structure is preceded by a natively disordered region at the N-terminus. Sequence comparisons indicate that the core folds of the other plasmid-encoded virulence-associated proteins from R. equi strains are similar to that of VapD. It is further shown that sequences encoding putative R. equi Vap-like proteins occur in diverse bacterial species. Finally, the functional implications of the structure are discussed in the light of the unique structural features of VapD and its partial structural similarity to other β-barrel proteins

  10. Structure of the virulence-associated protein VapD from the intracellular pathogen Rhodococcus equi

    Energy Technology Data Exchange (ETDEWEB)

    Whittingham, Jean L.; Blagova, Elena V. [University of York, Heslington, York YO10 5DD (United Kingdom); Finn, Ciaran E.; Luo, Haixia; Miranda-CasoLuengo, Raúl [University College Dublin, Dublin (Ireland); Turkenburg, Johan P.; Leech, Andrew P.; Walton, Paul H. [University of York, Heslington, York YO10 5DD (United Kingdom); Murzin, Alexey G. [MRC Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge Biomedical Campus, Cambridge CB2 0QH (United Kingdom); Meijer, Wim G. [University College Dublin, Dublin (Ireland); Wilkinson, Anthony J., E-mail: tony.wilkinson@york.ac.uk [University of York, Heslington, York YO10 5DD (United Kingdom)

    2014-08-01

    VapD is one of a set of highly homologous virulence-associated proteins from the multi-host pathogen Rhodococcus equi. The crystal structure reveals an eight-stranded β-barrel with a novel fold and a glycine rich ‘bald’ surface. Rhodococcus equi is a multi-host pathogen that infects a range of animals as well as immune-compromised humans. Equine and porcine isolates harbour a virulence plasmid encoding a homologous family of virulence-associated proteins associated with the capacity of R. equi to divert the normal processes of endosomal maturation, enabling bacterial survival and proliferation in alveolar macrophages. To provide a basis for probing the function of the Vap proteins in virulence, the crystal structure of VapD was determined. VapD is a monomer as determined by multi-angle laser light scattering. The structure reveals an elliptical, compact eight-stranded β-barrel with a novel strand topology and pseudo-twofold symmetry, suggesting evolution from an ancestral dimer. Surface-associated octyl-β-d-glucoside molecules may provide clues to function. Circular-dichroism spectroscopic analysis suggests that the β-barrel structure is preceded by a natively disordered region at the N-terminus. Sequence comparisons indicate that the core folds of the other plasmid-encoded virulence-associated proteins from R. equi strains are similar to that of VapD. It is further shown that sequences encoding putative R. equi Vap-like proteins occur in diverse bacterial species. Finally, the functional implications of the structure are discussed in the light of the unique structural features of VapD and its partial structural similarity to other β-barrel proteins.

  11. Identification of a novel Babesia sp. from a sable antelope (Hippotragus niger Harris, 1838).

    Science.gov (United States)

    Oosthuizen, Marinda C; Zweygarth, Erich; Collins, Nicola E; Troskie, Milana; Penzhorn, Banie L

    2008-07-01

    Babesiosis in a sable antelope (Hippotragus niger Harris, 1838) was first reported in 1930; the parasite was named Babesia irvinesmithi. Recently, specimens from an adult sable that presented with a sudden onset of disease and that subsequently died during immobilization were submitted for molecular characterization. Microscopic examination of thin blood smears revealed the presence of small piroplasms. DNA was extracted from blood samples; the V4 variable region of the 18S rRNA gene was amplified and analyzed using the reverse line blot (RLB) assay. Amplicons did not hybridize with any of the Babesia or Theileria species-specific probes present on the blot and hybridized only with a Babesia or Theileria genus-specific probe, suggesting the presence of a novel species. The full-length 18S rRNA gene sequence was obtained and aligned with published sequences of related genera, and phylogenetic trees were constructed. Sequence similarity analyses indicated that a Babesia species, designated Babesia sp. (sable), was present. The sequence showed its highest similarity to B. orientalis and to an unnamed Babesia species previously detected in bovine samples. The latter was later established to be Babesia occultans. A Babesia sp. (sable)-specific RLB oligonucleotide probe was designed and used to screen 200 South African sable samples, but so far, no other sample has been found to be positive for the presence of Babesia sp. (sable) DNA. In summary, we identified a novel piroplasm parasite from a sable antelope that died from an unknown illness. While the parasite was observed in blood smears, there is no direct evidence that it was the cause of death.

  12. The effect of surface treatment of silicone hydrogel contact lenses on the attachment of Acanthamoeba castellanii trophozoites.

    Science.gov (United States)

    Beattie, Tara K; Tomlinson, Alan

    2009-11-01

    To determine if plasma surface treatment of Focus Night & Day silicone hydrogel contact lenses affects the attachment of Acanthamoeba. Unworn lotrafilcon A contact lenses with (Focus Night & Day) and without surface treatment and Acuvue, conventional hydrogel lenses, were quartered before 90-min incubation with Acanthamoeba castellanii trophozoites. After incubation and rinsing, the trophozoites attached to one surface of each quarter were counted by direct light microscopy. Sixteen replicates were observed for each lens type. Logarithmic transformation of data allowed the use of parametric analysis of variance. No significant difference in attachment was established between the untreated lotrafilcon A and the conventional hydrogel lenses (Ptreatment of the native Focus Night & Day material produced a significant increase in attachment (Ptreatment to reduce lens hydrophobicity; however, this procedure results in an enhanced acanthamoebal attachment. It is possible that the silicone hydrogel lens could be at a greater risk of promoting Acanthamoeba infection if exposed to the organism because of the enhanced attachment characteristic of this material. Eye care professionals should be aware of the enhanced affinity that Acanthamoeba show for this lens and accordingly emphasise to patients the significance of appropriate lens hygiene. This is particularly important where lenses are worn in a regime that could increase the chance of exposure to the organism, i.e., 6 nights/7 days extended wear or daily wear, where lenses will be stored in a lens case, or where lenses are worn when in contact with potentially contaminated water sources, i.e., swimming or showering.

  13. Molecular detection and characterization of Babesia bovis, Babesia bigemina, Theileria species and Anaplasma marginale isolated from cattle in Kenya.

    Science.gov (United States)

    Adjou Moumouni, Paul Franck; Aboge, Gabriel Oluga; Terkawi, Mohamad Alaa; Masatani, Tatsunori; Cao, Shinuo; Kamyingkird, Ketsarin; Jirapattharasate, Charoonluk; Zhou, Mo; Wang, Guanbo; Liu, Mingming; Iguchi, Aiko; Vudriko, Patrick; Ybanez, Adrian Patalinghug; Inokuma, Hisashi; Shirafuji-Umemiya, Rika; Suzuki, Hiroshi; Xuan, Xuenan

    2015-09-30

    Infections with Babesia bovis, Babesia bigemina, Theileria species and Anaplasma marginale are endemic in Kenya yet there is a lack of adequate information on their genotypes. This study established the genetic diversities of the above tick-borne hemoparasites infecting cattle in Kenya. Nested PCR and sequencing were used to determine the prevalence and genetic diversity of the above parasites in 192 cattle blood samples collected from Ngong and Machakos farms. B. bovis spherical body protein 4, B. bigemina rhoptry-associated protein 1a, A. marginale major surface protein 5, Theileria spp. 18S rRNA, T. parva p104 and T. orientalis major piroplasm surface protein were used as the marker genes. B. bovis, B. bigemina, T. parva, T. velifera, T. taurotragi, T. mutans and A. marginale were prevalent in both farms, whereas T. ovis, Theileria sp. (buffalo) and T. orientalis were found only in Ngong farm. Co-infections were observed in more than 50 % of positive samples in both farms. Babesia parasites and A. marginale sequences were highly conserved while T. parva and T. orientalis were polymorphic. Cattle-derived T. parva was detected in Machakos farm. However, cattle and buffalo-derived Theileria were detected in Ngong farm suggesting interactions between cattle and wild buffaloes. Generally, the pathogens detected in Kenya were genetically related to the other African isolates but different from the isolates in other continents. The current findings reaffirm the endemicity and co-infection of cattle with tick-borne hemoparasites, and the role of wildlife in pathogens transmission and population genetics in Kenya.

  14. Sensibilidad de trofozoítos de Entamoeba histolytica a ivermectina Sensibility of Entamoeba histolytica trophozoites to ivermectin

    Directory of Open Access Journals (Sweden)

    Francisco González-Salazar

    2009-06-01

    . histolytica has two phases: trophozoite and cyst. Trophozoites are the causal agent of disease. The effective treatment for the amebiasis includes drugs with serious collateral effects. Ivermectin is a macrolid with activity against endoparasites and ectoparasites causing strongiloidosis, filariasis, oncocercosis, scabiasis and pediculosis. The use of ivermectin has been extended almost worldwide; it is recognized as a safe drug. The main objective of this study was to determine in vitro sensibility of trophozoites of E. histolytica was to the treatment with ivermectin. To determine the sensibility of the parasites to the drug, E. histolytica was cultivated in PEHPS medium. During its logarithmic growth phase the trophozoites were exposed to different concentrations of ivermectin. As controls other antiparasitic drugs were used. For each drug, serial dilutions were prepared, and mixed in culture tubes with parasites (2 x 10(4 cells/ml. They were incubated for 72 h and then the percentage of growth inhibition was calculated by Probit analysis. Ivermectin showed activity against trophozoites of E. histolytica. The 50% of growth inhibition of ivermectin was 6.40 mg/ml. This dose was higher than for other anti parasitic drugs. Its activity in vivo in animal models remains to be demonstrated.

  15. Genetic characterization and molecular survey of Babesia sp. Xinjiang infection in small ruminants and ixodid ticks in China.

    Science.gov (United States)

    Niu, Qingli; Liu, Zhijie; Yang, Jifei; Gao, Shandian; Pan, Yuping; Guan, Guiquan; Luo, Jianxun; Yin, Hong

    2017-04-01

    Babesia sp. Xinjiang is a large ovine Babesia species that was recently isolated in China. Compared with other ovine Babesia species, it has different morphological features, pathogenicity and vector tick species. The known transmitting vector is Hyalomma anatolicum. In this study, the distribution and the presence of Babesia sp. Xinjiang in small ruminants and ixodid ticks in China were assessed by specific nested-PCR assay based on the rap-1a gene. A total of 978 blood samples from sheep or goats from 15 provinces and 797 tick specimens from vegetation from 10 provinces were collected and analysed for the presence of the Babesia sp. Xinjiang. Full-length and partial rap-1a of Babesia sp. Xinjiang were amplified from field samples. The PCR results were further confirmed by DNA sequencing. Overall, 38 (3.89%) blood samples and 51 (6.4%) tick samples were positive for Babesia sp. Xinjiang infection. The highest presence (26.92%) was found in blood samples from Yunnan province, while H. qinghaiensis ticks with the highest presence of infection (21.3%) were from Gansu province. This study identified for the first time Babesia sp. Xinjiang infection in H. longicornis tick species. The rap-1a sequences of Babesia sp. Xinjiang from field blood and tick samples indicated 100% identity. The presence of Babesia sp. Xinjiang infection may increase in China. Novel potential transmitting vectors might be more extensive than previously thought. Copyright © 2017. Published by Elsevier B.V.

  16. A member of the HSP90 family from ovine Babesia in China: molecular characterization, phylogenetic analysis and antigenicity.

    Science.gov (United States)

    Guan, Guiquan; Liu, Junlong; Liu, Aihong; Li, Youquan; Niu, Qingli; Gao, Jinliang; Luo, Jianxun; Chauvin, Alain; Yin, Hong; Moreau, Emmanuelle

    2015-09-01

    Heat shock protein 90 (HSP90) is a key component of the molecular chaperone complex essential for activating many signalling proteins involved in the development and progression of pathogenic cellular transformation. A Hsp90 gene (BQHsp90) was cloned and characterized from Babesia sp. BQ1 (Lintan), an ovine Babesia isolate belonging to Babesia motasi-like group, by screening a cDNA expression library and performing rapid amplification of cDNA ends. The full-length cDNA of BQHsp90 is 2399 bp with an open reading frame of 2154 bp encoding a predicted 83 kDa polypeptide with 717 amino acid residues. It shows significant homology and similar structural characteristics to Hsp90 of other apicomplex organisms. Phylogenetic analysis, based on the HSP90 amino acid sequences, showed that the Babesia genus is clearly separated from other apicomplexa genera. Five Chinese ovine Babesia isolates were divided into 2 phylogenetic clusters, namely Babesia sp. Xinjiang (previously designated a new species) cluster and B. motasi-like cluster which could be further divided into 2 subclusters (Babesia sp. BQ1 (Lintan)/Babesia sp. Tianzhu and Babesia sp. BQ1 (Ningxian)/Babesia sp. Hebei). Finally, the antigenicity of rBQHSP90 protein from prokaryotic expression was also evaluated using western blot and enzyme-linked immunosorbent assay (ELISA).

  17. Activation of persistent Streptococcus equi subspecies zooepidemicus in mares with subclinical endometritis

    DEFF Research Database (Denmark)

    Petersen, Morten Rønn; Skive, Bolette; Christoffersen, Mette

    2015-01-01

    Endometritis in horses caused by Streptococcus equi subspecies zooepidemicus (S. zooepidemicus) may be underdiagnosed due to traditional diagnostic methods lacking sensitivity and specificity. We serendipitously identified a bacterial growth medium (bActivate) that appeared capable of inducing...

  18. Experimental in vitro transmission of Babesia sp. (EU1) by Ixodes ricinus.

    Science.gov (United States)

    Bonnet, Sarah; Brisseau, Nadine; Hermouet, Axelle; Jouglin, Maggy; Chauvin, Alain

    2009-01-01

    Babesia sp. (EU1), first characterized in 2003, has been implicated in human cases of babesiosis in Italy, Austria and Germany. It has been identified in roe deer and in its suspected tick vector, Ixodes ricinus, in several European countries. The aim of the present study was to validate the competence of I. ricinus as a vector of Babesia sp. (EU1) via experimental infections. For this purpose, a parasite strain isolated from roe deer was cloned in sheep erythrocytes. After experimental infections, parasite DNA was successfully amplified by PCR in both eggs and larvae originating from infected I. ricinus females and in the salivary glands of females exposed to Babesia sp. (EU1) as nymphs. We also demonstrate that infected females were able to transmit parasite DNA during a new blood meal. Together with previous epidemiological studies, these results validate I. ricinus as a competent vector for Babesia sp. (EU1).

  19. Babesia sp. BQ1 (Lintan): molecular evidence of experimental transmission to sheep by Haemaphysalis qinghaiensis and Haemaphysalis longicornis.

    Science.gov (United States)

    Guan, Guiquan; Moreau, Emmanuelle; Liu, Junlong; Hao, Xuefen; Ma, Miling; Luo, Jianxun; Chauvin, Alain; Yin, Hong

    2010-06-01

    Ovine babesiosis is an economically important disease induced by tick transmitted haemoparasites throughout the world. In China, several ovine Babesia strains have been isolated from field-collected ticks or sheep blood during the last two decades but little is known about the vector ticks and transmission pattern. Babesia sp. BQ1 (Lintan) is a Babesia strain infective for sheep and goats, isolated from blood of sheep experimentally infested with Haemaphysalis qinghaiensis collected in field. In the present study, we explored the experimental transmission of Babesia sp. BQ1 (Lintan) to sheep by H. qinghaiensis and Haemaphysalis longicornis. Based on the evidence from nested PCR, it suggested that H. qinghaiensis and H. longicornis are the potential vector ticks of Babesia sp. BQ1 (Lintan) and that larvae, nymphs and adults of both tick species were able to transmit Babesia sp. BQ1 (Lintan) to sheep. Parasites could be detected in the blood, by specific nested PCR, for one month post-infestation.

  20. Babesia spp. in European wild ruminant species: parasite diversity and risk factors for infection

    OpenAIRE

    Michel , Adam O; Mathis , Alexander; Ryser-Degiorgis , Marie-Pierre

    2014-01-01

    International audience; Babesia are tick-borne parasites that are increasingly considered as a threat to animal and public health. We aimed to assess the role of European free-ranging wild ruminants as maintenance mammalian hosts for Babesia species and to determine risk factors for infection. EDTA blood was collected from 222 roe deer (Capreolus c. capreolus), 231 red deer (Cervus e. elaphus), 267 Alpine chamois (Rupicapra r. rupicapra) and 264 Alpine ibex (Capra i. ibex) from all over Switz...

  1. Babesia (Theileria) annae in a red fox (Vulpes vulpes) from Prince Edward Island, Canada.

    Science.gov (United States)

    Clancey, Noel; Horney, Barbara; Burton, Shelley; Birkenheuer, Adam; McBurney, Scott; Tefft, Karen

    2010-04-01

    A 4-6-mo-old female red fox (Vulpes vulpes) was presented to the Atlantic Veterinary College (AVC) Teaching Hospital, Prince Edward Island, Canada. On presentation, the fox was weak and had pale mucous membranes. A complete blood count and a serum biochemistry profile were performed. Blood smear examination revealed low numbers of erythrocytes containing centrally to paracentrally located, single, rarely multiple, approximately 1 x 2 microm, oval to round organisms with morphology similar to Babesia microti. Polymerase chain reaction testing and DNA sequencing of the Babesia species 18S rRNA gene were performed on DNA extracted from whole blood. Results were positive for a Babesia microti-like parasite genetically identical to Babesia (Theileria) annae. The fox was euthanized due to poor prognosis for recovery. Necropsy examination revealed multifocal to locally extensive subacute nonsuppurative meningoencephalitis, an eosinophilic broncho-pneumonia, a moderate diffuse vacuolar hepatopathy, and lesions associated with blunt trauma to the left abdominal region. This is the first reported case of a red fox in Canada infected with a piroplasm. It remains uncertain whether the presence of this hemoparasite in this fox was pathogenic or an incidental finding. The potential for competent vectors of Babesia species on Prince Edward Island, the potential for this Babesia microti-like parasite to infect other wild and domestic canids, and the significance of this parasite to the health of infected individuals are yet to be determined.

  2. Detection and molecular identification of Hepatozoon canis and Babesia vogeli from domestic dogs in Palestine.

    Science.gov (United States)

    Azmi, Kifaya; Al-Jawabreh, Amer; Nasereddin, Abedelmajeed; Abdelkader, Ahmad; Zaid, Taher; Ereqat, Suheir; Sawalha, Samer S; Baneth, Gad; Abdeen, Ziad

    2017-04-01

    Dogs serve as hosts for a great number of parasites, which may affect their health and wellbeing. This study aimed to observe tick borne pathogens in dogs from Palestine including Hepatozoon canis and Babesia species. The prevalence of both H. canis and Babesia species infections in apparently healthy dogs, from ten districts of the West Bank was surveyed. DNA was extracted from blood samples obtained from dogs (n = 362) and ticks (n = 213) collected from dogs (n = 77). A primer set that amplifies a partial sequence of the Babesia and Hepatozoon 18S rRNA gene was used for PCR and the DNA sequences of the PCR products of all samples were determined. Twenty-nine (8·0%) of the dogs were found infected including 20 with H. canis (5·5%), seven with Babesia vogeli (1·9%) and two with undefined Babesia spp. (0·6%). Twelve Rhipicephalus sanguineus s.l ticks were pathogen-positive, including ten with H. canis (4·7%), one with B. vogeli (0·5%), and one with Hepatozoon felis (0·5%). The results indicated that a wide range of tick borne pathogens is circulating in the canine population in the surveyed region. This study is the first report on the prevalence of H. canis, B. vogeli and Babesia spp. in dogs in Palestine and its results will assist in the management of diseases associated with these blood parasites.

  3. Multiple Co-infections of Rodents with Hantaviruses, Leptospira, and Babesia in Croatia

    Science.gov (United States)

    Turk, Nenad; Korva, Miša; Margaletić, Josip; Beck, Relja; Vucelja, Marko; Habuš, Josipa; Svoboda, Petra; Županc, Tatjana Avšič; Henttonen, Heikki; Markotić, Alemka

    2012-01-01

    Abstract Hantaviruses, Leptospira spp., and Babesia spp. are rodent-borne pathogens present worldwide. We studied multiple co-infections of small rodents in Croatia with all three pathogens. Twenty-eight Apodemus flavicollis and 16 Myodes glareolus were tested for the presence of hantavirus RNA by real-time RT-PCR, Leptospira strains by renoculture method and Babesia DNA by PCR. Anti-hantavirus antibodies and anti-Leptospira antibodies were detected by serological methods. Very high infection rates with each pathogen were found in A. flavicollis: 20 of 28 rodents (71%) were infected with Dobrava virus, 13 rodents (46%) were infected with Leptospira, and 5 rodents (18%) were infected with Babesia. Multiple co-infections with all three pathogens were found in 3 of 28 (11%) A. flavicollis animals, suggesting that the same rodent host can be infected with several pathogens at the same time. Dual infections with both hantaviruses and Leptospira were found in 7 of 44 rodents (16%), with hantaviruses and Babesia in 2 rodents (5%), and double infection with both Leptospira and Babesia were found in 1 rodent (2%). Since hantaviruses, Leptospira, and Babesia have similar geographical distributions, it is to be expected that in other parts of the world multiple co-infections, representing a serious threat to public health, can be found. PMID:22217170

  4. The first detection of Babesia species DNA from Japanese black bears (Ursus thibetanus japonicus) in Japan.

    Science.gov (United States)

    Ikawa, Kazuhito; Aoki, Mikiko; Ichikawa, Madoka; Itagaki, Tadashi

    2011-06-01

    In this study, we tried to detect protozoan blood parasites from the liver or blood of 156 Japanese black bears (Ursus thibetanus japonicus) in Iwate Prefecture of Japan by polymerase chain reaction. Two amplicons (approximately 540 bp and 480 bp) were detected by amplification for V4 hyper-variable regions of the 18S rRNA gene. Approximately 540-bp products were obtained in 119 samples (76.3%) and were considered to be DNA of Hepatozoon ursi. Approximately 480-bp products were obtained in 22 samples (14.1%) and were considered to be DNA of Babesia species. The nucleotide sequences (1635 bp) of the 18S rRNA gene of Babesia sp. were very similar (99.3%) to those (AY190123, AY190124) of Babesia sp. detected previously from Ixodes ovatus. Phylogenetic analysis showed that Babesia sp. detected in this study closely related to Babesia sp. derived from raccoons in Japan and the U.S.A. This is the first report of Babesia species detected from Japanese black bears. Copyright © 2011 Elsevier Ireland Ltd. All rights reserved.

  5. Evaluation of Veterinary-Specific Interpretive Criteria for Susceptibility Testing of Streptococcus equi Subspecies with Trimethoprim-Sulfamethoxazole and Trimethoprim-Sulfadiazine.

    Science.gov (United States)

    Sadaka, Carmen; Kanellos, Theo; Guardabassi, Luca; Boucher, Joseph; Watts, Jeffrey L

    2017-01-01

    Antimicrobial susceptibility test results for trimethoprim-sulfadiazine with Streptococcus equi subspecies are interpreted based on human data for trimethoprim-sulfamethoxazole. The veterinary-specific data generated in this study support a single breakpoint for testing trimethoprim-sulfamethoxazole and/or trimethoprim-sulfadiazine with S. equi This study indicates trimethoprim-sulfamethoxazole as an acceptable surrogate for trimethoprim-sulfadiazine with S. equi. Copyright © 2016 Sadaka et al.

  6. Successful treatment of Rhodococcus equi pulmonary infection in a renal transplant recipient.

    OpenAIRE

    Marsh, H. P.; Bowler, I. C.; Watson, C. J.

    2000-01-01

    The rhodococcus is a mycobacterium-like organism which is normally a pathogen in foals. It usually spreads by direct contact or by aerosol from horse faeces and causes pyogranulomatous pulmonary infections. Occasionally, it acts opportunistically to infect immuno-compromised human hosts, most commonly those with the acquired immune deficiency syndrome (AIDS). Here we report a pulmonary infection by Rhodococcus equi in a renal transplant recipient who was successfully treated. The literature o...

  7. A-equi-2 influenza in horses in the Republic of South Africa.

    Science.gov (United States)

    Rogers, A L

    1988-06-01

    In early December 1986 A-equi-2 influenza virus was isolated for the first time in the Republic of South Africa. All horses were susceptible to the highly contagious aerosol-borne orthomyxovirus resulting in widespread outbreaks of equine influenza with typical primary respiratory symptoms. Treatment consisted of rest, anti-inflammatory drugs, antibiotics and good nursing. Future protection can be obtained by vaccination.

  8. Seroepidemiological survey of Rhodococcus equi infection in asymptomatic horses and donkeys from southeast Turkey.

    Science.gov (United States)

    Tel, O Y; Arserim, N B; Keskin, O

    2011-12-01

    In order to assess the level of Rhodococcus equi infection in southeast Turkey, 679 sera from healthy foals and adult horses and 78 sera from donkeys were tested by indirect ELISA using a R. equi reference strain (ATCC 33701) as antigen. Eighty (11.7%) sera from horses and 9 (11.5%) sera from donkeys with titres >0.85 were positive. The prevalence of seropositive horses in Sanliurfa Province was higher than in Diyarbakir Province; 56 (13.9%) horses in Sanliurfa Province and 24 (8.7%) horses in Diyarbakir Province were defined as seropositive. In Sanliurfa Province 14.5% of female (n=343) and 10.1% of male (n = 59) horses tested were defined as seropositive, while in Diyarbakir Province more males (11.4%, n=114) were seropositive than females (6.7%, n=163). Horses 1 to 5 years of age were found to have the highest seropositivity rate in both provinces. A total of 78 sera from donkeys were investigated in Sanliurfa Province, of which 9 (11.5%) were positive by ELISA. Among the 9 positive sera, 6 (12.8%) were from donkeys 1-5 years old and 3 (13.6%) were from donkeys >5 years of age. No positive sera were found in donkeys less than 1 year old. Five (12.5%) sera of females and 4 (10.5%) sera of males tested were positive. These results indicate the existence of R. equi in the horse populations in Sanliurfa and Diyarbakir Provinces. Similar infection rates were found for donkeys in Sanliurfa. This suggests the importance of serological surveys to diagnose R. equi infection in the region and to prevent the zoonotic risk.

  9. Seroepidemiological survey of Rhodococcus equi infection in asymptomatic horses and donkeys from southeast Turkey

    Directory of Open Access Journals (Sweden)

    O. Y. Tel

    2011-05-01

    Full Text Available n order to assess the level of Rhodococcus equi infection in southeast Turkey, 679 sera from healthy foals and adult horses and 78 sera from donkeys were tested by indirect ELISA using a R. equi reference strain (ATCC 33701 as antigen. Eighty (11.7 % sera from horses and 9 (11.5 % sera from donkeys with titres >0.85 were positive. The prevalence of seropositive horses in Sanliurfa Province was higher than in Diyarbakir Province; 56 (13.9 % horses in Sanliurfa Province and 24 (8.7 % horses in Diyarbakir Province were defined as seropositive. In Sanliurfa Province 14.5 % of female (n = 343 and 10.1 % of male (n = 59 horses tested were defined as seropositive, while in Diyarbakir Province more males (11.4 %, n = 114 were seropositive than females (6.7 %, n = 163. Horses 1 to 5 years of age were found to have the highest seropositivity rate in both provinces. A total of 78 sera from donkeys were investigated in Sanliurfa Province, of which 9 (11.5 % were positive by ELISA. Among the 9 positive sera, 6 (12.8 % were from donkeys 1-5 years old and 3 (13.6 % were from donkeys >5years of age. Nopositive sera were found in donkeys less than 1 year old. Five(12.5 % sera of females and 4(10.5 % sera of males tested were positive. These results indicate the existence of R. equi in the horse populations in Sanliurfa and Diyarbakir Provinces. Similar infection rates were found for donkeys in Sanliurfa. This suggests the importance of serological surveys to diagnose R. equi infection in the region and to prevent the zoonotic risk.

  10. Molecular detection of Hepatozoon canis and Babesia canis vogeli in domestic dogs from Cuiabá, Brazil

    OpenAIRE

    Spolidorio,Mariana Granziera; Torres,Mariana de Medeiros; Campos,Wilma Neres da Silva; Melo,Andréia Lima Tomé; Igarashi,Michelle; Amude,Alexandre Mendes; Labruna,Marcelo Bahia; Aguiar,Daniel Moura

    2011-01-01

    The objective of this study was to report for the first time infection by Hepatozoon spp. and Babesia spp. in 10 dogs from the city of Cuiabá, State of Mato Grosso, central-western Brazil. A pair of primers that amplifies a 574 bp fragment of the 18S rRNA of Hepatozoon spp., and a pair of primers that amplifies a 551 bp fragment of the gene 18S rRNA for Babesia spp. were used. Six dogs were positive for Babesia spp., and 9 were positive for Hepatozoon spp. Co-infection of Babesia spp. and Hep...

  11. Molecular characterization and phylogenetic analysis of Babesia sp. NV-1 detected from wild American Mink ( Neovison vison ) in Hokkaido, Japan.

    Science.gov (United States)

    Hirata, Haruyuki; Ishinabe, Satoki; Jinnai, Michio; Asakawa, Mitsuhiko; Ishihara, Chiaki

    2013-04-01

    Babesiosis is a tick-borne protozoan disease affecting many mammalian species worldwide, caused by the intraerythrocytic multiplication of Babesia spp. The present study aimed to detect the presence of Babesia sp. in 13 American mink from Hokkaido, Japan. One of 13 animals was positive, as indicated by nested PCR targeting the 18S ribosomal RNA (SSU rDNA) and subunit 7 (eta) of the chaperonin-containing t-complex polypeptide 1 (CCT7) genes from species of Babesia and Theileria. Sequencing of the PCR product of SSU rDNA revealed 99% homology to the isolates of Babesia sp. SAP#131 found in raccoons in Hokkaido, whereas that of the CCT7 gene showed 80% homology to the isolates of Babesia gibsoni in dogs as determined by BLAST analysis. We refer to the cognate sequence as Babesia sp. NV-1. Phylogenetic analyses of SSU rDNA and CCT7 genes from Babesia sp. NV-1 revealed them to be most closely related to the Babesia sp. SAP#131 from a raccoon in Hokkaido and to canine B. gibsoni, respectively. Here, we provide the first molecular evidence of the Babesia sp. NV-1 parasite in feral American mink ( Neovison vison ) in Hokkaido, Japan.

  12. Prevalence and diversity of Babesia, Hepatozoon, Ehrlichia, and Bartonella in wild and domestic carnivores from Zambia, Africa.

    Science.gov (United States)

    Williams, Brianna M; Berentsen, Are; Shock, Barbara C; Teixiera, Maria; Dunbar, Michael R; Becker, Matthew S; Yabsley, Michael J

    2014-03-01

    A molecular survey was conducted for several hemoparasites of domestic dogs and three species of wild carnivores from two sites in Zambia. Three Babesia spp. were detected including Babesia felis and Babesia leo in lions (Panthera leo) and a Babesia sp. (similar to Babesia lengau) in spotted hyenas (Crocuta crocuta) and a single lion. All wild dogs (Lycaon pictus) and domestic dogs were negative for Babesia. High prevalences for Hepatozoon were noted in all three wild carnivores (38-61%) and in domestic dogs (13%). Significantly higher prevalences were noted in hyenas and wild dogs compared with domestic dogs and lions. All carnivores were PCR negative for Ehrlichia canis, Ehrlichia ewingii, and Bartonella spp. Overall, high prevalences and diversity of Babesia and Hepatozoon were noted in wild carnivores from Zambia. This study is the first molecular characterization of Babesia from any hyena species and is the first report of a Babesia sp. closely related to B. lengau, a parasite previously only reported from cheetahs (Acinonyx jubatus), in lions and hyenas. Although usually benign in wild carnivores, these hemoparasites can be pathogenic under certain circumstances. Importantly, data on vectors for these parasites are lacking, so studies are needed to identify vectors as well as determine transmission routes, infection dynamics, and host specificity of these hemoparasites in wildlife in Africa and also the risk of transmission between domestic animals and wildlife.

  13. Infecção pulmonar por "Rhodococcus equi": relato dos dois primeiros casos brasileiros Pulmonary Rhodococcus equi infection: report of the first two Brazilian cases

    Directory of Open Access Journals (Sweden)

    LUIZ CARLOS SEVERO

    2001-05-01

    Full Text Available O Rhodococcus equi, principal agente da rodococose, é um cocobacilo pleomórfico, gram-positivo e aeróbio, que infecta humanos por via inalatória ou transcutânea e se manifesta clinicamente como abscesso pulmonar. Relatam-se os dois primeiros casos brasileiros da doença. Ambos os pacientes eram imunocomprometidos e apresentavam quadro infeccioso pulmonar. O primeiro tinha AIDS e apresentava pneumonia cavitada em lobo superior esquerdo, que teve evolução fatal. O segundo tinha doença de Goodpasture, insuficiência renal crônica e fazia uso de corticosteróides. Apresentava uma lesão pulmonar escavada no lobo superior direito, que foi tratada com sulfametoxazol-trimetoprim, com resolução do processo.Rhodococcus equi, the principal agent of rhodococcosis, is a pleomorphic, gram-positive, aerobic coccus bacillus that infects humans by inhalation or through a transcutaneous route. It is clinically manifested as a pulmonary abscess. The first two Brazilian cases of rhodococcosis are reported on. Both patients were immunocompromised and showed pulmonary infection. The first patient had AIDS and cavitating pneumonia in the left upper lobe, that was fatal. The second case presented Goodpasture syndrome and was under chronic corticotherapy. He displayed a cavitating nodular lesion in the right upper lobe, that was successfully treated with sulfametoxazol-trimethoprim.

  14. In vitro effect of josamycin in strains of Rhodococcus equi isolated from pulmonar infections in foals / Efeito in vitro da josamicina em cepas de rhodocaccus equi isoladas de afecções pulmonares em potros

    Directory of Open Access Journals (Sweden)

    Jaime Galvão Dias Júnior

    2000-12-01

    Full Text Available Antmicrobial susceptibility test was performed in thirty-one samples of Rhodococcus equi isolated from iung infections in foals. Among the antimicrobial tosted, erythromycin (100,0 %, rifampin (96.3%. neomycin (93,6 %Josamycin (90,4% and gentamicin (90,4 %presented the highest sensitivity against R- equi- The most-common occurrence of resistance was observed from cephalexim (100,0 %}, lincomycin (100,0 %, cephalothin (96,3 %. oxacillin (96,8 %, penicillin G (96,8 %, amoxicillin (90,3 % and trimethoprim/sulfamethoxazole (83,8 %. The highest susceptibility of the R. equi to josamycin, suggest the drug as alternative for therapy of R. equi infections in foals.Procedeu-se o teste de sensibilidade microbiana in vitro pelo teste de difusão com discos em 31 cepas de Rhodococcus equi. isoladas de afecções pulmonares cm potros. frente a 20 antimicrobianos. Os maioires índices de sensibilidade de R. equi foram constatados para entromicina (100,0 %, níampicina (96.8 %}t ncomicina (93. 6 %. josamicina (90.4% e gentamicina (90A %. Os maiores índices de resistência do agente foram verificados para cefalexina (100,0%, lincomicina (100.0%, cefalotina (96.8 %, oxacilino (96,8 %}, penicilina G (96,3 %, amoxicilina (90.3 % e sulfametoxazol/trimetoprim (83,8 %. A alta sensibilidade das cepas de R. equi para a josamicina, sugere a possibilidade de utilização da droga como alternativa no tratamento da rodococose em potros.

  15. The effects of irradiation on Babesia maintained in vitro

    International Nuclear Information System (INIS)

    Irvin, A.D.; Young, E.R.; Adams, P.J.V.

    1979-01-01

    Blood infected with Babesia rodhaini, B major or B divergens was irradiated to different absorbed doses between 0 and 120 krad, and then maintained in vitro in the presence of 3 H hypoxanthine for 24 h. The effects of irradiation were measured by the ability of the parasites to incorporate 3 H hypoxanthine and, in the case of B rodhaini, by the ability of the parasite to infect mice. B major and B divergens were slightly more radioresistant than B rodhaini, but all showed a progressive fall in ability to incorporate 3 H hypoxanthine with increasing does of irradiation when there was increased uptake of 3 H hypoxanthine. In the case of B rodhaini there was close correlation between the ability of the parasites to incorporate 3 H hypoxanthine and their infectivity for mice. Both types of activity were abolished at doses of 40 krad and above. (author)

  16. Novel foci of Dermacentor reticulatus ticks infected with Babesia canis and Babesia caballi in the Netherlands and in Belgium.

    Science.gov (United States)

    Jongejan, Frans; Ringenier, Moniek; Putting, Michael; Berger, Laura; Burgers, Stefan; Kortekaas, Reinier; Lenssen, Jesse; van Roessel, Marleen; Wijnveld, Michiel; Madder, Maxime

    2015-04-17

    Autochthonous populations of Dermacentor reticulatus ticks in the Netherlands were discovered after fatal cases of babesiosis occurred in resident dogs in 2004. The presence of D. reticulatus in the Netherlands has also linked with the emergence of piroplasmosis in the resident horse population. The aim of this study was to put together results of continued surveillance of field sites and hosts for this tick in the Netherlands and also in Belgium and determine their infection status for Babesia and Theileria species. Ticks were collected from the vegetation at 11 locations between 2011 and 2013. D. reticulatus ticks were also collected from different hosts between 2007 and 2013. Ticks were screened by PCR and reverse line blot (RLB). A total of 1368 D. reticulatus ticks were collected from 4 previously known field locations and from 5 new locations in the Netherlands and from 2 sites in Belgium (one old and one new location). A total of 855 ticks collected from 8 locations in the Netherlands and 2 locations in Belgium were tested. Fourteen ticks (1,64%) collected at 4 field locations (Dintelse Gorzen, Rozenburg, Slikken van de Heen and St. Philipsland) were positive for Babesia canis, whereas two ticks were positive for Babesia caballi, one tick in the Dintelse Gorzen in the Netherlands and one tick was found positive in De Panne in Belgium. A further 1092 D. reticulatus ticks were collected between 2007 and 2013 from 40 dogs (132 ticks), two ticks from two humans, 51 ticks from 15 horses, two ticks from two cats, one tick from a roe deer, whereas most ticks (904) were collected from cattle (n = 25). Ticks were found throughout the year on dogs in nearly all provinces of the Netherlands. None of the ticks collected from these hosts were infected. D. reticulatus is continuing its spread into novel areas. The finding that some autochthonous ticks are infected with B. canis and B. caballi poses a threat to the resident dog and horse population and justifies year

  17. Babesia microti, human babesiosis, and Borrelia burgdorferi in Connecticut.

    Science.gov (United States)

    Anderson, J F; Mintz, E D; Gadbaw, J J; Magnarelli, L A

    1991-12-01

    Babesia microti was isolated from a white-footed mouse (Peromyscus leucopus) that was captured in southeastern Connecticut in 1988, when the first human case of babesiosis acquired in Connecticut was recognized. To date, 13 cases of babesiosis have been reported in Connecticut, the largest number of human cases reported on the mainland United States. Two of nine patients quiried remembered a prior tick bite. Since Babesia parasites are known to be vectored only by ticks, we surmise that 12 of these infections were acquired via tick bites; 1 was obtained by blood transfusion (the patient was 46 years of age) from an endemically infected donor. The ages of the patients with tick-acquired babesiosis ranged from 61 to 95 years. Two patients died with active infections, and one patient died from chronic obstructive pulmonary disease soon after treatment with clindamycin and quinine. Indirect fluorescent-antibody titers of blood samples drawn at the time of hospitalization for 11 patients and at the time of active infection for 1 asymptomatic person ranged from 1:1,024 to 1:4,096. Five of eight patients with babesiosis also had significant immunoglobulin G or immunoglobulin M titers (1:640 to 1:5,120) to Borrelia burgdorferi. B. microti was isolated in Syrian hamsters inoculated with blood from 7 of 12 patients tested and was also isolated from mice captured in six towns. The peridomestic nature of the disease was demonstrated by isolating the parasite from white-footed mice captured in or near the yards of eight different patients. Of 59 mice tested, 27 were positive and 25 were coinfected with B. burgdorferi. The isolation of B. microti from a white-footed mouse captured in north-central Connecticut (West Hartford), away from the focus of human infections in southeastern Connecticut, suggests that this pathogen may spread into other areas where Ixodes dammini, the tick vector, becomes established.

  18. The apicoplast genomes of two taxonomic units of Babesia from sheep.

    Science.gov (United States)

    Wang, Tao; Guan, Guiquan; Korhonen, Pasi K; Koehler, Anson V; Hall, Ross S; Young, Neil D; Yin, Hong; Gasser, Robin B

    2017-01-15

    The apicoplast (ap) is a unique, non-photosynthetic organelle found in most apicomplexan parasites. Due to the essential roles that this organelle has, it has been widely considered as target for drugs against diseases caused by apicomplexans. Exploring the ap genomes of such parasites would provide a better understanding of their systematics and their basic molecular biology for therapeutics. However, there is limited information available on the ap genomes of apicomplexan parasites. In the present study, the ap genomes of two operational taxonomic units of Babesia (known as Babesia sp. Lintan [Bl] and Babesia sp. Xinjiang [Bx]) from sheep were sequenced, assembled and annotated using a massive parallel sequencing-based approach. Then, the gene content and gene order in these ap genomes (∼30.7kb in size) were defined and compared, and the genetic differences were assessed. In addition, a phylogenetic analysis of ap genomic data sets was carried out to assess the relationships of these taxonomic units with other apicomplexan parasites for which complete ap genomic data sets were publicly available. The results showed that the ap genomes of Bl and Bx encode 59 and 57 genes, respectively, including 2 ribosomal RNA genes, 25 transfer RNA genes and 30-32 protein-encoding genes, being similar in content to those of Babesia bovis and B. orientalis. Ap gene regions that might serve as markers for future epidemiological and population genetic studies of Babesia species were identified. Using sequence data for a subset of six protein-encoding genes, a close relationship of Bl and Bx with Babesia bovis from cattle and B. orientalis from water buffalo was inferred. Although the focus of the present study was on Babesia, we propose that the present sequencing-bioinformatic approach should be applicable to organellar genomes of a wide range of apicomplexans of veterinary importance. Copyright © 2016 Elsevier B.V. All rights reserved.

  19. Immunity to Babesia in mice I. Adoptive transfer of immunity to Babesia rodhaini with immune spleen cells and the effect of irradiation on the protection of immune mice

    NARCIS (Netherlands)

    Kuil, H.; Zivkovic, D.; Seinen, W.; Albers-van Bemmel, C.M.G.; Speksnijder, J.E.

    1984-01-01

    Immunisation of Balb/c mice against Babesia rodhaini by an amicarbalide- controlled infection resulted in a solid immunity which lasted for 216 days. With spleen cells of immune mice protection could be transferred both to naive mice pretreated with cyclophosphamide. Treatment of naive mice with

  20. Study of the effect of an equi-biaxial loading on the fatigue lifetime of austenitic stainless steel

    International Nuclear Information System (INIS)

    Bradai, Soumaya

    2014-01-01

    Fatigue lifetime assessment is essential in the design of structures. Under-estimated predictions may result in unnecessary in service inspections. Conversely, over-estimated predictions may have serious consequences on the integrity of structures.In some nuclear power plant components, the fatigue loading may be equi-biaxial because of thermal fatigue. So the potential impact of multiaxial loading on the fatigue life of components is a major concern. Meanwhile, few experimental data are available on austenitic stainless steels. It is essential to improve the fatigue assessment methodologies to take into account the potential equi-biaxial fatigue damage. Hence this requires obtaining experimental data on the considered material with a strain tensor in equi-biaxial tension. The aim of this study is to present the experimental and numerical results obtained with a device 'FABIME2' developed in the LISN in collaboration with EDF and AREVA. The association of the experimental results, obtained on the new experimental fatigue device FABIME2, with the numerical analyses obtained by FEM simulation with Cast3M code, has enabled to define the aggravating effect of the equi-biaxial fatigue loading. However, this effect is covered by the Design fatigue curve defined from the nuclear industry. For the crack propagation, a first simplified approach enables to study the kinetic behavior of crack propagation in equi-biaxial fatigue. (author) [fr

  1. A survey of Babesia spp. and Hepatozoon spp. in wild canids in Israel.

    Science.gov (United States)

    Margalit Levi, Maayan; Nachum-Biala, Yaarit; King, Roni; Baneth, Gad

    2018-03-20

    Babesia spp. and Hepatozoon spp. are apicomplexan parasites that infect a variety of animals, including canids. Their life-cycle includes an invertebrate hematophagous vector as a definitive host and vertebrates as intermediate hosts. The aims of this study were to investigate the prevalence and risk factors for Babesia spp. and Hepatozoon spp. infections in wild golden jackals (Canis aureus) and red foxes (Vulpes vulpes) in Israel and to compare spleen with blood sample polymerase chain reaction (PCR) for the detection of infection. Blood and spleen samples from 109 golden jackals and 21 red foxes were tested by PCR for the detection of Babesia spp. and Hepatozoon spp. using primers for the 18S ribosomal (r) RNA gene. Hepatozoon canis was detected in 50/109 (46%) of the jackals and 9/21 (43%) of the foxes. "Babesia vulpes" (the Babesia microti-like piroplasm) was detected in 4/21 (19%) of the foxes and in none of the jackals. A previously unknown genotype termed Babesia sp. MML related to Babesia lengau (96-97% identity) was detected in 1/109 (1%) of the jackals and 4/21 (19%) of the foxes. Further characterization of this genotype carried out by PCR of the rRNA internal transcribed spacer 2 (ITS2) indicated that it had only 87% identity with the B. lengau ITS2. Sex (male or female), age (juvenile or adult) and geographic zone (North, Central or South Israel) were not found to be significant risk factors for these protozoan infections. The prevalence of "B. vulpes" and Babesia sp. MML infections was significantly higher in foxes compared to jackals (χ 2  = 15.65, df = 1, P < 0.005), while there was no statistically significant difference in the rate of H. canis infection between these two canid species. A fair agreement beyond chance between identification in the blood and spleen of H. canis was found in 21 animals from which both blood and spleen samples were available (k = 0.33). This study describes a high prevalence of H. canis infection in

  2. Pathogenicity of Rhodococcus equi in mice, isolated from environment, human and horse clinical samples Patogenicidade atogenicidade em camundongos de isolados clí- clínicos, nicos, ambientais e humanos de Rhodococcus equi

    Directory of Open Access Journals (Sweden)

    Mateus M. Costa

    2006-09-01

    Full Text Available Rhodococcus equi is a facultative intracellular pathogen associated with bronchopneumonia, mesenteric lymphadenitis and enterocolitis in foals. Although R. equi is likely to be found in every horse-breeding farm, the clinical disease is unrecognized in most of them. Capsule components, equi factor, micolic acid and some products encoded by the large 85-90Kb plasmid were described as virulence factors. However, the pathogenesis of R. equi infections and the sensibility of foals are not completely understood. The aim of this study was evaluate the virulence of R. equi isolated from human, horses and environment for mices. Nine strains carrying the 85-90Kb plasmid isolated from foal clinical specimens, one from immunodeficient human patient and six plasmidless strains (four isolated from feces, one from pasture and one from immunodeficient human patient were inoculated in cyclophosphamide immunossuppressed mice. The pathological changes and viability of R. equi cells in the liver of mice was verified after the 3rd, 6th an 10th day after inoculation for horse and environmental isolates and for R. equi isolates from human patients on the 1st, 3rd and 6th day. During the necropsy procedures, infiltrate of macrophages and pyogranulomatous lesions were detected after the sixth pos-inoculation day in the liver and spleen. In horse isolates, only plasmid positive strains were virulent, but in human isolates both strains (plasmid positive e plasmid negative were virulent. Both groups of the immunossupressed mice inoculated with R. equi isolated from environment showed pathological changes. All R. equi strains were unable to kill non imunossuppressed mice.Rhodococ-cus equi é um patógeno intracelular facultativo associado com broncopneumonia, linfadenite mesentérica e enterocolite em potros. Apesar do patógeno ser amplamente distribuído no ambiente equino, a doença não é encontrada em todos os criatórios. Componentes capsulares, "fator equi",

  3. Wide Distribution and Genetic Diversity of Babesia microti in Small Mammals from Yunnan Province, Southwestern China.

    Science.gov (United States)

    Gao, Zi-Hou; Huang, Tao-Hua; Jiang, Bao-Gui; Jia, Na; Liu, Zheng-Xiang; Shao, Zong-Ti; Jiang, Rui-Ruo; Liu, Hong-Bo; Wei, Ran; Li, Yu-Qiong; Yao, Hong-Wu; von Fricken, Michael E; Jiang, Jia-Fu; Du, Chun-Hong; Cao, Wu-Chun

    2017-10-01

    Babesia, usually found in wild and domestic mammals worldwide, have recently been responsible for emerging malaria-like zoonosis in infected patients. Human B. microti infection has been identified in China, primarily in the Southwest along the Myanmar border but little direct surveillance of B. microti infection in rodents has been carried out here (Yunnan province). In this region, a diverse topographic range combined with tropical moisture sustains a high biodiversity of small mammals, which might play important role on Babesia transmission. Small mammals were captured in 141 sample locations from 18 counties located Yunnan Province, and screened for B. microti-like parasites infection by a nested PCR to target 18S rRNA gene of Babesia, plus directly sequencing for positive samples. Univariate and multivariate forward stepwise logistic regression analysis was used to access the association between infections and some related risk factors. Infection with Babesia microti was confirmed in 2.4% (53/ 2204) of small mammals. Significant differences in prevalence rates of B. microti were observed based on variations in forest, agricultural, and residential landscapes. Furthermore, adult small mammals had higher prevalence rates than younger, pubertal mammals. The near full-length 18S rRNA gene revealed that there were two types of B. microti, Kobe and Otsu, which demonstrate the genetic diversity and regional distribution. There exists a wide distribution and genetic diversity of endemic B. microti in Southwestern China, warranting further investigations and monitoring of clinical disease in individuals presenting with Babesia like symptoms in these areas.

  4. Genetic diversity of Streptococcus equi subsp. zooepidemicus and doxycycline resistance in kennelled dogs.

    Science.gov (United States)

    Chalker, Victoria J; Waller, Andrew; Webb, Katy; Spearing, Emma; Crosse, Patricia; Brownlie, Joe; Erles, Kerstin

    2012-06-01

    The genetic diversity and antibiotic resistance profiles of 38 Streptococcus equi subsp. zooepidemicus isolates were determined from a kennelled canine population during two outbreaks of hemorrhagic pneumonia (1999 to 2002 and 2007 to 2010). Analysis of the szp gene hypervariable region and the 16S-23S rRNA intergenic spacer region and multilocus sequence typing (MLST) indicated a predominant tetO-positive, doxycycline-resistant ST-10 strain during 1999 to 2002 and a predominant tetM-positive doxycycline-resistant ST-62 strain during 2007 to 2010.

  5. Pulmonary Rhodococcus equi infection: report of the first two Brazilian cases

    OpenAIRE

    SEVERO, LUIZ CARLOS; RITTER, PATRICIA; PETRILLO, VICTOR FLÁVIO; DIAS, CÍCERO ARMÍDIO GOMES; PORTO, NELSON DA SILVA

    2001-01-01

    O Rhodococcus equi, principal agente da rodococose, é um cocobacilo pleomórfico, gram-positivo e aeróbio, que infecta humanos por via inalatória ou transcutânea e se manifesta clinicamente como abscesso pulmonar. Relatam-se os dois primeiros casos brasileiros da doença. Ambos os pacientes eram imunocomprometidos e apresentavam quadro infeccioso pulmonar. O primeiro tinha AIDS e apresentava pneumonia cavitada em lobo superior esquerdo, que teve evolução fatal. O segundo tinha doença de Goodpas...

  6. The effect of mutation on Rhodococcus equi virulence plasmid gene expression and mouse virulence.

    Science.gov (United States)

    Ren, Jun; Prescott, John F

    2004-11-15

    An 81 kb virulence plasmid containing a pathogenicity island (PI) plays a crucial role in the pathogenesis of Rhodococcus equi pneumonia in foals but its specific function in virulence and regulation of plasmid-encoded virulence genes is unclear. Using a LacZ selection marker developed for R. equi in this study, in combination with an apramycin resistance gene, an efficient two-stage homologous recombination targeted gene mutation procedure was used to mutate three virulence plasmid genes, a LysR regulatory gene homologue (ORF4), a ResD-like two-component response regulator homologue (ORF8), and a gene (ORF10) of unknown function that is highly expressed by R. equi inside macrophages, as well as the chromosomal gene operon, phoPR. Virulence testing by liver clearance after intravenous injection in mice showed that the ORF4 and ORF8 mutants were fully attenuated, that the phoPR mutant was hypervirulent, and that virulence of the ORF10 mutant remained unchanged. A virulence plasmid DNA microarray was used to compare the plasmid gene expression profile of each of the four gene-targeted mutants against the parental R. equi strain. Changes were limited to PI genes and gene induction was observed for all mutants, suggesting that expression of virulence plasmid genes is dominated by a negative regulatory network. The finding of attenuation of ORF4 and ORF8 mutants despite enhanced transcription of vapA suggests that factors other than VapA are important for full expression of virulence. ORF1, a putative Lsr antigen gene, was strongly and similarly induced in all mutants, implying a common regulatory pathway affecting this gene for all four mutated genes. ORF8 is apparently the centre of this common pathway. Two distinct highly correlated gene induction patterns were observed, that of the ORF4 and ORF8 mutants, and that of the ORF10 and phoPR mutants. The gene induction pattern distinguishing these two groups paralleled their virulence in mice.

  7. Performance Assessment of the Wave Dragon Wave Energy Converter Based on the EquiMar Methodology

    DEFF Research Database (Denmark)

    Parmeggiani, Stefano; Chozas, Julia Fernandez; Pecher, Arthur

    2011-01-01

    At the present pre-commercial phase of the wave energy sector, device developers are called to provide reliable estimates on power performance and production at possible deployment locations. The EU EquiMar project has proposed a novel approach, where the performance assessment is based mainly...... on experimental data deriving from sea trials rather than solely on numerical predictions. The study applies this methodology to evaluate the performance of Wave Dragon at two locations in the North Sea, based on the data acquired during the sea trials of a 1:4.5 scale prototype. Indications about power...

  8. Course of infection by Babesia sp. BQ1 (Lintan) and B. divergens in sheep depends on the production of IFNgamma and IL10.

    Science.gov (United States)

    Guan, G; Chauvin, A; Yin, H; Luo, J; Moreau, E

    2010-02-01

    Ovine babesiosis is an important disease in China and responsible for economic losses. Several Babesia strains are involved, but Babesia sp. BQ1 (Lintan) and Babesia sp. BQ1 (Ningxian) are particularly prevalent in the Guansu region. Babesia divergens, in contrast, can experimentally infect spleen-intact sheep, but does not induce clinical signs. The immune response of spleen-intact sheep to Babesia sp. BQ1 (Lintan) and to B. divergens was therefore compared to identify the immune mechanisms involved in pathogenicity. The greater pathogenicity of Babesia sp. BQ1 (Lintan) than that of B. divergens was confirmed: sheep infected with Babesia sp. BQ1 (Lintan), but not with B. divergens, developed hyperthermia and showed patent parasitaemia in Giemsa-stained blood smears from the ear vein. Furthermore, more parasites were also detected in the blood from the jugular vein of Babesia sp. BQ1 (Lintan)-infected sheep. Pathogenicity of Babesia spp. involved cellular responses, but not humoral responses. Interferon-gamma was produced only by specifically activated PBMC from B. divergens-infected sheep and interleukin-10 only by specifically activated PBMC from Babesia sp. BQ1 (Lintan)-infected sheep. The role of these cytokines in the course of infection by Babesia spp. is discussed.

  9. Babesia, Theileria, and Hepatozoon species in ticks infesting animal hosts in Romania.

    Science.gov (United States)

    Andersson, Martin O; Tolf, Conny; Tamba, Paula; Stefanache, Mircea; Radbea, Gabriel; Rubel, Franz; Waldenström, Jonas; Dobler, Gerhard; Chițimia-Dobler, Lidia

    2017-08-01

    Babesia spp., Theileria spp., and Hepatozoon spp. are tick-transmitted apicomplexan parasites that cause several important diseases in animals. To increase current knowledge about the diversity of tick-transmitted pathogens in Romania, we investigated the occurrence of Babesia spp., Theileria spp., and Hepatozoon spp. in a wide range of tick species infesting animal hosts. We collected 852 ticks from 10 different animal species from 20 counties in Romania. The assessment was based on detection of parasite DNA by PCR. Five different apicomplexan parasite species were detected; among them three different species of Babesia: B. canis, B. microti, and B. ovis. Hepatozoon canis was the most frequently detected parasite, found predominately in Ixodes ricinus ticks collected from domestic dogs. It was also detected in I. ricinus collected from goat, fox, and cat. Furthermore, H. canis was found in Haemaphysalis punctata and Haemaphysalis concinna ticks. In addition, Theileria buffeli was detected in Rhipicephalus bursa ticks collected from cattle.

  10. A molecular epidemiological survey of Babesia, Hepatozoon, Ehrlichia and Anaplasma infections of dogs in Japan

    Science.gov (United States)

    KUBO, Shotaro; TATENO, Morihiro; ICHIKAWA, Yasuaki; ENDO, Yasuyuki

    2015-01-01

    Tick-borne diseases are often encountered in canine clinical practice. In the present study, a molecular epidemiological survey of dogs in Japan was conducted to understand the prevalence and geographical distribution of Babesia spp., Hepatozoon spp., Ehrlichia spp. and Anaplasma spp. Pathogen-derived DNA in blood samples obtained from 722 dogs with a history of exposure to ticks and/or fleas was examined by PCR. The prevalence of Babesia gibsoni, Babesia odocoilei-like species, Hepatozoon canis and Ehrlichia spp./Anaplasma spp. was 2.4% (16/722), 0.1% (1/722), 2.5% (18/722) and 1.5% (11/722), respectively. While B. gibsoni and Ehrlichia spp./Anaplasma spp. were detected in the western part of Japan, H. canis was detected in Tohoku area in addition to western and central parts of Japan. PMID:25947226

  11. Molecular characterization of Babesia and Cytauxzoon species in wild South-African meerkats.

    Science.gov (United States)

    Leclaire, Sarah; Menard, Sandie; Berry, Antoine

    2015-04-01

    Piroplasms, including Babesia, Cytauxzoon and Theileria species, frequently infect domestic and wild mammals. At present, there is no information on the occurrence and molecular identity of these tick-borne blood parasites in the meerkat, one of South Africa's most endearing wildlife celebrities. Meerkats live in territorial groups, which may occur on ranchland in close proximity to humans, pets and livestock. Blood collected from 46 healthy meerkats living in the South-African Kalahari desert was screened by microscopy and molecular methods, using PCR and DNA sequencing of 18S rRNA and ITS1 genes. We found that meerkats were infected by 2 species: one species related to Babesia sp. and one species related to Cytauxzoon sp. Ninety one percent of the meerkats were infected by the Cytauxzoon and/or the Babesia species. Co-infection occurred in 46% of meerkats. The pathogenicity and vectors of these two piroplasm species remains to be determined.

  12. vanO, a new glycopeptide resistance operon in environmental Rhodococcus equi isolates

    DEFF Research Database (Denmark)

    Gudeta, Dereje Dadi; Moodley, Arshnee; Bortolaia, Valeria

    2014-01-01

    We describe sequence and gene organization of a new glycopeptide resistance operon (vanO) in Rhodococcus equi from soil. The vanO operon has low homology to enterococccal van operons and harbors a vanHOX cluster transcribed in opposite direction to the vanS-vanR regulatory system and comprised be...... between three open reading frames with unknown function. This finding has clinical interest since glycopeptides are used to treat R. equi infections and resistance has been reported in clinical isolates....

  13. The steroid catabolic pathway of the intracellular pathogen Rhodococcus equi is important for pathogenesis and a target for vaccine development.

    Directory of Open Access Journals (Sweden)

    R van der Geize

    2011-08-01

    Full Text Available Rhodococcus equi causes fatal pyogranulomatous pneumonia in foals and immunocompromised animals and humans. Despite its importance, there is currently no effective vaccine against the disease. The actinobacteria R. equi and the human pathogen Mycobacterium tuberculosis are related, and both cause pulmonary diseases. Recently, we have shown that essential steps in the cholesterol catabolic pathway are involved in the pathogenicity of M. tuberculosis. Bioinformatic analysis revealed the presence of a similar cholesterol catabolic gene cluster in R. equi. Orthologs of predicted M. tuberculosis virulence genes located within this cluster, i.e. ipdA (rv3551, ipdB (rv3552, fadA6 and fadE30, were identified in R. equi RE1 and inactivated. The ipdA and ipdB genes of R. equi RE1 appear to constitute the α-subunit and β-subunit, respectively, of a heterodimeric coenzyme A transferase. Mutant strains RE1ΔipdAB and RE1ΔfadE30, but not RE1ΔfadA6, were impaired in growth on the steroid catabolic pathway intermediates 4-androstene-3,17-dione (AD and 3aα-H-4α(3'-propionic acid-5α-hydroxy-7aβ-methylhexahydro-1-indanone (5α-hydroxy-methylhexahydro-1-indanone propionate; 5OH-HIP. Interestingly, RE1ΔipdAB and RE1ΔfadE30, but not RE1ΔfadA6, also displayed an attenuated phenotype in a macrophage infection assay. Gene products important for growth on 5OH-HIP, as part of the steroid catabolic pathway, thus appear to act as factors involved in the pathogenicity of R. equi. Challenge experiments showed that RE1ΔipdAB could be safely administered intratracheally to 2 to 5 week-old foals and oral immunization of foals even elicited a substantial protective immunity against a virulent R. equi strain. Our data show that genes involved in steroid catabolism are promising targets for the development of a live-attenuated vaccine against R. equi infections.

  14. Occurrence of Babesia species in captive reindeer (Rangifer tarandus) in Germany.

    Science.gov (United States)

    Wiegmann, Lisa; Silaghi, Cornelia; Obiegala, Anna; Karnath, Carolin; Langer, Sandra; Ternes, Kerstin; Kämmerling, Jens; Osmann, Christine; Pfeffer, Martin

    2015-06-30

    Two cases of acute babesiosis in captive reindeer (Rangifer tarandus) in two German zoos in 2009 and 2012 triggered this study to investigate the occurrence and species diversity of Babesia parasites infecting reindeer in different zoos and deer parks in Germany. Between June and December 2013, blood samples were taken from 123 clinically inapparent reindeer from 16 different facilities. Samples were tested for the presence of Babesia species DNA by conventional PCR and sequence analysis of part of the 18S rRNA gene. Also, Giemsa-stained smears of reindeer blood samples were examined for parasitaemia by light microscopy. The overall PCR-prevalence in blood samples was 23.6% (n=29). Comparison of sequenced amplicons with GenBank entries possibly revealed up to five different Babesia species: B. venatorum (n=19), B. capreoli (n=2) and B. capreoli-like (n=4), B. odocoilei-like (n=2) and B. divergens (n=1), while one sample turned out to be a Theileria sp. Out of the 16 facilities in the study, 12 housed at least one positive animal. In Giemsa-stained blood smears, intra-erythrocytic Babesia parasites were detected in samples of three reindeer from three locations. The high prevalence of Babesia infections implicates babesiosis to be a relevant infectious disease threat for captive reindeer in Germany. Consequently, reindeer with clinical signs compatible to those of acute babesiosis should either be tested for the presence of Babesia spp. DNA or blood smears should be examined for parasitaemia. Copyright © 2015 Elsevier B.V. All rights reserved.

  15. Expression analysis and biological characterization of Babesia sp. BQ1 (Lintan) (Babesia motasi-like) rhoptry-associated protein 1 and its potential use in serodiagnosis via ELISA.

    Science.gov (United States)

    Niu, Qingli; Liu, Zhijie; Yang, Jifei; Yu, Peifa; Pan, Yuping; Zhai, Bintao; Luo, Jianxun; Moreau, Emmanuelle; Guan, Guiquan; Yin, Hong

    2016-05-31

    In China, ovine babesiosis is one of the most important tick-borne haemoparasitic diseases of small ruminants. It has a significant economic impact, and several Babesia motasi-like isolates have been recently shown to be responsible for ovine babesiosis in this country. Full-length and C-terminal-truncated forms of the rap-1a61-1 gene of Babesia sp. BQ1 (Lintan) were cloned into the pET-30a plasmid and subsequently expressed as His-fusion proteins. The resulting recombinant RAP-1a proteins (rRAP-1a61-1 and rRAP-1a61-1/CT) were purified and evaluated as diagnostic antigens using Western blot analysis and ELISA. The native Babesia sp. BQ1 (Lintan) RAP-1 protein was recognized using Western blots and IFAT by antibodies that were raised in rabbits against rRAP-1a61-1/CT. The specificity, sensitivity and positive threshold values for rRAP-1a61-1/CT in ELISA were evaluated. Cross-reactivity was observed between rRAP-1a61-1/CT and positive sera for Babesia sp. BQ1 (Lintan), Babesia sp. BQ1 (Ningxian) and Babesia sp. Tianzhu isolates obtained from infected sheep. At one week post-inoculation, a significant increase was observed in the amount of antibodies produced against RAP-1a, and high levels of antibodies against RAP-1a were observed for 3 months (at 84 days p.i.). A total of 3198 serum samples were collected from small ruminants in 54 different regions in 23 provinces of China. These samples were tested using ELISA based on the rRAP-1a61-1/CT protein. The results indicated that the average positive rate was 36.02 %. The present study suggests that rRAP-1a61-1/CT might be a potential diagnostic antigen for detecting several isolates of B. motasi-like parasites infection.

  16. A PCR method targeting internal transcribed spacers: the simultaneous detection of Babesia bigemina and Babesia bovis in cattle.

    Science.gov (United States)

    Liu, Junlong; Guan, Guiquan; Liu, Aihong; Li, Youquan; Yin, Hong; Luo, Jianxun

    2014-03-01

    In this study, two pairs of oligonucleotide primers were designed according to the nucleotide sequence of the internal transcribed spacers (ITSs) of Babesia bigemina and B. bovis isolates from China. The primers were used in a multiplex PCR to detect parasite DNA in blood samples from cattle. There was no cross reactions with B. ovata, B. major, B. sp. Kashi, Theileria annulata, T. sergenti, T. sinensis or normal bovine DNA. The sensitivity of multiplex PCR assay was 1 pg and 10 pg DNA for B. bigemina and B. bovis, respectively. A total of 260 field blood samples collected from cattle in five provinces of China were analyzed by multiplex PCR and light microscopy. PCR testing revealed that 7.3% (19/260) and 5.8% (15/260) of cattle were positive for B. bigemina and B. bovis and 1.2% (3/260) of cattle were co-infected with B. bigemina and B. bovis. Using light microscopy, 2.3% (6/260) and 1.5% (4/260) of cattle were infected by B. bigemina and B. bovis, respectively, and no co-infection was found. The results showed that the multiplex PCR developed in the present study could be an alternative diagnostic tool for the detection of B. bovis and B. bigemina infection in cattle.

  17. A new equi-dimensional fracture model using polyhedral cells for microseismic data sets

    KAUST Repository

    Al-Hinai, Omar

    2017-04-09

    We present a method for modeling flow in porous media in the presence of complex fracture networks. The approach utilizes the Mimetic Finite Difference (MFD) method. We employ a novel equi-dimensional approach for meshing fractures. By using polyhedral cells we avoid the common challenge in equi-dimensional fracture modeling of creating small cells at the intersection point. We also demonstrate how polyhedra can mesh complex fractures without introducing a large number of cells. We use polyhedra and the MFD method a second time for embedding fracture boundaries in the matrix domain using a “cut-cell” paradigm. The embedding approach has the advantage of being simple and localizes irregular cells to the area around the fractures. It also circumvents the need for conventional mesh generation, which can be challenging when applied to complex fracture geometries. We present numerical results confirming the validity of our approach for complex fracture networks and for different flow models. In our first example, we compare our method to the popular dual-porosity technique. Our second example compares our method with directly meshed fractures (single-porosity) for two-phase flow. The third example demonstrates two-phase flow for the case of intersecting ellipsoid fractures in three-dimensions, which are typical in microseismic analysis of fractures. Finally, we demonstrate our method on a two-dimensional fracture network produced from microseismic field data.

  18. Clonal Streptococcus equi subsp. zooepidemicus post breeding endometritis in thoroughbred broodmares

    DEFF Research Database (Denmark)

    Christoffersen, Mette; Söderlind, Maja; Rydemann Rudefalk, Sofia

    Streptococcus equi subsp. zooepidemicus is one of the most commonly isolated pathogens from the uterus of mares with infectious endometritis. Its ability to cause chronic latent infection by residing deep within the endometrial tissue has previously been described. The aim of the study was to inv......Streptococcus equi subsp. zooepidemicus is one of the most commonly isolated pathogens from the uterus of mares with infectious endometritis. Its ability to cause chronic latent infection by residing deep within the endometrial tissue has previously been described. The aim of the study...... was to investigate whether clonal or genetically distinct S. zooepidemicus strains isolated from mares with endometritis were associated with mare risk factors and the outcome of natural cover. Uterine swabs were obtained from mares with intrauterine fluid after natural cover (n=31) at thoroughbred stud farms...... in Australia. Fifty two percent of the mares (n=16) were diagnosed with infectious endometritis, and S.zooepidemicus was isolated in 81% (n=13) of these mares. Up to four S. zooepidemicus isolates were selected from each mare with growth of S. zooepidemicus and isolates from an additional five mares were...

  19. Cholesterol oxidase (ChoE) is not important in the virulence of Rhodococcus equi.

    Science.gov (United States)

    Pei, Yanlong; Dupont, Chris; Sydor, Tobias; Haas, Albert; Prescott, John F

    2006-12-20

    To analyze further the role in virulence of the prominent cholesterol oxidase (ChoE) of Rhodococcus equi, an allelic exchange choE mutant from strain 103+ was constructed and assessed for virulence in macrophages, in mice, and in foals. There was no difference between the mutant and parent strain in cytotoxic activity for macrophages or in intra-macrophage multiplication. No evidence of attenuation was obtained in macrophages and in mice, but there was slight attenuation apparent in four intra-bronchially infected foals compared to infection of four foals with the virulent parent strain, based on a delayed rise in temperature of the choE-mutant infected foals. However, bacterial colony counts in the lung 2 weeks after infection were not significantly different, although there was a slight but non-significant (P=0.12) difference in lung:body weight ratio of the choE mutant versus virulent parent infected foals (mean 2.67+/-0.25% compared to 4.58+/-0.96%). We conclude that the cholesterol oxidase is not important for the virulence of R. equi.

  20. A new equi-dimensional fracture model using polyhedral cells for microseismic data sets

    KAUST Repository

    Al-Hinai, Omar; Dong, Rencheng; Srinivasan, Sanjay; Wheeler, Mary F.

    2017-01-01

    We present a method for modeling flow in porous media in the presence of complex fracture networks. The approach utilizes the Mimetic Finite Difference (MFD) method. We employ a novel equi-dimensional approach for meshing fractures. By using polyhedral cells we avoid the common challenge in equi-dimensional fracture modeling of creating small cells at the intersection point. We also demonstrate how polyhedra can mesh complex fractures without introducing a large number of cells. We use polyhedra and the MFD method a second time for embedding fracture boundaries in the matrix domain using a “cut-cell” paradigm. The embedding approach has the advantage of being simple and localizes irregular cells to the area around the fractures. It also circumvents the need for conventional mesh generation, which can be challenging when applied to complex fracture geometries. We present numerical results confirming the validity of our approach for complex fracture networks and for different flow models. In our first example, we compare our method to the popular dual-porosity technique. Our second example compares our method with directly meshed fractures (single-porosity) for two-phase flow. The third example demonstrates two-phase flow for the case of intersecting ellipsoid fractures in three-dimensions, which are typical in microseismic analysis of fractures. Finally, we demonstrate our method on a two-dimensional fracture network produced from microseismic field data.

  1. Reproductive efficiency of asymptomatic Theileria equi carriers mares submitted to an embryo transfer program

    Directory of Open Access Journals (Sweden)

    Luciana L. Bezerra

    2015-03-01

    Full Text Available This study aimed to assess and evaluate the effects of Theileria equi infection on embryonic recovery, gestation and early embryonic loss. Thirteen Mangalarga Marchador Theileria equi positive donors (diagnosed through nested-PCR and 40 embryos receptors were used. Donors were submitted to two embryo collections in two consecutive estrous cycles (GId; after, the same mares were treated with imidocarb dipropionate (1.2mg/kg IM. in order to collect more embryos in two more estrous cycles (GIId. Receptors were divided into two groups (control and with treated with 20 animals each, where one group was the control (GIr and the other one (GIIr treated with 1.2mg/kg IM of imidocarb dipropionate assessing the gestation rate at 15, 30, 45 and 60 days. After 52 embryo collections, the embryonic recovery rates were 53.84% (14/26 and 65.38% (17/26 (p> 0.05 for GId and GIId, respectively. The gestation rate was 70% (14/20 (p>0.05 at 15, 30, 45 and 60 days in group GIr and for GIIr was 85% (17/20 (p>0.05 at 15 days, 80% (16/20 (p>0.05 at 30, 45 and 60 days. The treatment with imidocarb dipropionate did not cause significant improvement in the reproductive efficiency at an ET program.

  2. Immunogenicity and antigenicity of the recombinant EMA-1 protein of Theileria equi expressed in the yeast Pichia pastoris Imunogenicidade e antigenicidade da proteína recombinante EMA-1 de Theileria equi expressa em Pichia pastoris

    Directory of Open Access Journals (Sweden)

    Leandro Q. Nizoli

    2009-06-01

    Full Text Available The equine piroplasmosis caused by Theileria equi is one of the most important parasitic diseases of the equine, causing damage to animal health and economic losses. In T. equi, 2 merozoite surface proteins, equi merozoite antigen EMA-1 and EMA-2, have been identified as the most immunodominant antigens. This suggests that these antigens might be used as immunobiological tools. The EMA-1 of Theileria equi was cloned and expressed in the yeast Pichia pastoris. The transformed yeast was grown at high cell density, expressing up to 389 mg.L-1 of recombinant protein. The protein was concentrated and detected in Dot blot. The recombinant product was antigenically similar to the native protein as determined using monoclonal antibodies, and polyclonal antibodies obtained from equines naturally infected with T. equi. The immunogenicity of rEMA-1 protein was demonstrated by IFAT using sera from recombinant-protein-immunized mice using aluminum hydroxide as adjuvant. All animals vaccinated with rEMA-1 developed a high specific antibody response. This results suggest that rEMA-1expressed in P. pastoris might be a strong candidate to be used as an antigen for immune diagnostics as well as a vaccine antigen.A piroplasmose equina causada por Theileria equi é uma das mais importantes doenças parasitárias de equídeos, causando danos a saúde animal e perdas econômicas. Em T. equi, 2 proteínas de superfície de merozoítos, equi merozoite antigen EMA-1 e EMA-2, têm sido identificadas como antígenos imunodominantes. Sugerindo que estes antígenos possam ser usados como produtos imunobiológicos. O gene EMA-1 de T. equi foi clonado e expressado na levedura Pichia pastoris. As leveduras transformadas foram cultivadas a altas densidades celulares expressando 389 mg.L-1 de proteína recombinante. A proteína foi concentrada e detectada em Dot blot. O produto recombinante foi antigenicamente similar à proteína nativa quando determinado usando anticorpo

  3. Detection of Babesia and Theileria species infection in cattle from Portugal using a reverse line blotting method.

    Science.gov (United States)

    Silva, M G; Marques, P X; Oliva, A

    2010-12-15

    Babesiosis and Theileriosis are tick-borne diseases widespread in tropical and sub-tropical regions with high economic impact worldwide. In Portugal there are at least 4 tick vectors known to be competent for the transmission of Babesia and Theileria sp. identified: Rhipicephalus bursa, Rhipicephalus (Boophilus) annulatus, Ixodes ricinus and Haemaphysalis punctata. All these potential Babesia and Theileria tick vectors are widely distributed in Portugal, although they are predominant in the Southern region. In this study, 1104 cattle blood samples were randomly collected from Central and Southern regions of Portugal and analyzed by PCR-reverse line blotting (RLB) for the detection of Babesia and Theileria sp. Testing indicated that 74.7% of the bovines tested were positive for either Babesia and/or Theileria sp. In addition, five different apicomplexan species, namely, Theileria buffeli, Theileria annulata, Babesia divergens, Babesia bovis, and Babesia bigemina were detected by RLB among the bovines tested. T. buffeli was the most frequently found species, being present in 69.9% of the positive samples either as single infections (52.4%), or as mixed infections (17.5%). The Babesia specie most frequently found was B. divergens, detected in 4.2% of the infected bovines. Overall, infected bovines were found in all regions tested; however the highest number of infected bovines was observed in Évora district (96.2%) and in cattle from Limousin breeds (81.7%). The results indicate widespread Babesia and Theileria infections in Portuguese bovines, suggesting the need for improved control of ticks and tick-borne diseases. Copyright © 2010 Elsevier B.V. All rights reserved.

  4. Genome sequence of Corynebacterium pseudotuberculosis biovar equi strain 258 and prediction of antigenic targets to improve biotechnological vaccine production

    DEFF Research Database (Denmark)

    Soares, Siomar C; Trost, Eva; Ramos, Rommel T J

    2013-01-01

    Corynebacterium pseudotuberculosis is the causative agent of several veterinary diseases in a broad range of economically important hosts, which can vary from caseous lymphadenitis in sheep and goats (biovar ovis) to ulcerative lymphangitis in cattle and horses (biovar equi). Existing vaccines ag...

  5. Draft genome sequence of Streptococcus equi subsp. zooepidemicus strain S31A1, isolated from equine infectious endometritis

    DEFF Research Database (Denmark)

    da Piedade, Isabelle; Skive, Bolette; Christensen, Henrik

    2013-01-01

    We present the draft genome sequence of Streptococcus equi subsp. zooepidemicus S31A1, a strain isolated from equine infectious endometritis in Denmark. Comparative analyses of this genome were done with four published reference genomes: S. zooepidemicus strains MGCS10565, ATCC 35246, and H70 and S...

  6. Diagnosis of theileria equi infections in horses in the Azores using cELISA and nested PCR

    Science.gov (United States)

    Equine piroplasmosis is a tick-borne disease of equids that is often caused by the parasite Theileria equi. We applied competitive ELISA (cELISA) and nested PCR diagnostic methods to detect this parasite in horses by screening 162 samples from mainland Portugal where the parasite is endemic, and 143...

  7. 75 FR 36456 - Channel America Television Network, Inc., EquiMed, Inc., Kore Holdings, Inc., Robotic Vision...

    Science.gov (United States)

    2010-06-25

    ... SECURITIES AND EXCHANGE COMMISSION [File No. 500-1] Channel America Television Network, Inc., EquiMed, Inc., Kore Holdings, Inc., Robotic Vision Systems, Inc. (n/k/a Acuity Cimatrix, Inc.), Security... accurate information concerning the securities of Robotic Vision Systems, Inc. (n/k/a Acuity Cimatrix, Inc...

  8. Bovine babesiosis: Cattle protected in the field with a frozen vaccine containing Babesia bovis and Babesia bigemina cultured in vitro with a serum-free medium.

    Science.gov (United States)

    Rojas-Martínez, Carmen; Rodríguez-Vivas, Roger Iván; Millán, Julio Vicente Figueroa; Bautista-Garfias, Carlos Ramón; Castañeda-Arriola, Roberto Omar; Lira-Amaya, José Juan; Urióstegui, Patricia Vargas; Carrasco, Juan José Ojeda; Martínez, Jesús Antonio Álvarez

    2018-04-01

    An attenuated live vaccine containing Babesia bovis and B. bigemina cultured in vitro with a serum-free medium was assessed for its clinical protection conferred of naïve cattle, under natural tick-challenge in a high endemicity zone to Babesia spp. Three groups of six animals were treated as follows: group I (GI) received a vaccine derived from parasites cultured with a free-serum medium; group II (GII) were immunized with the standard vaccine, with parasites cultured in a medium supplemented with 40% (v/v) bovine serum; and a control group (GIII) inoculated with non-infected bovine erythrocytes. Inocula were administered by IM route. Experimental animals were kept during 23days after vaccination in a cattle farm free of ticks and Babesia spp. Thereafter, cattle were moved to a high endemicity farm for natural exposure to Babesia spp. transmitted by Rhipicephalus microplus ticks. Protection against clinical babesiosis was observed in bovines belonging to GI (100%) and GII (83.33%), while the control animals (GIII) were not protected, and showed severe clinical signs, closely related to babesiosis, were observed for at least three consecutive days during the challenge. These were fever, anemia, which were measured simultaneously, and circulating parasites were detected by optic light microscopy. All cattle showed B. bovis and B. bigemina in stained blood films during the challenge; B. bovis antibody titers were higher than those to B. bigemina in GI and GII, and lower titers were determined in GIII. The protective capacity of the vaccine derived from B. bovis and B. bigemina cultured in vitro in a serum-free medium was demonstrated. Copyright © 2017 Elsevier B.V. All rights reserved.

  9. Nested PCR detection and phylogenetic analysis of Babesia bovis and Babesia bigemina in cattle from Peri-urban localities in Gauteng Province, South Africa.

    Science.gov (United States)

    Mtshali, Phillip Senzo; Tsotetsi, Ana Mbokeleng; Thekisoe, Matlhahane Molifi Oriel; Mtshali, Moses Sibusiso

    2014-01-01

    Babesia bovis and Babesia bigemina are tick-borne hemoparasites causing babesiosis in cattle worldwide. This study was aimed at providing information about the occurrence and geographical distribution of B. bovis and B. bigemina species in cattle from Gauteng province, South Africa. A total of 268 blood samples collected from apparently healthy animals in 14 different peri-urban localities were tested using previously established nested PCR assays for the detection of B. bovis and B. bigemina species-specific genes encoding rhoptry-associated protein 1 (RAP-1) and SpeI-AvaI restriction fragment, respectively. Nested PCR assays revealed that the overall prevalence was 35.5% (95% confidence interval [CI]=± 5.73) and 76.1% (95% CI=± 5.11) for B. bovis and B. bigemina, respectively. PCR results were corroborated by sequencing amplicons of randomly selected samples. The neighbor-joining trees were constructed to study the phylogenetic relationship between B. bovis and B. bigemina sequences of randomly selected isolates. Analysis of phylogram inferred with B. bovis RAP-1 sequences indicated a close relationship between our isolates and GenBank strains. On the other hand, a tree constructed with B. bigemina gp45 sequences revealed a high degree of polymorphism among the B. bigemina isolates investigated in this study. Taken together, the results presented in this work indicate the high incidence of Babesia parasites in cattle from previously uncharacterised peri-urban areas of the Gauteng province. These findings suggest that effective preventative and control measures are essential to curtail the spread of Babesia infections among cattle populations in Gauteng.

  10. Seroprevalence of Babesia microti in Individuals with Lyme Disease.

    Science.gov (United States)

    Curcio, Sabino R; Tria, Laurel P; Gucwa, Azad L

    2016-12-01

    Babesiosis is an emerging tick-borne disease (TBD) caused by Babesia microti, an intracellular parasite of red blood cells. Currently, it is the highest ranked pathogen transmitted by blood transfusion. Most healthy individuals infected with B. microti are asymptomatic, but may be at risk for chronic infection. Similar to Lyme disease transmitted by Borrelia burgdorferi, B. microti is spread by Ixodes scapularis ticks. The rate of coinfection with these TBDs in humans is unclear as most studies have focused their prevalence in ticks or rodent reservoirs. In this study, we aimed to determine the seroprevalence of B. microti infection in individuals who tested positive for Lyme disease. Serum samples obtained from 130 subjects in New York were tested by immunofluorescence assay (IFA) for the presence of IgM and IgG antibodies against B. microti. Overall, 26.9% of the serum samples tested were positive for IgM and IgG antibodies against B. microti, suggesting exposure to TBD. Individuals who tested positive for Lyme disease as determined by two-tiered serological testing and the presence of both IgM and IgG antibodies directed against B. burgdorferi, were significantly increased for antibodies directed against B. microti (28.6%; p Lyme disease-negative control group had only 6.7% of samples seropositive for B. microti. These findings suggest the need for more extensive studies investigating infection rates with multiple TBDs in areas where they are endemic and further support for the need to implement an FDA-approved screening test for blood products to help prevent transfusion-transmitted babesiosis.

  11. Lymphocytes and macrophages are infected by Theileria equi, but T cells and B cells are not required to establish infection in vivo.

    Directory of Open Access Journals (Sweden)

    Joshua D Ramsay

    Full Text Available Theileria equi has a biphasic life cycle in horses, with a period of intraleukocyte development followed by patent erythrocytic parasitemia that causes acute and sometimes fatal hemolytic disease. Unlike Theileria spp. that infect cattle (Theileria parva and Theileria annulata, the intraleukocyte stage (schizont of Theileria equi does not cause uncontrolled host cell proliferation or other significant pathology. Nevertheless, schizont-infected leukocytes are of interest because of their potential to alter host cell function and because immune responses directed against this stage could halt infection and prevent disease. Based on cellular morphology, Theileria equi has been reported to infect lymphocytes in vivo and in vitro, but the specific phenotype of schizont-infected cells has yet to be defined. To resolve this knowledge gap in Theileria equi pathogenesis, peripheral blood mononuclear cells were infected in vitro and the phenotype of infected cells determined using flow cytometry and immunofluorescence microscopy. These experiments demonstrated that the host cell range of Theileria equi was broader than initially reported and included B lymphocytes, T lymphocytes and monocyte/macrophages. To determine if B and T lymphocytes were required to establish infection in vivo, horses affected with severe combined immunodeficiency (SCID, which lack functional B and T lymphocytes, were inoculated with Theileria equi sporozoites. SCID horses developed patent erythrocytic parasitemia, indicating that B and T lymphocytes are not necessary to complete the Theileria equi life cycle in vivo. These findings suggest that the factors mediating Theileria equi leukocyte invasion and intracytoplasmic differentiation are common to several leukocyte subsets and are less restricted than for Theileria annulata and Theileria parva. These data will greatly facilitate future investigation into the relationships between Theileria equi leukocyte tropism and pathogenesis

  12. Platelet activation and platelet-leukocyte interaction in dogs naturally infected with Babesia rossi

    DEFF Research Database (Denmark)

    Goddard, Amelia; Leisewitz, Andrew L; Kristensen, Annemarie Thuri

    2015-01-01

    EDTA as anticoagulant. Activated platelets and PLA formation were detected by measuring surface expression of P-selectin (CD62P) on platelets, monocytes and neutrophils. Of the Babesia-infected dogs, 29 survived and seven died. The percentage of CD62P-positive monocytes was significantly higher (P = 0.......036) in the Babesia-infected dogs (54%) than in healthy control dogs (35.3%). However, there were no significant differences between the Babesia-infected and control groups for CD62P-positive platelets (4.9% and 1.2%, respectively) and CD62P-positive neutrophils (28.3% and 17.9%, respectively). The percentage of CD62...... groups for the percentage of CD62P-positive platelets (survivors 4.8%; non-survivors 5.3%; controls 1.2%) or CD62P-positive neutrophils (survivors 31.6%; non-survivors 5.6%; controls 17.9%). In conclusion, Babesia-infected dogs, specifically dogs that survived, had a significantly increased percentage...

  13. Targeted surface expression of an exogenous antigen in stably transfected babesia bovis

    Science.gov (United States)

    Babesia bovis is a tick-borne intraerythocytic protozoan responsible for acute disease in cattle which can be controlled by vaccination with attenuated B. bovis strains. Emerging B. bovis transfection technologies may increase the usefulness of these live vaccines. Here we propose using transfected ...

  14. Molecular prevalence of Bartonella, Babesia, and hemotropic Mycoplasma sp. in dogs with splenic disease.

    Science.gov (United States)

    Varanat, M; Maggi, R G; Linder, K E; Breitschwerdt, E B

    2011-01-01

    Among diseases that cause splenomegaly in dogs, lymphoid nodular hyperplasia (LNH), splenic hemangiosarcoma (HSA), and fibrohistiocytic nodules (FHN) are common diagnoses. The spleen plays an important role in the immunologic control or elimination of vector-transmitted, blood-borne pathogens, including Bartonella sp., Babesia sp., and hemotropic Mycoplasma sp. To compare the prevalence of Bartonella sp., Babesia sp., and hemotropic Mycoplasma sp. DNA in spleens from dogs with LNH, HSA, and FHN. Paraffin-embedded, surgically obtained biopsy tissues from LNH (N = 50), HSA (N = 50), and FHN (N = 37) were collected from the anatomic pathology archives. Spleens from specific pathogen-free (SPF) dogs (N = 8) were used as controls. Bartonella sp., Babesia sp., and Mycoplasma sp. DNA was amplified by PCR, followed by DNA sequencing. Bartonella sp. DNA was more prevalent in FHN (29.7%) and HSA (26%) as compared to LNH (10%) (P = .019, .0373, respectively) or control spleens (0.0%). The prevalence of Babesia sp. and hemotropic Mycoplasma sp. DNA was significantly lower than Bartonella sp. DNA in HSA (P = .0005, .006, respectively) and FHN (P = .003, .0004, respectively). There was no statistically significant difference in DNA prevalence among the 3 genera in the LNH group. The higher prevalence of Bartonella sp. in FHN and HSA warrants future investigations to determine if this bacterium plays a role in the development of these splenic diseases. Copyright © 2011 by the American College of Veterinary Internal Medicine.

  15. Molecular and serological detection of Ehrlichia canis and Babesia vogeli in dogs in Colombia.

    Science.gov (United States)

    Vargas-Hernández, G; André, M R; Faria, J L M; Munhoz, T D; Hernandez-Rodriguez, M; Machado, R Z; Tinucci-Costa, M

    2012-05-25

    Ehrlichiosis and babesiosis are tick-borne diseases, caused mainly by Ehrlichia canis and Babesia canis, respectively, with a worldwide occurrence in dogs, whose main vector is the brown-dog tick, Rhipicephalus sanguineus. The present work aimed to detect the presence of E. canis and Babesia sp. in 91 dog blood samples in Colombia, by molecular and serological techniques. We also performed sequence alignment to indicate the identity of the parasite species infecting these animals. The present work shows the first molecular detection of E. canis and B. vogeli in dogs from Colombia. Immunoglobulin-G (IgG) antibodies to E. canis and Babesia vogeli were found in 75 (82.4%) and 47 (51.6%) sampled dogs, respectively. Thirty-seven (40.6%) and 5 (5.5%) dogs were positive in PCR for E. canis and Babesia sp., respectively. After sequencing, amplicons showed 99% of identity with isolates of E. canis and B. vogeli. The phylogenetic trees based on 16S rRNA-Anaplasmataceae sequences and 18S rRNA-piroplasmid sequences supported the identity of the found E. canis and B. vogeli DNAs, respectively. The present work shows the first molecular detection of E. canis and B. vogeli in dogs in Colombia. Copyright © 2011 Elsevier B.V. All rights reserved.

  16. Epidemiology and molecular phylogeny of Babesia sp. in Little Penguins Eudyptula minor in Australia

    Directory of Open Access Journals (Sweden)

    Ralph Eric Thijl Vanstreels

    2015-08-01

    Full Text Available Blood parasites are potential threats to the health of penguins and to their conservation and management. Little penguins Eudyptula minor are native to Australia and New Zealand, and are susceptible to piroplasmids (Babesia, hemosporidians (Haemoproteus, Leucocytozoon, Plasmodium and kinetoplastids (Trypanosoma. We studied a total of 263 wild little penguins at 20 sites along the Australian southeastern coast, in addition to 16 captive-bred little penguins. Babesia sp. was identified in seven wild little penguins, with positive individuals recorded in New South Wales, Victoria and Tasmania. True prevalence was estimated between 3.4% and 4.5%. Only round forms of the parasite were observed, and gene sequencing confirmed the identity of the parasite and demonstrated it is closely related to Babesia poelea from boobies (Sula spp. and B. uriae from murres (Uria aalge. None of the Babesia-positive penguins presented signs of disease, confirming earlier suggestions that chronic infections by these parasites are not substantially problematic to otherwise healthy little penguins. We searched also for kinetoplastids, and despite targeted sampling of little penguins near the location where Trypanosoma eudyptulae was originally reported, this parasite was not detected.

  17. Babesia sp. EU1 from Roe Deer and Transmission within Ixodes ricinus

    Science.gov (United States)

    Jouglin, Maggy; L’Hostis, Monique; Chauvin, Alain

    2007-01-01

    We report in vitro culture of zoonotic Babesia sp. EU1 from blood samples of roe deer in France. This study provides evidence of transovarial and transstadial transmission of the parasite within Ixodes ricinus, which suggests that this tick could be a vector and reservoir of EU1. PMID:17953093

  18. In vitro cultivation of a newly recognized Babesia sp. in dogs in North Carolina.

    Science.gov (United States)

    Lehtinen, Lauren E; Birkenheuer, Adam J; Droleskey, Robert E; Holman, Patricia J

    2008-02-14

    A novel large Babesia sp. from an infected dog was cultivated in vitro by microaerophilous stationary phase culture methodology. A primary culture initiated in enriched RPMI-1640 medium supplemented with 40% canine serum and incubated in a 2% oxygen environment supported parasite growth in vitro. Subsequent subcultures into enriched HL-1 medium with 20% fetal bovine serum also supported parasite propagation. Cultures were successfully introduced to 5% carbon dioxide in air atmosphere at passage 4. To date, the parasites have been continuously cultured through 35 passages, although the parasitemias are low, ranging from 0.2 to 0.3%. Parasites cultured in RPMI with canine serum were cryopreserved and successfully recovered from liquid nitrogen storage. The small subunit ribosomal rRNA gene sequence was identical in blood-derived and culture-derived parasites, differing in a single base position from the previously reported sequence for this Babesia sp. The ultrastructure of the parasite was consistent with that of other large Babesia spp., except that the spherical body contained numerous round particles unlike the inclusions previously described in Babesia spp.

  19. Epidemiology and molecular phylogeny of Babesia sp. in Little Penguins Eudyptula minor in Australia

    Science.gov (United States)

    Vanstreels, Ralph Eric Thijl; Woehler, Eric J.; Ruoppolo, Valeria; Vertigan, Peter; Carlile, Nicholas; Priddel, David; Finger, Annett; Dann, Peter; Herrin, Kimberly Vinette; Thompson, Paul; Ferreira Junior, Francisco C.; Braga, Érika M.; Hurtado, Renata; Epiphanio, Sabrina; Catão-Dias, José Luiz

    2015-01-01

    Blood parasites are potential threats to the health of penguins and to their conservation and management. Little penguins Eudyptula minor are native to Australia and New Zealand, and are susceptible to piroplasmids (Babesia), hemosporidians (Haemoproteus, Leucocytozoon, Plasmodium) and kinetoplastids (Trypanosoma). We studied a total of 263 wild little penguins at 20 sites along the Australian southeastern coast, in addition to 16 captive-bred little penguins. Babesia sp. was identified in seven wild little penguins, with positive individuals recorded in New South Wales, Victoria and Tasmania. True prevalence was estimated between 3.4% and 4.5%. Only round forms of the parasite were observed, and gene sequencing confirmed the identity of the parasite and demonstrated it is closely related to Babesia poelea from boobies (Sula spp.) and B. uriae from murres (Uria aalge). None of the Babesia-positive penguins presented signs of disease, confirming earlier suggestions that chronic infections by these parasites are not substantially problematic to otherwise healthy little penguins. We searched also for kinetoplastids, and despite targeted sampling of little penguins near the location where Trypanosoma eudyptulae was originally reported, this parasite was not detected. PMID:25853053

  20. Comparative Genomics of Rhodococcus equi Virulence Plasmids Indicates Host-Driven Evolution of the vap Pathogenicity Island.

    Science.gov (United States)

    MacArthur, Iain; Anastasi, Elisa; Alvarez, Sonsiray; Scortti, Mariela; Vázquez-Boland, José A

    2017-05-01

    The conjugative virulence plasmid is a key component of the Rhodococcus equi accessory genome essential for pathogenesis. Three host-associated virulence plasmid types have been identified the equine pVAPA and porcine pVAPB circular variants, and the linear pVAPN found in bovine (ruminant) isolates. We recently characterized the R. equi pangenome (Anastasi E, et al. 2016. Pangenome and phylogenomic analysis of the pathogenic actinobacterium Rhodococcus equi. Genome Biol Evol. 8:3140-3148.) and we report here the comparative analysis of the virulence plasmid genomes. Plasmids within each host-associated type were highly similar despite their diverse origins. Variation was accounted for by scattered single nucleotide polymorphisms and short nucleotide indels, while larger indels-mostly in the plasticity region near the vap pathogencity island (PAI)-defined plasmid genomic subtypes. Only one of the plasmids analyzed, of pVAPN type, was exceptionally divergent due to accumulation of indels in the housekeeping backbone. Each host-associated plasmid type carried a unique PAI differing in vap gene complement, suggesting animal host-specific evolution of the vap multigene family. Complete conservation of the vap PAI was observed within each host-associated plasmid type. Both diversity of host-associated plasmid types and clonality of specific chromosomal-plasmid genomic type combinations were observed within the same R. equi phylogenomic subclade. Our data indicate that the overall strong conservation of the R. equi host-associated virulence plasmids is the combined result of host-driven selection, lateral transfer between strains, and geographical spread due to international livestock exchanges. © The Author(s) 2017. Published by Oxford University Press on behalf of the Society for Molecular Biology and Evolution.

  1. The use of different diagnostic tools for Babesia and Theileria parasites in cattle in Menofia, Egypt.

    Science.gov (United States)

    Nayel, Mohamed; El-Dakhly, Khaled Mohamed; Aboulaila, Mahmoud; Elsify, Ahmed; Hassan, Hany; Ibrahim, Elsayed; Salama, Akram; Yanai, Tokuma

    2012-09-01

    Bovine piroplasmosis is caused by tick-borne hemoprotozoans of the genera Babesia and Theileria and is the most prevalent in tropical and subtropical countries, causing a major economic impact worldwide. In the current study, a total of 405 cattle of different ages, sexes, and breeds were randomly sampled for surveying and diagnosis of babesiosis and theileriosis using three methods: direct microscopy (blood smears), indirect fluorescent antibody test (IFAT) and polymerase chain reaction (PCR). Giemsa-stained blood smears revealed that, out of 405 examined cattle, 33 (8.15 %) were infected with Babesia sp. and 65 (16.05 %) with Theileria sp. (total number of infected cattle was 98). Mixed infection was seen in 11 (2.72 %) animals. Moreover, application of the three diagnostic assays on 158 randomly sampled cattle indicated that 17 (10.76 %) and 33 (20.89 %) were positive for Babesia and Theileria spp. by the direct smear technique, 25 (15.82 %) and 33 (20.89 %) by IFAT (fluorescence was greenish yellow for Babesia and yellowish for Theileria), and 20 (12.66 %) and 38 (24.05 %) by PCR. Using primers specific for Babesia and Theileria spp., we found that diagnostic bands appeared at ~350 and ~370 bp, respectively indicating the presence of these piroplasms. Statistically, there was a non-significant difference of the positivity in response to the three techniques; thus, any of these methods can be described as useful for diagnosing blood parasites in both domesticated animals and birds. On the basis of the obtained results, it could be concluded that direct microscopy can be used in acute infections, whereas IFAT and PCR are useful in chronicity.

  2. Continuous in vitro cultivation of a recently identified Babesia that infects small ruminants in China.

    Science.gov (United States)

    Guan, Guiquan; Ma, Miling; Liu, Aihong; Du, Pengfei; Ren, Qiaoyun; Li, Youquan; Wang, Jinming; Liu, Zhijie; Yin, Hong; Luo, Jianxun

    2012-07-06

    Babesia sp. Xinjiang was isolated from a splenectomised sheep infested by Rhipicephalus sanguineus and Hylomma anatolicum anatolicum, collected from sheep and cattle in Xinjiang province. It was considered to be a novel ovine Babesia species on the basis of its morphology, pathogenicity, vector tick species and alignments of 18S ribosomal RNA (18S rRNA) and internal transcribed spacers (ITS) gene sequences. Continuous in vitro cultures of the ovine parasite were established using infected sheep blood. In RPMI 1640 medium with 7.5% sheep red blood cells (RBCs) maintained in an incubator at 37 °C and 5% CO(2), the percentage of parasitized erythrocytes (PPE) peaked at 10% in 24- and 6-well plates. It increased to 20-50% with the same culture medium but with 2.5% RBC in 75 cm(2) flasks. Two clonal lines of Babesia sp. Xinjiang were screened using the limiting dilution method. Growth characteristics of these lines in vitro were measured by a microtiter-based spectrophotometric method and from the PPE. The generation time in sheep erythrocytes was between 15.20 h and 16.27 h. Furthermore, the host range of parasite was identified with in vitro culture and in vivo infection. Erythrocytes of sheep, cattle, sika deer and humans could be invaded into by lines in vitro, but the parasites could not propagate in human erythrocytes. The parasites could not enter erythrocytes from goats in vitro. However, in vivo, only sheep could be infected by lines. Finally, a Babesia sp. Xinjiang-like parasite (which shared 99.5% identity with the original strain of Babesia sp. Xinjiang) was isolated using this in vitro culture system from 1 of 19 sheep blood samples collected from western Gansu province, China. Copyright © 2012 Elsevier B.V. All rights reserved.

  3. Molecular detection of Hepatozoon canis and Babesia canis vogeli in domestic dogs from Cuiabá, Brazil.

    Science.gov (United States)

    Spolidorio, Mariana Granziera; Torres, Mariana de Medeiros; Campos, Wilma Neres da Silva; Melo, Andréia Lima Tomé; Igarashi, Michelle; Amude, Alexandre Mendes; Labruna, Marcelo Bahia; Aguiar, Daniel Moura

    2011-01-01

    The objective of this study was to report for the first time infection by Hepatozoon spp. and Babesia spp. in 10 dogs from the city of Cuiabá, State of Mato Grosso, central-western Brazil. A pair of primers that amplifies a 574 bp fragment of the 18S rRNA of Hepatozoon spp., and a pair of primers that amplifies a 551 bp fragment of the gene 18S rRNA for Babesia spp. were used. Six dogs were positive for Babesia spp., and 9 were positive for Hepatozoon spp. Co‑infection of Babesia spp. and Hepatozoon spp. was seen in 5 dogs. Sequenced samples revealed 100% identity with B. canis vogeli, and H. canis. This is the first molecular detection of H. canis in domestic dogs from Cuiabá. Additionally, it is described for the first time the presence of B. canis vogeli circulating among dogs in Cuiabá.

  4. Characterization of SeseC_01411 as a surface protective antigen of Streptococcus equi ssp. zooepidemicus.

    Science.gov (United States)

    Xie, Honglin; Wei, Zigong; Ma, Chunquan; Li, Shun; Liu, Xiaohong; Fu, Qiang

    2018-06-01

    Streptococcus equi ssp. zooepidemicus (Streptococcus zooepidemicus, SEZ) is a commensal bacterium related to opportunistic infections of many species, including humans, dogs, cats, and pigs. SeseC_01411 has been proven to be immunogenic. However, its protective efficacy remained to be evaluated. In the present study, the purified recombinant SeseC_01411 could elicit a strong humoral antibody response and protect against lethal challenge with virulent SEZ in mice. Our finding confirmed that SeseC_01411 distributes on the surface of SEZ. In addition, the hyperimmune sera against SeseC_01411 could efficiently kill the bacteria in the phagocytosis test. The present study identified the immunogenic protein, SeseC_01411, as a novel surface protective antigen of SEZ. Copyright © 2018 Elsevier Ltd. All rights reserved.

  5. Piroplasmosis in wildlife: Babesia and Theileria affecting free-ranging ungulates and carnivores in the Italian Alps.

    Science.gov (United States)

    Zanet, Stefania; Trisciuoglio, Anna; Bottero, Elisa; de Mera, Isabel Garcia Fernández; Gortazar, Christian; Carpignano, Maria Grazia; Ferroglio, Ezio

    2014-02-17

    Piroplasmosis are among the most relevant diseases of domestic animals. Babesia is emerging as cause of tick-borne zoonosis worldwide and free-living animals are reservoir hosts of several zoonotic Babesia species. We investigated the epidemiology of Babesia spp. and Theileria spp. in wild ungulates and carnivores from Northern Italy to determine which of these apicomplexan species circulate in wildlife and their prevalence of infection. PCR amplification of the V4 hyper-variable region of the 18S rDNA of Babesia sp./Theileria sp was carried out on spleen samples of 1036 wild animals: Roe deer Capreolus capreolus (n = 462), Red deer Cervus elaphus (n = 52), Alpine Chamois Rupicapra rupicapra (n = 36), Fallow deer Dama dama (n = 17), Wild boar Sus scrofa (n = 257), Red fox Vulpes vulpes (n = 205) and Wolf Canis lupus (n = 7). Selected positive samples were sequenced to determine the species of amplified Babesia/Theileria DNA. Babesia/Theileria DNA was found with a mean prevalence of 9.94% (IC95% 8.27-11.91). The only piroplasms found in carnivores was Theileria annae, which was detected in two foxes (0.98%; IC95% 0.27-3.49). Red deer showed the highest prevalence of infection (44.23%; IC95% 31.6-57.66), followed by Alpine chamois (22.22%; IC95% 11.71-38.08), Roe deer (12.55%; IC95% 9.84-15.89), and Wild boar (4.67%; IC95% 2.69-7.98). Genetic analysis identified Babesia capreoli as the most prevalent piroplasmid found in Alpine chamois, Roe deer and Red deer, followed by Babesia bigemina (found in Roe deer, Red deer and Wild boar), and the zoonotic Babesia venatorum (formerly Babesia sp. EU1) isolated from 2 Roe deer. Piroplasmids of the genus Theileria were identified in Wild boar and Red deer. The present study offers novel insights into the role of wildlife in Babesia/Theileria epidemiology, as well as relevant information on genetic variability of piroplasmids infecting wild ungulates and carnivores.

  6. Hap2, a novel gene in Babesia bigemina is expressed in tick stages, and specific antibodies block zygote formation

    OpenAIRE

    Camacho-Nuez, Minerva; Hernández-Silva, Diego Josimar; Castañeda-Ortiz, Elizabeth Jacqueline; Paredes-Martínez, María Elena; Rocha-Martínez, Marisol Karina; Alvarez-Sánchez, María Elizbeth; Mercado-Curiel, Ricardo Francisco; Aguilar-Tipacamu, Gabriela; Mosqueda, Juan

    2017-01-01

    Background Bovine babesiosis is a tick-borne disease caused by the protozoan parasites of the genus Babesia. In their host vector, Babesia spp. undergo sexual reproduction. Therefore, the development of sexual stages and the subsequent formation of the zygote are essential for the parasite to invade the intestinal cells of the vector tick and continue its life-cycle. HAP2/GCS1 is a protein identified in plants, protozoan parasites and other organisms that has an important role during membrane...

  7. Molecular phylogenetic studies on an unnamed bovine Babesia sp. based on small subunit ribosomal RNA gene sequences.

    Science.gov (United States)

    Luo, Jianxun; Yin, Hong; Liu, Zhijie; Yang, Dongying; Guan, Guiquan; Liu, Aihong; Ma, Miling; Dang, Shengzhi; Lu, Bingyi; Sun, Caiqin; Bai, Qi; Lu, Wenshun; Chen, Puyan

    2005-10-10

    The 18S small subunit ribosomal RNA (18S rRNA) gene of an unnamed Babesia species (designated B. U sp.) was sequenced and analyzed in an attempt to distinguish it from other Babesia species in China. The target DNA segment was amplified by polymerase chain reaction (PCR). The PCR product was ligated to the pGEM-T Easy vector for sequencing. It was found that the length of the 18S rRNA gene of all B. U sp. Kashi 1 and B. U sp. Kashi 2 was 1699 bp and 1689 bp. Two phylogenetic trees were, respectively, inferred based on 18S rRNA sequence of the Chinese bovine Babesia isolates and all of Babesia species available in GenBank. The first tree showed that B. U sp. was situated in the branch between B. major Yili and B. bovis Shannxian, and the second tree revealed that B. U sp. was confined to the same group as B. caballi. The percent identity of B. U sp. with other Chinese Babesia species was between 74.2 and 91.8, while the percent identity between two B. U sp. isolates was 99.7. These results demonstrated that this B. U sp. is different from other Babesia species, but that two B. U sp. isolates obtained with nymphal and adultal Hyalomma anatolicum anatolicum tick belong to the same species.

  8. Evaluation of Veterinary-Specific Interpretive Criteria for Susceptibility Testing of Streptococcus equi Subspecies with Trimethoprim-Sulfamethoxazole and Trimethoprim-Sulfadiazine

    DEFF Research Database (Denmark)

    Sadaka, Carmen; Kanellos, Theo; Guardabassi, Luca

    2017-01-01

    Antimicrobial susceptibility test results for trimethoprim-sulfadiazine with Streptococcus equi subspecies are interpreted based on human data for trimethoprim-sulfamethoxazole. The veterinary-specific data generated in this study support a single breakpoint for testing trimethoprim-sulfamethoxaz......Antimicrobial susceptibility test results for trimethoprim-sulfadiazine with Streptococcus equi subspecies are interpreted based on human data for trimethoprim-sulfamethoxazole. The veterinary-specific data generated in this study support a single breakpoint for testing trimethoprim...

  9. Differences in Rhodococcus equi Infections Based on Immune Status and Antibiotic Susceptibility of Clinical Isolates in a Case Series of 12 Patients and Cases in the Literature

    Science.gov (United States)

    Suzuki, Yasuhiro; Ribes, Julie A.; Thornton, Alice

    2016-01-01

    Rhodococcus equi is an unusual zoonotic pathogen that can cause life-threatening diseases in susceptible hosts. Twelve patients with R. equi infection in Kentucky were compared to 137 cases reported in the literature. Although lungs were the primary sites of infection in immunocompromised patients, extrapulmonary involvement only was more common in immunocompetent patients (P antibiotics, preferably selected from vancomycin, imipenem, clarithromycin/azithromycin, ciprofloxacin, rifampin, or cotrimoxazole. Local antibiograms should be checked prior to using cotrimoxazole due to developing resistance. PMID:27631004

  10. Tracing outbreaks of Streptococcus equi infection (strangles) in horses using sequence variation in the seM gene and pulsed-field gel electrophoresis.

    Science.gov (United States)

    Lindahl, Susanne; Söderlund, Robert; Frosth, Sara; Pringle, John; Båverud, Viveca; Aspán, Anna

    2011-11-21

    Strangles is a serious respiratory disease in horses caused by Streptococcus equi subspecies equi (S. equi). Transmission of the disease occurs by direct contact with an infected horse or contaminated equipment. Genetically, S. equi strains are highly homogenous and differentiation of strains has proven difficult. However, the S. equi M-protein SeM contains a variable N-terminal region and has been proposed as a target gene to distinguish between different strains of S. equi and determine the source of an outbreak. In this study, strains of S. equi (n=60) from 32 strangles outbreaks in Sweden during 1998-2003 and 2008-2009 were genetically characterized by sequencing the SeM protein gene (seM), and by pulsed-field gel electrophoresis (PFGE). Swedish strains belonged to 10 different seM types, of which five have not previously been described. Most were identical or highly similar to allele types from strangles outbreaks in the UK. Outbreaks in 2008/2009 sharing the same seM type were associated by geographic location and/or type of usage of the horses (racing stables). Sequencing of the seM gene generally agreed with pulsed-field gel electrophoresis profiles. Our data suggest that seM sequencing as a epidemiological tool is supported by the agreement between seM and PFGE and that sequencing of the SeM protein gene is more sensitive than PFGE in discriminating strains of S. equi. Copyright © 2011 Elsevier B.V. All rights reserved.

  11. Detecção molecular de Hepatozoon canis e Babesia canis vogeli em cães domésticos de Cuiabá, Brasil

    OpenAIRE

    Spolidorio, Mariana Granziera; Torres, Mariana de Medeiros; Campos, Wilma Neres da Silva; Melo, Andréia Lima Tomé; Igarashi, Michelle; Amude, Alexandre Mendes; Labruna, Marcelo Bahia; Aguiar, Daniel Moura

    2011-01-01

    The objective of this study was to report for the first time infection by Hepatozoon spp. and Babesia spp. in 10 dogs from the city of Cuiabá, State of Mato Grosso, central-western Brazil. A pair of primers that amplifies a 574 bp fragment of the 18S rRNA of Hepatozoon spp., and a pair of primers that amplifies a 551 bp fragment of the gene 18S rRNA for Babesia spp. were used. Six dogs were positive for Babesia spp., and 9 were positive for Hepatozoon spp. Co-infection of Babesia spp. and Hep...

  12. Detection of Babesia annae DNA in lung exudate samples from Red foxes (Vulpes vulpes) in Great Britain.

    Science.gov (United States)

    Bartley, Paul M; Hamilton, Clare; Wilson, Cari; Innes, Elisabeth A; Katzer, Frank

    2016-02-12

    This study aimed to determine the prevalence of Babesia species DNA in lung exudate samples collected from red foxes (Vulpes vulpes) from across Great Britain. Babesia are small piroplasmid parasites which are mainly transmitted through the bite of infected ticks of the family Ixodidae. Babesia can cause potentially fatal disease in a wide-range of mammalian species including humans, dogs and cattle, making them of significant economic importance to both the medical and veterinary fields. DNA was extracted from lung exudate samples of 316 foxes. A semi-nested PCR was used to initially screen samples, using universal Babesia-Theileria primers which target the 18S rRNA gene. A selection of positive PCR amplicons were purified and sequenced. Subsequently specific primers were designed to detect Babesia annae and used to screen all 316 DNA samples. Randomly selected positive samples were purified and sequenced (GenBank accession KT580786). Clones spanning a 1717 bp region of the 18S rRNA gene were generated from 2 positive samples, the resultant consensus sequence was submitted to GenBank (KT580785). Sequence KT580785 was used in the phylogenetic analysis Babesia annae DNA was detected in the fox samples, in total 46/316 (14.6%) of samples tested positive for the presence of Babesia annae DNA. The central region of England had the highest prevalence at 36.7%, while no positive samples were found from Wales, though only 12 samples were tested from this region. Male foxes were found to have a higher prevalence of Babesia annae DNA than females in all regions of Britain. Phylogenetic and sequence analysis of the GenBank submissions (Accession numbers KT580785 and KT580786) showed 100% identity to Babesia sp.-'Spanish Dog' (AY534602, EU583387 and AF188001). This is the first time that Babesia annae DNA has been reported in red foxes in Great Britain with positive samples being found across England and Scotland indicating that this parasite is well established within the

  13. IDENTIFIKASI PROFIL DASAR LAUT MENGGUNAKAN INSTRUMEN SIDE SCAN SONAR DENGAN METODE BEAM PATTERN DISCRETE-EQUI-SPACED UNSHADED LINE ARRAY

    Directory of Open Access Journals (Sweden)

    Muhammad Zainuddin Lubis

    2017-05-01

    Full Text Available Laut Punggur merupakan laut yang terletak di Batam, Kepulauan Riau yang mempunyai beragam habitat objek,dan bentuk struktur bawah laut yang memiliki dinamika laut yang sangat tinggi. Side scan sonar (SSS merupakan instrumen pengembangan sistem sonar yang mampu menunjukkan dalam gambar dua dimensional permukaan dasar laut dengan kondisi kontur, topografi, dan target secara bersamaan. Metode Beam Pattern Discrete-Equi-Spaced Unshaded Line Array digunakan untuk menghitung beam pattern dua dimensi yang tergantung pada sudut dari gelombang suara yang masuk dari sumbu array yang diterima tergantung pada sudut di mana sinar suara pada array. Penelitian ini dilakukan pada Desember 2016 di laut Punggur,Batam, Kepulauan Riau-Indonesia, dengan koordinat 104 ° 08,7102 E dan 1° 03,2448 N sampai 1 ° 03.3977 N dan 104 ° 08,8133 E, menggunakan instrumen Side Scan Sonar C-Max CM2 Tow fish dengan frekuensi 325 kHz. Hasil yang diperoleh dari perekaman terdapat 7 target, dan Beam pattern dari metode Beam Discrete-Equi-Spaced Unshaded Line Array target 4 memiliki nilai tertinggi pada directivity Pattern yaitu 21.08 dB. Hasil model beam pattern ini memiliki nilai pusat pada incidence angle (o terhadap Directivity pattern (dB tidak berada di nilai 0 ataupun pada pusat beam pattern yang dihasilkan pada target 6 dengan nilai incident angle -1.5 o dan 1.5o mengalami penurunan hingga -40 dB. Karakteristik sedimen dasar perairan di laut punggur ditemukan lebih banyak pasir. Hasil metode Beam Discrete-Equi-Spaced Unshaded Line Array ditemukan bangkai kapal tenggelam.Kata Kunci: Side Scan Sonar, Beam Pattern Discrete-Equi-Spaced Unshaded Line Array, Incidence angle, Directivity pattern IDENTIFICATION OF SEABED PROFILE USING SIDE SCAN SONAR INSTRUMENT WITH PATTERN DISCRETE-EQUI-SPACED UNSHADED LINE ARRAY METHODRiau Islands is an island that has a variety of habitat objects, and forms of submarine structures that have a very high ocean dynamics, Punggur Sea is the sea

  14. Babesia bovis and Babesia bigemina infection levels estimated by qPCR in Angus cattle from an endemic area of São Paulo state, Brazil.

    Science.gov (United States)

    Giglioti, R; Oliveira, H N; Santana, C H; Ibelli, A M G; Néo, T A; Bilhassi, T B; Rabelo, M D; Machado, R Z; Brito, L G; Oliveira, M C S

    2016-07-01

    The levels of infection by Babesia bovis and Babesia bigemina were estimated by absolute quantification through the quantitative PCR technique (qPCR). Fifty-one contemporaneous Angus cattle were evaluated on two occasions. The number of standard female Rhipicephalus microplus ticks present on the left side of the body was counted and blood samples were drawn from the tail vein into tubes containing the anticoagulant EDTA. The blood samples were submitted to DNA extraction and used to quantify the number of copies (NC) of DNA from B. bovis and B. bigemina by qPCR. The data on tick count and number of DNA copies were transformed for normalization and analyzed by a mixed model method. A multivariate model with repeated measures of the same animal, including the effects of collection, parasite species and their interaction, was used. The repeatability values were obtained from the matrix of (co)variances and were expressed for each species. The correlations between the counts of different species on the same animal, in the same collection or different collections, were also estimated. The results showed the qPCR could distinguish the two between infection by the two Babesia species. Infection levels by B. bovis and B. bigemina were detected in 100% and 98% of the animals, respectively. Significant differences were found (Phemoparasites did not depend on the tick infestation levels at the moment of each collection. The repeatability values estimated indicate that under the study conditions, the variations in the tick infestation levels and of parasitemia by B. bovis and B. bigemina depend more on factors related to each collection than on intrinsic factors of the animal. Copyright © 2016 Elsevier GmbH. All rights reserved.

  15. Molecular cloning, phylogenetic analysis and heat shock response of Babesia gibsoni heat shock protein 90.

    Science.gov (United States)

    Yamasaki, Masahiro; Tsuboi, Yoshihiro; Taniyama, Yusuke; Uchida, Naohiro; Sato, Reeko; Nakamura, Kensuke; Ohta, Hiroshi; Takiguchi, Mitsuyoshi

    2016-09-01

    The Babesia gibsoni heat shock protein 90 (BgHSP90) gene was cloned and sequenced. The length of the gene was 2,610 bp with two introns. This gene was amplified from cDNA corresponding to full length coding sequence (CDS) with an open reading frame of 2,148 bp. A phylogenetic analysis of the CDS of HSP90 gene showed that B. gibsoni was most closely related to B. bovis and Babesia sp. BQ1/Lintan and lies within a phylogenetic cluster of protozoa. Moreover, mRNA transcription profile for BgHSP90 exposed to high temperature were examined by quantitative real-time reverse transcription-polymerase chain reaction. BgHSP90 levels were elevated when the parasites were incubated at 43°C for 1 hr.

  16. Molecular phylogeny of Babesia poelea from brown boobies (Sula leucogaster) from Johnston Atoll, Central Pacific

    Science.gov (United States)

    Yabsley, Michael J.; Work, Thierry M.; Rameyer, Robert A.

    2006-01-01

    The phylogenetic relationship of avian Babesia with other piroplasms remains unclear, mainly because of a lack of objective criteria such as molecular phylogenetics. In this study, our objective was to sequence the entire 18S, ITS-1, 5.8S, and ITS-2 regions of the rRNA gene and partial ß-tubulin gene of B. poelea, first described from brown boobies (Sula leucogaster) from the central Pacific, and compare them to those of other piroplasms. Phylogenetic analyses of the entire 18S rRNA gene sequence revealed that B. poelea belonged to the clade of piroplasms previously detected in humans, domestic dogs, and wild ungulates in the western United States. The entire ITS-1, 5.8S, ITS-2, and partial ß-tubulin gene sequence shared conserved regions with previously described Babesia and Theileria species. The intron of the ß-tubulin gene was 45 bp. This is the first molecular characterization of an avian piroplasm.

  17. Streptococcus equi subsp zooepidemicus pleuropneumonia and peritonitis in a dromedary camel (Camelus dromedarius) calf in North America.

    Science.gov (United States)

    Stoughton, William B; Gold, Jenifer

    2015-08-01

    A 12-week-old female dromedary camel (Camelus dromedarius) calf was evaluated because of acute (blood, peritoneal fluid, and pleural fluid samples indicated Streptococcus equi subsp zooepidemicus septicemia as the etiology for the polyserositis (ie, alpaca fever). Treatment with IV broad-spectrum antimicrobials, an NSAID, and pleural drainage was initiated. Clinical signs of pleuropneumonia, peritonitis, and systemic infection improved rapidly 24 hours after initiation of medical treatment. The calf was discharged from the hospital after 11 days, and antimicrobial treatment continued for 2 weeks after discharge. At follow-up approximately 4 weeks after hospital discharge (6 weeks after the initial examination), there were no clinical signs suggestive of relapse or any reported complications. S equi subsp zooepidemicus may cause polyserositis in Old World camelids (eg, dromedary camels) with signs similar to those seen in New World camelids (eg, alpaca and llama). The rapid response to medical treatment for the patient described suggested that S equi subsp zooepidemicus-induced polyserositis (alpaca fever) in dromedary camels may respond favorably to appropriate treatment. Reducing stress, reducing overcrowding, and separate housing of equids and camelids are suggested. Further studies are needed to better assess the epidemiology of alpaca fever in dromedary camels in North America.

  18. High-resolution melting PCR assay, applicable for diagnostics and screening studies, allowing detection and differentiation of several Babesia spp. infecting humans and animals.

    Science.gov (United States)

    Rozej-Bielicka, Wioletta; Masny, Aleksander; Golab, Elzbieta

    2017-10-01

    The goal of the study was to design a single tube PCR test for detection and differentiation of Babesia species in DNA samples obtained from diverse biological materials. A multiplex, single tube PCR test was designed for amplification of approximately 400 bp region of the Babesia 18S rRNA gene. Universal primers were designed to match DNA of multiple Babesia spp. and to have low levels of similarity to DNA sequences of other intracellular protozoa and Babesia hosts. The PCR products amplified from Babesia DNA isolated from human, dog, rodent, deer, and tick samples were subjected to high-resolution melting analysis for Babesia species identification. The designed test allowed detection and differentiation of four Babesia species, three zoonotic (B. microti, B. divergens, B. venatorum) and one that is generally not considered zoonotic-Babesia canis. Both detection and identification of all four species were possible based on the HRM curves of the PCR products in samples obtained from the following: humans, dogs, rodents, and ticks. No cross-reactivity with DNA of Babesia hosts or Plasmodium falciparum and Toxoplasma gondii was observed. The lack of cross-reactivity with P. falciparum DNA might allow using the assay in endemic malaria areas. The designed assay is the first PCR-based test for detection and differentiation of several Babesia spp. of medical and veterinary importance, in a single tube reaction. The results of the study show that the designed assay for Babesia detection and identification could be a practical and inexpensive tool for diagnostics and screening studies of diverse biological materials.

  19. Changing epidemiology of the tick-borne bovine parasite, Babesia divergens

    OpenAIRE

    Zintl, A; McGrath, G; O'Grady, L; Fanning, J; Downing, K; Roche, D; Casey, M; Gray, JS

    2014-01-01

    Bovine babesiosis is caused by the tick-borne blood parasite, Babesia divergens. A survey of veterinary practitioners and farmers in Ireland in the 1980’s revealed an annual incidence of 1.7% associated with considerable economic losses. However, two subsequent surveys in the 1990’s indicated a decline in clinical babesiosis. In order to determine whether any such changes have affected the incidence of bovine babesiosis in Ireland, a questionnaire survey of farmers and veterinarians was carri...

  20. Neglected intravascular pathogens, Babesia vulpes and haemotropic Mycoplasma spp. in European red fox (Vulpes vulpes) population.

    Science.gov (United States)

    Koneval, Martina; Miterpáková, Martina; Hurníková, Zuzana; Blaňarová, Lucia; Víchová, Bronislava

    2017-08-30

    Wild animals, especially canids, are important reservoirs of vector-borne pathogens, that are transmitted by the ticks and other bloodsucking arthropods. In total, 300 red foxes (Vulpes vulpes), shot by the hunters in eastern and northern Slovakia, were screened for the presence of vector-borne pathogens by PCR-based methods Blood samples were obtained from nine red foxes and tissue samples originated from 291 animals (the liver tissue samples from 49 foxes and spleen samples from 242 red foxes). Babesia vulpes and haemotropic Mycoplasma species were identified by amplification and sequencing of 18S rRNA and 16S rRNA gene fragments, respectively. Overall, the presence of these pathogens was recorded in 12.3% of screened DNA samples. Altogether 9.7% (29/300) of investigated foxes carried DNA of Babesia spp. In total, 12 out of 29 Babesia spp. PCR - positive amplicons were further sequenced and identified as B. vulpes (41.4%; 12/29), remaining 17 samples are referred as Babesia sp. (58.6%; 17/29). Overall prevalence of B. vulpes reached 4.0% (n=300). Thirteen (4.3%) samples tested positive for distinct Mycoplasma species. To the best of our knowledge, this study brings the first information on B. vulpes infection in red foxes in Slovakia, and the first data on the prevalence and diversity of haemotropic Mycoplasma spp. in European red fox population. Moreover, co-infections with B. vulpes and Mycoplasma spp. were confirmed in 1.7% of tested DNA samples. The relatively high rates of blood pathogen' prevalence and species diversity in wild foxes indicate the role of the fox population in the maintenance of the parasites in sylvatic cycles and strengthen the assumption that foxes play an important role in spreading of infectious microorganisms within and outside the natural foci. Copyright © 2017 Elsevier B.V. All rights reserved.

  1. Babesia microti (Piroplasmida: Babesiidae) in nymphal Ixodes ricinus (Acari: Ixodidae) in the Czech Republic

    Czech Academy of Sciences Publication Activity Database

    Rudolf, Ivo; Golovchenko, Maryna; Šikutová, Silvie; Rudenko, Natalia; Grubhoffer, Libor; Hubálek, Zdeněk

    2005-01-01

    Roč. 52, č. 3 (2005), s. 274-276 ISSN 0015-5683 R&D Projects: GA ČR(CZ) GA206/03/0726 Institutional research plan: CEZ:AV0Z60930519; CEZ:AV0Z60220518 Keywords : Babesia microti * Ixodes ricinus Subject RIV: EE - Microbiology, Virology Impact factor: 1.138, year: 2005 http://folia.paru.cas.cz/pdfs/showpdf.php?pdf=20766

  2. Entomologic and Serologic Evidence of Zoonotic Transmission of Babesia microti, Eastern Switzerland

    OpenAIRE

    Foppa, Ivo M.; Krause, Peter J.; Spielman, Andrew; Goethert, Heidi; Gern, Lise; Brand, Brigit; Telford, Sam R.

    2002-01-01

    We evaluated human risk for infection with Babesia microti at a site in eastern Switzerland where several B. microti–infected nymphal Ixodes ricinus ticks had been found. DNA from pooled nymphal ticks amplified by polymerase chain reaction was highly homologous to published B. microti sequences. More ticks carried babesial infection in the lower portion of the rectangular 0.7-ha grid than in the upper (11% vs. 0.8%). In addition, we measured seroprevalence of immunoglobulin (Ig) G antibodies ...

  3. First report on occurrence of Babesia infection in Nilgai Boselaphus tragocamelus from central India

    Directory of Open Access Journals (Sweden)

    B. Baviskar

    2009-03-01

    Full Text Available A male Nilgai Boselaphus tragocamelus of approximately 6-7 years was presented for post mortem examination to the Nagpur Veterinary College, Nagpur (Maharashtra. At necropsy, the lesions observed were enlarged spleen, congestion of liver and kidney and pale mucous membranes indicating severe anaemia. Blood smears were prepared, stained with leishman’s stain which revealed Babesia sp. Organisms in the erythrocytes, which seems to be the first report in Nilgai from Central India.

  4. Genomic resources for a unique, low-virulence Babesia taxon from China

    Directory of Open Access Journals (Sweden)

    Guiquan Guan

    2016-10-01

    Full Text Available Abstract Background Babesiosis is a socioeconomically important tick-borne disease of animals (including humans caused by haemoprotozoan parasites. The severity of babesiosis relates to host and parasite factors, particularly virulence/pathogenicity. Although Babesia bovis is a particularly pathogenic species of cattle, there are species of Babesia of ruminants that have limited pathogenicity. For instance, the operational taxonomic unit Babesia sp. Xinjiang (abbreviated here as Bx of sheep from China is substantially less virulent/pathogenic than B. bovis is in cattle. Although the reason for this distinctiveness is presently unknown, it is possible that Bx has a reduced ability to adhere to cells or evade/suppress immune responses, which might relate to particular proteins, such as the variant erythrocyte surface antigens (VESAs. Results We sequenced and annotated the 8.4 Mb nuclear draft genome of Bx and compared it with those of B. bovis and B. bigemina by synteny analysis; we also investigated the genetic relationship of Bx with selected Babesia species and related apicomplexans for which genomic datasets are available, and explored the VESA complement in Bx. Conclusions The availability of the Bx genome now provides unique opportunities to elucidate aspects of the molecular biology, biochemistry and physiology of Bx, and to explore the reason(s for its limited virulence and/or apparent ability to evade immune attack by the host animal. Moreover, the present genomic resource and an in vitro culture system for Bx raises the prospect of establishing a functional genomic platform to explore essential genes as new intervention targets against babesiosis.

  5. Genomic resources for a unique, low-virulence Babesia taxon from China.

    Science.gov (United States)

    Guan, Guiquan; Korhonen, Pasi K; Young, Neil D; Koehler, Anson V; Wang, Tao; Li, Youquan; Liu, Zhijie; Luo, Jianxun; Yin, Hong; Gasser, Robin B

    2016-10-27

    Babesiosis is a socioeconomically important tick-borne disease of animals (including humans) caused by haemoprotozoan parasites. The severity of babesiosis relates to host and parasite factors, particularly virulence/pathogenicity. Although Babesia bovis is a particularly pathogenic species of cattle, there are species of Babesia of ruminants that have limited pathogenicity. For instance, the operational taxonomic unit Babesia sp. Xinjiang (abbreviated here as Bx) of sheep from China is substantially less virulent/pathogenic than B. bovis is in cattle. Although the reason for this distinctiveness is presently unknown, it is possible that Bx has a reduced ability to adhere to cells or evade/suppress immune responses, which might relate to particular proteins, such as the variant erythrocyte surface antigens (VESAs). We sequenced and annotated the 8.4 Mb nuclear draft genome of Bx and compared it with those of B. bovis and B. bigemina by synteny analysis; we also investigated the genetic relationship of Bx with selected Babesia species and related apicomplexans for which genomic datasets are available, and explored the VESA complement in Bx. The availability of the Bx genome now provides unique opportunities to elucidate aspects of the molecular biology, biochemistry and physiology of Bx, and to explore the reason(s) for its limited virulence and/or apparent ability to evade immune attack by the host animal. Moreover, the present genomic resource and an in vitro culture system for Bx raises the prospect of establishing a functional genomic platform to explore essential genes as new intervention targets against babesiosis.

  6. Simultaneous Detection of Bovine Theileria and Babesia Species by Reverse Line Blot Hybridization

    Science.gov (United States)

    Gubbels, J. M.; de Vos, A. P.; van der Weide, M.; Viseras, J.; Schouls, L. M.; de Vries, E.; Jongejan, F.

    1999-01-01

    A reverse line blot (RLB) assay was developed for the identification of cattle carrying different species of Theileria and Babesia simultaneously. We included Theileria annulata, T. parva, T. mutans, T. taurotragi, and T. velifera in the assay, as well as parasites belonging to the T. sergenti-T. buffeli-T. orientalis group. The Babesia species included were Babesia bovis, B. bigemina, and B. divergens. The assay employs one set of primers for specific amplification of the rRNA gene V4 hypervariable regions of all Theileria and Babesia species. PCR products obtained from blood samples were hybridized to a membrane onto which nine species-specific oligonucleotides were covalently linked. Cross-reactions were not observed between any of the tested species. No DNA sequences from Bos taurus or other hemoparasites (Trypanosoma species, Cowdria ruminantium, Anaplasma marginale, and Ehrlichia species) were amplified. The sensitivity of the assay was determined at 0.000001% parasitemia, enabling detection of the carrier state of most parasites. Mixed DNAs from five different parasites were correctly identified. Moreover, blood samples from cattle experimentally infected with two different parasites reacted only with the corresponding species-specific oligonucleotides. Finally, RLB was used to screen blood samples collected from carrier cattle in two regions of Spain. T. annulata, T. orientalis, and B. bigemina were identified in these samples. In conclusion, the RLB is a versatile technique for simultaneous detection of all bovine tick-borne protozoan parasites. We recommend its use for integrated epidemiological monitoring of tick-borne disease, since RLB can also be used for screening ticks and can easily be expanded to include additional hemoparasite species. PMID:10325324

  7. Successful vaccination against Boophilus microplus and Babesia bovis using recombinat antigens

    OpenAIRE

    Willadsen,P.; Kemp,D. H.; Cobon,G. S.; Wright,I. G.

    1992-01-01

    Current methods for the control of the cattle tick Boophils microplus and the agent of bovine babesiosis, Babesia bovis are unsatisfactory. Effective immunological control of both parasites would have great advantages. However, naturally acquired immunity to the tick is generally unable to prevent serious production losses. A vaccine against the tick, based on a novel form of immunization, is being developed. A protective antigen has been isolated from the tick, characterized and produced as ...

  8. A study on ovine tick-borne hemoprotozoan parasites (Theileria and Babesia) in the East Black Sea Region of Turkey.

    Science.gov (United States)

    Altay, Kursat; Dumanli, Nazir; Aktas, Munir

    2012-07-01

    In this study, the frequency of Theileria and Babesia species was assessed via reverse line blotting and blood smear-based diagnostic methods in small ruminants. A total of 201 apparently healthy animals from 26 randomly selected herds located in 4 locations (Artvin, Giresun, Gumushane, and Tokat) of East Black Sea Region of Turkey were investigated for the blood protozoans. In a polymerase chain reaction (PCR), the hypervariable V4 region of the 18S ribosomal RNA gene was amplified with a set of general primers specific for all Theileria and Babesia species. The PCR products were hybridized against catchall and species-specific (Theileria spp., Theileria lestoquardi, Theileria ovis, Theileria sp. OT1, Theileria sp., OT3, Theileria sp., MK, Theileria luwenshuni, Theileria uilenbergi, Babesia spp., Babesia ovis, Babesia motasi, and Babesia crassa) probes. Theileria piroplasms were identified in nine (4.47%) samples by microscopic examination. Reverse line blotting (RLB) detected the infection in 19.90% of the samples. The infection rate of sheep (28.90%) was higher than goats (4.10%). T. ovis, Theileria sp., MK, and Theileria sp. OT3 were detected by RLB. The most prevalent Theileria species was T. ovis (18.90%) followed by Theileria sp. MK (0.99%). Theileria sp. OT3 was detected in one sample (0.43%). A single animal was infected as mix with T. ovis and Theileria sp. MK. The other Theileria (T. lestoquardi, Theileria sp. OT1, T. luwenshuni, and T. uilenbergi) and Babesia (B. ovis, B. motasi, and B. crassa) species were not detected. This study is the first molecular survey on ovine tick-borne protozoans in East Black Sea Region of Turkey.

  9. A PCR-based survey of selected Babesia and Theileria parasites in cattle in Sri Lanka.

    Science.gov (United States)

    Sivakumar, Thillaiampalam; Kothalawala, Hemal; Abeyratne, Sembukutti Arachchige Eranga; Vimalakumar, Singarayar Caniciyas; Meewewa, Asela Sanjeewa; Hadirampela, Dilhani Thilanka; Puvirajan, Thamotharampillai; Sukumar, Subramaniyam; Kuleswarakumar, Kulanayagam; Chandrasiri, Alawattage Don Nimal; Igarashi, Ikuo; Yokoyama, Naoaki

    2012-11-23

    Hemoprotozoan parasites are responsible for significant economic losses in cattle. We screened Sri Lankan cattle populations for the presence of Babesia bovis, Babesia bigemina, Theileria annulata, and Theileria orientalis, using species-specific PCR assays. Out of 316 samples collected from animals in four different districts of Sri Lanka (Nuwara Eliya, Polonnaruwa, Ampara, and Jaffna), 231 (73.1%) were positive for at least one parasite species. All four parasite species were detected among the study groups from all of the districts surveyed. The first and second commonest hemoprotozoan parasites identified were T. orientalis (53.5%) and B. bigemina (30.1%), respectively. We found that the dry zones (Polonnaruwa, Ampara, and Jaffna) had more Babesia-positive animals than the hill country wet zone (Nuwara Eliya). In contrast, T. orientalis was the predominant species detected in Nuwara Eliya, while infection with T. annulata was more common in the dry zones. In addition, 81 (35.1%) of the 231 positive samples were infected with more than one parasite species. The presence of multiple parasite species among the different cattle populations is of clinical and economic significance. Therefore, island-wide control and prevention programs against bovine babesiosis and theileriosis are needed to minimize the financial burden caused by these parasites. Copyright © 2012 Elsevier B.V. All rights reserved.

  10. Prevalence of Babesia spp. and Anaplasma marginale in cattle in the municipality of Palma, MG

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    Marluce Aparecida Mattos Paula

    2015-12-01

    Full Text Available ABSTRACT. Paula M.A.M., Oliveira F.C.R., Melo Jr O.A. & Frazão-Teixeira E. [Prevalence of Babesia spp. and Anaplasma marginale in cattle in the municipality of Palma, MG.] Prevalência de Babesia spp. e Anaplasma marginale em bovinos no município de Palma, MG. Revista Brasileira de Medicina Veterinária, 37(4: 359-365, 2015. Laboratório de Biologia Estrutural, Instituto Oswaldo Cruz/ Fiocruz, Avenida Brasil, 4365, Manguinhos, Rio de Janeiro, RJ 21040-361, Brasil. E-mail: edwards.teixeira@ioc.fiocruz.br We verified the prevalence of hemoparasites in 40 cattle with ages varying from one month to 12 years old, in two farms of the Municipality of Palma, Minas Gerais state, Brazil. Two blood smear samples were collected from each animal: one from the tail tip and another from the ear tip. The smears were fixed, stained and observed under 100X lighted microscope magnifying glass. Twenty- -seven out of 40 animals studied (67.5% had at least one species of hemoparasite. Among these, 21 (52.5% were infected with Babesia spp., 10 (25% with Anaplasma marginale and four (10% parasitized with both hemoparasites. The studied region is potentially enzootic for the detected parasites and there is high risk for clinical cases of tick-borne disease. Both anatomic points, tail and ear tips, are good spots for blood collection and smear confection for hemoparasite investigation.

  11. Babesia canis vogeli infection in dogs and ticks in the semiarid region of Pernambuco, Brazil

    Directory of Open Access Journals (Sweden)

    Andreina C. Araujo

    2015-05-01

    Full Text Available Abstract:This study aimed to report the prevalence of Babesia canis vogeli in dogs and ticks in the urban and rural areas of Petrolina, Pernambuco. Serum and peripheral blood samples of 404 dogs were tested by indirect immunofluorescence assay (IFA and by blood smears, respectively. The presence of tick infestation was evaluated, and some specimens were submitted to DNA amplification by polymerase chain reaction (PCR. The presence of antibodies anti-B. canis vogeli was determinate in 57.9% (234/404 of dogs. The direct detection of Babesia spp was obtained in 0.5% (2/404 dogs by visualization of intraerythrocytic forms. Infestation by Rhipicephalus sanguineus sensu lato was observed in 54.5% (220/404 of dogs in both urban and rural areas. DNA of Babesia canis vogeli were obtained by PCR in 6% individual (3/50 and 8.7% of pool of ticks (7/80. The risk factors for the presence of anti-B. canis vogeli antibodies, as determined through the application of logistic regression models (P<0.05, were the following: medium breed size variables (P<0.001; contact with areas of forest (P=0.021; and access on the street (P=0.046. This study describes, for the first time, the confirmation of infection of B. canis vogeli in dogs and ticks in the semiarid region of Pernambuco, Brazil.

  12. A specific DNA probe which identifies Babesia bovis in whole blood.

    Science.gov (United States)

    Petchpoo, W; Tan-ariya, P; Boonsaeng, V; Brockelman, C R; Wilairat, P; Panyim, S

    1992-05-01

    A genomic library of Babesia bovis DNA from the Mexican strain M was constructed in plasmid pUN121 and cloned in Escherichia coli. Several recombinants which hybridized strongly to radioactively labeled B. bovis genomic DNA in an in situ screening were selected and further analyzed for those which specifically hybridized to B. bovis DNA. It was found that pMU-B1 had the highest sensitivity, detecting 25 pg of purified B. bovis DNA, and 300 parasites in 10 microliters of whole infected blood, or 0.00025% parasitemia. pMU-B1 contained a 6.0 kb B. bovis DNA insert which did not cross-hybridize to Babesia bigemina, Trypanosoma evansi, Plasmodium falciparum, Anaplasma marginale, Boophilus microplus and cow DNA. In the Southern blot analysis of genomic DNA, pMU-B1 could differentiate between two B. bovis geographic isolates, Mexican strain M and Thai isolate TS4. Thus, the pMU-B1 probe will be useful in the diagnosis of Babesia infection in cattle and ticks, and in the differentiation of B. bovis strains.

  13. Validation of an indirect ELISA for the diagnosis of Babesia bovis in cattle in Yucatan, Mexico

    International Nuclear Information System (INIS)

    Dominguez, A.J.L.; Rodriguez, V.R.I.; Oura, C.; Cob, G.L.A.

    1998-01-01

    The ELISA kit provided by the FAO/IAEA for the diagnosis of Babesia bovis was validated. In order to determine the appropriate ELISA cut-off point that would serve as the threshold between positive and negative samples, 119 serum samples from a Mexican Babesia-free zone were analyzed. The optimal cut-off point chosen was at 12% of the reactivity of the high positive control serum sample (PP) which resulted in a specificity of 97%. One hundred and ninety-six cattle from Wisconsin, USA, were introduced into Yucatan, Mexico, of which 181 were vaccinated with an attenuated live Babesia bovis vaccine; 15 animals remained as unvaccinated controls. Before and after vaccination all animals were bled and tested by enzyme linked immunosorbent assay (ELISA) and indirect fluorescence antibody test (IFAT). Both tests showed a high degree of correlation in their results. To evaluate an immune response to vaccination the optimal cut-off point chosen was 12% PP resulting in a sensitivity 99% and a specificity 95%. We concluded that the ELISA test has proved to be useful in Yucatan, Mexico for serological surveys and monitoring the efficiency o vaccination programmes. (author)

  14. Animal level risk factors associated with Babesia and Theileria infections in cattle in Egypt.

    Science.gov (United States)

    Rizk, Mohamed Abdo; Salama, Akram; El-Sayed, Shimaa Abd-El-Salam; Elsify, Ahmed; El-Ashkar, Maged; Ibrahim, Hussam; Youssef, Mohamed; El-Khodery, Sabry

    2017-12-20

    In present study, blood samples were collected randomly from 439 cows at three main regions of Egypt (northern, central and southern). Molecular diagnosis of Babesia and Theileria infections by PCR amplification of DNA (gene) fragments, then cloning and sequencing of the positive samples were conducted. A questionnaire was created to imply the assumed risk factors and logistic regression statistical analysis was carried out to appraise the potential factors on the animal level. The results revealed that 49 (11.16%) and 45 (10.25%) cattle were infected with Babesia and Theileria parasites, respectively. B. bigemina (7.97%) and T. annulata (9.56%) were the most prevalent parasites. For Babesia sp., final multivariate logistic regression analysis showed a significant association between the infection and irregular use of antiprotozoal drugs (P = 0.003; OR: 0.28; 95% CI: 0.12-0.65), management practice (P = 0.029; OR: 6.66; 95% CI: 1.21-36.59) and ecology area (P = 0.006; OR: 5.62; 95% CI: 1.63-19.31). However, for Theileria sp. infection, animal breed (P = 0.003; OR: 0.44; 95% CI: .45-1.00) and irregular use of antiprotozoal drugs (PBabesia and Theileria sp. in Egypt based on molecular description. An impression on the potential risk factors associated with infections was obtained. Recognition of the potential risk factors associated with tick borne disease may be helpful to construct the best preventive measures.

  15. Effect of ionizing radiation on the virulence and the immunogenic properties of Babesia ovis

    International Nuclear Information System (INIS)

    Khalacheva, M.; Kararizova, L.

    1977-01-01

    The effect was followed up of ionizing radiation of 20, 25, 30, 32, 35, 40 and 50 krad on the virulence and the immunigenic properties of B.ovis. The experiments were carried out with 22 sheep (20 test and 2 donor) according to the rate of treatment into 7 test groups with a total of 17 animals, and 1 control with 3 sheep. One month after infection with irradiated blood reinfection was carried out of 8 animals (of different groups) that had recovered, using virulent untrated blood. The titer of the antibodies in the remaining sheep was followed up by means of the complement fixation test. It was found that irradiation at 20 - 25 krad lowered the virulence of the Babesia organisms, but these could provoke the disease in an acute form and cause death. The animals that survived after being infected with the indicated doses developed a comparatively good immunity. It is considered that the most appropriate dose of irradiation is 30 krad. 32 and 35 krad do not kill Babesiae; the parasites thus irradiated, however, cannot protect the animals ar reinfection. It has been found that Babesiae irradiated at the rates of 40 and 450 krad do not cause an infection process, and at reinfection the animals respond almost in the same way as the controls. (author)

  16. Prevalence of Streptococcus dysgalactiae subsp. equisimilis and S. equi subsp. zooepidemicus in a sample of healthy dogs, cats and horses.

    Science.gov (United States)

    Acke, E; Midwinter, A C; Lawrence, K; Gordon, S J G; Moore, S; Rasiah, I; Steward, K; French, N; Waller, A

    2015-09-01

    To estimate the prevalence of β-haemolytic Lancefield group C streptococci in healthy dogs, cats and horses; to determine if frequent contact with horses was associated with isolation of these species from dogs and cats; and to characterise recovered S. equi subsp. zooepidemicus isolates by multilocus sequence typing. Oropharyngeal swabs were collected from 197 dogs and 72 cats, and nasopharyngeal swabs from 93 horses. Sampling was carried out at the Massey University Veterinary Teaching Hospital, on sheep and beef farms or on premises where horses were present. All animals were healthy and were categorised as Urban dogs and cats (minimal contact with horses or farm livestock), Farm dogs (minimal contact with horses) and Stable dogs and cats (frequent contact with horses). Swabs were cultured for β-haemolytic Streptococcus spp. and Lancefield group C streptococcal subspecies were confirmed by phenotypic and molecular techniques. Of the 197 dogs sampled, 21 (10.7 (95% CI= 4.0-25.4)%) tested positive for S. dysgalactiae subsp. equisimilis and 4 (2.0 (95% CI=0.7-5.5)%) tested positive for S. equi subsp. zooepidemicus. All these isolates, except for one S. dysgalactiae subsp. equisimilis isolate in an Urban dog, were from Stable dogs. S. dysgalactiae subsp. equisimilis was isolated from one Stable cat. Of the 93 horses, 22 (23.7 (95% CI=12.3-40.6)%) and 6 (6.5 (95% CI=2.8-14.1)%) had confirmed S. dysgalactiae subsp. equisimilis and S. equi subsp. zooepidemicus isolation respectively. Isolation of S. dysgalactiae subsp. equisimilis from dogs was associated with frequent contact with horses (OR=9.8 (95% CI=2.6-72.8)). Three different multilocus sequence type profiles of S. equi subsp. zooepidemicus that have not been previously reported in dogs were recovered. Subclinical infection or colonisation by S. equi subsp. zooepidemicus and S. dysgalactiae subsp. equisimilis occurs in dogs and further research on inter-species transmission and the pathogenic potential of these

  17. Critical analysis of vector-borne infections in dogs: Babesia vogeli, Babesia gibsoni, Ehrlichia canis and Hepatozoon canis in Punjab, India.

    Science.gov (United States)

    Singla, Lachhman Das; Sumbria, Deepak; Mandhotra, Ajay; Bal, M S; Kaur, Paramjit

    2016-12-01

    There are few published studies on various vector borne diseases of dogs in India and most depict clinical infection in dogs, diagnosed by observation of the haemopathogens in stained blood smears. This study provides the first report regarding molecular confirmation and ancestral relationship analysis of blood smears positive cases of assorted haemopathogens in Punjab province of India. On blood smear examination, haemopathogens were observed in 124 out of 778 (15.95%, 95% CI: 13.53- 18.68) blood smears. Further polymerase chain reactions (PCR) was used on bloods smear positive cases to validate the results. Out of 778 blood samples, Babesia gibsoni was most common parasite infecting dogs (15.04%, 95% CI: 12.7-17.72), followed by Ehrlichia canis (0.39%, 95% CI: 0.0-1.13), infection of Babesia vogeli and Hepatozoon canis was same (0.26%, 95% CI: 0.0-0.9). Among various risk factors studied (age, sex, season), prevalence of infection was non-significantly higher in 1-2 year of age group (19.88%, 95% CI: 14.45-26.71), regarding sex same prevalence was recorded (15.94%), and chances of infection was highest in pre-monsoon i.e. summer (18.26%, 95% CI: 14.49-22.76). Phylogenetic analysis revealed ancestral background of Ludhiana isolates of B. vogeli, B. gibsoni, H. canis, and E. canis with the isolates of Philippines, Mongolia and Tunisia.

  18. Detection of Babesia canis rossi, B. canis vogeli, and Hepatozoon canis in Dogs in a Village of Eastern Sudan by Using a Screening PCR and Sequencing Methodologies

    Science.gov (United States)

    Oyamada, Maremichi; Davoust, Bernard; Boni, Mickaël; Dereure, Jacques; Bucheton, Bruno; Hammad, Awad; Itamoto, Kazuhito; Okuda, Masaru; Inokuma, Hisashi

    2005-01-01

    Babesia and Hepatozoon infections of dogs in a village of eastern Sudan were analyzed by using a single PCR and sequencing. Among 78 dogs, 5 were infected with Babesia canis rossi and 2 others were infected with B. canis vogeli. Thirty-three dogs were positive for Hepatozoon. Hepatozoon canis was detected by sequence analysis. PMID:16275954

  19. Detection of Babesia canis rossi, B. canis vogeli, and Hepatozoon canis in dogs in a village of eastern Sudan by using a screening PCR and sequencing methodologies.

    Science.gov (United States)

    Oyamada, Maremichi; Davoust, Bernard; Boni, Mickaël; Dereure, Jacques; Bucheton, Bruno; Hammad, Awad; Itamoto, Kazuhito; Okuda, Masaru; Inokuma, Hisashi

    2005-11-01

    Babesia and Hepatozoon infections of dogs in a village of eastern Sudan were analyzed by using a single PCR and sequencing. Among 78 dogs, 5 were infected with Babesia canis rossi and 2 others were infected with B. canis vogeli. Thirty-three dogs were positive for Hepatozoon. Hepatozoon canis was detected by sequence analysis.

  20. Detection of Babesia canis rossi, B. canis vogeli, and Hepatozoon canis in Dogs in a Village of Eastern Sudan by Using a Screening PCR and Sequencing Methodologies

    OpenAIRE

    Oyamada, Maremichi; Davoust, Bernard; Boni, Mickaël; Dereure, Jacques; Bucheton, Bruno; Hammad, Awad; Itamoto, Kazuhito; Okuda, Masaru; Inokuma, Hisashi

    2005-01-01

    Babesia and Hepatozoon infections of dogs in a village of eastern Sudan were analyzed by using a single PCR and sequencing. Among 78 dogs, 5 were infected with Babesia canis rossi and 2 others were infected with B. canis vogeli. Thirty-three dogs were positive for Hepatozoon. Hepatozoon canis was detected by sequence analysis.

  1. Babesia bicornis sp. nov. and Theileria bicornis sp. nov.: Tick-Borne Parasites Associated with Mortality in the Black Rhinoceros (Diceros bicornis)

    Science.gov (United States)

    Nijhof, Ard M.; Penzhorn, Banie L.; Lynen, Godelieve; Mollel, Johnson O.; Morkel, Pete; Bekker, Cornelis P. J.; Jongejan, Frans

    2003-01-01

    A novel Babesia species, designated Babesia bicornis sp. nov., was identified in three black rhinoceroses (Diceros bicornis) that died in wildlife areas in Tanzania and South Africa. Screening of black rhinoceroses in South Africa revealed, in addition to B. bicornis, a second parasite, designated Theileria bicornis sp. nov. PMID:12734294

  2. Babesia bicornis sp. nov. and Theileria bicornis sp. nov.: Tick-Borne Parasites Associated with Mortality in the Black Rhinoceros (Diceros bicornis)

    OpenAIRE

    Nijhof, Ard M.; Penzhorn, Banie L.; Lynen, Godelieve; Mollel, Johnson O.; Morkel, Pete; Bekker, Cornelis P. J.; Jongejan, Frans

    2003-01-01

    A novel Babesia species, designated Babesia bicornis sp. nov., was identified in three black rhinoceroses (Diceros bicornis) that died in wildlife areas in Tanzania and South Africa. Screening of black rhinoceroses in South Africa revealed, in addition to B. bicornis, a second parasite, designated Theileria bicornis sp. nov.

  3. Anaplasma phagocytophilum and Babesia spp. in roe deer (Capreolus capreolus), fallow deer (Dama dama) and mouflon (Ovis musimon) in Germany.

    Science.gov (United States)

    Kauffmann, Melanie; Rehbein, Steffen; Hamel, Dietmar; Lutz, Walburga; Heddergott, Mike; Pfister, Kurt; Silaghi, Cornelia

    2017-02-01

    Infections with the tick-borne pathogens Anaplasma phagocytophilum and Babesia spp. can cause febrile disease in several mammalian species, including humans. Wild ruminants in Europe are suggested to serve as reservoir hosts for particular strains or species of these pathogens. The aims of this study were to investigate the occurrence of A. phagocytophilum and Babesia spp. in roe deer (Capreolus capreolus), fallow deer (Dama dama) and mouflon (Ovis musimon orientalis) in Germany, and the diversity and host association of genetic variants of A. phagocytophilum and Babesia species. From 2009 to 2010, 364 spleen samples from 153 roe deer, 43 fallow deer and 168 mouflon from 13 locations in Germany were tested for DNA of A. phagocytophilum and Babesia spp. by real-time PCR or conventional PCR, respectively. Variants of A. phagocytophilum were investigated with a nested PCR targeting the partial 16S rRNA gene, and species of piroplasms were identified by sequencing. DNA of A. phagocytophilum was detected in 303 (83.2%) samples: roe deer, 96.1% (147/153); fallow deer, 72.1% (31/43); and mouflon, 74.4% (125/168). Sequence analysis of 16S rRNA-PCR products revealed the presence of nine different genetic variants. DNA of Babesia spp. was found in 113 (31.0%) samples: roe deer, 62.8% (96/153); fallow deer, 16.3% (6/43); and mouflon, 6.5% (11/168). Babesia capreoli, Babesia sp. EU1 (referred to also as B. venatorum), B. odocoilei-like and a Theileria species were identified. Co-infections with A. phagocytophilum and Babesia spp. were detected in 30.0% of the animals which were tested positive for A. phagocytophilum and/or Babesia spp. Roe deer had a significantly higher percentage of co-infections (60.8%), followed by fallow deer (14.0%) and mouflon (6.5%). Thus, the results suggest that roe deer plays a key role in the endemic cycles of the pathogens investigated. Copyright © 2016 Elsevier Ltd. All rights reserved.

  4. Equi-biaxial loading effect on austenitic stainless steel fatigue life

    Directory of Open Access Journals (Sweden)

    C. Gourdin

    2016-10-01

    Full Text Available Fatigue lifetime assessment is essential in the design of structures. Under-estimated predictions may result in unnecessary in service inspections. Conversely, over-estimated predictions may have serious consequences on the integrity of structures. In some nuclear power plant components, the fatigue loading may be equibiaxial because of thermal fatigue. So the potential impact of multiaxial loading on the fatigue life of components is a major concern. Meanwhile, few experimental data are available on austenitic stainless steels. It is essential to improve the fatigue assessment methodologies to take into account the potential equi-biaxial fatigue damage. Hence this requires obtaining experimental data on the considered material with a strain tensor in equibiaxial tension. Two calibration tests (with strain gauges and image correlation were used to obtain the relationship between the imposed deflection and the radial strain on the FABIME2 specimen. A numerical study has confirmed this relationship. Biaxial fatigue tests are carried out on two austenitic stainless steels for different values of the maximum deflection, and with a load ratio equal to -1. The interpretation of the experimental results requires the use of an appropriate definition of strain equivalent. In nuclear industry, two kinds of definition are used: von Mises and TRESCA strain equivalent. These results have permitted to estimate the impact of the equibiaxiality on the fatigue life of components

  5. Interaction between M-like protein and macrophage thioredoxin facilitates antiphagocytosis for Streptococcus equi ssp. zooepidemicus.

    Directory of Open Access Journals (Sweden)

    Zhe Ma

    Full Text Available Streptococcus equi ssp. zooepidemicus (S. zooepidemicus, S.z is one of the common pathogens that can cause septicemia, meningitis, and mammitis in domesticated species. M-like protein (SzP is an important virulence factor of S. zooepidemicus and contributes to bacterial infection and antiphagocytosis. The interaction between SzP of S. zooepidemicus and porcine thioredoxin (TRX was identified by the yeast two-hybrid and further confirmed by co-immunoprecipitation. SzP interacted with both reduced and the oxidized forms of TRX without inhibiting TRX activity. Membrane anchored SzP was able to recruit TRX to the surface, which would facilitate the antiphagocytosis of the bacteria. Further experiments revealed that TRX regulated the alternative complement pathway by inhibiting C3 convertase activity and associating with factor H (FH. TRX alone inhibited C3 cleavage and C3a production, and the inhibitory effect was additive when FH was also present. TRX inhibited C3 deposition on the bacterial surface when it was recruited by SzP. These new findings indicated that S. zooepidemicus used SzP to recruit TRX and regulated the alternative complement pathways to evade the host immune phagocytosis.

  6. Interaction between M-Like Protein and Macrophage Thioredoxin Facilitates Antiphagocytosis for Streptococcus equi ssp. zooepidemicus

    Science.gov (United States)

    Ma, Zhe; Zhang, Hui; Zheng, Junxi; Li, Yue; Yi, Li; Fan, Hongjie; Lu, Chengping

    2012-01-01

    Streptococcus equi ssp. zooepidemicus (S. zooepidemicus, S.z) is one of the common pathogens that can cause septicemia, meningitis, and mammitis in domesticated species. M-like protein (SzP) is an important virulence factor of S. zooepidemicus and contributes to bacterial infection and antiphagocytosis. The interaction between SzP of S. zooepidemicus and porcine thioredoxin (TRX) was identified by the yeast two-hybrid and further confirmed by co-immunoprecipitation. SzP interacted with both reduced and the oxidized forms of TRX without inhibiting TRX activity. Membrane anchored SzP was able to recruit TRX to the surface, which would facilitate the antiphagocytosis of the bacteria. Further experiments revealed that TRX regulated the alternative complement pathway by inhibiting C3 convertase activity and associating with factor H (FH). TRX alone inhibited C3 cleavage and C3a production, and the inhibitory effect was additive when FH was also present. TRX inhibited C3 deposition on the bacterial surface when it was recruited by SzP. These new findings indicated that S. zooepidemicus used SzP to recruit TRX and regulated the alternative complement pathways to evade the host immune phagocytosis. PMID:22384152

  7. Recovery and subsequent characterization of polyhydroxybutyrate from Rhodococcus equi cells grown on crude palm kernel oil

    Directory of Open Access Journals (Sweden)

    Nadia Altaee

    2016-07-01

    Full Text Available The gram-positive bacterium Rhodococcus equi was isolated from fertile soil, and mineral salt media (MM and trace elements were used to provide the necessary elements for its growth and PHB production in addition to using crude palm kernel oil (CPKO 1% as the carbon source. Gas chromatography (GC demonstrated that the composition of the recovered biopolymer was homopolymer polyhydroxybutyrate (PHB. The strain of the present study has a dry biomass of 1.43 (g/l with 38% PHB, as determined by GC. The recovered PHB was characterized by NMR to study the chemical structure. In addition, DSC and TGA were used to study the thermal properties of the recovered polymer, where the melting temperature (Tm was 173 °C, the glass transition temperature (Tg was 2.79 °C, and the decomposition temperature (Td was 276 °C. Gel permeation chromatography (GPC was used to study the molecular mass of the recovered PHB in addition to comparing the results with other studies using different bacteria and substrates, where the molecular weight was 642 kDa, to enable its usage in many applications. The present study demonstrated the use of an inexpensive substrate for PHB production, i.e., using gram-positive bacteria to produce PHB polymer with characterization.

  8. Identification and characterization of a novel protective antigen, Sec_205 of Streptococcus equi ssp. Zooepidemicus.

    Science.gov (United States)

    Liang, Huihuang; Tang, Bin; Zhao, Pengpeng; Deng, Mingyong; Yan, Lili; Zhai, Pan; Wei, Zigong

    2018-02-01

    Streptococcus equi ssp. zooepidemicus (SEZ) is an important pathogen of swine streptococcal diseases and can infect a wide range of animals as well as human beings. The absence of effective vaccine confounds the control of SEZ infection. Sec_205, a novel protein identified in the previous study, was inducibly over-expressed in Escherichia coli in the present study. The purified recombinant protein could elicit a significant humoral antibody response and provide efficient protection against lethal challenge of SEZ C55138 in mouse model. The protection against SEZ infection was mediated by specific antibodies to Sec_205 to some extent and was identified by the passive protection assay. The Sec_205 was an in vivo-induced antigen confirmed by the real-time PCR and could adhere to the Hep-2 cells by the inhibition assay. These suggest that Sec_205 may play a vital role in pathogenicity and serve as a new vaccine candidate against SEZ infection. Copyright © 2018 Elsevier Ltd. All rights reserved.

  9. CD44 deficiency enhanced Streptococcus equi ssp. zooepidemicus dissemination and inflammation response in a mouse model.

    Science.gov (United States)

    Fu, Qiang; Xiao, Pingping; Chen, Yaosheng; Wei, Zigong; Liu, Xiaohong

    2017-12-01

    Streptococcus equi ssp. zooepidemicus (S. zooepidemicus) is responsible for peritonitis, septicemia, meningitis, arthritis and several other serious diseases in various species. Recent studies have demonstrated that CD44 is implicated in the process of host defense against pathogenic microorganisms. In the present study, the role of CD44 in the host response to S. zooepidemicus infection was investigated in a mouse model. Upon intraperitoneal infection with S. zooepidemicus, the expression of CD44 on the peritoneal exudate cells from wild-type (WT) mice was increased. CD44 deficiency accelerated mortality, which was accompanied by increased peritoneal bacterial growth and dissemination to distant body sites. CD44 knock-out (KO) mice showed enhanced early inflammatory cell recruitment into the peritoneal fluid on S. zooepidemicus infection. In line with this, the expression of proinflammatory cytokines, chemokines in peritoneal exudate cells and peritoneal macrophages of CD44 KO mice were increased compared with those of WT mice. In addition, CD44 deficiency was associated with reduced expression of A20, a negative regulator in TLR signaling. Overall, the present study suggests that CD44 plays a protective role in antibacterial defense against S. zooepidemicus in mice. Copyright © 2017. Published by Elsevier Ltd.

  10. Rangelia vitalii, Babesia spp. and Ehrlichia spp. in dogs in Passo Fundo, state of Rio Grande do Sul, Brazil

    Directory of Open Access Journals (Sweden)

    Juliana Gottlieb

    2016-06-01

    Full Text Available Abstract Pathogens transmitted by ticks are an emerging problem worldwide, this study aimed to diagnose the causal agents of infection in dogs presenting suspected hemoparasitoses. Fifty-eight dogs with clinical signs such as depression, hemorrhagic diathesis and fever were evaluated regarding clinical presentation, hemogram, blood smears and serological tests, using the indirect immunofluorescence method for the agents Babesia vogeli and Ehrlichia canis and conventional PCR for Babesia spp. (gene 18S rRNA, Rangelia vitalii (gene 18S rRNA and Ehrlichia spp. (gene dsb. Five (8.6% of the 58 dogs were serologically positive for Babesia spp. and three (5.1% for E. canis. Four dogs (6.8% were positive for R. vitalii through the molecular diagnosis. The PCR products were sequenced and the DNA from R. vitalii was found to be 99% genetically identical to samples of R. vitalii that had been isolated in Brazil. No presence of Babesia spp. or E. canis was observed through PCR on the dogs evaluated here. The results indicate the presence of R. vitalii and exposure to Babesia spp. and Ehrlichia spp. among the dogs analyzed.

  11. Molecular identification of badger-associated Babesia sp. DNA in dogs: updated phylogeny of piroplasms infecting Caniformia.

    Science.gov (United States)

    Hornok, Sándor; Horváth, Gábor; Takács, Nóra; Kontschán, Jenő; Szőke, Krisztina; Farkas, Róbert

    2018-04-11

    Piroplasms are unicellular, tick-borne parasites. Among them, during the past decade, an increasing diversity of Babesia spp. has been reported from wild carnivores. On the other hand, despite the known contact of domestic and wild carnivores (e.g. during hunting), and a number of ixodid tick species they share, data on the infection of dogs with babesiae from other families of carnivores are rare. In this study blood samples were collected from 90 dogs and five road-killed badgers. Ticks were also removed from these animals. The DNA was extracted from all blood samples, and from 33 ticks of badgers, followed by molecular analysis for piroplasms with PCR and sequencing, as well as by phylogenetic comparison of detected genotypes with piroplasms infecting carnivores. Eleven of 90 blood DNA extracts from dogs, and all five samples from badgers were PCR-positive for piroplasms. In addition to the presence of B. canis DNA in five dogs, sequencing identified the DNA of badger-associated "Babesia sp. Meles-Hu1" in six dogs and in all five badgers. The DNA of "Babesia sp. Meles-Hu1" occurred significantly more frequently in dogs often taken to forests (i.e. the preferred habitat of badgers in Hungary), than in dogs without this characteristic. Moreover, detection of DNA from this Babesia sp. was significantly associated with hunting dogs in comparison with dogs not used for hunting. Two PCR-positive dogs (in one of which the DNA of the badger-associated Babesia sp. was identified, whereas in the other the DNA of B. canis was present) showed clinical signs of babesiosis. Engorged specimens of both I. canisuga and I. hexagonus were collected from badgers with parasitaemia, but only I. canisuga contained the DNA of "Babesia sp. Meles-Hu1". This means a significant association of the DNA from "Babesia sp. Meles-Hu1" with I. canisuga. Phylogenetically, "Babesia sp. Meles-Hu1" belonged to the "B. microti" group. This is the first detection of the DNA from a badger

  12. Molecular characterisation of Babesia gibsoni infection from a Pit-bull terrier pup recently imported into South Africa

    Directory of Open Access Journals (Sweden)

    P.T. Matjila

    2007-05-01

    Full Text Available Canine babesiosis caused by Babesia gibsoni was diagnosed in a 3-month-old Pit-bull pup during a routine clinical examination. Diagnosis was confirmed by way of smear examination, PCR, Reverse Line Blot (RLB and sequence analysis which showed 100% homology with B. gibsoni (Japan AB118032 and Babesia sp. (Oklahoma (AF205636. Haematology showed moderate anaemia and severe thrombocytopenia. Treatment was initiated with diminazene aceturate (Berenil RTU(R followed by 2 doses of imidocarb diproprionate (Forray-65(R 3 days and 14 days later, respectively. Babesia gibsoni DNA was still detectable 2 weeks post-treatment on the PCR/RLB test. A 10-day course of combination drug therapy using atovaquone and azithromycin was initiated. Blood samples taken on Day 1 and Day 40 after completion of treatment were negative for B. gibsoni DNA on PCR/RLB test. The implications of a possible introduction of B. gibsoni into South Africa are discussed.

  13. Identification of novel Babesia and Theileria genotypes in the endangered marsupials, the woylie (Bettongia penicillata ogilbyi) and boodie (Bettongia lesueur).

    Science.gov (United States)

    Paparini, Andrea; Ryan, Una M; Warren, Kris; McInnes, Linda M; de Tores, Paul; Irwin, Peter J

    2012-05-01

    Piroplasms, which include the genera Theileria and Babesia, are blood-borne parasites transmitted mainly by tick vectors. Relatively little is known about their prevalence and clinical impact in Australian marsupials. In the present study the occurrence and molecular phylogeny of these parasites were studied in both wild and captive marsupials from Western Australia (WA) and Queensland (QLD). Blood samples were screened by microscopy and molecular methods, using PCR and DNA sequencing of the 18S ribosomal RNA gene (18S rDNA). Overall, 7.1% of the blood samples (8/113) were positive for piroplasm 18S rDNA. Theileria and Babesia rDNA was detected in 0.9% (1/113) and 6.2% (7/113) of the animals, respectively. The single Theileria positive was identified in one of three boodies (Bettongia lesueur) screened from a wildlife rehabilitation centre in WA, while all seven Babesia positives were detected in WA in wild captured woylies (Bettongia penicillata ogilbyi). Small intraerythrocytic inclusions were observed in blood films made from six of these individuals. This is the first report of a Babesia sp. in woylies, and Theileria sp. in boodies. Phylogenetic analysis indicated that the woylie-derived Babesia was genetically distinct and most closely related to Babesia occultans, the causative agent of a benign form of cattle babesiosis (genetic similarity 98.4%). The Theileria identified was most closely related to the marsupial-derived species Theileria penicillata from the woylie, Theileria brachyuri from the quokka (Setonix brachyurus), and Theileria sp. from the long-nosed potoroo (Potorous tridactylus). Copyright © 2012 Elsevier Inc. All rights reserved.

  14. Evaluation of a nested PCR test and bacterial culture of swabs from the nasal passages and from abscesses in relation to diagnosis of Streptococcus equi infection (strangles)

    DEFF Research Database (Denmark)

    Grønbæk, L.M.; Angen, Øystein; Vigre, Håkan

    2006-01-01

    . Methods: Two herds with natural outbreaks of strangles were visited over a period of 15 weeks and 323 samples originating from 35 horses investigated. The diagnostic use of a nested PCR test was evaluated using a collection of 165 isolates of Lancefield group C streptococci (species specificity) and swabs...... from nasal passages or from abscesses from horses infected with S. equi (diagnostic sensitivity). Results: All 45 S. equi isolates tested positive in the nested PCR, whereas no amplicon was formed when testing the other 120 Lancefield group C isolates. A total of 43 samples were collected from 11...... horses with and without clinical signs. Conclusions and potential relevance: The nested PCR test represents a species-specific and -sensitive method for diagnosis of S. equi from clinical samples. It may, however, be desirable in future to develop detection methods with high diagnostic sensitivity...

  15. Development of multiplex polymerase chain reaction for detection of Ehrlichia canis, Babesia spp and Hepatozoon canis in canine blood.

    Science.gov (United States)

    Kledmanee, Kan; Suwanpakdee, Sarin; Krajangwong, Sakranmanee; Chatsiriwech, Jarin; Suksai, Parut; Suwannachat, Pongpun; Sariya, Ladawan; Buddhirongawatr, Ruangrat; Charoonrut, Phingphol; Chaichoun, Kridsada

    2009-01-01

    A multiplex polymerase chain reaction (PCR) has been developed for simultaneous detection of canine blood parasites, Ehrlichia canis, Babesia spp and Hepatozoon canis, from blood samples in a single reaction. The multiplex PCR primers were specific to E. canis VirB9, Babesia spp 16S rRNA and H. canis 16S rRNA genes. Specificity of the amplicons was confirmed by DNA sequencing. The assay was evaluated using normal canine and infected blood samples, which were detected by microscopic examination. This multiplex PCR offers scope for simultaneous detection of three important canine blood parasites and should be valuable in monitoring parasite infections in dogs and ticks.

  16. Equi-axed and columnar grain growth in UO{sub 2}

    Energy Technology Data Exchange (ETDEWEB)

    White, R J [Berkely Technology Centre, Nuclear Electric plc, Berkeley (United Kingdom)

    1997-08-01

    The grain size of UO{sub 2} is an important parameter in the actual performance and the modelling of the performance of reactor fuel elements. Many processes depend critically on the grain size, for example, the degree of initial densification, the evolution rate of stable fission gases, the release rates of radiologically hazardous fission products, the fission gas bubble swelling rates and the fuel creep. Many of these processes are thermally activated and further impact on the fuel thermal behavior thus creating complex feedback processes. In order to model the fuel performance accurately it is necessary to model the evolution of the fuel grain radius. When UO{sub 2} is irradiated, the fission gases xenon and krypton are created from the fissioning uranium nucleus. At high temperatures these gases diffuse rapidly to the grain boundaries where they nucleate immobile lenticular shaped fission gas bubbles. In this paper the Hillert grain growth model is adapted to account for the inhibiting ``Zener`` effects of grain boundary fission gas porosity on grain boundary mobility and hence grain growth. It is shown that normal grain growth ceases at relatively low levels of irradiation. At high burnups, high temperatures and in regions of high temperature gradients, columnar grain growth is often observed, in some cases extending over more than fifty percent of the fuel radius. The model is further extended to account for the de-pinning of grains in the radial direction by the thermal gradient induced force on a fission gas grain boundary bubble. The observed columnar/equi-axed boundary is in fair agreement with the predictions of an evaporation/condensation model. The grain growth model described in this paper requires information concerning the scale of grain boundary porosity, the local fuel temperature and the local temperature gradient. The model is currently used in the Nuclear Electric version of the ENIGMA fuel modelling code. (author). 14 refs, 3 figs, 1 tab.

  17. EquiMoves: A Wireless Networked Inertial Measurement System for Objective Examination of Horse Gait

    Directory of Open Access Journals (Sweden)

    Stephan Bosch

    2018-03-01

    Full Text Available In this paper, we describe and validate the EquiMoves system, which aims to support equine veterinarians in assessing lameness and gait performance in horses. The system works by capturing horse motion from up to eight synchronized wireless inertial measurement units. It can be used in various equine gait modes, and analyzes both upper-body and limb movements. The validation against an optical motion capture system is based on a Bland–Altman analysis that illustrates the agreement between the two systems. The sagittal kinematic results (protraction, retraction, and sagittal range of motion show limits of agreement of ± 2.3 degrees and an absolute bias of 0.3 degrees in the worst case. The coronal kinematic results (adduction, abduction, and coronal range of motion show limits of agreement of − 8.8 and 8.1 degrees, and an absolute bias of 0.4 degrees in the worst case. The worse coronal kinematic results are most likely caused by the optical system setup (depth perception difficulty and suboptimal marker placement. The upper-body symmetry results show no significant bias in the agreement between the two systems; in most cases, the agreement is within ±5 mm. On a trial-level basis, the limits of agreement for withers and sacrum are within ±2 mm, meaning that the system can properly quantify motion asymmetry. Overall, the bias for all symmetry-related results is less than 1 mm, which is important for reproducibility and further comparison to other systems.

  18. Molecular characterization of Rhodococcus equi isolates from horses in Poland: pVapA characteristics and plasmid new variant, 85-kb type V.

    Science.gov (United States)

    Witkowski, Lucjan; Rzewuska, Magdalena; Takai, Shinji; Chrobak-Chmiel, Dorota; Kizerwetter-Świda, Magdalena; Feret, Małgorzata; Gawryś, Marta; Witkowski, Maciej; Kita, Jerzy

    2017-01-26

    Rhodococcus equi is one of the most significant bacterial pathogens affecting foals up to 6 months of age worldwide. Rhodococcosis is present in Poland however information about molecular characterization of R. equi isolates is scarce. This study describes molecular characterization of Rhodococcus equi infection on 13 horse breeding farms in Poland between 2001 and 2012. Samples were collected by tracheobronchial aspiration from pneumonic foals or during necropsy. The R. equi isolates were genotyped by plasmid profiling and pulsed-field gel electrophoresis. Totally, 58 R. equi isolates were investigated. One isolate lost its plasmid. Among the 57 VapA-positive isolates, 48 contained 85-kb type I plasmid (82.8%), 8 contained 87-kb type I plasmid (13.8%). One isolate (1.7%) had a unique restriction cleavage pattern and the 2nd fragment of EcoRI digests of this plasmid DNA was about 2600 bases smaller than that of the 85 kb type I. This new plasmid variant was designated as the "85-kb type V". Among the 58 isolates typeable with VspI-PFGE, ten PFGE clusters were detected. The majority of foals were infected mostly with isolates of low genetic diversity. Most of clinical isolates of R. equi from foals in Poland contain pVapA 85-kb type I and 87-kb type I similarly to the other European countries and the United States. However, the new variant of pVapA 85-kb type V was identified. The chromosomal variability was detected among some of the investigated isolates and the presence of farm-specific isolates might be possible.

  19. Characterization of Virulence Plasmids and Serotyping of Rhodococcus equi Isolates from Submaxillary Lymph Nodes of Pigs in Hungary

    OpenAIRE

    Makrai, László; Takayama, Saki; Dénes, Béla; Hajtós, István; Sasaki, Yukako; Kakuda, Tsutomu; Tsubaki, Shiro; Major, Andrea; Fodor, László; Varga, János; Takai, Shinji

    2005-01-01

    The plasmid types and serotypes of 164 Rhodococcus equi strains obtained from submaxillary lymph nodes of swine from different piggeries in 28 villages and towns located throughout the country were examined. The strains were tested by PCR for the presence of 15- to 17-kDa virulence-associated protein antigen (VapA) and 20-kDa virulence-associated protein antigen (VapB) genes. Plasmid DNAs were isolated and analyzed by digestion with restriction endonucleases to estimate size and compare their...

  20. PREVALENCE OF BABESIA SPP., EHRLICHIA SPP., AND TICK INFESTATIONS IN OKLAHOMA BLACK BEARS (URSUS AMERICANUS).

    Science.gov (United States)

    Skinner, Delaina; Mitcham, Jessica R; Starkey, Lindsay A; Noden, Bruce H; Fairbanks, W Sue; Little, Susan E

    2017-10-01

    American black bears (Ursus americanus) are commonly infested with ticks throughout their range, but there are few surveys for tick-borne disease agents in bears. To characterize tick infestations and determine the prevalence of current infection with Babesia spp. and past or current infection with Ehrlichia spp. in newly re-established populations of black bears in east central and southeastern Oklahoma, US, we identified adult (n=1,048) and immature (n=107) ticks recovered from bears (n=62). We evaluated serum and whole blood samples from a subset (n=49) for antibodies reactive to, and characteristic DNA fragments of, Ehrlichia spp., as well as characteristic DNA fragments of Babesia spp. Amblyomma americanum, the most common tick identified, was found on a majority (56/62; 90%) of bears and accounted for 697/1,048 (66.5%) of all ticks recovered. Other ticks included Dermacentor variabilis (338/1,048; 32.3%) from 36 bears, Amblyomma maculatum (9/1,048; 0.9%) from three bears, and Ixodes scapularis (4/1,048; 0.4%) from three bears. Antibodies reactive to Ehrlichia spp. were detected in every bear tested (49/49; 100%); maximum inverse titers to Ehrlichia chaffeensis ranged from 64-4,096 (geometric mean titer 1,525). However, PCR failed to identify active infection with E. chaffeensis, Ehrlichia ewingii, or an Ehrlichia ruminantium-like agent. Infection with Babesia spp. was detected by PCR in 3/49 (6%) bears. Together these data confirm that tick infestations and infection with tick-borne disease agents are common in bears in the southern US. The significance of these infestations and infections to the health of bears, if any, and the identity of the Ehrlichia spp. responsible for the antibody reactivity seen, warrant further evaluation.

  1. Molecular detection of Theileria, Babesia, and Hepatozoon spp. in ixodid ticks from Palestine.

    Science.gov (United States)

    Azmi, Kifaya; Ereqat, Suheir; Nasereddin, Abedelmajeed; Al-Jawabreh, Amer; Baneth, Gad; Abdeen, Ziad

    2016-07-01

    Ixodid ticks transmit various infectious agents that cause disease in humans and livestock worldwide. A cross-sectional survey on the presence of protozoan pathogens in ticks was carried out to assess the impact of tick-borne protozoa on domestic animals in Palestine. Ticks were collected from herds with sheep, goats and dogs in different geographic districts and their species were determined using morphological keys. The presence of piroplasms and Hepatozoon spp. was determined by PCR amplification of a 460-540bp fragment of the 18S rRNA gene followed by RFLP or DNA sequencing. A PCR-RFLP method based on the 18S rRNA was used in order to detect and to identify Hepatozoon, Babesia and Theileria spp. A total of 516 ticks were collected from animals in six Palestinian localities. Five tick species were found: Rhipicephalus sanguineus sensu lato, Rhipicephalus turanicus, Rhipicephalus bursa, Haemaphysalis parva and Haemaphysalis adleri. PCR-based analyses of the ticks revealed Theileria ovis (5.4%), Hepatozoon canis (4.3%), Babesia ovis (0.6%), and Babesia vogeli (0.4%). Theileria ovis was significantly associated with ticks from sheep and with R. turanicus ticks (p<0.01). H. canis was detected only in R. sanguineus s.l. and was significantly associated with ticks from dogs (p<0.01). To our knowledge, this is the first report describing the presence of these pathogens in ticks collected from Palestine. Communicating these findings with health and veterinary professionals will increase their awareness, and contribute to improved diagnosis and treatment of tick-borne diseases. Copyright © 2016. Published by Elsevier GmbH.

  2. Distribution patterns of Babesia gibsoni infection in hunting dogs from nine Japanese islands.

    Science.gov (United States)

    El-Dakhly, Khaled Mohamed; Goto, Minami; Noishiki, Kaori; El-Nahass, El-Shaymaa; Sakai, Hiroki; Yanai, Tokuma; Takashima, Yasuhiro

    2015-04-01

    Canine babesiosis constitutes a major global veterinary medical problem caused by tick-borne hemoparasites Babesia gibsoni and Babesia canis. Babesia gibsoni induces more severe clinical signs and is mainly transmitted by the ixodid Haemaphysalis longicornis. In Japan, B. gibsoni is primarily found in the western districts, with few records in the eastern parts. The aim of the current investigation was to evaluate distribution patterns of B. gibsoni infection in 9 Japanese islands and peninsulas using direct microscopy and PCR. Therefore, 196 hunting dogs were randomly sampled during the period from March to September 2011. Ages and sexes of dogs were identified. Direct microscopy of Giemsa-stained blood smear revealed pear-shaped piroplasms of B. gibsoni in 3 (1.6%) dogs. PCR was done initially with the universal primer set (B18S-F and B18S-R) amplifying the 1,665-bp portion of the 18S rRNA gene, followed by the specific primer set (Bg18F1 and Bg18R2) amplifying 2,363-bp fragments of the same gene. Accordingly, 84 (42.9%) and 8 (4.1%) dogs were positive, respectively. The current investigation shows that canine babesiosis was recorded in all islands except for Sado Island, Atsumi Peninsula, and Tanegashima Island. The highest infection rate was detected in the main island of Okinawa, while the lowest was on Ishigaki Island. Both sexes were non-significantly infected. However, the diversity of infection in islands was significantly different (P < 0.05). Although B. gibsoni has been previously found in western and eastern Japan, the present work highlights the prevalence of infection in many Japanese districts, including islands and peninsulas, giving realistic data that can facilitate treatment and control.

  3. Dimetilsulfóxido - DMSO no teste de sensibilidade microbiana in vitro em cepas de Rhodococcus equi isoladas de afecções pulmonares em potros

    Directory of Open Access Journals (Sweden)

    Ribeiro Márcio Garcia

    2001-01-01

    Full Text Available Comparou-se a sensibilidade microbiana in vitro de isolados de Rhodococcus equi pelo teste padrão de difusão com discos, com o modificado, pela adição de 5% de dimetilsulfóxido-DMSO. Observou-se aumento da sensibilidade do R. equi no teste com DMSO, frente a aminoglicosídeos (canamicina, amicacina, estreptomicina e ao cloranfenicol, enquanto para a eritromicina e derivados ß-lactâmicos (penicilina G, cefalosporinas, amoxicilina, oxacilina, constatou-se redução da sensibilidade do agente.

  4. The diagnosis of bovine basesiosis (babesia bovis) by means of the test of ELISA

    International Nuclear Information System (INIS)

    Leon Arenas, Edgar; Guillen, Ana Teresa; Silva, Maglene

    1997-01-01

    Between 1994 and 1996 a kit ELISA, developed by the FAO - IAEA for the diagnose of bovine babesiosis produced by Babesia bovis, was validated. There were processed a total of 547 blood serums from bovine between 9 and 18 months old, coming from high and low risk to illness areas. The point obtained for the test was 0.178 (DO) and the resulting percentages inside the population studied was 48% animal positive and 52% bovine negative. These results confirm that bovine population in Venezuela is in enzootic uncertainty areas for bovine babesiosis [es

  5. Inhibitory effect of apicidin on in vitro and in vivo growth of Babesia parasites

    OpenAIRE

    Munkhjargal, T; Aboulaila, M.R.A; Sivakumar, T; Yokoyama, Naoaki; Igarashi, Ikuo

    2009-01-01

    Apicidin, a histone deacetylase inhibitor, has a broad spectrum of anti-protozoal activities against apicomplexan parasites. In the present study, we evaluated the inhibitory effect of apicidin on the asexual growth of bovine Babesia parasites (B. bovis and B. bigemina) in vitro, as well as on the in vivo growth of B. microti in mice. The growth of B. bovis and B. bigemina was significantly inhibited in the presence of 3 ng/ml apicidin. Complete inhibition of B. bovis and B. bigemina growth w...

  6. Validation of an indirect ELISA for the diagnosis of Babesia bovis in El Salvador

    International Nuclear Information System (INIS)

    Molina, G.; Cardona, D.A.

    1998-01-01

    Validation and a preliminary serological study of Babesia bovis was made in El Salvador, using the indirect ELISA kit provided by the Joint FAO/IAEA Division of the International Atomic Energy Agency. Sera were collected from 545 cattle involving 10 regions of the country and various ages of cattle between 8 and 16 months. These were tested from May 1993 to February 1994. A 79.5% prevalence was found, but with a wide range from (5.8-100%), explained by different farm managing systems and different breeds. (author)

  7. Validate or falsify: Lessons learned from a microscopy method claimed to be useful for detecting Borrelia and Babesia organisms in human blood.

    Science.gov (United States)

    Aase, Audun; Hajdusek, Ondrej; Øines, Øivind; Quarsten, Hanne; Wilhelmsson, Peter; Herstad, Tove K; Kjelland, Vivian; Sima, Radek; Jalovecka, Marie; Lindgren, Per-Eric; Aaberge, Ingeborg S

    2016-01-01

    A modified microscopy protocol (the LM-method) was used to demonstrate what was interpreted as Borrelia spirochetes and later also Babesia sp., in peripheral blood from patients. The method gained much publicity, but was not validated prior to publication, which became the purpose of this study using appropriate scientific methodology, including a control group. Blood from 21 patients previously interpreted as positive for Borrelia and/or Babesia infection by the LM-method and 41 healthy controls without known history of tick bite were collected, blinded and analysed for these pathogens by microscopy in two laboratories by the LM-method and conventional method, respectively, by PCR methods in five laboratories and by serology in one laboratory. Microscopy by the LM-method identified structures claimed to be Borrelia- and/or Babesia in 66% of the blood samples of the patient group and in 85% in the healthy control group. Microscopy by the conventional method for Babesia only did not identify Babesia in any samples. PCR analysis detected Borrelia DNA in one sample of the patient group and in eight samples of the control group; whereas Babesia DNA was not detected in any of the blood samples using molecular methods. The structures interpreted as Borrelia and Babesia by the LM-method could not be verified by PCR. The method was, thus, falsified. This study underlines the importance of doing proper test validation before new or modified assays are introduced.

  8. Molecular and Parasitological Survey of Bovine Piroplasms in the Black Sea Region, Including the First Report of Babesiosis Associated with Babesia divergens in Turkey.

    Science.gov (United States)

    Aktas, M; Ozubek, S

    2015-11-01

    Clinical cases of babesiosis were evaluated, and the frequency of bovine Babesia and Theileria parasites was determined in cattle. Blood samples and thin blood smears were collected from 23 cattle exhibiting clinical signs of babesiosis. In addition, tick and blood samples were collected from 100 apparently healthy cattle cograzing from the same area. Egg masses obtained from fully engorged female ticks were included. DNA isolated from blood and tick samples was screened for Babesia and Theileria by reverse line blot assay. Piroplasms compatible with Babesia spp. were observed microscopically for symptomatic cattle as circular, oval, elongated, or pear-shaped bodies. Parasitemia ranged from 0.08 to 0.9% for Babesia bovis, 2.5 to 15.4% for Babesia bigemina, and 7.4% for Babesia divergens. Reverse line blot showed positivity in 13 (13%) of the sampled clinically normal cattle and revealed the presence of three Babesia species. Babesia bovis was the most prevalent (9/100, 9%), followed by Babesia occultans (3/100, 3%) and B. bigemina (1/100, 1%). One animal infected with B. bigemina was also infected with B. bovis. The single animal infected with B. divergens showed symptoms of babesiosis. Ticks were identified as Rhipicephalus annulatus, Rhipicephalus turanicus, and Ixodes ricinus. One female R. annulatus and its egg mass were infected with B. bigemina. Neither Theileria annulata nor Theileria buffeli/orientalis infections were observed in cattle or ticks. This is the first report of clinical babesiosis caused by B. divergens in cattle from Turkey. © The Authors 2015. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  9. Genetic characterization and phylogenetic relationships based on 18S rRNA and ITS1 region of small form of canine Babesia spp. from India.

    Science.gov (United States)

    Mandal, M; Banerjee, P S; Garg, Rajat; Ram, Hira; Kundu, K; Kumar, Saroj; Kumar, G V P P S Ravi

    2014-10-01

    Canine babesiosis is a vector borne disease caused by intra-erythrocytic apicomplexan parasites Babesia canis (large form) and Babesia gibsoni (small form), throughout the globe. Apart from few sporadic reports on the occurrence of B. gibsoni infection in dogs, no attempt has been made to characterize Babesia spp. of dogs in India. Fifteen canine blood samples, positive for small form of Babesia, collected from northern to eastern parts of India, were used for amplification of 18S rRNA gene (∼1665bp) of Babesia sp. and partial ITS1 region (∼254bp) of B. gibsoni Asian genotype. Cloning and sequencing of the amplified products of each sample was performed separately. Based on sequences and phylogenetic analysis of 18S rRNA and ITS1 sequences, 13 were considered to be B. gibsoni. These thirteen isolates shared high sequence identity with each other and with B. gibsoni Asian genotype. The other two isolates could not be assigned to any particular species because of the difference(s) in 18S rRNA sequence with B. gibsoni and closer identity with Babesiaoccultans and Babesiaorientalis. In the phylogenetic tree, all the isolates of B. gibsoni Asian genotype formed a separate major clade named as Babesia spp. sensu stricto clade with high bootstrap support. The two unnamed Babesia sp. (Malbazar and Ludhiana isolates) clustered close together with B. orientalis, Babesia sp. (Kashi 1 isolate) and B. occultans of bovines. It can be inferred from this study that 18S rRNA gene and ITS1 region are highly conserved among 13 B. gibsoni isolates from India. It is the maiden attempt of genetic characterization by sequencing of 18S rRNA gene and ITS1 region of B. gibsoni from India and is also the first record on the occurrence of an unknown Babesia sp. of dogs from south and south-east Asia. Copyright © 2014 Elsevier B.V. All rights reserved.

  10. Optimization of a Fluorescence-Based Assay for Large-Scale Drug Screening against Babesia and Theileria Parasites.

    Science.gov (United States)

    Rizk, Mohamed Abdo; El-Sayed, Shimaa Abd El-Salam; Terkawi, Mohamed Alaa; Youssef, Mohamed Ahmed; El Said, El Said El Shirbini; Elsayed, Gehad; El-Khodery, Sabry; El-Ashker, Maged; Elsify, Ahmed; Omar, Mosaab; Salama, Akram; Yokoyama, Naoaki; Igarashi, Ikuo

    2015-01-01

    A rapid and accurate assay for evaluating antibabesial drugs on a large scale is required for the discovery of novel chemotherapeutic agents against Babesia parasites. In the current study, we evaluated the usefulness of a fluorescence-based assay for determining the efficacies of antibabesial compounds against bovine and equine hemoparasites in in vitro cultures. Three different hematocrits (HCTs; 2.5%, 5%, and 10%) were used without daily replacement of the medium. The results of a high-throughput screening assay revealed that the best HCT was 2.5% for bovine Babesia parasites and 5% for equine Babesia and Theileria parasites. The IC50 values of diminazene aceturate obtained by fluorescence and microscopy did not differ significantly. Likewise, the IC50 values of luteolin, pyronaridine tetraphosphate, nimbolide, gedunin, and enoxacin did not differ between the two methods. In conclusion, our fluorescence-based assay uses low HCT and does not require daily replacement of culture medium, making it highly suitable for in vitro large-scale drug screening against Babesia and Theileria parasites that infect cattle and horses.

  11. Optimization of a Fluorescence-Based Assay for Large-Scale Drug Screening against Babesia and Theileria Parasites.

    Directory of Open Access Journals (Sweden)

    Mohamed Abdo Rizk

    Full Text Available A rapid and accurate assay for evaluating antibabesial drugs on a large scale is required for the discovery of novel chemotherapeutic agents against Babesia parasites. In the current study, we evaluated the usefulness of a fluorescence-based assay for determining the efficacies of antibabesial compounds against bovine and equine hemoparasites in in vitro cultures. Three different hematocrits (HCTs; 2.5%, 5%, and 10% were used without daily replacement of the medium. The results of a high-throughput screening assay revealed that the best HCT was 2.5% for bovine Babesia parasites and 5% for equine Babesia and Theileria parasites. The IC50 values of diminazene aceturate obtained by fluorescence and microscopy did not differ significantly. Likewise, the IC50 values of luteolin, pyronaridine tetraphosphate, nimbolide, gedunin, and enoxacin did not differ between the two methods. In conclusion, our fluorescence-based assay uses low HCT and does not require daily replacement of culture medium, making it highly suitable for in vitro large-scale drug screening against Babesia and Theileria parasites that infect cattle and horses.

  12. Functional genomics studies of Rhipicephalus (Boophilus) annulatus ticks in response to infection with the cattle protozoan parasite, Babesia bigemina

    Czech Academy of Sciences Publication Activity Database

    Antunes, S.; Galindo, R. C.; Almazán, C.; Rudenko, Natalia; Golovchenko, Maryna; Grubhoffer, Libor; Shkap, V.; do Rosário, A.; de la Fuente, J.; Domingos, A.

    2012-01-01

    Roč. 42, č. 2 (2012), s. 187-195 ISSN 0020-7519 Institutional research plan: CEZ:AV0Z60220518 Keywords : Tick * Genomics * Babesia * Rhipicephalus * Boophilus * RNA interference * Vaccine Subject RIV: EC - Immunology Impact factor: 3.637, year: 2012 http://www.sciencedirect.com/science/article/pii/S0020751912000033

  13. Occurrence of Theileria and Babesia species in water buffalo (Bubalus babalis, Linnaeus, 1758) in the Hubei province, South China.

    Science.gov (United States)

    He, Lan; Feng, Hui-Hui; Zhang, Wen-Jie; Zhang, Qing-Li; Fang, Rui; Wang, Li-Xia; Tu, Pan; Zhou, Yan-Qin; Zhao, Jun-Long; Oosthuizen, Marinda C

    2012-05-25

    The presence and prevalence of tick-borne haemoparasites in water buffalo from the Hubei province, south China was investigated using the reverse line blot (RLB) hybridization assay and phylogenetic analysis of the parasite 18S rRNA gene. Theileria buffeli (19.1%) was the most frequently found species in all of the locations, followed by Babesia orientalis (8.9%), Babesia bovis (1.0%) and Babesia bigemina (0.7%). Only 12 (3.9%) of the samples had mixed infections. Eleven samples with single infections were selected for further characterization using 18S rRNA gene sequence analysis. Phylogenetic analysis showed that the eight T. buffeli 18S rRNA gene sequences obtained grouped into four clusters, of which three grouped with the known T. buffeli types B and D. The remaining five grouped separately from the previously describe T. buffeli types, constituting new T. buffeli types. The two B. bigemina 18S rRNA gene sequences obtained grouped closely with B. bigemina Kunming; this serves as the first report of B. bigemina in the Hubei province. The B. orientalis Daye 18S rRNA gene sequence obtained grouped closely with the previously reported B. orientalis Wuhan strain and with Babesia sp. Kashi 1 and Kashi 2. Copyright © 2011 Elsevier B.V. All rights reserved.

  14. Molecular detection and characterization of potentially new Babesia and Theileria species/variants in wild felids from Kenya.

    Science.gov (United States)

    Githaka, Naftaly; Konnai, Satoru; Kariuki, Edward; Kanduma, Esther; Murata, Shiro; Ohashi, Kazuhiko

    2012-10-01

    Piroplasms frequently infect domestic and wild carnivores. At present, there is limited information on the occurrence and molecular identity of these tick-borne parasites in wild felids in Kenya. In 2009, a pair of captive lions (Panthare leo) was diagnosed with suspected babesiosis and mineral deficiency at an animal orphanage on the outskirts of Nairobi, Kenya. Blood smears indicated presences of haemoparasites in the erythrocytes, however, no further investigations were conducted to identify the infecting agent. The animals recovered completely following diet supplementation and treatment with anti-parasite drug. In this report, we extracted and detected parasite DNA from the two lions and seven other asymptomatic feline samples; two leopards (Panthera pardus) and five cheetahs (Acinonyx jubatus). Reverse line blot with probes specific for Babesia spp. of felines indicated the presence of new Babesia species or genotypes in the lions and leopards, and unknown Theileria sp. in the cheetahs. Phylogenetic analyses using partial sequences of 18S ribosomal RNA (18S rRNA) gene showed that the parasite infecting the lions belong to the Babesia canis complex, and the parasite variant detected in the leopards clusters in a clade bearing other Babesia spp. reported in wild felids from Africa. The cheetah isolates falls in the Theileria sensu stricto group. Our findings indicate the occurrence of potentially new species or genotypes of piroplams in all three feline species. Copyright © 2012 Elsevier B.V. All rights reserved.

  15. Clinical outbreak of babesiosis caused by Babesia capreoli in captive reindeer (Rangifer tarandus tarandus) in the Netherlands

    NARCIS (Netherlands)

    Bos, Jan H; Klip, Fokko C; Sprong, Hein; Broens, Els M|info:eu-repo/dai/nl/314627723; Kik, Marja J L|info:eu-repo/dai/nl/080432565

    2017-01-01

    From a herd of captive reindeer (Rangifer tarandus tarandus) consisting of two males and seven females with five calves, three calves were diagnosed on post mortem examination with a Babesia capreoli infection. The diagnosis was indicated by PCR and when the other reindeer were examined two adult

  16. Clinical outbreak of babesiosis caused by Babesia capreoli in captive reindeer (Rangifer tarandus tarandus) in the Netherlands.

    NARCIS (Netherlands)

    Bos, Jan H; Klip, Fokko C; Sprong, Hein; Broens, Els M; Kik, Marja J L

    From a herd of captive reindeer (Rangifer tarandus tarandus) consisting of two males and seven females with five calves, three calves were diagnosed on post mortem examination with a Babesia capreoli infection. The diagnosis was indicated by PCR and when the other reindeer were examined two adult

  17. Morphological and Molecular Descriptors of the Developmental Cycle of Babesia divergens Parasites in Human Erythrocytes.

    Directory of Open Access Journals (Sweden)

    Ingrid Rossouw

    2015-05-01

    Full Text Available Human babesiosis, especially caused by the cattle derived Babesia divergens parasite, is on the increase, resulting in renewed attentiveness to this potentially life threatening emerging zoonotic disease. The molecular mechanisms underlying the pathophysiology and intra-erythrocytic development of these parasites are poorly understood. This impedes concerted efforts aimed at the discovery of novel anti-babesiacidal agents. By applying sensitive cell biological and molecular functional genomics tools, we describe the intra-erythrocytic development cycle of B. divergens parasites from immature, mono-nucleated ring forms to bi-nucleated paired piriforms and ultimately multi-nucleated tetrads that characterizes zoonotic Babesia spp. This is further correlated for the first time to nuclear content increases during intra-erythrocytic development progression, providing insight into the part of the life cycle that occurs during human infection. High-content temporal evaluation elucidated the contribution of the different stages to life cycle progression. Moreover, molecular descriptors indicate that B. divergens parasites employ physiological adaptation to in vitro cultivation. Additionally, differential expression is observed as the parasite equilibrates its developmental stages during its life cycle. Together, this information provides the first temporal evaluation of the functional transcriptome of B. divergens parasites, information that could be useful in identifying biological processes essential to parasite survival for future anti-babesiacidal discoveries.

  18. Coinfection of sheep with Anaplasma, Theileria and Babesia species in the Kurdistan Region, Iraq.

    Science.gov (United States)

    Renneker, S; Abdo, J; Bakheit, M A; Kullmann, B; Beyer, D; Ahmed, J; Seitzer, U

    2013-11-01

    Infections of small ruminants with Anaplasma, Theileria and Babesia species are widely distributed in the old world and are of great economic impact. In Iraq, data on disease occurrence in sheep caused by above-mentioned infectious agents are scarce. This study provides information on various haemoparasitic agents infecting sheep in the Kurdistan Region, Iraq, using molecular diagnostic tools. Altogether, 195 samples originating from three governorates in the Kurdistan Region, namely Duhok, Erbil and Sulaimaniya, were analysed. The following pathogens were identified: Anaplasma ovis (62.6%), Theileria ovis (14.35%), T. lestoquardi (7.7%), T. uilenbergi (5.6%) and Babesia ovis (1.5%). T. uilenbergi is detected for the first time in Iraq. Coinfection of sheep with different pathogens could be observed in this study, and it was found that 45 of 195 (23%) of the samples contained more than one pathogen. Even triple-positive samples were identified in 3% of the investigated animals. In conclusion, we confirm the coinfection of sheep with various haemoparasitic pathogen species in the Kurdistan Region of Iraq. Further investigations are needed to reveal the epidemiology of the diseases, the respective tick vectors, and, in the case of coinfection, pathogens' interaction and possible cross-protection. © 2013 Blackwell Verlag GmbH.

  19. Molecular detection and genetic diversity of Babesia gibsoni in dogs in Bangladesh.

    Science.gov (United States)

    Terao, Masashi; Akter, Shirin; Yasin, Md Golam; Nakao, Ryo; Kato, Hirotomo; Alam, Mohammad Zahangir; Katakura, Ken

    2015-04-01

    Babesia gibsoni is a tick-borne hemoprotozoan parasite of dogs that often causes fever and hemolytic illness. Detection of B. gibsoni has been predominantly reported in Asian countries, including Japan, Korea, Taiwan, Malaysia, Bangladesh and India. The present study shows the first molecular characterization of B. gibsoni detected from dogs in Bangladesh. Blood samples were collected on FTA® Elute cards from 50 stray dogs in Mymensingh District in Bangladesh. DNA eluted from the cards was subjected to nested PCR for the 18S rRNA gene of Babesia species. Approximately 800bp PCR products were detected in 15 of 50 dogs (30%). Based on restriction fragment length polymorphism (RFLP) and direct sequencing of the PCR products, all parasite isolates were identified as B. gibsoni. Furthermore, the BgTRAP (B. gibsoni thrombospondin-related adhesive protein) gene fragments were detected in 13 of 15 18S rRNA gene PCR positive blood samples. Phylogenetic analysis of the BgTRAP gene revealed that B. gibsoni parasites in Bangladesh formed a cluster, which was genetically different from other Asian B. gibsoni isolates. In addition, tandem repeat analysis of the BgTRAP gene clearly showed considerable genetic variation among Bangladeshi isolates. These results suggested that B. gibsoni parasites in a different genetic clade are endemic in dogs in Bangladesh. Further studies are required to elucidate the origin, distribution, vector and pathogenesis of B. gibsoni parasites circulating in dogs in Bangladesh. Copyright © 2015 Elsevier B.V. All rights reserved.

  20. Ticks circulate Anaplasma, Ehrlichia, Babesia and Theileria parasites in North of Iran.

    Science.gov (United States)

    Bekloo, Ahmad Jafar; Bakhshi, Hasan; Soufizadeh, Ayoub; Sedaghat, Mohammad Mehdi; Bekloo, Romina Jafar; Ramzgouyan, Maryam Roya; Chegeni, Asadollah Hosseini; Faghihi, Faezeh; Telmadarraiy, Zakkyeh

    2017-12-15

    Ticks serve as important vectors of some pathogens of medical importance all over the world and identification of their rate of infection plays an important role for further control of diseases. In the current study, we investigated on ticks collected from north of Iran where raising and caring livestock are the main task of the people in order to find evidences of infection of Babesia, Theileria, Anaplasma and Ehrlichia microbial agents. Totally, 609 hard tick species from two genera Hyalomma and Rhipicephalus including; Hy. scupense, Hy. dromedarii, Hy. rufipes, Hy. marginatum, Hy. asiaticum, Hy. anatolicum, R. bursa, R. sanguineus and R. turanicus were identified. Molecular analysis revealed the presence of Anaplasma, Ehrlichia, Babesia and Theileria microorganism agents in all collected tick species except Hy. asiaticum and R. turanicus. To the best of our knowledge, this is the first report on identification of B. occultans in Hyalomma anatolicum and B. ovis in Hyalomma sp in Iran. Copyright © 2017 Elsevier B.V. All rights reserved.

  1. First Molecular Detection of Babesia gibsoni in Dogs from Wuhan, China

    Directory of Open Access Journals (Sweden)

    Lan He

    2017-08-01

    Full Text Available Canine piroplasmosis is a significant disease in dogs caused by Babesia and Theileria parasites. The clinical manifestations range from mild illness to serious disease depending on the parasite species and the physical condition of the infected dog. Canine piroplasmosis has been reported to be prevalent in China. However, no molecular evidence of the disease has been reported in pet dogs from Wuhan. In this study, 118 blood samples were randomly collected from pet dogs in veterinary clinics. The blood samples were subjected to both microscopic examination and reverse line blot (RLB hybridization assays to detect piroplasm infection. Parasites were observed in 10 blood samples via microscopic examination, whereas there were 14 Babesia gibsoni-positive RLB tests. Phylogenetic analysis was performed after the 18S rRNA and ITS gene sequences from the 14 positive samples were cloned and sequenced. The results confirmed the existence of B. gibsoni in this area. This is the first molecular report of canine babesiosis in pet dogs from Wuhan, China. Pet dogs are companion animals, and the prevalence of babesiosis will be of concern in daily life. This study will help veterinarians better understand the prevalence of canine babesiosis and provide a guide for disease control in pet dogs.

  2. Evaluation of an indirect ELISA for the diagnosis of Babesia bovis in Uruguay

    International Nuclear Information System (INIS)

    Cardozo, H.; Solari, M.A.; Etchebarne, J.

    1992-01-01

    In initially establishing the FAO/IAEA indirect ELISA for the detection of antibodies to Babesia bovis, the optical density (OD) values of sera from known positive or negative local cattle were compared to the OD values obtained from the negative and positive reference sera provided with the ELISA kit. The percentage of false positive and negative sera were 2.53% and 2.97% respectively. The cut-off values for the negative reference serum in the kit were compared with those of a local negative population. These values were found to be similar. The specificity of the test was evaluated by testing 30 sera from animals experimentally infected with Anaplasma marginale and 30 sera from animals infected with Babesia bigemina. These were no cross-reaction either between A. marginale and B. bovis or between B. bigemina and B. bovis. A serological survey using this ELISA kit was carried out on animals from an enzootic area and an area free from the vecot Boophilus microplus. 53 out of 282 animals (18.8%) in the enzootic area were positive whilst all the animals (113) from the free area were negative. This study would indicate that the FAO/IAEA ELISA kit has a sensitivity of around 98% and specificity of 97%. (author). 8 refs, 2 figs, 2 tabs

  3. Seroepidemiological study of Babesia bovis in support of the Uruguayan Boophilus microplus control programme

    International Nuclear Information System (INIS)

    Cardozo, H.; Solari, M.A.; Etchebarne, J.; Larrauri, J.H.

    1998-01-01

    Bovine blood samples were collected from a region endemic for Boophilus microplus and consisting of 125 ranches with a cattle population of 76,918. A total of 1,728 cattle were bled (1,485 adults and 243 calves less from 1 year of age) from 27 ranches. This sample size was determined to provide incidence and prevalence values with a precision of ±10% at a confidence level of 95%. The FAO/IAEA ELISA kit was used to detect antibody to Babesia bovis. Dispersion (proportion of ranches with babesia infection) was estimated to be 70.5% ± 8.8 (SD). A positive ranch was defined as having one or more test-positive animals. Apparent prevalence (proportion of cattle with a positive test result) within the region was estimated to be 3.5% ± 0.3, with a range from 0 to 18.5%. Incidence based on apparent prevalence in calves less than 1 year of age was estimated to be 2.8%. The dispersion, apparent prevalence, and incidence data for this region of Uruguay will be compared through a repeated sampling of cattle in this area during a three-year period to assess the effectiveness of the eradication/control campaign. (author)

  4. High prevalence of small Babesia species in canines of Kerala, South India.

    Science.gov (United States)

    Jain, Kollannur Jose; Lakshmanan, Bindu; Syamala, Karunakaran; Praveena, Jose E; Aravindakshan, Thazhathuveetil

    2017-11-01

    Canine babesiosis is an important vector-borne hemoparasitic disease caused by Babesia canis vogeli and Babesia gibsoni , in India. The communication places on record the salient findings of the study directed to detect and characterize the pathogenic B. gibsoni isolates of Kerala state. A total of 150 dogs were examined for the presence of hemoparasites by light microscopy as well as by PCR targeting the 18S rRNA gene of B. gibsoni . Hematological parameters were also analysed. Phylogenetic tree was constructed based on Tamura kei model adopting ML method. A sensitive and specific polymerase chain reaction assay was developed with newly designed primer pair BAGI-F/BAGI-R for the amplification of 488 bp fragment of 18S rRNA gene of B. gibsoni . Out of the 150 dogs examined, molecular evidence of B. gibsoni was recorded in 47.3% animals, while light microscopy detected the infection in 26.67% cases. The phylogenetic analyses revealed that B. gibsoni , Kerala, isolate was closest and occurred together with Bareilly isolate. Anemia and thrombocytopenia were the significant hematological alterations in chronic B. gibsoni infection. A high prevalence of natural infection of B. gibsoni was detected among the study population. The affected animals showed anaemia and thrombocytopenia. Phylogenetic analysis of this pathogenic isolate from south India revealed the closest similarity with Bareilly isolates.

  5. Babesia vesperuginis, a neglected piroplasmid: new host and geographical records, and phylogenetic relations.

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    Corduneanu, Alexandra; Hrazdilová, Kristýna; Sándor, Attila D; Matei, Ioana Adriana; Ionică, Angela Monica; Barti, Levente; Ciocănău, Marius-Alexandru; Măntoiu, Dragoş Ștefan; Coroiu, Ioan; Hornok, Sándor; Fuehrer, Hans-Peter; Leitner, Natascha; Bagó, Zoltán; Stefke, Katharina; Modrý, David; Mihalca, Andrei Daniel

    2017-12-06

    Babesia spp. are hemoparasites which infect the red blood cells of a large variety of mammals. In bats, the only known species of the genus is Babesia vesperuginis. However, except a few old reports, the host range and geographical distribution of this bat parasite have been poorly studied. This study aimed to investigate the presence of piroplasms in tissues of bats collected in four different countries from eastern and central Europe: Austria, Czech Republic, Hungary and Romania. A total of 461 bat carcasses (24 species) were collected between 2001 and 2016 from caves, mines and buildings. PCR was performed using specific primers targeting a portion of the 18S rDNA nuclear gene and cytochrome c oxidase subunit 1 mitochondrial gene, followed by sequencing. The results of this study show for the first time the presence of B. vesperuginis in bats in central and eastern Europe. The phylogenetic analysis of the 18S rDNA nuclear gene revealed no variability between the sequences and the phylogenetic analysis of the cox1 mitochondrial gene proved that B. vesperuginis could be divided into two subclades. Our study showed a broad geographical distribution of B. vesperuginis in European bats, reporting its presence in five new host species (M. cf. alcathoe, M. bechsteinii, M. myotis, Pi. nathusii and V. murinus) and three new countries.

  6. High prevalence of small Babesia species in canines of Kerala, South India

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    Kollannur Jose Jain

    2017-11-01

    Full Text Available Aim: Canine babesiosis is an important vector-borne hemoparasitic disease caused by Babesia canis vogeli and Babesia gibsoni, in India. The communication places on record the salient findings of the study directed to detect and characterize the pathogenic B. gibsoni isolates of Kerala state. Materials and Methods: A total of 150 dogs were examined for the presence of hemoparasites by light microscopy as well as by PCR targeting the 18S rRNA gene of B. gibsoni. Hematological parameters were also analysed. Phylogenetic tree was constructed based on Tamura kei model adopting ML method. Results: A sensitive and specific polymerase chain reaction assay was developed with newly designed primer pair BAGI-F/ BAGI-R for the amplification of 488 bp fragment of 18S rRNA gene of B. gibsoni. Out of the 150 dogs examined, molecular evidence of B. gibsoni was recorded in 47.3% animals, while light microscopy detected the infection in 26.67% cases. The phylogenetic analyses revealed that B. gibsoni, Kerala, isolate was closest and occurred together with Bareilly isolate. Anemia and thrombocytopenia were the significant hematological alterations in chronic B. gibsoni infection. Conclusion: A high prevalence of natural infection of B. gibsoni was detected among the study population. The affected animals showed anaemia and thrombocytopenia. Phylogenetic analysis of this pathogenic isolate from south India revealed the closest similarity with Bareilly isolates.

  7. Prevalence, genetic identity and vertical transmission of Babesia microti in three naturally infected species of vole, Microtus spp. (Cricetidae).

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    Tołkacz, Katarzyna; Bednarska, Małgorzata; Alsarraf, Mohammed; Dwużnik, Dorota; Grzybek, Maciej; Welc-Falęciak, Renata; Behnke, Jerzy M; Bajer, Anna

    2017-02-06

    Vertical transmission is one of the transmission routes for Babesia microti, the causative agent of the zoonotic disease, babesiosis. Congenital Babesia invasions have been recorded in laboratory mice, dogs and humans. The aim of our study was to determine if vertical transmission of B. microti occurs in naturally-infected reservoir hosts of the genus Microtus. We sampled 124 common voles, Microtus arvalis; 76 root voles, M. oeconomus and 17 field voles, M. agrestis. In total, 113 embryos were isolated from 20 pregnant females. Another 11 pregnant females were kept in the animal house at the field station in Urwitałt until they had given birth and weaned their pups (n = 62). Blood smears and/or PCR targeting the 550 bp 18S rRNA gene fragment were used for the detection of B. microti. Selected PCR products, including isolates from females/dams and their embryos/pups, were sequenced. Positive PCR reactions were obtained for 41% (89/217) of the wild-caught voles. The highest prevalence of B. microti was recorded in M. arvalis (56/124; 45.2%), then in M. oeconomus (30/76; 39.5%) and the lowest in M. agrestis (3/17; 17.7%). Babesia microti DNA was detected in 61.4% (27/44) of pregnant females. Vertical transmission was confirmed in 81% (61/75) of the embryos recovered from Babesia-positive wild-caught pregnant females. The DNA of B. microti was detected in the hearts, lungs and livers of embryos from 98% of M. arvalis, 46% of M. oeconomus and 0% of M. agrestis embryos from Babesia-positive females. Of the pups born in captivity, 90% were born to Babesia-positive dams. Babesia microti DNA was detected in 70% (35/50) of M. arvalis and 83% (5/6) of M. oeconomus pups. Congenitally acquired infections had no impact on the survival of pups over a 3-week period post partum. Among 97 B. microti sequences, two genotypes were found. The IRU1 genotype (Jena-like) was dominant in wild-caught voles (49/53; 92%), pregnant females (9/11; 82%) and dams (3/5; 60%). The IRU2

  8. Occurrence of Babesia spp., Rickettsia spp. and Bartonella spp. in Ixodes ricinus in Bavarian public parks, Germany

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    Mahling Monia

    2011-07-01

    Full Text Available Abstract Background Only limited information is available about the occurrence of ticks and tick-borne pathogens in public parks, which are areas strongly influenced by human beings. For this reason, Ixodes ricinus were collected in public parks of different Bavarian cities in a 2-year survey (2009 and 2010 and screened for DNA of Babesia spp., Rickettsia spp. and Bartonella spp. by PCR. Species identification was performed by sequence analysis and alignment with existing sequences in GenBank. Additionally, coinfections with Anaplasma phagocytophilum were investigated. Results The following prevalences were detected: Babesia spp.: 0.4% (n = 17, including one pool of two larvae in 2009 and 0.5 to 0.7% (n = 11, including one pool of five larvae in 2010; Rickettsia spp.: 6.4 to 7.7% (n = 285, including 16 pools of 76 larvae in 2009. DNA of Bartonella spp. in I. ricinus in Bavarian public parks could not be identified. Sequence analysis revealed the following species: Babesia sp. EU1 (n = 25, B. divergens (n = 1, B. divergens/capreoli (n = 1, B. gibsoni-like (n = 1, R. helvetica (n = 272, R. monacensis IrR/Munich (n = 12 and unspecified R. monacensis (n = 1. The majority of coinfections were R. helvetica with A. phagocytophilum (n = 27, but coinfections between Babesia spp. and A. phagocytophilum, or Babesia spp. and R. helvetica were also detected. Conclusions I. ricinus ticks in urban areas of Germany harbor several tick-borne pathogens and coinfections were also observed. Public parks are of particularly great interest regarding the epidemiology of tick-borne pathogens, because of differences in both the prevalence of pathogens in ticks as well as a varying species arrangement when compared to woodland areas. The record of DNA of a Babesia gibsoni-like pathogen detected in I. ricinus suggests that I. ricinus may harbor and transmit more Babesia spp. than previously known. Because of their high recreational value for human beings, urban green

  9. Prevalence of ticks and tick-borne pathogens: Babesia and Borrelia species in ticks infesting cats of Great Britain.

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    Davies, Saran; Abdullah, Swaid; Helps, Chris; Tasker, Séverine; Newbury, Hannah; Wall, Richard

    2017-09-15

    In a study of tick and tick-borne pathogen prevalence, between May and October 2016, 278 veterinary practices in Great Britain examined 1855 cats. Six-hundred and one cats were found to have attached ticks. The most frequently recorded tick species was Ixodes ricinus (57.1%), followed by Ixodes hexagonus (41.4%) and Ixodes trianguliceps (1.5%). Male cats, 4-6 years of age living in rural areas were most likely to be carrying a tick; hair length and tick treatment history had no significant association with attachment. For cats that were parasitized by ticks in large urban areas, I. hexagonus was the most frequent species recorded. Molecular analysis was possible for 541 individual tick samples, others were too damaged for analysis; Babesia spp., and Borrelia burgdorferi sensu lato were identified in 1.1% (n=6) and 1.8% (n=10) of these, respectively. Babesia spp. included Babesia vulpes sp. nov./Babesia microti-like (n=4) in I. hexagonus and Babesia venatorum (n=2) in I. ricinus. Borrelia burgdorferi s.l. species included Borrelia garinii (n=6) and Borrelia afzelii (n=4). The majority of B. burgorferi s.l. cases were found in I. ricinus, with B. afzelii in one I. hexagonus nymph. No Borrelia or Babesia spp. were present in I. trianguliceps. To determine a true prevalence for ticks on cats, practices that only submitted questionnaires from cats with ticks and practices that submitted fewer than 5 returns per week were removed; amongst those considered to have adhered strictly to the collection protocol, feline tick prevalence amongst cats that had access to the outdoors was 6.6%. These results show that ticks can be found on cats throughout Great Britain, which harbour a range of species of Babesia and B. burgdorferi s.l. and that cats, particularly in green spaces within urban areas, may form an important host for I. hexagonus, a known vector of pathogens. Copyright © 2017 Elsevier B.V. All rights reserved.

  10. Occurrence of Babesia spp., Rickettsia spp. and Bartonella spp. in Ixodes ricinus in Bavarian public parks, Germany.

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    Schorn, Sabine; Pfister, Kurt; Reulen, Holger; Mahling, Monia; Silaghi, Cornelia

    2011-07-15

    Only limited information is available about the occurrence of ticks and tick-borne pathogens in public parks, which are areas strongly influenced by human beings. For this reason, Ixodes ricinus were collected in public parks of different Bavarian cities in a 2-year survey (2009 and 2010) and screened for DNA of Babesia spp., Rickettsia spp. and Bartonella spp. by PCR. Species identification was performed by sequence analysis and alignment with existing sequences in GenBank. Additionally, coinfections with Anaplasma phagocytophilum were investigated. The following prevalences were detected: Babesia spp.: 0.4% (n = 17, including one pool of two larvae) in 2009 and 0.5 to 0.7% (n = 11, including one pool of five larvae) in 2010; Rickettsia spp.: 6.4 to 7.7% (n = 285, including 16 pools of 76 larvae) in 2009. DNA of Bartonella spp. in I. ricinus in Bavarian public parks could not be identified. Sequence analysis revealed the following species: Babesia sp. EU1 (n = 25), B. divergens (n = 1), B. divergens/capreoli (n = 1), B. gibsoni-like (n = 1), R. helvetica (n = 272), R. monacensis IrR/Munich (n = 12) and unspecified R. monacensis (n = 1). The majority of coinfections were R. helvetica with A. phagocytophilum (n = 27), but coinfections between Babesia spp. and A. phagocytophilum, or Babesia spp. and R. helvetica were also detected. I. ricinus ticks in urban areas of Germany harbor several tick-borne pathogens and coinfections were also observed. Public parks are of particularly great interest regarding the epidemiology of tick-borne pathogens, because of differences in both the prevalence of pathogens in ticks as well as a varying species arrangement when compared to woodland areas. The record of DNA of a Babesia gibsoni-like pathogen detected in I. ricinus suggests that I. ricinus may harbor and transmit more Babesia spp. than previously known. Because of their high recreational value for human beings, urban green areas are likely to remain in the research focus on

  11. Genetic diversity and molecular characterization of Babesia motasi-like in small ruminants and ixodid ticks from China.

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    Niu, Qingli; Liu, Zhijie; Yang, Jifei; Yu, Peifa; Pan, Yuping; Zhai, Bintao; Luo, Jianxun; Yin, Hong

    2016-07-01

    Ovine babesioses, an important tick-borne disease of sheep and goats in China, is caused by the reproduction of intraerythrocytic protozoa of the Babesia genus. Babesia motasi-like is a Babesia parasite that infects small ruminant in China, and two sub-groups of B. motasi-like can be subdivided based on differences in the rhoptry-associated-protein-1 gene. This study aimed to characterize the distribution, epidemiology and genetics of B. motasi-like in animals and ticks. A molecular investigation was carried out from 2009 to 2015 in 16 provinces in China. In total, 1081 blood samples were collected from sheep and goats originating from 27 different regions, and 778 ixodid tick samples were collected from 8 regions; the samples were tested for the presence of B. motasi-like using a specific nested PCR assay based on the rap-1b gene. The results indicated that 139 (12.9%), 91 (8.4%), 48 (4.4%) and 6 (0.7%) of the blood samples were positive for general B. motasi-like, Babesia sp. BQ1 (Lintan and Ningxian), Babesia sp. Tianzhu and Babesia sp. Hebei sub-groups, mixed infections, respectively. Among the collected 778 ixodid ticks (including Haemaphysalis longicornis, Haemaphysalis qinghaiensis, Dermacentor silvarum, Ixodes persulcatus, Rhipicephalus sanguineus and Rhipicephalus (Boophilus) microplus), the most frequently infected with Babesia were D. silvarum and I. persulcatus (35.7%), followed by H. longicornis (26.8%), H. qinghaiensis (24.8%) and R. sanguineus (9.3%). The PCR results were confirmed by DNA sequencing. The positive rates of B. motasi-like infection in ticks were found to be higher in China, compared with previous studies in other countries. B. motasi-like infections have not previously been reported in D. silvarum, I. persulcatus or R. sanguineus. The findings obtained in this study could be used for planning effective control strategies against babesiosis in China. Copyright © 2016 Elsevier B.V. All rights reserved.

  12. Hemoparasites in a wild primate: Infection patterns suggest interaction of Plasmodium and Babesia in a lemur species

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    Andrea Springer

    2015-12-01

    Full Text Available Hemoparasites can cause serious morbidity in humans and animals and often involve wildlife reservoirs. Understanding patterns of hemoparasite infections in natural populations can therefore inform about emerging disease risks, especially in the light of climate change and human disruption of natural ecosystems. We investigated the effects of host age, sex, host group size and season on infection patterns of Plasmodium sp., Babesia sp. and filarial nematodes in a population of wild Malagasy primates, Verreaux's sifakas (Propithecus verreauxi, as well as the effects of these infections on hematological variables. We tested 45 blood samples from 36 individuals and identified two species of Plasmodium, one species of Babesia and two species of filarial nematodes. Plasmodium spp. and Babesia sp. infections showed opposite patterns of age-dependency, with babesiosis being prevalent among young animals, while older animals were infected with Plasmodium sp. In addition, Babesia sp. infection was a statistically significant negative predictor of Plasmodium sp. infection. These results suggest that Plasmodium and Babesia parasites may interact within the host, either through cross-immunity or via resource competition, so that Plasmodium infections can only establish after babesiosis has resolved. We found no effects of host sex, host group size and season on hemoparasite infections. Infections showed high prevalences and did not influence hematological variables. This preliminary evidence supports the impression that the hosts and parasites considered in this study appear to be well-adapted to each other, resulting in persistent infections with low pathogenic and probably low zoonotic potential. Our results illustrate the crucial role of biodiversity in host-parasite relationships, specifically how within-host pathogen diversity may regulate the abundance of parasites.

  13. Hap2, a novel gene in Babesia bigemina is expressed in tick stages, and specific antibodies block zygote formation

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    Minerva Camacho-Nuez

    2017-11-01

    Full Text Available Abstract Background Bovine babesiosis is a tick-borne disease caused by the protozoan parasites of the genus Babesia. In their host vector, Babesia spp. undergo sexual reproduction. Therefore, the development of sexual stages and the subsequent formation of the zygote are essential for the parasite to invade the intestinal cells of the vector tick and continue its life-cycle. HAP2/GCS1 is a protein identified in plants, protozoan parasites and other organisms that has an important role during membrane fusion in fertilization processes. The identification and characterization of HAP-2 protein in Babesia would be very significant to understand the biology of the parasite and to develop a transmission-blocking vaccine in the future. Results To isolate and sequence the hap2 gene DNA from an infected bovine with Babesia bigemina was purified. The hap2 gene was amplified, cloned and sequenced. The sequences of hap2 from four geographically different strains showed high conservation at the amino acid level, including the typical structure with a signal peptide and the HAP2/GSC domain. Antisera anti-HAP2 against the conserved extracellular region of the HAP2 amino acid sequence were obtained from rabbits. The expression of hap2 in the host and vector tissues was analyzed by using semi-quantitative RT-PCR, and the protein was examined by western blot and immunofluorescence. Based on the RT-PCR and WB results, HAP2 is expressed in both, sexual stages induced in vitro, and in infected ticks as well. We did not detect any expression in asexual erythrocytic stages of B. bigemina, relevantly anti-HAP2 specific antibodies were able to block zygotes formation in vitro. Conclusion Babesia bigemina HAP2 is expressed only in tick-infecting stages, and specific antibodies block zygote formation. Further studies regarding the function of HAP2 during tick infection may provide new insights into the molecular mechanisms of sexual reproduction of the parasite.

  14. Leishmania, Babesia and Ehrlichia in urban pet dogs: co-infection or cross-reaction in serological methods?

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    Felipe da Silva Krawczak

    2015-02-01

    Full Text Available INTRODUCTION: The present study was designed to assess the occurrence of co-infection or cross-reaction in the serological techniques used for detecting the anti-Leishmania spp., -Babesia canis vogeli and -Ehrlichia canis antibodies in urban dogs from an area endemic to these parasites. METHODS: The serum samples from dogs were tested for the Babesia canis vogeli strain Belo Horizonte antigen and Ehrlichia canis strain São Paulo by immunofluorescence antibody test (IFAT and by anti-Leishmania immunoglobulin G (IgG antibody detection to assess Leishmania infection. We used the following four commercial kits for canine visceral leishmaniasis: ELISA, IFAT, Dual Path Platform (DPP (Bio Manguinhos(r/FIOCRUZ/MS and a rK39 RDT (Kalazar Detect Canine Rapid Test; Inbios. RESULTS : Of 96 serum samples submitted to serological assays, 4 (4.2% were positive for Leishmania as determined by ELISA; 12 (12.5%, by IFAT; 14 (14.6% by rK39 RDT; and 20 (20.8%, by DPP. Antibodies against Ehrlichia and Babesia were detected in 23/96 (23.9% and 30/96 (31.2% samples, respectively. No significant association was identified between the results of tests for detecting Babesia or Ehrlichia and those for detecting Leishmania (p-value>0.05. CONCLUSIONS: In the present study, we demonstrated co-infection with Ehrlichia or Babesia and Leishmania in dogs from Minas Gerais (Brazil; we also found that the serological tests that were used did not cross-react.

  15. Molecular detection of Theileria spp. and Babesia spp. in sheep and ixodid ticks from the northeast of Iran.

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    Razmi, Gholamreza; Pourhosseini, Moslem; Yaghfouri, Saeed; Rashidi, Ahmad; Seidabadi, Mohsen

    2013-02-01

    Theilerioses and babesioses are important diseases in Iranian sheep. The present study was undertaken to identify and classify/specify Theileria spp. and Babesia spp. in sheep and vector ticks. Investigation was carried out from 2009 to 2011 in the Khorasan Razavi Province, Iran. In total, 302 sheep originating from 60 different flocks were clinically examined and their blood collected. In addition, from the same flocks, ixodid ticks were sampled. Stained blood smears were microscopically examined for the presence of Theileria and Babesia organisms, and a semi-nested PCR was used for subsequent molecular specification. From the ticks, salivary glands and uterus were isolated and subsequently analyzed by semi-nested PCR. Piroplasm organisms were observed in 29% of the blood smears with low parasitemia, whereas 65% of the blood samples yielded positive PCR findings. The presence of Theileria ovis (55.6%), Theileria lestoquardi, and mixed infection with Theileria spp. and Babesia ovis were detected by semi-nested PCR in 0.3%, 5.6%, and 0.99%, respectively. In total, 429 ixodid ticks were collected from different areas of the province. The most prevalent ticks were Rhipicephalus turanicus (n = 376; 87.6% of the total), followed by Hyalomma marginatum turanicum (n = 30; 7.0%), Dermacentor raskemensis (n = 12; 2.8%), Hyalomma anatolicum anatolicum (n = 7; 1.6%), Dermacentor marginatus (n = 2; 0.5%), Rhipicephalus bursa (n = 1; 0.2%), and Haemaphysalis sp. (n = 1; 0.2%). Of the positive R. turanicus samples, 5 (5.7%) were infected with T. ovis and 2 (2.9%) with T. lestoquardi. Neither Babesia ovis nor Babesia motasi infection was detected in salivary glands or uterine samples of the ticks. The results also suggest that R. turanicus could be the vector responsible for transmission of the 2 Theileria species.

  16. Diversity of Babesia and Rickettsia species in questing Ixodes ricinus: a longitudinal study in urban, pasture, and natural habitats.

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    Overzier, Evelyn; Pfister, Kurt; Thiel, Claudia; Herb, Ingrid; Mahling, Monia; Silaghi, Cornelia

    2013-08-01

    In a previous study, our group investigated the Babesia spp. prevalence in questing Ixodes ricinus ticks from nine city parks in South Germany in the years 2009 and 2010. We showed predominant prevalence of B. venatorum (in previous literature also known as Babesia sp. EU1), especially in those parks in a more natural condition and with occurrence of large wild animals, such as roe deer. To obtain longitudinal data and to broaden the knowledge about this pathogen, further investigations were carried out in 2011 and 2012 in four of those city parks. Two additional habitat types were chosen for comparison of prevalence data and species analysis focusing on occurrence of potential reservoir hosts. A total of 10,303 questing I. ricinus were collected in four city parks, a pasture, and a natural area in Bavaria, and a representative number of samples were investigated for prevalence of DNA of Babesia spp. (n=4381) and Rickettsia spp. (n=2186) by PCR. In the natural and pasture area, a significantly higher Babesia spp. prevalence compared to the urban area was detected. The natural area revealed sequences of B. microti, B. venatorum, and B. capreoli. In the pasture and urban habitat, predominantly B. venatorum was found, whereas B. capreoli was less frequent and only one B. microti-infected tick was found. All B. microti sequences were 100% identical to the zoonotic Jena/Germany strain. For Rickettsia spp., the significantly highest prevalence was also detected in the natural and pasture areas, whereas lower prevalence was found in the urban area. Sequence analysis revealed R. helvetica (98%) and R. monacensis (2%). Prevalence rates and occurrence of Babesia spp. and Rickettsia spp. differed in urban, pasture and natural sites, most likely depending on the habitat structure (natural or cultivated) and therefore on the appearance and availability of reservoir hosts like roe deer or small mammals.

  17. Molecular identification of Theileria and Babesia in sheep and goats in the Black Sea Region in Turkey.

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    Aydin, Mehmet Fatih; Aktas, Munir; Dumanli, Nazir

    2013-08-01

    This study was carried out to investigate presence and distribution of Theileria and Babesia species via microscopic examination and reverse line blotting (RLB) techniques in sheep and goats in the Black Sea region of Turkey. For this purpose, 1,128 blood samples (869 sheep and 259 goats) were collected by active surveillance from sheep and goats in different provinces of various cities in the region in the years 2010 and 2011. Smears were prepared from the blood samples, stained with Giemsa, and examined under the light microscope for Theileria and Babesia piroplasms. The genomic DNAs were extracted from blood samples. The length of 360-430-bp fragment in the variable V4 region of 18S SSU rRNA gene of Theileria and Babesia species was amplified using the gDNAs. The polymerase chain reaction products were hybridized to the membrane-connected species-specific probes. A total of 38 animals (3.37%) including 34 sheep (3.91%) and 4 goats (1.54%) were found to be positive for Theileria spp. piroplasms in microscopic examination of smears while Babesia spp. piroplasm could not detected. Infection rates were 34.64% in sheep, 10.04% in goats, and totally 28.99% for Theileria ovis while 0.58% in sheep and totally 0.44% for Babesia ovis. However, Theileria sp. OT3 was detected in 2.65% of sheep and 2.04% of all animals; besides Theileria sp., MK had 0.58% prevalence in sheep and 0.77% in goats, with a total 0.62% with RLB. Although T. ovis and Theileria sp. MK were determined in both sheep and goats, B. ovis and Theileria sp. OT3 were observed only in the sheep. These results provide the first detailed molecular data for sheep and goat theileriosis and babesiosis in the region.

  18. Longitudinal field study on bovine Babesia spp. and Anaplasma phagocytophilum infections during a grazing season in Belgium.

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    Lempereur, Laetitia; Lebrun, Maude; Cuvelier, Pascale; Sépult, Géraldine; Caron, Yannick; Saegerman, Claude; Shiels, Brian; Losson, Bertrand

    2012-04-01

    Anaplasmosis and babesiosis are major tick-borne diseases with a high economic impact but are also a public health concern. Blood samples collected in the spring, summer, and autumn of 2010 from 65 cows in seven different farms in Belgium were monitored with an indirect immunofluorescence antibody test to assess seroprevalence against these pathogens. Seroprevalences to Babesia spp. were measured as 10.7%, 20%, and 12.3% in spring, summer, and autumn, respectively, whereas seroprevalences to Anaplasma phagocytophilum were 30.8%, 77%, and 56.9%, respectively. A total of 805 Ixodes ricinus ticks were collected at the same time from both cattle (feeding ticks) and grazed pastures (questing ticks). The infection level of ticks, assessed by PCR assay, for Babesia spp. DNA was 14.6% and 7.9% in feeding and questing ticks, respectively, whereas 21.7% and 3% of feeding and questing ticks were found be positive for A. phagocytophilum cDNA. Fifty-five PCR-positive samples were identified by sequencing as Babesia sp. EU1, of which five from feeding ticks were positive for both A. phagocytophilum and Babesia sp. EU1. The high density of wild cervids in the study area could explain these observations, as deer are considered to be the main hosts for adults of I. ricinus. However, the absence of Babesia divergens both in feeding and questing ticks is surprising, as the study area is known to be endemic for cattle babesiosis. Increasing cervid populations and comorbidity could play an import role in the epidemiology of these tick-borne diseases.

  19. Hemoparasites in a wild primate: Infection patterns suggest interaction of Plasmodium and Babesia in a lemur species.

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    Springer, Andrea; Fichtel, Claudia; Calvignac-Spencer, Sébastien; Leendertz, Fabian H; Kappeler, Peter M

    2015-12-01

    Hemoparasites can cause serious morbidity in humans and animals and often involve wildlife reservoirs. Understanding patterns of hemoparasite infections in natural populations can therefore inform about emerging disease risks, especially in the light of climate change and human disruption of natural ecosystems. We investigated the effects of host age, sex, host group size and season on infection patterns of Plasmodium sp., Babesia sp. and filarial nematodes in a population of wild Malagasy primates, Verreaux's sifakas (Propithecus verreauxi), as well as the effects of these infections on hematological variables. We tested 45 blood samples from 36 individuals and identified two species of Plasmodium, one species of Babesia and two species of filarial nematodes. Plasmodium spp. and Babesia sp. infections showed opposite patterns of age-dependency, with babesiosis being prevalent among young animals, while older animals were infected with Plasmodium sp. In addition, Babesia sp. infection was a statistically significant negative predictor of Plasmodium sp. infection. These results suggest that Plasmodium and Babesia parasites may interact within the host, either through cross-immunity or via resource competition, so that Plasmodium infections can only establish after babesiosis has resolved. We found no effects of host sex, host group size and season on hemoparasite infections. Infections showed high prevalences and did not influence hematological variables. This preliminary evidence supports the impression that the hosts and parasites considered in this study appear to be well-adapted to each other, resulting in persistent infections with low pathogenic and probably low zoonotic potential. Our results illustrate the crucial role of biodiversity in host-parasite relationships, specifically how within-host pathogen diversity may regulate the abundance of parasites.

  20. Hemoparasites in a wild primate: Infection patterns suggest interaction of Plasmodium and Babesia in a lemur species☆

    Science.gov (United States)

    Springer, Andrea; Fichtel, Claudia; Calvignac-Spencer, Sébastien; Leendertz, Fabian H.; Kappeler, Peter M.

    2015-01-01

    Hemoparasites can cause serious morbidity in humans and animals and often involve wildlife reservoirs. Understanding patterns of hemoparasite infections in natural populations can therefore inform about emerging disease risks, especially in the light of climate change and human disruption of natural ecosystems. We investigated the effects of host age, sex, host group size and season on infection patterns of Plasmodium sp., Babesia sp. and filarial nematodes in a population of wild Malagasy primates, Verreaux's sifakas (Propithecus verreauxi), as well as the effects of these infections on hematological variables. We tested 45 blood samples from 36 individuals and identified two species of Plasmodium, one species of Babesia and two species of filarial nematodes. Plasmodium spp. and Babesia sp. infections showed opposite patterns of age-dependency, with babesiosis being prevalent among young animals, while older animals were infected with Plasmodium sp. In addition, Babesia sp. infection was a statistically significant negative predictor of Plasmodium sp. infection. These results suggest that Plasmodium and Babesia parasites may interact within the host, either through cross-immunity or via resource competition, so that Plasmodium infections can only establish after babesiosis has resolved. We found no effects of host sex, host group size and season on hemoparasite infections. Infections showed high prevalences and did not influence hematological variables. This preliminary evidence supports the impression that the hosts and parasites considered in this study appear to be well-adapted to each other, resulting in persistent infections with low pathogenic and probably low zoonotic potential. Our results illustrate the crucial role of biodiversity in host-parasite relationships, specifically how within-host pathogen diversity may regulate the abundance of parasites. PMID:26767166

  1. Pneumonia por Rhodococcus equi em doente VIH (+: A propósito de uma associação rara

    Directory of Open Access Journals (Sweden)

    Paula Esteves

    2007-09-01

    Full Text Available Resumo: A infecção humana por Rhodococcus equi, mesmo no contexto da infecção pelo VIH, é relativamente rara. Os autores apresentam o caso clínico de um indivíduo, infectado pelo VIH, que apresentava um quadro clínico e imagiológico compatível com pneumonia, cujos exames iniciais revelaram a presença de Pneumocystis jiroveci. Após realização de terapêutica específica, perante a manutenção do quadro clínico-imagiológico, procedeu-se à realização de novos exames complementares, tendo sido identificado a presença de Rhodococcus equi no lavado broncoalveolar. Com instituição de terapêutica dirigida a este agente, verificou--se progressiva melhoria clínica e imagiológica. Os autores efectuam uma revisão bibliográfica da pneumonia por Rhodococcus equi, salientando a raridade desta associação e a elevada sobrevida observada. Abstract: The human infection by Rhodococcus equi, even in the presence of HIV infection, remains a rare disease. The authors present a case report of pneumonia, occurring in a HIV (+ man. After identifying Pneumocystis jiroveci in the BAL, despite proper medication, the patient didn’t improve. Another BAL was performed and a Rhodoccus equi isolated. The therapeutic regimen was changed according to this finding and the patient improved. The authors make a review of the literature, focusing on the rarity of this association and the high survival observed. Palavras-chave: Rhodococcus equi, Pneumocystis jiroveci, VIH, Key-words: Rhodococcus equi, Pneumocystis jiroveci, HIV

  2. Changes in intestinal fluid and mucosal immune responses to cholera toxin in Giardia muris infection and binding of cholera toxin to Giardia muris trophozoites.

    Science.gov (United States)

    Ljungström, I; Holmgren, J; Svennerholm, A M; Ferrante, A

    1985-10-01

    The effect of Giardia muris infection on the diarrheal response and gut mucosal antibody response to cholera toxin was examined in mice. The results obtained showed that the fluid accumulation in intestinal loops exposed to cholera toxin was increased in mice infected with a low number (5 X 10(4) ) of G. muris cysts compared with the response in noninfected mice. This effect was associated with a marked reduction in absorption of oral rehydration fluid from the intestine. In contrast, mice infected with a high dose (2 X 10(5) ) of cysts showed a marked decrease in fluid accumulation in response to the toxin. This decrease might be related to the finding that both G. muris and Giardia lamblia trophozoites can bind significant amounts of cholera toxin. Evidence is presented which suggests that the gut mucosal antibody response, mainly immunoglobulin A but also immunoglobulin G, to an immunization course with perorally administered cholera toxin was depressed in mice infected with G. muris. The reduction in antibody levels was particularly evident when the primary immunization was made very early after infection. The serum antitoxin antibodies to the oral immunization with cholera toxin were, however, not affected. Likewise, the delayed-type hypersensitivity response against sheep erythrocytes in animals primed subcutaneously with sheep erythrocytes was not modified during the course of G. muris infection.

  3. A Study of Naturally Acquired Canine Babesiosis Caused by Single and Mixed Babesia Species in Zambia: Clinicopathological Findings and Case Management

    Directory of Open Access Journals (Sweden)

    King Shimumbo Nalubamba

    2015-01-01

    Full Text Available A retrospective and prospective analysis of clinical records of dogs diagnosed with Babesia infections was carried out for the years 2000 to 2013 from practices in Lusaka, Zambia. Records of 363 dogs with confirmed Babesia infections were analysed using demographic factors including sex, breed, age, and clinical signs in relation to haematological findings and Babesia species. The clinical and laboratory findings observed are described as well as Babesia species identification. The study included 18 breeds and the highest proportion were mongrels (32.2%, males representing 64.5% of the population. The most common presenting problems were anorexia (65.3% and lethargy/weakness (65.3%. The most common clinical signs were fever (87.3%, pallor (52.3%, lymphadenopathy (47.4%, and presence of ticks (44.9%. Anaemia (96.4% and nucleated erythrocytes (42.2% were the most common laboratory findings. A mixed infection of Babesia rossi and Babesia gibsoni was present in 59.7% of dogs, whilst 8% and 32.2% had B. rossi and B. gibsoni as a single infection, respectively. Case management mainly involved therapy with tetracyclines and imidocarb and was usually accompanied by clinical improvement. This study highlights, for the first time, the presence of B. gibsoni in natural dog populations in Zambia, where previously only B. rossi was reported.

  4. Detection of Babesia Sp. EU1 and members of spotted fever group rickettsiae in ticks collected from migratory birds at Curonian Spit, North-Western Russia.

    Science.gov (United States)

    Movila, Alexandru; Reye, Anna L; Dubinina, Helen V; Tolstenkov, Oleg O; Toderas, Ion; Hübschen, Judith M; Muller, Claude P; Alekseev, Andrey N

    2011-01-01

    To reveal the prevalence of spotted fever group (SFG) rickettsiae and Babesia sp. in Ixodes ricinus (L.) ticks from migratory birds, 236 specimens represented 8 species of Passeriformes and were collected at Curonian Spit in Kaliningrad enclave of North-Western Russia. The ticks (total 126) being detached from four bird species, Turdus philomelos, Fringilla coelebs, Parus major, and Sturnus vulgaris, were investigated by PCR using the primers Rp CS.877p/Rp CS.1258n for the detection of Rickettsia and BJ1/BN2 for Babesia spp. Babesia spp. were detected in 2 of 126 (1.6%) ticks. The partial sequence of 18S rDNA had 100% similarity to human pathogenic Babesia sp. EU1. The SFG rickettsiae were detected in 19 of 126 (15.1%) ticks collected from the above-mentioned bird species. BLAST analysis of SFG rickettsia gltA assigned sequences to human pathogenic Rickettsia helvetica (10.3%), Rickettsia monacensis (3.9%), and Rickettsia japonica (0.8%) with 98%-100% sequence similarity. The SFG rickettsiae and Babesia sp. EU1 in ticks collected from the passerines in Russia were detected for the first time. The survey indicates that migratory birds may become a reservoir for Babesia spp. and SFG rickettsiae. Future investigations need to characterize the role of birds in the epidemiology of these human pathogens in the region.

  5. An epidemiological survey of bovine Babesia and Theileria parasites in cattle, buffaloes, and sheep in Egypt.

    Science.gov (United States)

    Elsify, Ahmed; Sivakumar, Thillaiampalam; Nayel, Mohammed; Salama, Akram; Elkhtam, Ahmed; Rizk, Mohamed; Mosaab, Omar; Sultan, Khaled; Elsayed, Shimaa; Igarashi, Ikuo; Yokoyama, Naoaki

    2015-02-01

    Cattle, buffaloes, and sheep are the main sources of meat and milk in Egypt, but their productivity is thought to be greatly reduced by hemoprotozoan parasitic diseases. In this study, we analyzed the infection rates of Babesia bovis, Babesia bigemina, Theileria annulata, and Theileria orientalis, using parasite-specific PCR assays in blood-DNA samples sourced from cattle (n=439), buffaloes (n=50), and sheep (n=105) reared in Menoufia, Behera, Giza, and Sohag provinces of Egypt. In cattle, the positive rates of B. bovis, B. bigemina, T. annulata, and T. orientalis were 3.18%, 7.97%, 9.56%, and 0.68%, respectively. On the other hand, B. bovis and T. orientalis were the only parasites detected in buffaloes and each of these parasites was only found in two individual DNA samples (both 2%), while one (0.95%) and two (1.90%) of the sheep samples were positive for B. bovis and B. bigemina, respectively. Sequence analysis showed that the B. bovis Rhoptry Associated Protein-1 and the B. bigemina Apical Membrane Antigen-1 genes were highly conserved among the samples, with 99.3-100% and 95.3-100% sequence identity values, respectively. In contrast, the Egyptian T. annulata merozoite surface antigen-1 gene sequences were relatively diverse (87.8-100% identity values), dispersing themselves across several clades in the phylogenetic tree containing sequences from other countries. Additionally, the T. orientalis Major Piroplasm Surface Protein (MPSP) gene sequences were classified as types 1 and 2. This is the first report of T. orientalis in Egypt, and of type 2 MPSP in buffaloes. Detection of MPSP type 2, which is considered a relatively virulent genotype, suggests that T. orientalis infection may have veterinary and economic significance in Egypt. In conclusion, the present study, which analyzed multiple species of Babesia and Theileria parasites in different livestock animals, may shed an additional light on the epidemiology of hemoprotozoan parasites in Egypt. Copyright

  6. Evaluation of immunochromatographic test (ICT) strips for the serological detection of Babesia bovis and Babesia bigemina infection in cattle from Western Java, Indonesia.

    Science.gov (United States)

    Guswanto, Azirwan; Allamanda, Puttik; Mariamah, Euis Siti; Munkjargal, Tserendorf; Tuvshintulga, Bumduuren; Takemae, Hitoshi; Sivakumar, Thillaiampalam; AbouLaila, Mahmoud; Terkawi, Mohamad Alaa; Ichikawa-Seki, Madoka; Nishikawa, Yoshifumi; Yokoyama, Naoaki; Igarashi, Ikuo

    2017-05-30

    Three types of immunochromatographic test (ICT) strips were prepared for the detection of an antibody response against spherical body protein 4 (SBP-4) of Babesia bovis (bovICT), C-terminal-truncated rhoptry-associated protein 1 (rRAP1/CT17) of B. bigemina (bigICT), and the combination of both proteins (dual-ICT). The evaluation of their performance was conducted using a confirmed positive and negative serum panel for B. bovis and B. bigemina. Together with ELISA, the ICT strips were applied to determine the seroprevalence of bovine babesiosis in Western Java, Indonesia. Among 991 serum samples, 28.4%, 25.3%, and 24.5% of cattle were detected to be seropositive to B. bovis infection using ELISA, bovICT, and dual-ICT, respectively. B. bigemina seropositive was detected in 27.1%, 24.2%, and 22.8% of samples using ELISA, bigICT, and dual-ICT, respectively. The comparison of ICT strips and ELISA results using field serum samples showed good agreement with Kappa values >0.7 between all methods The application of ICT strips is preferable in the field situations where rapid diagnosis is required. Furthermore, the data showed the current seroprevalence of bovine babesiosis in Western Java, Indonesia, and efficient control strategies are needed to reduce economic losses due to the disease. Copyright © 2017 Elsevier B.V. All rights reserved.

  7. EquiMar : Equitable Testing and Evaluation of Marine Energy Extraction Devices in Terms of Performance, Cost and Environmental Impact

    DEFF Research Database (Denmark)

    Kofoed, Jens Peter; Pecher, Arthur; Margheritini, Lucia

    The Sea Trial Manual (D4.1) describes the type of operations required to advance an ocean energy conversion device (wave and tide) from an intermediate scaled sub-systems proving machine (circa 1:4) to a full size solo prototype pre-production unit and on towards a pre-commercial device ready for...... of marine energy converters, according to Annex 1 – Description of Work of the EquiMar project, where task 4.2 is defined. Some slight modifications have been made to the original structure due to re-adjustments in accordance with the on-going research.......The Sea Trial Manual (D4.1) describes the type of operations required to advance an ocean energy conversion device (wave and tide) from an intermediate scaled sub-systems proving machine (circa 1:4) to a full size solo prototype pre-production unit and on towards a pre-commercial device ready...

  8. Molecular epidemiology of bovine Babesia spp. and Theileria orientalis parasites in beef cattle from northern and northeastern Thailand.

    Science.gov (United States)

    Jirapattharasate, Charoonluk; Adjou Moumouni, Paul Franck; Cao, Shinuo; Iguchi, Aiko; Liu, Mingming; Wang, Guanbo; Zhou, Mo; Vudriko, Patrick; Changbunjong, Tanasak; Sungpradit, Sivapong; Ratanakorn, Parntep; Moonarmart, Walasinee; Sedwisai, Poonyapat; Weluwanarak, Thekhawet; Wongsawang, Witsanu; Suzuki, Hiroshi; Xuan, Xuenan

    2016-02-01

    Beef cattle production represents the largest cattle population in Thailand. Their productivity is constrained by tick-borne diseases such as babesiosis and theileriosis. In this study, we determined the prevalence of Babesia bigemina, Babesia bovis and Theileria orientalis using polymerase chain reaction (PCR). The genetic markers that were used for detection of the above parasites were sequenced to determine identities and similarity for Babesia spp. and genetic diversity of T. orientalis. Furthermore the risk factors for the occurrence of the above protozoan parasites in beef cattle from northern and northeastern parts of Thailand were assessed. A total of 329 blood samples were collected from beef cattle in 6 provinces. The study revealed that T. orientalis was the most prevalent (30.1%) parasite in beef cattle followed by B. bigemina (13.1%) and B. bovis (5.5%). Overall, 78.7% of the cattle screened were infected with at least one of the above parasites. Co-infection with Babesia spp. and T. orientalis was 30.1%. B. bigemina and T. orientalis were the most prevalent (15.1%) co-infection although triple infection with the three parasites was observed in 3.0% of the samples. Sequencing analysis revealed that B. bigemina RAP1 gene and B. bovis SBP2 gene were conserved among the parasites from different cattle samples. Phylogenetic analysis showed that the T. orientalis MPSP gene from parasites isolated from cattle in north and northeast Thailand was classified into types 5 and 7 as reported previously. Lack of tick control program was the universal risk factor of the occurrence of Babesia spp. and T. orientalis infection in beef cattle in northern and northeastern Thailand. We therefore recommend training of farmers on appropriate tick control strategies and further research on potential vectors for T. orientalis and elucidate the effect of co-infection with Babesia spp. on the pathogenicity of T. orientalis infection on beef in northern and northeastern Thailand

  9. Entomologic and serologic evidence of zoonotic transmission of Babesia microti, eastern Switzerland.

    Science.gov (United States)

    Foppa, Ivo M; Krause, Peter J; Spielman, Andrew; Goethert, Heidi; Gern, Lise; Brand, Brigit; Telford, Sam R

    2002-07-01

    We evaluated human risk for infection with Babesia microti at a site in eastern Switzerland where several B. microti-infected nymphal Ixodes ricinus ticks had been found. DNA from pooled nymphal ticks amplified by polymerase chain reaction was highly homologous to published B. microti sequences. More ticks carried babesial infection in the lower portion of the rectangular 0.7-ha grid than in the upper (11% vs. 0.8%). In addition, we measured seroprevalence of immunoglobulin (Ig) G antibodies against B. microti antigen in nearby residents. Serum from 1.5% of the 396 human residents of the region reacted to B. microti antigen (>1:64), as determined by indirect immunofluorescence assay (IgG). These observations constitute the first report demonstrating B. microti in a human-biting vector, associated with evidence of human exposure to this agent in a European site.

  10. Urinary creatinine to serum creatinine ratio and renal failure index in dogs infected with Babesia canis.

    Science.gov (United States)

    Zygner, Wojciech; Gójska-Zygner, Olga; Wesołowska, Agnieszka; Wędrychowicz, Halina

    2013-09-01

    Urinary creatinine to serum creatinine (UCr/SCr) ratio and renal failure index (RFI) are useful indices of renal damage. Both UCr/SCr ratio and RFI are used in differentiation between prerenal azotaemia and acute tubular necrosis. In this work the authors calculated the UCr/SCr ratio and RFI in dogs infected with Babesia canis and the values of these indices in azotaemic dogs infected with the parasite. The results of this study showed significantly lower UCr/SCr ratio in dogs infected with B. canis than in healthy dogs. Moreover, in azotaemic dogs infected with B. canis the UCr/SCr ratio was significantly lower and the RFI was significantly higher than in non-azotaemic dogs infected with B. canis. The calculated correlation between RFI and duration of the disease before diagnosis and treatment was high, positive and statistically significant (r = 0.89, p caused by B. canis in Poland acute tubular necrosis may develop.

  11. Canine Babesiosis in China Caused by Babesia gibsoni: A Molecular Approach.

    Directory of Open Access Journals (Sweden)

    Da-Wei Yao

    2014-06-01

    Full Text Available To provide a point of reference to study the epidemiology and clinical expression of canine babesiosis in China.A total of 30 dogs infected with canine babesiosis were evaluated by mean of clinical history, physical examination, hematological, restriction fragment length polymorphism of PCR products (PCR-RFLP and sequencing analysis.The most prevalent clinical abnormalities were lethargy (100%, anorexia (100%, pale or icteric mucous membranes (80%, fever (70% and dark urine (70%. Hematology parameters revealed that anemia and thrombocytopenia were the major abnormalities in blood of dogs infected with canine babesia. The results of PCR-RFLP and sequencing analysis indicated that B. gibsoni was the main species responsible for canine babesiosis cases at the time of the study in Nanjing, China.The results provide valuable information for better understanding of the epidemiology of canine babesiosis in China.

  12. Konsentrasi Serum Anjing yang Optimum untuk Menumbuhkan dan Memelihara Babesia canis dalam Biakan

    Directory of Open Access Journals (Sweden)

    Tutuk Astyawati

    2010-12-01

    Full Text Available The use of cultivation system in vitro is very important in the future study of Babesia canis. Theaim of this study was to cultivate B. canis in vitro using RPMI media with different concentration of dogsera. B. canis infected erythrocytes were collected from splenectomized infected dog. Parasites werecultivated with RPMI 1640 medium supplemented with normal dog sera at the concentration of 10%, 20%and 40%, the culture were then incubated in 5% CO2 , 37oC temperature for 17 days and subcultured every48 hours. The Percentage of Parasitized Erythrocytes (PPE in culture with 10% dog serum was significantlylower than those 20%, and 40% The used of 20% and 40 % sera were better than 10%. It is recommendedthat 40 % serum can be used for initiation phase of cultivation, whereas 20% concentration were used formaintenance of the culture.

  13. Babesia canis vogeli, Ehrlichia canis, and Anaplasma platys infection in a dog.

    Science.gov (United States)

    Al Izzi, Salah; Martin, Donald S; Chan, Roxanne Y Y; Leutenegger, Christian M

    2013-12-01

    A 12-month-old male neutered mixed breed dog was presented with a history of diarrhea, lethargy, emaciation, polydypsia, and sniffling. Physical examination findings included pale mucous membranes, increased heart and respiratory rates, and normal rectal temperature (38°C). Hematologic abnormalities included anemia and thrombocytopenia. Biochemical abnormalities included hypoalbuminemia, hyperbilirubinemia, and elevated ALP and ALT activities. A SNAP 4Dx test result was positive for Ehrlichia canis. Babesia canis vogeli organisms were found in the peripheral blood films, while morulae of E canis were not seen. Real-time polymerase chain reaction testing confirmed the presence of both B c vogeli and E canis organisms, and also was positive for Anaplasma platys infection. The dog recovered following treatment with doxycycline and imidocarb dipropionate, with normal hematology and biochemical profiles. © 2013 American Society for Veterinary Clinical Pathology.

  14. Successful vaccination against Boophilus microplus and Babesia bovis using recombinat antigens

    Directory of Open Access Journals (Sweden)

    P. Willadsen

    1992-01-01

    Full Text Available Current methods for the control of the cattle tick Boophils microplus and the agent of bovine babesiosis, Babesia bovis are unsatisfactory. Effective immunological control of both parasites would have great advantages. However, naturally acquired immunity to the tick is generally unable to prevent serious production losses. A vaccine against the tick, based on a novel form of immunization, is being developed. A protective antigen has been isolated from the tick, characterized and produced as an effective, recombinant protein. A vaccine incorporating this antigen is currently undergoing field trials. In the Australian situation, improved tick control will probably increase endemic instability with respect to B. bovis. Fortunately, a trivalent, recombinant B. bovis vaccine has also been developed. This too is now undergoing pre-registration field trials.

  15. PERFORMANCE EVALUATION OF A PROTOTYPE ARCHITECT ANTIBODY ASSAY FOR BABESIA MICROTI.

    Science.gov (United States)

    Cheng, Kevin; Coller, Kelly E; Marohnic, Christopher C; Pfeiffer, Zachary A; Fino, James R; Elsing, Randee R; Bergsma, Janet; Marcinkus, Marilee A; Kar, Alak K; Gumbs, Orlando H; Otis, Kathy S; Fishpaugh, Jeffrey; Schultz, Phillip W; Pope, Mark R; Narvaez, Alfredo R; Wong, Susan J; Madison-Antenucci, Susan; Leary, Thomas P; Dawson, George J

    2018-05-09

    The tick-borne protozoan Babesia microti is responsible for more than 200 cases of transfusion-transmitted babesiosis (TTB) infection in the United States over the last 30 years. Measures to mitigate the risk of TTB include nucleic acid testing (NAT) and B. microti antibody testing. A fully automated prototype B. microti antibody test was developed on the ARCHITECT instrument. The specificity was determined to be 99.98% in volunteer blood donors (n=28,740) from areas considered as low endemic for B. microti The sensitivity of the prototype test was studied in experimentally-infected macaques; a total of 128 samples were detected compared to 125 with the indirect fluorescent antibody test (IFA), additionally, 83 (89.2%) of the PCR positive samples were detected compared to 81 (87.1%) using the IFA test. All PCR positive samples that tested negative in the prototype antibody test were pre-seroconversion period samples. Following seroconversion, periods of intermittent parasitemia occurred; 17 PCR negative samples drawn in-between PCR positive bleed dates, tested positive both by the prototype test (robust reactivity) and IFA (marginal reactivity) prior to the administration of therapeutic drugs, indicating that the PCR test failed to detect samples from persistently infected macaques. The prototype assay detected 56 of 58 (96.6%) human subjects diagnosed with clinical babesiosis by both PCR and IFA testing. Overall, the prototype anti-babesia assay provides a highly sensitive and specific test for the diagnosis of B. microti infection. While PCR is preferred for detection of window period parasitemia, antibody tests detect infected subjects during periods of low level parasitemia. Copyright © 2018 Cheng et al.

  16. Detection of Babesia bovis carrier cattle by using polymerase chain reaction amplification of parasite DNA.

    Science.gov (United States)

    Fahrimal, Y; Goff, W L; Jasmer, D P

    1992-01-01

    Carrier cattle infected with Babesia bovis are difficult to detect because of the low numbers of parasites that occur in peripheral blood. However, diagnosis of low-level infections with the parasite is important for evaluating the efficacies of vaccines and in transmission and epidemiological studies. We used the polymerase chain reaction (PCR) to amplify a portion of the apocytochrome b gene from the parasite and tested the ability of this method to detect carrier cattle. The target sequence is associated with a 7.4-kb DNA element in undigested B. bovis genomic DNA (as shown previously), and the amplified product was detected by Southern and dot blot hybridization. The assay was specific for B. bovis, since no amplification was detected with Babesia bigemina, Trypanosoma brucei, Anaplasma marginale, or leukocyte DNA. The target sequence was amplified in DNA from B. bovis Mexico, Texas, and Australia S and L strains, demonstrating the applicability of the method to strains from different geographic regions. The sensitivity of the method ranged from 1 to 10 infected erythrocytes extracted from 0.5 ml of blood. This sensitivity was about 1,000 times greater than that from the use of unamplified parasite DNA. By the PCR method, six B. bovis carrier cattle were detected 86% of the time (range, 66 to 100%) when they were tested 11 times, while with microscopic examination of thick blood smears, the same carrier cattle were detected only 36% of the time (range, 17 to 66%). The method provides a useful diagnostic tool for detecting B. bovis carrier cattle, and the sensitivity is significantly improved over that of current methods. The results also suggest that characteristics of the apocytchrome b gene may make this a valuable target DNA for PCR-based detection of other hemoparasites. Images PMID:1624551

  17. Epidemiology of Babesia, Anaplasma and Trypanosoma species using a new expanded reverse line blot hybridization assay.

    Science.gov (United States)

    Paoletta, Martina Soledad; López Arias, Ludmila; de la Fournière, Sofía; Guillemi, Eliana Carolina; Luciani, Carlos; Sarmiento, Néstor Fabián; Mosqueda, Juan; Farber, Marisa Diana; Wilkowsky, Silvina Elizabeth

    2018-02-01

    Vector-borne hemoparasitic infections are a major problem that affects livestock industries worldwide, particularly in tropical and subtropical regions. In this work, a reverse line blot (RLB) hybridization assay was developed for the simultaneous detection and identification of Anaplasma, Babesia and bovine trypanosomes, encompassing in this way the most relevant hemoparasites that affect cattle. A total of 186 bovine blood samples collected from two different ecoepidemiological regions of northeast Argentina, with and without tick control, were analyzed with this new RLB. High diversity of parasites, such as Babesia bovis, B. bigemina, Anaplasma marginale and three different Trypanosoma species, was found. High rates of coinfections were also detected, and significant differences were observed not only in the prevalence of parasites but also in the level of coinfections between the two analyzed areas. Regarding the Trypanosoma genus, we provide molecular evidence of the presence of T. vivax and T. theileri for the first time in Argentina. Besides, since the RLB is a prospective tool, it allowed the identification of a yet unknown bovine trypanosome which could not be assigned to any of the bovine species known so far. In the present study we provide new insights on the prevalence of several pathogens that directly impact on livestock production in Argentina. The RLB assay developed here allows to identify simultaneously numerous pathogenic species which can also be easily expanded to detect other blood borne pathogens. These characteristics make the RLB hybridization assay an essential tool for epidemiological survey of all vector-borne pathogens. Copyright © 2017 Elsevier GmbH. All rights reserved.

  18. An Evaluation of Quantitative PCR Assays (TaqMan® and SYBR Green for the Detection of Babesia bigemina and Babesia bovis, and a Novel Fluorescent-ITS1-PCR Capillary Electrophoresis Method for Genotyping B. bovis Isolates

    Directory of Open Access Journals (Sweden)

    Bing Zhang

    2016-09-01

    Full Text Available Babesia spp. are tick-transmitted haemoparasites causing tick fever in cattle. In Australia, economic losses to the cattle industry from tick fever are estimated at AUD$26 Million per annum. If animals recover from these infections, they become immune carriers. Here we describe a novel multiplex TaqMan qPCR targeting cytochrome b genes for the identification of Babesia spp. The assay shows high sensitivity, specificity and reproducibility, and allows quantification of parasite DNA from Babesia bovis and B. bigemina compared to standard PCR assays. A previously published cytochrome b SYBR Green qPCR was also tested in this study, showing slightly higher sensitivity than the Taqman qPCRs but requires melting curve analysis post-PCR to confirm specificity. The SYBR Green assays were further evaluated using both diagnostic submissions and vaccinated cattle (at 7, 9, 11 and 14 days post-inoculation showed that B. bigemina can be detected more frequently than B. bovis. Due to fewer circulating parasites, B. bovis detection in carrier animals requires higher DNA input. Preliminary data for a novel fluorescent PCR genotyping based on the Internal Transcribed Spacer 1 region to detect vaccine and field alleles of B. bovis are described. This assay is capable of detecting vaccine and novel field isolate alleles in a single sample.

  19. Clinical outbreak of babesiosis caused by Babesia capreoli in captive reindeer (Rangifer tarandus tarandus) in the Netherlands.

    Science.gov (United States)

    Bos, Jan H; Klip, Fokko C; Sprong, Hein; Broens, Els M; Kik, Marja J L

    2017-08-01

    From a herd of captive reindeer (Rangifer tarandus tarandus) consisting of two males and seven females with five calves, three calves were diagnosed on post mortem examination with a Babesia capreoli infection. The diagnosis was indicated by PCR and when the other reindeer were examined two adult females and a one-year-old male were Babesia-positive. Molecular characterization of the 18S rDNA of the parasite showed complete identity with known B. capreoli sequences. Ixodes ricinus has been demonstrated to be a competent vector for B. capreoli from infected roe deer (Capreolus capreolus), the natural host of B. capreoli. The B. capreoli infection in these reindeer may have been transmitted by infected ticks (Ixodes ricinus) originating from roe deer living in the forest and meadows surrounding the enclosure. Copyright © 2017 Elsevier GmbH. All rights reserved.

  20. Optimization of a Fluorescence-Based Assay for Large-Scale Drug Screening against Babesia and Theileria Parasites

    OpenAIRE

    Rizk, Mohamed Abdo; El-Sayed, Shimaa Abd El-Salam; Terkawi, Mohamed Alaa; Youssef, Mohamed Ahmed; El Said, El Said El Shirbini; Elsayed, Gehad; El-Khodery, Sabry; El-Ashker, Maged; Elsify, Ahmed; Omar, Mosaab; Salama, Akram; Yokoyama, Naoaki; Igarashi, Ikuo

    2015-01-01

    A rapid and accurate assay for evaluating antibabesial drugs on a large scale is required for the discovery of novel chemotherapeutic agents against Babesia parasites. In the current study, we evaluated the usefulness of a fluorescence-based assay for determining the efficacies of antibabesial compounds against bovine and equine hemoparasites in in vitro cultures. Three different hematocrits (HCTs; 2.5%, 5%, and 10%) were used without daily replacement of the medium. The results of a high-thr...

  1. Heterogeneity in cytokine profiles of Babesia bovis-specific bovine CD4+ T cells clones activated in vitro.

    OpenAIRE

    Brown, W C; Woods, V M; Dobbelaere, D A; Logan, K S

    1993-01-01

    The central role of T cells in the immune response against hemoprotozoan parasites, both as helper cells for T cell-dependent antibody production and as effector cells acting on intracellular parasites through the elaboration of cytokines, has prompted an investigation of the bovine cellular immune response against Babesia bovis antigens. CD4+ T helper (Th) cell clones generated from four B. bovis-immune cattle by in vitro stimulation with a soluble or membrane-associated merozoite antigen we...

  2. A comparative study of an elisa test and an indirect immunofluorescence test for serological diagnosis of Babesia bovis infection

    International Nuclear Information System (INIS)

    Martins, J.R.; Cheong, F.H.; Correa, B.L.; Radley, D.E.; Cereser, V.H.

    1998-01-01

    Detection of antibodies to Babesia bovis in cattle is essential for the understanding of the epidemiology of babesiosis and this study was concerned with comparing the indirect fluorescent antibody with the ELISA. Both assays gave rise to 100% sensitivity whilst the ELISA was shown to be marginally more specific at 98%. The ease of use and low cost of the ELISA would make it the more obvious choice in conducting future serological surveys for this parasite. (author)

  3. Babesia genotypes in Haemaphysalis concinna collected from birds in Hungary reflect phylogeographic connections with Siberia and the Far East.

    Science.gov (United States)

    Flaisz, Barbara; Sulyok, Kinga M; Kováts, Dávid; Kontschán, Jenő; Csörgő, Tibor; Csipak, Ármin; Gyuranecz, Miklós; Hornok, Sándor

    2017-06-01

    Haemaphysalis concinna is the second most common tick species attaching to birds in Hungary. Recently, Babesia genotypes, found in Siberia and the Far East, have been detected in this tick species collected from the vegetation in Hungary and Slovakia. The aim of this study was to molecularly investigate if these piroplasms also occur in H. concinna carried by migratory birds, which might explain their occurrence in the western Palaearctic. During a 2-year period, 321 H. concinna larvae and nymphs were collected from 121 passerine birds (of 19 species) in Hungary. These were molecularly investigated for the presence of piroplasm DNA with PCR and sequencing. The prevalence of PCR positive ticks was 15.9% (51 out of 321). Piroplasm PCR positivity of H. concinna ticks was significantly more frequent during the summer and autumn compared to spring, suggesting that migratory birds arriving in Hungary from the north or north east are the most important in the dispersal of H. concinna-associated piroplasms. Three genotypes, i.e. Babesia sp. "Irk-Hc133", "Irk-Hc130" (originally found in Irkutsk, Siberia) and "Kh-Hc222" (originally found in Khabarovsk, Far East) were detected. Phylogenetically all these belonged to the group formed by Babesia spp. of ruminants. Four bird species, which had 14-60% prevalence of PCR positive ticks, are known to be associated with northeast to southwest autumn migration. In conclusion, the presence of Central and East Asian Babesia genotypes in Central Europe are most likely related to bird species with known eastern migratory habit and/or phylogenetically substantiated connections between their eastern and western Eurasian populations. Copyright © 2017 Elsevier GmbH. All rights reserved.

  4. Hemoparasites in a wild primate: Infection patterns suggest interaction of Plasmodium and Babesia in a lemur species

    OpenAIRE

    Andrea Springer; Claudia Fichtel; Sébastien Calvignac-Spencer; Fabian H. Leendertz; Peter M. Kappeler

    2015-01-01

    Hemoparasites can cause serious morbidity in humans and animals and often involve wildlife reservoirs. Understanding patterns of hemoparasite infections in natural populations can therefore inform about emerging disease risks, especially in the light of climate change and human disruption of natural ecosystems. We investigated the effects of host age, sex, host group size and season on infection patterns of Plasmodium sp., Babesia sp. and filarial nematodes in a population of wild Malagasy pr...

  5. The development and evaluation of a loop-mediated isothermal amplification (LAMP) method for detection of Babesia spp. infective to sheep and goats in China.

    Science.gov (United States)

    Guan, Guiquan; Chauvin, Alain; Luo, Jianxun; Inoue, Noboru; Moreau, Emmanuelle; Liu, Zhijie; Gao, Jinliang; Thekisoe, Oriel M M; Ma, Miling; Liu, Aihong; Dang, Zhisheng; Liu, Junlong; Ren, Qiaoyun; Jin, Yurong; Sugimoto, Chihiro; Yin, Hong

    2008-09-01

    The loop-mediated isothermal amplification (LAMP) reaction is a method that amplifies with high sensitivity, efficiency, and rapidity, deoxyribonucleic acid (DNA) under isothermal condition in simple incubators. Two primer sets for the LAMP method were designed using the nucleotide sequences of 18S rRNA gene of Babesia sp. BQ1 (Lintan) and Babesia sp. Xinjiang-2005 isolated in China. The primers were used to detect parasite DNA extracted from infected blood and purified parasites by LAMP. The specific ladder bands were amplified from the autologous genomic DNA of two Babesia species, respectively, and did not cross-react with the genomic DNA of Theileria sp. China 1, Theileria sp. China 2, B. bovis, Theileria sp. (Japan) and sheep. The LAMP was sensitive enough to detect 0.02 pg and 0.2 pg genomic DNA of Babesia sp. BQ1 (Lintan) and Babesia sp. Xinjiang-2005, respectively, from 10-fold serially diluted samples corresponding to the amount of DNA present in 50 microl of 0.000002% and 0.00002% parasitemic erythrocytes. Furthermore, DNA extracted from blood of intact (non-splenectomized) sheep experimentally infected with Babesia sp. BQ1 (Lintan) and Babesia sp. Xinjiang-2005 was amplified by the LAMP from week 1 to 9 and week 2 and 3 post-infection, respectively, demonstrating the high sensitivity of these primers. Of 365 samples collected from Gansu province, 14.3% (52/365) were positively detected by the LAMP. Of 145 samples collected on filter papers (Whatman) from the grazing sheep in Xinjiang province, 3.5% (5/145) were positive. These results show that the LAMP could be an alternative diagnostic tool for the detection of babesial infection in sheep and goats.

  6. Determination of erythrocyte susceptibility of Chinese sheep (Tan mutton breed) and French sheep (Vendéen breed) to Babesia sp. BQ1 (Lintan) by in vitro culture.

    Science.gov (United States)

    Guan, Guiquan; Moreau, Emmanuelle; Brisseau, Nadine; Luo, Jianxun; Yin, Hong; Chauvin, Alain

    2010-05-28

    The Babesia species "BQ1 (Lintan)" is infective to sheep and goats. The species was isolated from Haemaphysalis qinghaiensis collected in the Gannan Tibet Autonomous Region, China in April 2000. In this study, an in vitro culture system was developed for the propagation of Babesia sp. BQ1 (Lintan). Continuous cultivation and 5.0% parasitemia was obtained in vitro in RPMI 1640 medium with sheep red blood cells (RBC) (7.5%) supplemented with Fetal Bovine Serum (FBS) (20%), Amphotericin B (0.5 microg/ml) and Gentamicin (50 microg/ml) in an incubator at 37 degrees C and 6% CO(2) in 24-well and 6-well plates. Parasitemia could attain 10% in 75 cm(2) flasks with the same culture medium but with 2.5% RBC. A clonal line of Babesia sp. BQ1 (Lintan) was screened using the limiting dilution method and designated G7. Growth of Babesia sp. BQ1 (Lintan) in vitro was measured by microtitre-based spectrophotometric method and from parasitemia counts. The generation time was between 20.57 h (based the A(405) of the culture supernatant) and 26.41 h (based on parasitemia). Three French sheep were successfully infected with the culture and the infectivity of the clonal line G7 was determined. Finally, this in vitro culture system was used to compare the susceptibility (capacity to sustain Babesia sp. growth in vitro) of RBC from French sheep (Vendéen breed) and Chinese sheep (Tan mutton breed) for Babesia sp. BQ1 (Lintan) and B. divergens. The lower susceptibility to B. divergens and Babesia sp. BQ1 (Lintan) of RBC from French sheep, compared to Chinese sheep, is discussed. (c) 2010 Elsevier B.V. All rights reserved.

  7. Survey of Ehrlichia canis, Babesia spp. and Hepatozoon spp. in dogs from a semiarid region of Brazil

    Directory of Open Access Journals (Sweden)

    Tereza Emmanuelle de Farias Rotondano

    Full Text Available This study assessed the occurrence of Ehrlichia spp., Babesia spp. and Hepatozoon spp. infections in 100 tick-harboring dogs from a semiarid region of the State of Paraíba, Northeastern Brazil. Blood samples and ticks were collected from the animals, and a questionnaire was submitted to dog owners to obtain general data. Blood samples were used to perform hemogram, direct blood smear and immunological and molecular hemoparasite detection. The 1,151 ticks collected were identified as Rhipicephalus sanguineus; direct smears revealed E. canis-like morulae in the monocytes of 4% (4/100 of the non-vaccinated female dogs, and 34% and 25% of the dogs tested positive for Ehrlichia canis by indirect immunofluorescence assay (IFA and polymerase chain reaction (PCR, respectively. Blood smear examination revealed Babesia-suggestive merozoites in the erythrocytes of 2% (2/100 of the animals. Babesia vogeli was detected by PCR in ten animals (10% and was correlated with young age (p = 0.007 and thrombocytopenia (p = 0.01. None of the animals showed Hepatozoon spp. positivity. These results indicate that E. canis is the main tick-borne canine pathogen in the study area and provide the first report of B. vogeli infection in dogs from Paraiba State.

  8. Survey of Ehrlichia canis, Babesia spp. and Hepatozoon spp. in dogs from a semiarid region of Brazil.

    Science.gov (United States)

    Rotondano, Tereza Emmanuelle de Farias; Almeida, Herta Karyanne Araújo; Krawczak, Felipe da Silva; Santana, Vanessa Lira; Vidal, Ivana Fernandes; Labruna, Marcelo Bahia; de Azevedo, Sérgio Santos; Ade lmeida, Alzira Maria Paiva; de Melo, Marcia Almeida

    2015-01-01

    This study assessed the occurrence of Ehrlichia spp., Babesia spp. and Hepatozoon spp. infections in 100 tick-harboring dogs from a semiarid region of the State of Paraíba, Northeastern Brazil. Blood samples and ticks were collected from the animals, and a questionnaire was submitted to dog owners to obtain general data. Blood samples were used to perform hemogram, direct blood smear and immunological and molecular hemoparasite detection. The 1,151 ticks collected were identified as Rhipicephalus sanguineus; direct smears revealed E. canis-like morulae in the monocytes of 4% (4/100) of the non-vaccinated female dogs, and 34% and 25% of the dogs tested positive for Ehrlichia canis by indirect immunofluorescence assay (IFA) and polymerase chain reaction (PCR), respectively. Blood smear examination revealed Babesia-suggestive merozoites in the erythrocytes of 2% (2/100) of the animals. Babesia vogeli was detected by PCR in ten animals (10%) and was correlated with young age (p = 0.007) and thrombocytopenia (p = 0.01). None of the animals showed Hepatozoon spp. positivity. These results indicate that E. canis is the main tick-borne canine pathogen in the study area and provide the first report of B. vogeli infection in dogs from Paraiba State.

  9. Babesia bovis and B. bigemina DNA detected in cattle and ticks from Zimbabwe by polymerase chain reaction

    Directory of Open Access Journals (Sweden)

    I. Smeenk

    2000-07-01

    Full Text Available From blood collected from 94 cattle at 12 locations in the eastern and northeastern areas of Zimbabwe, DNA was extracted and analysed by polymerase chain reaction with primers previously reported to be specific for Babesia bigemina and Babesia bovis. Overall, DNA of Babesia bigemina was detected in the blood of 33/94 (35 % cattle and DNA from B. bovis was detected in 27/58 (47 % of cattle. The prevalence of DNA of B. bigemina was significantly higher in young animals (<2 years (23/46 than in animals over 2 years of age (10/48; (chi2 = 8.77; P < 0.01 %. Although tick sampling was not thorough, Boophilus decoloratus could be collected at 7/9 sites sampled and Boophilus microplus at 4/9 sites. Of the 20 B. decoloratus allowed to oviposit before PCR analysis, 1 (5 % contained DNA that could be amplified with primers for B. bigemina while 12 (60 % were positive with primers for B. bovis. Of the B. microplus allowed to oviposit, 11/16 (69 % were positive for B. bovis DNAby PCR and 2/16 (12 % were positive for B. bigemina.

  10. The evolutionary dynamics of variant antigen genes in Babesia reveal a history of genomic innovation underlying host-parasite interaction

    KAUST Repository

    Jackson, Andrew P.

    2014-05-05

    Babesia spp. are tick-borne, intraerythrocytic hemoparasites that use antigenic variation to resist host immunity, through sequential modification of the parasite-derived variant erythrocyte surface antigen (VESA) expressed on the infected red blood cell surface. We identified the genomic processes driving antigenic diversity in genes encoding VESA (ves1) through comparative analysis within and between three Babesia species, (B. bigemina, B. divergens and B. bovis). Ves1 structure diverges rapidly after speciation, notably through the evolution of shortened forms (ves2) from 5? ends of canonical ves1 genes. Phylogenetic analyses show that ves1 genes are transposed between loci routinely, whereas ves2 genes are not. Similarly, analysis of sequence mosaicism shows that recombination drives variation in ves1 sequences, but less so for ves2, indicating the adoption of different mechanisms for variation of the two families. Proteomic analysis of the B. bigemina PR isolate shows that two dominant VESA1 proteins are expressed in the population, whereas numerous VESA2 proteins are co-expressed, consistent with differential transcriptional regulation of each family. Hence, VESA2 proteins are abundant and previously unrecognized elements of Babesia biology, with evolutionary dynamics consistently different to those of VESA1, suggesting that their functions are distinct. 2014 The Author(s) 2014.

  11. The evolutionary dynamics of variant antigen genes in Babesia reveal a history of genomic innovation underlying host-parasite interaction

    KAUST Repository

    Jackson, Andrew P.; Otto, Thomas D.; Darby, Alistair; Ramaprasad, Abhinay; Xia, Dong; Echaide, Ignacio Eduardo; Farber, Marisa; Gahlot, Sunayna; Gamble, John; Gupta, Dinesh; Gupta, Yask; Jackson, Louise; Malandrin, Laurence; Malas, Tareq B.; Moussa, Ehab; Nair, Mridul; Reid, Adam J.; Sanders, Mandy; Sharma, Jyotsna; Tracey, Alan; Quail, Mike A.; Weir, William; Wastling, Jonathan M.; Hall, Neil; Willadsen, Peter; Lingelbach, Klaus; Shiels, Brian; Tait, Andy; Berriman, Matt; Allred, David R.; Pain, Arnab

    2014-01-01

    Babesia spp. are tick-borne, intraerythrocytic hemoparasites that use antigenic variation to resist host immunity, through sequential modification of the parasite-derived variant erythrocyte surface antigen (VESA) expressed on the infected red blood cell surface. We identified the genomic processes driving antigenic diversity in genes encoding VESA (ves1) through comparative analysis within and between three Babesia species, (B. bigemina, B. divergens and B. bovis). Ves1 structure diverges rapidly after speciation, notably through the evolution of shortened forms (ves2) from 5? ends of canonical ves1 genes. Phylogenetic analyses show that ves1 genes are transposed between loci routinely, whereas ves2 genes are not. Similarly, analysis of sequence mosaicism shows that recombination drives variation in ves1 sequences, but less so for ves2, indicating the adoption of different mechanisms for variation of the two families. Proteomic analysis of the B. bigemina PR isolate shows that two dominant VESA1 proteins are expressed in the population, whereas numerous VESA2 proteins are co-expressed, consistent with differential transcriptional regulation of each family. Hence, VESA2 proteins are abundant and previously unrecognized elements of Babesia biology, with evolutionary dynamics consistently different to those of VESA1, suggesting that their functions are distinct. 2014 The Author(s) 2014.

  12. Discrimination between ovine Babesia and Theileria species in China based on the ribosomal protein S8 (RPS8) gene.

    Science.gov (United States)

    Tian, Zhancheng; Liu, Guangyuan; Yin, Hong; Luo, Jianxun; Guan, Guiquan; Luo, Jin; Xie, Junren; Zheng, Jinfeng; Yuan, Xiaosong; Wang, Fangfang; Shen, Hui; Tian, Meiyuan

    2013-10-18

    Ovine babesiosis and theileriosis are important hemoprotozoal diseases of sheep and goats in tropical and subtropical regions that lead to economic losses in these animals. PCR-restriction fragment length polymorphism (PCR-RFLP) is a reliable molecular diagnostic tool for discriminating Theileria or Babesia species in the same host. In this study, the DNA sequences of a ribosomal protein S8 (RPS8) gene from four species of piroplasms in China were used to develop a species-specific PCR-RFLP diagnostic tool. The sensitivity of the PCR assays was 0.1 pg DNA for B. motasi and 1 pg DNA for T. uilenbergi and 10 pg DNA for Babesia sp. Xinjiang-2005 and T. luwenshuni. The clear size difference of the PCR products allowed for a direct discrimination for B. motasi, Babesia sp. Xinjiang-2005 and ovine Theileria species (T. uilenbergi and T. luwenshuni), except that the mixed infection between T. uilenbergi and T. luwenshuni may be difficult to distinguish, simply after the electrophoretic separation of the amplification products. Further T. uilenbergi and T. luwenshuni diagnoses were made by digesting the PCR product with SacI. The established method could be applicable for the survey of parasite dynamics, and epidemiological studies as well as prevention and control of the disease. Copyright © 2013 Elsevier B.V. All rights reserved.

  13. Detection of Babesia spp. in free-ranging Pukus, Kobus vardonii, on a game ranch in Zambia.

    Science.gov (United States)

    Munang'andu, Hetron Mweemba; Munyeme, Musso; Nambota, Andrew Mubila; Nalubamba, King Shimumbo; Siamudaala, Victor M

    2011-12-01

    Babesia spp. were detected from 4 asymptomatic pukus captured on a game ranch in central Zambia in October 2008. Blood smears were examined in 4 species of aymptomatic free-ranging antelopes, namely the puku (Kobus vordanii), reedbuck (Redunca arundinum), bushbuck (Tragelaphus sylvaticus), and kudu (Tragelaphus strepsiceros), and showed the presence of Babesia parasites only in the puku. In the puku, the prevalence of babesiosis was estimated at 33.3% (n = 12), while the overall prevalence in all examined animals was 8.5% (n = 47). The parasites showed morphological characteristics of paired ring-like stages with the length varying between 1.61 µm and 3.02 µm (mean = 2.12 µm, n = 27; SD = 0.76 µm). Both the infected and non-infected pukus showed good body condition scores (BCS), while the dominant tick species detected from all animals were Rhipicephalus appendiculatus, Rhipicephalus spp., and Boophilus spp. To our knowledge this is the first report of Babesia spp. infection in pukus in Zambia. These findings suggest that wildlife could play an important role in the epidemiology of babesiosis in Zambia.

  14. The evolutionary dynamics of variant antigen genes in Babesia reveal a history of genomic innovation underlying host–parasite interaction

    Science.gov (United States)

    Jackson, Andrew P.; Otto, Thomas D.; Darby, Alistair; Ramaprasad, Abhinay; Xia, Dong; Echaide, Ignacio Eduardo; Farber, Marisa; Gahlot, Sunayna; Gamble, John; Gupta, Dinesh; Gupta, Yask; Jackson, Louise; Malandrin, Laurence; Malas, Tareq B.; Moussa, Ehab; Nair, Mridul; Reid, Adam J.; Sanders, Mandy; Sharma, Jyotsna; Tracey, Alan; Quail, Mike A.; Weir, William; Wastling, Jonathan M.; Hall, Neil; Willadsen, Peter; Lingelbach, Klaus; Shiels, Brian; Tait, Andy; Berriman, Matt; Allred, David R.; Pain, Arnab

    2014-01-01

    Babesia spp. are tick-borne, intraerythrocytic hemoparasites that use antigenic variation to resist host immunity, through sequential modification of the parasite-derived variant erythrocyte surface antigen (VESA) expressed on the infected red blood cell surface. We identified the genomic processes driving antigenic diversity in genes encoding VESA (ves1) through comparative analysis within and between three Babesia species, (B. bigemina, B. divergens and B. bovis). Ves1 structure diverges rapidly after speciation, notably through the evolution of shortened forms (ves2) from 5′ ends of canonical ves1 genes. Phylogenetic analyses show that ves1 genes are transposed between loci routinely, whereas ves2 genes are not. Similarly, analysis of sequence mosaicism shows that recombination drives variation in ves1 sequences, but less so for ves2, indicating the adoption of different mechanisms for variation of the two families. Proteomic analysis of the B. bigemina PR isolate shows that two dominant VESA1 proteins are expressed in the population, whereas numerous VESA2 proteins are co-expressed, consistent with differential transcriptional regulation of each family. Hence, VESA2 proteins are abundant and previously unrecognized elements of Babesia biology, with evolutionary dynamics consistently different to those of VESA1, suggesting that their functions are distinct. PMID:24799432

  15. The evolutionary dynamics of variant antigen genes in Babesia reveal a history of genomic innovation underlying host-parasite interaction.

    Science.gov (United States)

    Jackson, Andrew P; Otto, Thomas D; Darby, Alistair; Ramaprasad, Abhinay; Xia, Dong; Echaide, Ignacio Eduardo; Farber, Marisa; Gahlot, Sunayna; Gamble, John; Gupta, Dinesh; Gupta, Yask; Jackson, Louise; Malandrin, Laurence; Malas, Tareq B; Moussa, Ehab; Nair, Mridul; Reid, Adam J; Sanders, Mandy; Sharma, Jyotsna; Tracey, Alan; Quail, Mike A; Weir, William; Wastling, Jonathan M; Hall, Neil; Willadsen, Peter; Lingelbach, Klaus; Shiels, Brian; Tait, Andy; Berriman, Matt; Allred, David R; Pain, Arnab

    2014-06-01

    Babesia spp. are tick-borne, intraerythrocytic hemoparasites that use antigenic variation to resist host immunity, through sequential modification of the parasite-derived variant erythrocyte surface antigen (VESA) expressed on the infected red blood cell surface. We identified the genomic processes driving antigenic diversity in genes encoding VESA (ves1) through comparative analysis within and between three Babesia species, (B. bigemina, B. divergens and B. bovis). Ves1 structure diverges rapidly after speciation, notably through the evolution of shortened forms (ves2) from 5' ends of canonical ves1 genes. Phylogenetic analyses show that ves1 genes are transposed between loci routinely, whereas ves2 genes are not. Similarly, analysis of sequence mosaicism shows that recombination drives variation in ves1 sequences, but less so for ves2, indicating the adoption of different mechanisms for variation of the two families. Proteomic analysis of the B. bigemina PR isolate shows that two dominant VESA1 proteins are expressed in the population, whereas numerous VESA2 proteins are co-expressed, consistent with differential transcriptional regulation of each family. Hence, VESA2 proteins are abundant and previously unrecognized elements of Babesia biology, with evolutionary dynamics consistently different to those of VESA1, suggesting that their functions are distinct. © The Author(s) 2014. Published by Oxford University Press on behalf of Nucleic Acids Research.

  16. Molecular and serologic evidence for Babesia bovis-like parasites in white-tailed deer (Odocoileus virginianus) in south Texas.

    Science.gov (United States)

    Ramos, Christina M; Cooper, Susan M; Holman, Patricia J

    2010-09-20

    The current study was undertaken to determine if white-tailed deer in south Texas harbor Babesia bovis, a causative agent of bovine babesiosis. Blood samples from free-ranging white-tailed deer (Odocoileus virginianus) on two ranches in LaSalle and Webb Counties were screened for B. bovis and other hemoparasites by the polymerase chain reaction (PCR) to detect the piroplasm 18S rDNA. Serology was conducted on selected samples to detect antibody activity to B. bovis by the immunofluorescent antibody test (IFAT). PCR revealed that 16% of the LaSalle County samples and 4% of the Webb County samples were positive for B. bovis. Five of the LaSalle County and the two Webb County B. bovis 18S rDNA amplicons were cloned and sequenced. The resulting clones shared 99% identity to B. bovis 18S rRNA gene sequences derived from cattle isolates. Weak seroreactivity to B. bovis was shown by the IFAT. The samples were also screened for additional hemoparasites of deer including Theileria cervi, Babesia odocoilei and other Babesia spp. A genotypically unique Theileria sp. was found, along with T. cervi and B. odocoilei. The finding of putative B. bovis in white-tailed deer necessitates further study to determine if deer may act as a transient host or even a reservoir of infection for B. bovis pathogenic to cattle.

  17. In vitro culture, PCR , and nested PCR for the detection of Theileria equi in horses submitted to exercise Cultivo in vitro, PCR e nested PCR na detecção de Theileria equi em eqüinos submetidos a exercícios

    Directory of Open Access Journals (Sweden)

    C.D. Baldani

    2008-06-01

    Full Text Available This study compared the usefulness of in vitro culture, PCR, and nested PCR for the diagnosis of Theileria equi in horses submitted to stress during exercise. Blood samples from 15 apparently healthy horses, previously conditioned to a high-speed equine treadmill, were taken prior to and after exercise. The animals were divided into two experimental groups: 30-day training schedule (G1 and 90-day training schedule (G2. Statistical analysis was performed using a chi-square test and kappa statistic was used in order to assess agreement. No significant difference was observed between samples collected at resting or after exercise. In G1, merozoites of T. equi were detected in the blood smears of four horses before in vitro culture, whereas 14 samples were positive, confirmed by culture. In G2, five and 11 horses were positive before and after culture, respectively. No PCR amplified product was observed in any of the tested animals although the PCR system based on the 16S rRNA gene of T. equi detected DNA in blood with an equivalent 8x10-5% parasitaemia. The nested PCR based on the T. equi merozoite antigen gene (EMA-1 allowed the visualization of amplified products in all the horses. Therefore, nested PCR should be considered as a means of detection of sub-clinical T. equi infections and in vitro culture could be used as a complement to other methods of diagnosis.Comparou-se a utilização do cultivo in vitro, PCR e nested PCR no diagnóstico de Theileria equi em eqüinos submetidos ao estresse induzido por exercícios. Amostras de sangue foram obtidas de 15 eqüinos submetidos a treinamento em esteira rolante de alto desempenho, sendo as amostras colhidas antes e após os exercícios. Os animais foram divididos em dois grupos experimentais: 30 dias de treinamento (G1 e 90 dias de treinamento (G2. O teste do qui-quadrado foi empregado para as análises estatísticas e o índice kappa utilizado para avaliar a concordância. Não houve diferen

  18. Infecção pulmonar por rhodococcus equi em doente vih+ - revisão baseada num caso clínico

    Directory of Open Access Journals (Sweden)

    João Faria

    2013-03-01

    Full Text Available Apesar do aumento no número de casos de infecção humana por Rhodococcus equi (R. equi registado nas últimas décadas, sobretudo em indivíduos infectados pelo vírus da imunodeficiência humana (VIH, esse diagnóstico permanece uma raridade. Os autores apresentam um caso de infecção pulmonar por R. equi num homem de 36 anos com síndrome de imunodeficiência adquirida (SIDA. A doença manifestou-se de forma insidiosa, apresentando-se radiologicamente sob a forma de lesão pulmonar esquerda com cavitação e nível hidroaéreo, tendo sido inicialmente colocada a hipótese diagnóstica de tuberculose pulmonar (TP dada a pesquisa de bacilos álcool-ácido resistentes (BAAR na expectoração ter sido positiva. Posteriormente, foi possível isolar o R. equi nas culturas de expectoração e lavado broncoalveolar (LBA. O estudo anátomo-patológico confirmou a presença concomitante de malacoplaquia pulmonar. O doente cumpriu antibioterapia dirigida ao agente e teve boa evolução clínica, analítica e radiológica. A propósito deste caso, os autores fazem uma revisão teórica do tema à luz dos conhecimentos actuais.

  19. Associations between physical examination, laboratory, and radiographic findings and outcome and subsequent racing performance of foals with Rhodococcus equi infection: 115 cases (1984-1992)

    International Nuclear Information System (INIS)

    Ainsworth, D.M.; Eicker, S.W.; Yeager, A.E.; Sweeney, C.R.; Viel, L.; Tesarowski, D.; Lavoie, J.P.; Hoffman, A.; Paradis, M.R.; Reed, S.M.

    1998-01-01

    Objective-To determine whether physical examination, laboratory, or radiographic abnormalities in foals with Rhodococcus equi infection were associated with survival, ability to race at least once after recovery, or, for foals that survived and went on to race, subsequent Facing performance. Design-Retrospective study. Animals-49 Thoroughbreds and 66 Standardbreds admitted to 1 of 6 veterinary teaching hospitals between 1984 and 1992 in which R equi infection was positively diagnosed. Procedure-Results of physical examination, laboratory testing, and thoracic radiography were reviewed. Indices of Facing performance were obtained for feats that recovered and eventually raced and compared with values for the US racing population. Results-83 (72%) feats survived. Foals that did not survive were more likely to have extreme tachycardia (heart rate > 100 beats/min), be in respiratory distress, and have severe radiographic abnormalities on thoracic radiographs at the time of initial examination than were foals that survived. Clinicopathologic abnormalities were not associated with whether feats did or did not survive. Forty-five of the 83 surviving foals (54%) eventually raced at least once, but none of the factors examined was associated with whether foals went on to race. Racing performance of foals that raced as adults was not significantly different from that of the US racing population. Clinical Implications-R equi infection in foals is associated with a decreased chance of racing as an adult, however, foals that eventually go on to race perform comparably to the US racing population

  20. Immunization by intrabronchial administration to 1-week-old foals of an unmarked double gene disruption strain of Rhodococcus equi strain 103+.

    Science.gov (United States)

    Pei, Yanlong; Nicholson, Vivian; Woods, Katharine; Prescott, John F

    2007-11-15

    Rhodococcus equi causes fatal granulomatous pneumonia in foals and immunocompromised animals and humans. However, there is no effective vaccine against this infection. In this study, the chromosomal genes isocitrate lyase (icl) and cholesterol oxidase (choE) were chosen as targets for mutation and assessment of the double mutant as an intrabronchial vaccine in 1-week-old foals. Using a modification of a suicide plasmid previously developed in this laboratory, we developed a choE-icl unmarked deletion mutant of R. equi strain 103+. Five 1-week-old foals were infected intrabronchially with the mutant and challenged intrabronchially with the parent, virulent, strain 2 weeks later. Three of the foals were protected against pneumonia caused by the virulent strain, but the other two foals developed pneumonia caused by the mutant strain during the post-challenge period. Since infection of 3-week-old foals by an icl mutant in an earlier study had shown complete attenuation of the strain, we conclude that a proportion of foals in the 1st week or so of life are predisposed to developing R. equi pneumonia because of an inability to mount an effective immune response. This has been suspected previously but this is the first time that this has been demonstrated experimentally.

  1. Improved molecular detection of Babesia infections in animals using a novel quantitative real-time PCR diagnostic assay targeting mitochondrial DNA.

    Science.gov (United States)

    Qurollo, Barbara A; Archer, Nikole R; Schreeg, Megan E; Marr, Henry S; Birkenheuer, Adam J; Haney, Kaitlin N; Thomas, Brittany S; Breitschwerdt, Edward B

    2017-03-07

    Babesiosis is a protozoal, tick transmitted disease found worldwide in humans, wildlife and domesticated animals. Commonly used approaches to diagnose babesiosis include microscopic examination of peripheral blood smears, detection of circulating antibodies and PCR. To screen and differentiate canine Babesia infections many PCR assays amplify the 18S rRNA gene. These sequences contain hypervariable regions flanked by highly conserved regions allowing for amplification of a broad-range of Babesia spp. However, differences in the 18S rRNA gene sequence of distantly related clades can make it difficult to design assays that will amplify all Babesia species while excluding the amplification of other eukaryotes. By targeting Babesia mitochondrial genome (mtDNA), we designed a novel three primer qPCR with greater sensitivity and broader screening capabilities to diagnose and differentiate Babesia spp. Using 13 Babesia mtDNA sequences, a region spanning two large subunit rRNA gene fragments (lsu5-lsu4) was aligned to design three primers for use in a qPCR assay (LSU qPCR) capable of amplifying a wide range of Babesia spp. Plasmid clones were generated and used as standards to determine efficiency, linear dynamic range and analytical sensitivity. Animals naturally infected with vector-borne pathogens were tested retrospectively and prospectively to determine relative clinical sensitivity and specificity by comparing the LSU qPCR to an established 18S rDNA qPCR. The LSU qPCR efficiencies ranged between 92 and 100% with the limit of detection at five copies/reaction. The assay did not amplify mammalian host or other vector-borne pathogen gDNA except Cytauxzoon felis (a feline protozoal pathogen). The LSU qPCR assay amplified 12 different Babesia. sp. and C. felis from 31/31 (100%) archived samples, whereas the 18S qPCR amplified only 26/31 (83.9%). By prospective analysis, 19/394 diagnostic accessions (4.8%) were LSU qPCR positive, compared to 11/394 (2.8%) 18S rDNA q

  2. Expression of sheep pathogen Babesia sp. Xinjiang rhoptry-associated protein 1 and evaluation of its diagnostic potential by enzyme-linked immunosorbent assay.

    Science.gov (United States)

    Niu, Qingli; Liu, Zhijie; Yang, Jifei; Yu, Peifa; Pan, Yuping; Zhai, Bintao; Luo, Jianxun; Guan, Guiquan; Yin, Hong

    2016-12-01

    Ovine babesiosis is one of the most important tick-borne haemoparasitic diseases of small ruminants. The ovine parasite Babesia sp. Xinjiang is widespread in China. In this study, recombinant full-length XJrRAP-1aα2 (rhoptry-associated protein 1aα2) and C-terminal XJrRAP-1aα2 CT of Babesia sp. Xinjiang were expressed and used to evaluate their diagnostic potential for Babesia sp. Xinjiang infections by indirect enzyme-linked immunosorbent assay (ELISA). Purified XJrRAP-1aα2 was tested for reactivity with sera from animals experimentally infected with Babesia sp. Xinjiang and other haemoparasites using Western blotting and ELISA. The results showed no cross-reactivities between XJrRAP-1aα2 CT and sera from animals infected by other pathogens. High level of antibodies against RAP-1a usually lasted 10 weeks post-infection (wpi). A total of 3690 serum samples from small ruminants in 23 provinces located in 59 different regions of China were tested by ELISA. The results indicated that the average positive rate was 30·43%, and the infections were found in all of the investigated provinces. This is the first report on the expression and potential use of a recombinant XJrRAP-1aα2 CT antigen for the development of serological assays for the diagnosis of ovine babesiosis, caused by Babesia sp. Xinjiang.

  3. Babesia microti Infection Changes Host Spleen Architecture and Is Cleared by a Th1 Immune Response

    Directory of Open Access Journals (Sweden)

    Vitomir Djokic

    2018-01-01

    Full Text Available Babesia microti is a malaria-like parasite, which infects ∼2000 people annually, such that babesiosis is now a notifiable disease in the United States. Immunocompetent individuals often remain asymptomatic and are tested only after they feel ill. Susceptible C3H/HeJ mice show several human-like disease manifestations and are ideal to study pathogenesis of Babesia species. In this study, we examined parasitemia of B. microti at different time points and assessed its impact on hemoglobin levels in blood, on spleen pathology and overall immune response in C3H/HeJ mice. Peak parasitemia of 42.5% was immediately followed by diminished hemoglobin level. Parasitemia at 21 days of infection was barely detectable by microscopy presented 5.7 × 108 to 5.9 × 109B. microti DNA copies confirming the sensitivity of our qPCR. We hypothesize that qPCR detects DNA released from recently lysed parasites or from extracellular B. microti in blood, which are not easily detected in blood smears and might result in under-diagnosis of babesiosis in patients. Splenectomized patients have been reported to show increased babesiosis severity and result in high morbidity and mortality. These results emphasize the importance of splenic immunity in resolution of B. microti infection. Splenomegaly in infected mice associated with destruction of marginal zone with lysed erythrocytes and released B. microti life forms in our experiments support this premise. At conclusion of the experiment at 21 days post-infection, significant splenic B and T cells depletion and increase in macrophages levels were observed in B. microti infected mice suggesting a role of macrophage in disease resolution. Infected mice also showed significantly higher plasmatic concentration of CD4 Th1 cells secreted cytokines such as IL-2 and IFN-γ while cytokines such as IL-4, IL-5, and IL-13 secreted by Th2 cells increase was not always significant. Thus, Th1 cells-mediated immunity appears to be

  4. The Equine Antimicrobial Peptide eCATH1 Is Effective against the Facultative Intracellular Pathogen Rhodococcus equi in Mice

    Science.gov (United States)

    Schlusselhuber, Margot; Torelli, Riccardo; Martini, Cecilia; Leippe, Matthias; Cattoir, Vincent; Leclercq, Roland; Laugier, Claire; Grötzinger, Joachim; Sanguinetti, Maurizio

    2013-01-01

    Rhodococcus equi, the causal agent of rhodococcosis, is a major pathogen of foals and is also responsible for severe infections in immunocompromised humans. Of great concern, strains resistant to currently used antibiotics have emerged. As the number of drugs that are efficient in vivo is limited because of the intracellular localization of the bacterium inside macrophages, new active but cell-permeant drugs will be needed in the near future. In the present study, we evaluated, by in vitro and ex vivo experiments, the ability of the alpha-helical equine antimicrobial peptide eCATH1 to kill intracellular bacterial cells. Moreover, the therapeutic potential of the peptide was assessed in experimental rhodococcosis induced in mice, while the in vivo toxicity was evaluated by behavioral and histopathological analysis. The study revealed that eCATH1 significantly reduced the number of bacteria inside macrophages. Furthermore, the bactericidal potential of the peptide was maintained in vivo at doses that appeared to have no visible deleterious effects for the mice even after 7 days of treatment. Indeed, daily subcutaneous injections of 1 mg/kg body weight of eCATH1 led to a significant reduction of the bacterial load in organs comparable to that obtained after treatment with 10 mg/kg body weight of rifampin. Interestingly, the combination of the peptide with rifampin showed a synergistic interaction in both ex vivo and in vivo experiments. These results emphasize the therapeutic potential that eCATH1 represents in the treatment of rhodococcosis. PMID:23817377

  5. Equi-frequency contour of photonic crystals with the extended Dirichlet-to-Neumann wave vector eigenvalue equation method

    International Nuclear Information System (INIS)

    Jiang Bin; Zhang Yejing; Wang Yufei; Liu Anjin; Zheng Wanhua

    2012-01-01

    We present the extended Dirichlet-to-Neumann wave vector eigenvalue equation (DtN-WVEE) method to calculate the equi-frequency contour (EFC) of square lattice photonic crystals (PhCs). With the extended DtN-WVEE method and Snell's law, the effective refractive index of the mode with a circular EFC can be obtained, which is further validated with the refractive index weighted by the electric field or magnetic field. To further verify the EFC calculated by the DtN-WVEE method, the finite-difference time-domain method is also used. Compared with other wave vector eigenvalue equation methods that calculate EFC directly, the size of the eigenmatrix used in the DtN-WVEE method is much smaller, and the computation time is significantly reduced. Since the DtN-WVEE method solves wave vectors for given arbitrary frequencies, it can also find applications in studying the optical properties of a PhC with dispersive, lossy and magnetic materials. (paper)

  6. Virulent and Vaccine Strains of Streptococcus equi ssp. zooepidemicus Have Different Influences on Phagocytosis and Cytokine Secretion of Macrophages.

    Science.gov (United States)

    Jie, Peng; Zhe, Ma; Chengwei, Hua; Huixing, Lin; Hui, Zhang; Chengping, Lu; Hongjie, Fan

    2017-01-06

    Swine streptococcosis is a significant threat to the Chinese pig industry, and Streptococcus equi ssp. zooepidemicus (SEZ) is one of the major pathogens. SEZ ATCC35246 is a classical virulent strain, while SEZ ST171 is a Chinese attenuated vaccine strain. In this study, we employed stable isotope labeling by amino acids in cell culture and liquid chromatography-mass spectrometry (LC-MS) to determine the differential response of macrophages to infection by these two strains. Eighty-seven upregulated proteins and 135 downregulated proteins were identified. The proteomic results were verified by real-time polymerase chain reaction for 10 chosen genes and Western blotting for three proteins. All differentially abundant proteins were analyzed for their Gene Ontology and Kyoto Encyclopedia of Genes and Genomes annotations. Certain downregulated proteins were associated with immunity functions, and the upregulated proteins were related to cytomembrane and cytoskeleton regulation. The phagocytosis rate and cytokine genes transcription in Raw264.7 cells during SEZ ATCC35246 and ST171 infection were detected to confirm the bioinformatics results. These results showed that different effects on macrophage phagocytosis and cytokine expression might explain the different phenotypes of SEZ ATCC35246 and ST171 infection. This research provided clues to the mechanisms of host immunity responses to SEZ ST171and SEZ ATCC35246, which could identify potential therapy and vaccine development targets.

  7. The horse pinworm (Oxyuris equi) in archaeology during the Holocene: Review of past records and new data.

    Science.gov (United States)

    Dufour, Benjamin; Hugot, Jean-Pierre; Lepetz, Sébastien; Le Bailly, Matthieu

    2015-07-01

    This paper focuses on the horse pinworm, Oxyuris equi, in archaeology during the Holocene period, and presents an overview of past published occurrences, early mentions in texts, and new data from our paleoparasitology research. This original compilation shows that the most ancient record of the horse pinworm dates to ca. 2500 years before present (ybp) in Central Asia and to ca. 2020 ybp in Western Europe. It also shows that the parasite is not detected on the American continent until contemporary periods. The role of European migrations from 1492 (Christopher Columbus) is discussed to explain the transfer of the horse pinworm from the Old World to the Americas. The absence of any record of this parasite before ca. 2500 ybp in Eurasia could be explained by parasite ecology, unfavorable sampling and scarcity of horse archeological remains. For the Americas, the absence of horse for long periods can be an additional explanation for the absence of the parasite. Copyright © 2015 Elsevier B.V. All rights reserved.

  8. Serial haematology results in transfused and non-transfused dogs naturally infected with Babesia rossi

    Directory of Open Access Journals (Sweden)

    E. Scheepers

    2011-04-01

    Full Text Available This prospective longitudinal study investigated the progression of haematological changes in 32 transfused and 54 non-transfused dogs naturally infected with Babesia rossi over the 1st 6 days following diagnosis and treatment. The effect of patient age on the results of complete blood counts was determined. Haematology data were analysed at presentation and at 24 hours, 3 days and 6 days after presentation. Dogs were treated with diminazene aceturate at diagnosis and a blood transfusion was given if deemed clinically required. Mildly to moderately regenerative normocytic normochromic anaemia was observed in all dogs throughout the study period. Transfused dogs more often had an inflammatory leukogram at presentation and at 24 hours, than dogs that were not transfused. In dogs with a left shift, a concurrent normal or decreased segmented neutrophil count was found more commonly than neutrophilia. Severe thrombocytopenia that resolved within a week was common. Blood transfusion alleviated the anaemia, but had no significant effect on white blood cell or platelet responses. Blood cell responses were not significantly influenced by age. In conclusion, the red blood cell and white blood cell responses were less than expected in dogs with babesiosis, given the degree of anaemia and inflammation present. The magnitude of thrombocytopenia and rapid return of the platelet count to normal suggested a possible immune-mediated mechanism for the thrombocytopenia.

  9. Field evaluation of an exoantigen-containing Babesia vaccine in Venezuela

    Directory of Open Access Journals (Sweden)

    S. Montenegro-James

    1992-01-01

    Full Text Available Bovine babesiosis is endemic in Venezuela, causing significant losses in highly susceptible imported cattle. Current immunoprphylatic methods include the less desirable use of live parasites. Inactivated vaccines derived from exoantigen-containing supernatant fluids of in vitro Babesia bovis and B. bigemina cultures have been developed and constitute a major improvement in vaccine safety, stability and ease of handling. Vaccination trials conducted under field conditions provide the final evaluation of a culture-derived B. bovis-B. bigemina vaccine. During a 5-year period, approximately 8,000 cattle were vaccinated and 16 clinical trials carried out in. 7 states of Venezuela Clinical, serologic and parasitologic data were collected monthly from 10% of the animals over a 2-year period. Data were also collected from a similar number of nonvaccinated control cattle. Analysis of results from these trials demonstrated a reduction in the incidence of clinical disease among vaccinated animals and complete protection against mortality among vaccinated and nonvaccinated cattle. Use of this inactivated vaccine offers the best combination od safety, potency and efficacy for thew immunoprophylatic control of bovine babesiosis.

  10. Evaluation of an ELISA kit in the serological diagnosis of Babesia bovis for epidemiological studies

    International Nuclear Information System (INIS)

    Martins, J.R.; Correa, B.L.; Cereser, V.H.; Artiles, J.M.; Alves-Branco, F.P.J.

    1998-01-01

    An enzyme linked immunosorbent assay (ELISA) kit for detect antibodies to Babesia bovis, an intraerytrocitic bovine parasite was evaluated using known negative and positive samples and the results were compared with an indirect immunofluorescent antibody test (IFAT). Results obtained with field samples were used to estimate seroprevalence of B. bovis in an endemic area to the cattle tick (Boophilus microplus) vector of bovine babesiosis. Percentage of positivity (PP) values (optical density of tested serum/mean optical density of positive control) on 274 negative samples, had major values ranged in the frequency of 4.0 to 7.0 PP. Comparison between ELISA and IFAT showed an agreement of 93.3% on field sera samples, collected in areas of low, good and high soil fertility in the region of Bage (31 degree 20 min 13 sec S, 54 degree 06 min 21 sec W), RS, Brazil. From 5,082 tested sera, 3,751 (73%) were positive for B. bovis antibodies. No significant difference (P>0.05) was observed between results from calves living in areas of low and good soil fertility (80 and 82% of seroprevalence, respectively). However, calves living in soil of high fertility showed a minor inoculation rate for B. bovis (63% of seroprevalence), indicating needs of measures to prevent losses due to babesiosis. (author)

  11. Field evaluation of an exoantigen-containing Babesia vaccine in Venezuela.

    Science.gov (United States)

    Montenegro-James, S; Toro, M; Leon, E; Guillen, A T

    1992-01-01

    Bovine babesiosis is endemic in Venezuela, causing significant losses in highly susceptible imported cattle. Current immunoprophylactic methods include the less desirable use of live parasites. Inactivated vaccines derived from exoantigen-containing supernatant fluids of in vitro Babesia bovis and B. bigemina cultures have been developed and constitute a major improvement in vaccine safety, stability and ease of handling. Vaccination trials conducted under field conditions provide the final evaluation of a culture-derived B. bovis-B. bigemina vaccine. During a 5-year period, approximately 8,000 cattle were vaccinated and 16 clinical trials carried out in 7 states of Venezuela. Clinical, serologic and parasitologic data were collected monthly from 10% of the animals over a 2-year period. Data were also collected from a similar number of nonvaccinated control cattle. Analysis of results from these trials demonstrated a reduction in the incidence of clinical disease among vaccinated animals and complete protection against mortality caused by babesiosis. Vaccine efficacy was measured calculating the incidence rates of disease and mortality among vaccinated and nonvaccinated cattle. Use of this inactivated vaccine offers the best combination of safety, potency and efficacy for the effective immunoprophylactic control of bovine babesiosis.

  12. Babesia major: protection of intact calves against homologous challenge by the injection of irradiated piroplasms

    International Nuclear Information System (INIS)

    Purnell, R.E.; Lewis, D.; Brocklesby, D.W.

    1979-01-01

    Blood from a splenectomized calf infected with Babesia major was divided into 20 ml aliquots which were γ-irradiated at doses of 0, 23.3, 27.3, 31.4, 35.4 and 39.5 krad and then inoculated into groups of three intact calves. Animals receiving non-irradiated blood had typical mild B. major reactions, but those receiving blood irradiated at 23.3, 27.3 and 31.4 krad and 2 of 3 receiving blood irradiated at 35.4 krad had minimal reactions. The remaining 4 animals had no detectable parasitaemic reactions. When the calves were challenged with a similar number (6.0 x 10 9 ) of homologous parasites, they were all immune with the exception of the 4 animals which had not reacted initially. The immune status of individual cattle was reflected accurately in the results of the micro-ELISA test, which detected a significant rise in serum antibody titre of the 4 susceptible animals 7 days after challenge. (author)

  13. Babesia major: protection of intact calves against homologous challenge by the injection of irradiated piroplasms

    Energy Technology Data Exchange (ETDEWEB)

    Purnell, R E; Lewis, D; Brocklesby, D W [Agricultural Research Council, Compton (UK). Inst. for Research on Animal Diseases

    1979-02-01

    Blood from a splenectomized calf infected with Babesia major was divided into 20 ml aliquots which were ..gamma..-irradiated at doses of 0, 23.3, 27.3, 31.4, 35.4 and 39.5 krad and then inoculated into groups of three intact calves. Animals receiving non-irradiated blood had typical mild B. major reactions, but those receiving blood irradiated at 23.3, 27.3 and 31.4 krad and 2 of 3 receiving blood irradiated at 35.4 krad had minimal reactions. The remaining 4 animals had no detectable parasitaemic reactions. When the calves were challenged with a similar number (6.0 x 10/sup 9/) of homologous parasites, they were all immune with the exception of the 4 animals which had not reacted initially. The immune status of individual cattle was reflected accurately in the results of the micro-ELISA test, which detected a significant rise in serum antibody titre of the 4 susceptible animals 7 days after challenge.

  14. Babesia divergens: protection of intact calves against heterologous challenge by the injection of irradiated piroplasms

    Energy Technology Data Exchange (ETDEWEB)

    Lewis, D; Purnell, R E; Brocklesby, D W [Agricultural Research Council, Compton (UK). Inst. for Research on Animal Diseases

    1980-03-01

    Blood from a splenectomized calf infected with Babesia divergens was divided into 20 ml aliquots which were ..gamma..-irradiated at doses of 0, 24, 28, 32, 36 and 40 krad and then inoculated into groups of three intact calves. Animals receiving non-irradiated blood had typical mild B. divergens reactions, but those receiving irradiated blood had either very mild reactions or no overt reaction. When the calves were challenged with a similar number (7.5 X 10/sup 9/) of heterologous parasites of a recently-isolated field strain, those which had received blood irradiated at 0, 24, 28 and 32 krad were all immune whereas four of the five surviving animals which had received blood irradiated at 36 or 40 krad were susceptible. The immune status of individual cattle was reflected accurately in the results of the micro-ELISA test, which detected a significant rise in serum antibody titre of the four susceptible animals by day 21 after challenge.

  15. Investigation of hematological and biochemical parameters in small ruminants naturally infected with Babesia ovis

    Directory of Open Access Journals (Sweden)

    Bijan Esmaeilnejad

    2012-03-01

    Full Text Available Babesia ovis plays an important role in causing anemia and kidney dysfunction in affected animals. There are few extensive studies about hematological and biochemical findings of small ruminants' babesiosis caused by B. ovis. The aim of this study was to evaluate the effect of babesiosis on some hematological and biochemical parameters in infected small ruminants with B. ovis. A total of 280 sheep and 122 goats from 40 herds were randomly examined for the presence of B. ovis in blood samples. Of 402 samples, 67 animals (16.7% were positive for B. ovis of which 52 (18.5% were sheep and 15 (12.2% goats, respectively. The infected animals were divided into four subgroups according to parasitemia rates (<1%, 1%, 2%, and 3%. As a control group, 67 uninfected animals were also selected from the same farms. With increase in parasitemia rates, hemoglobin concentration (Hb, packed cell volume (PCV, red blood cells (RBCs, mean corpuscular volume (MCV and mean corpuscular hemoglobin concentration (MCHC significantly decreased (P < 0.05, while, total leukocyte count, number of lymphocyte, monocyte, neutrophil and eosinophil showed a significant increase (P < 0.05. Infected animals presented a significant elevation (P < 0.05 of total proteins and significantly lower level (P < 0.05 of albumin compared to non-infected animals. Significant elevation (P < 0.05 of BUN, creatinine, cholestrol, triglyceride, HDL and LDL level were found with parasitemia progression.

  16. Human Babesiosis Caused by Babesia duncani Has Widespread Distribution across Canada

    Directory of Open Access Journals (Sweden)

    John D. Scott

    2018-05-01

    Full Text Available Human babesiosis caused by Babesia duncani is an emerging infectious disease in Canada. This malaria-like illness is brought about by a protozoan parasite infecting red blood cells. Currently, controversy surrounds which tick species are vectors of B. duncani. Since the availability of a serological or molecular test in Canada for B. duncani has been limited, we conducted a seven-year surveillance study (2011–2017 to ascertain the occurrence and geographic distribution of B. duncani infection country-wide. Surveillance case data for human B. duncani infections were collected by contacting physicians and naturopathic physicians in the United States and Canada who specialize in tick-borne diseases. During the seven-year period, 1119 cases were identified. The presence of B. duncani infections was widespread across Canada, with the highest occurrence in the Pacific coast region. Patients with human babesiosis may be asymptomatic, but as this parasitemia progresses, symptoms range from mild to fatal. Donors of blood, plasma, living tissues, and organs may unknowingly be infected with this piroplasm and are contributing to the spread of this zoonosis. Our data show that greater awareness of human babesiosis is needed in Canada, and the imminent threat to the security of the Canadian blood supply warrants further investigation. Based on our epidemiological findings, human babesiosis should be a nationally notifiable disease in Canada. Whenever a patient has a tick bite, health practitioners must watch for B. duncani infections, and include human babesiosis in their differential diagnosis.

  17. Human Babesiosis Caused by Babesia duncani Has Widespread Distribution across Canada.

    Science.gov (United States)

    Scott, John D; Scott, Catherine M

    2018-05-17

    Human babesiosis caused by Babesia duncani is an emerging infectious disease in Canada. This malaria-like illness is brought about by a protozoan parasite infecting red blood cells. Currently, controversy surrounds which tick species are vectors of B. duncani. Since the availability of a serological or molecular test in Canada for B. duncani has been limited, we conducted a seven-year surveillance study (2011⁻2017) to ascertain the occurrence and geographic distribution of B. duncani infection country-wide. Surveillance case data for human B. duncani infections were collected by contacting physicians and naturopathic physicians in the United States and Canada who specialize in tick-borne diseases. During the seven-year period, 1119 cases were identified. The presence of B. duncani infections was widespread across Canada, with the highest occurrence in the Pacific coast region. Patients with human babesiosis may be asymptomatic, but as this parasitemia progresses, symptoms range from mild to fatal. Donors of blood, plasma, living tissues, and organs may unknowingly be infected with this piroplasm and are contributing to the spread of this zoonosis. Our data show that greater awareness of human babesiosis is needed in Canada, and the imminent threat to the security of the Canadian blood supply warrants further investigation. Based on our epidemiological findings, human babesiosis should be a nationally notifiable disease in Canada. Whenever a patient has a tick bite, health practitioners must watch for B. duncani infections, and include human babesiosis in their differential diagnosis.

  18. Molecular Confirmation of Trypanosoma evansi and Babesia bigemina in Cattle from Lower Egypt

    Directory of Open Access Journals (Sweden)

    Mahmoud M. Elhaig, Abdelfattah Selim, Mohamed M. Mahmoud and Eman K El-Gayar

    2016-11-01

    Full Text Available Trypanosomosis and babesiosis are economically important vector-borne diseases for animal health and productivity in developing countries. In Egypt, molecular epidemiological surveys on such diseases are scarce. In the present study, we examined 475 healthy and 25 clinically diagnosed cattle from three provinces in Lower Egypt, for Trypanosoma (T. and Babesia (B. infections using an ITS1 PCR assay that confirmed Trypanosoma species presence and an 18S rRNA assay that detected B. bigemina. Results confirmed Trypanosoma spp. and B. bigemina presence in 30.4% and 11% individuals, respectively, with eight animals (1.6% being co-infected with both hemoparasites. Subsequent type-specific PCRs revealed that all Trypanosoma PCR positive samples corresponded to T. evansi and that none of the animals harboured T. brucei gambiense or T. brucei rhodesiense. Nucleotide sequencing of the variable surface glycoprotein revealed the T. evansi cattle strain to be most closely related (99% nucleotide sequence identity to strains previously detected in dromedary camels in Egypt, while the 18S rRNA gene phylogeny confirmed the presence of a unique B. bigemina haplotype closely related to strains from Turkey and Brazil. Statistically significant differences in PCR prevalence were noted with respect to gender, clinical status and locality. These results confirm the presence of high numbers of carrier animals and signal the need for expanded surveillance and control efforts.

  19. Babesia divergens: protection of intact calves against heterologous challenge by the injection of irradiated piroplasms

    International Nuclear Information System (INIS)

    Lewis, D.; Purnell, R.E.; Brocklesby, D.W.

    1980-01-01

    Blood from a splenectomized calf infected with Babesia divergens was divided into 20 ml aliquots which were γ-irradiated at doses of 0, 24, 28, 32, 36 and 40 krad and then inoculated into groups of three intact calves. Animals receiving non-irradiated blood had typical mild B. divergens reactions, but those receiving irradiated blood had either very mild reactions or no overt reaction. When the calves were challenged with a similar number (7.5 X 10 9 ) of heterologous parasites of a recently-isolated field strain, those which had received blood irradiated at 0, 24, 28 and 32 krad were all immune whereas four of the five surviving animals which had received blood irradiated at 36 or 40 krad were susceptible. The immune status of individual cattle was reflected accurately in the results of the micro-ELISA test, which detected a significant rise in serum antibody titre of the four susceptible animals by day 21 after challenge. (Auth.)

  20. In vitro cultivation and cryopreservation of Babesia bigemina sporokinetes in hemocytes of Rhipicephalus microplus.

    Science.gov (United States)

    de Rezende, Jania; Rangel, Charles P; McIntosh, Douglas; Silveira, Júlia A G; Cunha, Nathalie C; Ramos, Carlos A N; Fonseca, Adivaldo H

    2015-09-15

    Cultures of tick hemocytes represent alternative cell lines for the isolation and cultivation of a variety of hemoparasites. The present study reports the development and evaluation of methods for the in vitro culture and maintenance of sporokinetes of Babesia bigemina in association with hemocytes of the tick Rhipicephalus microplus. Hemolymph, from engorged females infected with B. bigemina sporokinetes, was incubated at 28 °C in L15 culture medium supplemented with 40% fetal bovine serum. Adherence of hemocytes to flask surfaces and the development of B. bigemina sporokinetes commenced on the first day of cultivation. The protozoa demonstrated clear motility and the capacity to adhere to hemocyte membranes for up to 25 days, at which time the hemocytes began to show signs of degeneration. Examination of Giemsa stained hemocyte cultures, revealed the presence of pyriformis forms, as well as mature and immature sporokinetes with dark red nuclei, centralized or near the apical extremities. Sporokinetes harvested from culture supernatants were cryopreserved in liquid nitrogen. Inoculation of parasite-free hemocyte cultures with defrosted sporokinetes, demonstrated the viability and interaction of the protozoa with the hemocytes over 21 days. Cultured hemocytes of R. microplus hold potential for development as a tool in the study of host parasite interactions and as a substrate for the in vitro maintenance of B. bigemina sporokinetes. Copyright © 2015 Elsevier B.V. All rights reserved.

  1. Some aspects of the epidemiology of Babesia bovis in Santana do Livramento, Southern Brazil

    International Nuclear Information System (INIS)

    Martins, J.R.; Correa, B.L.; Cereser, V.H.; Arteche, C.C.P.; Guglielmone, A.A.

    1998-01-01

    Some aspects of the epidemiology of Babesia bovis were studied in Santana do Livramento, Rio Grande do Sul, Brazil by analysing cattle raising practices applied to 101 herds and by diagnosing B. bovis antibodies in cattle of about 11 months old using an enzyme linked immunosorbent assay. Herds with prevalence of antibodies ranging between 15% to 80% were considered at risk of babesiosis outbreaks of economic importance (enzootic instability). 53% of herds were found in enzootic instability to B. bovis. The proportion of Bos taurus and B. indicus x B. taurus herds in instability were similar (P=0.771, qui square) and the number of acaricides treatments applied yearly had no influence in the instability to B. bovis (P=0.866, chi square). Herds maintained along with sheep in a ratio < 1.5 (P=0.012, chi square); this probability was further increased in herds maintained on properties greater than 500 ha (P=0.057, chi square). High B. bovis antibody prevalence was found in B. taurus x B. indicus herds subjected to an average of 5.8 tick treatments yearly with long residual period acaricides, indicating misuse of the chemicals or tick resistance to them. The epidemiological situation to B. bovis seems to justify vaccination to avoid economic losses in herds in enzootic instability and those in enzootic stability due to low antibody prevalence. (author)

  2. FREQÜÊNCIA DE CÃES INFECTADOS POR Babesia spp. EM CAMPOS DOS GOYTACAZES, RJ FREQUENCY OF DOGS INFECTED BY Babesia spp. IN CAMPOS DOS GOYTACAZES, RJ

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    Orlando Augusto Melo JR

    2008-04-01

    Full Text Available A babesiose canina é transmitida pelo carrapato Rhipicephalus sanguineus e causada por protozoários intra-eritrocitários do gênero Babesia, sendo a B. canis e a B. gibsoni os agentes etiológicos. Comumente há quadros de anemia hemolítica, febre e letargia, anorexia, hematúria e esplenomegalia, com a patogenia relacionada principalmente à multiplicação destes parasitos nas hemácias dos hospedeiros. Tendo em vista a pesquisa de hemoparasitos, no presente trabalho foram realizados exames microscópicos de 2.031 esfregaços sangüíneos delgados, preparados com sangue capilar colhido por perfuração da orelha. Baseou-se o diagnóstico na pesquisa direta, visualizando-se os agentes etiológicos. Trinta cães (1,47% foram considerados infectados por Babesia spp., confirmando, assim, a presença destes parasitos na cidade de Campos dos Goytacazes, onde o clima é favorável ao desenvolvimento do vetor natural
    PALAVRAS-CHAVES: Babesia spp., cão, hemoparasito, ocorrência
    The canine babesiosis is transmitted by the Rhipicephalus sanguineus tick and caused by the intracytoplasmatic protozoa of the Babesia genus, being the B. canis and the B. gibsoni the etiologic agents. Normally there are hemolytic anemia, fever and lethargy, anorexy, hematuria and splenomegaly, with the pathogenesis related mainly to the multiplication of these parasites in the erythrocytes of the hosts. In the present work 2

  3. Molecular biological identification of Babesia, Theileria, and Anaplasma species in cattle in Egypt using PCR assays, gene sequence analysis and a novel DNA microarray.

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    El-Ashker, Maged; Hotzel, Helmut; Gwida, Mayada; El-Beskawy, Mohamed; Silaghi, Cornelia; Tomaso, Herbert

    2015-01-30

    In this preliminary study, a novel DNA microarray system was tested for the diagnosis of bovine piroplasmosis and anaplasmosis in comparison with microscopy and PCR assay results. In the Dakahlia Governorate, Egypt, 164 cattle were investigated for the presence of piroplasms and Anaplasma species. All investigated cattle were clinically examined. Blood samples were screened for the presence of blood parasites using microscopy and PCR assays. Seventy-one animals were acutely ill, whereas 93 were apparently healthy. In acutely ill cattle, Babesia/Theileria species (n=11) and Anaplasma marginale (n=10) were detected. Mixed infections with Babesia/Theileria spp. and A. marginale were present in two further cases. A. marginale infections were also detected in apparently healthy subjects (n=23). The results of PCR assays were confirmed by DNA sequencing. All samples that were positive by PCR for Babesia/Theileria spp. gave also positive results in the microarray analysis. The microarray chips identified Babesia bovis (n=12) and Babesia bigemina (n=2). Cattle with babesiosis were likely to have hemoglobinuria and nervous signs when compared to those with anaplasmosis that frequently had bloody feces. We conclude that clinical examination in combination with microscopy are still very useful in diagnosing acute cases of babesiosis and anaplasmosis, but a combination of molecular biological diagnostic assays will detect even asymptomatic carriers. In perspective, parallel detection of Babesia/Theileria spp. and A. marginale infections using a single microarray system will be a valuable improvement. Copyright © 2015 The Authors. Published by Elsevier B.V. All rights reserved.

  4. Detection of Theileria and Babesia in brown brocket deer (Mazama gouazoubira) and marsh deer (Blastocerus dichotomus) in the State of Minas Gerais, Brazil.

    Science.gov (United States)

    da Silveira, Júlia A G; Rabelo, Elida M L; Ribeiro, Múcio F B

    2011-04-19

    Intraerythrocytic protozoan species of the genera Theileria and Babesia are known to infect both wild and domestic animals, and both are transmitted by hard-ticks of the family Ixodidae. The prevalences of hemoprotozoa and ectoparasites in 15 free-living Mazama gouazoubira, two captive M. gouazoubira and four captive Blastocerus dichotomus from the State of Minas Gerais, Brazil, have been determined through the examination of blood smears and the use of nested polymerase chain reaction (nPCR). The cervid population was inspected for the presence of ticks and any specimens encountered were identified alive under the stereomicroscope. Blood samples were collected from all 21 animals, following which blood smears were prepared, subjected to quick Romanowsky staining and examined under the optical microscope. DNA was extracted with the aid of commercial kits from cervid blood samples and from tick salivary glands. The nPCR assay comprised two amplification reactions: the first was conducted using primers specific for a 1700 bp segment of the 18S rRNA gene of Babesia and Theileria species, whilst the second employed primers designed to amplify a common 420 bp Babesia 18S rRNA fragment identified by aligning sequences from Babesia spp. available at GenBank. The ticks Amblyomma cajennense, Rhipicephalus microplus and Dermacentor nitens were identified in various of the cervids examined. Of the animals investigated, 71.4% (15/21) were infected with hemoprotozoa, including Theileria cervi (47.6%), Theileria sp. (14.3%), Babesia bovis (4.8%) and Babesia bigemina (4.8%). However, only one of the infected wild cervids exhibited accentuated anaemia (PCV=17%). This is first report concerning the occurrence of Theileria spp. in Brazilian cervids. Copyright © 2010 Elsevier B.V. All rights reserved.

  5. Estudo genômico do nível de infecção por Babesia bovis em bovinos da raça angus

    OpenAIRE

    Santana, Clarissa Helena [UNESP

    2016-01-01

    A bovinocultura é um setor com importante destaque no agronegócio brasileiro. O carrapato Ripicephalus (Boophilus) microplus é responsável por perdas econômicas significativas aos pecuaristas e é vetor de hemoparasitoses como Anaplasma spp e Babesia spp. Sabe-se que os bovinos Bos taurus taurus são mais susceptíveis à infestação por carrapatos do que Bos taurus indicus. Acredita-se que o mesmo ocorra para a infecção por Babesia bovis. Neste trabalho, foram avaliados, em duas colheitas, 355 bo...

  6. Parasitological and molecular detection of Babesia canis vogeli in dogs of Recife, Pernambuco and evaluation of risk factors associated

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    Vanessa Carla Lima da Silva

    2016-02-01

    Full Text Available This work aims to detect the presence of Babesia canis vogeli in dogs from Recife, Pernambuco via molecular and parasitological detection methods, and to assess the risk factors associated with this parasite. A total of 146 dogs (male and female of varying breeds and ages that presented clinical symptoms of babesiosis were assessed at a clinical care center in the Veterinary School Hospital. Blood was obtained via venopuncture for hemoparasite detection and polymerase chain reaction (PCR. Using a commercial kit, DNA was extracted from blood samples. For the PCR reaction, an approximately 590 base pair long genetic sequence was used to detect the presence of B. canis vogeli. The forward primer, denoted as BAB1 (5’-GTG AAC CTT ATC ACT TAA AGG-3’, was specific for a conserved region on the 18S rRNA gene of Babesia spp., and the antisense primer was denoted as BAB4 (5’-CAA CTC CTC CAC GCA ATC G-3’. PCR results suggested that the percentage of Babesia canis vogeli infection was 4.8%. Through descriptive statistical analysis of the data, we observed that there was higher frequency of parasite infection associated with male dogs above two years of age, with a defined breed, from the countryside, are domiciled, and also suffer from tick infestation. We conclude that regardless of the type of risk factor, babesiosis can be found throughout Recife, Pernambuco, and its prevalence does not vary in most regions of Brazil. Our results indicate that PCR is a sensitive test for the detection of blood parasites, and should be performed as a clinical routine.

  7. Questing Dermacentor reticulatus harbouring Babesia canis DNA associated with outbreaks of canine babesiosis in the Swiss Midlands.

    Science.gov (United States)

    Schaarschmidt, Daniel; Gilli, Urs; Gottstein, Bruno; Marreros, Nelson; Kuhnert, Peter; Daeppen, Jérôme A; Rosenberg, Gertrud; Hirt, Didier; Frey, Caroline F

    2013-06-01

    In 2011 and 2012, outbreaks of clinical canine babesiosis were observed in 2 areas of the Swiss Midlands that had no history of this disease so far. In one area, cases of canine babesiosis occurred over 2 consecutive tick seasons. The outbreaks involved 29 dogs, 4 of which died. All dogs were infected with large Babesia sp. as diagnosed in Giemsa-stained blood smears and/or PCR. These were identified as B. canis (formerly known as B. canis canis) by subsequent partial sequencing of the 18S rRNA gene of Babesia sp. Interestingly, the sequence indicated either a genotype with heterogeneity in the ssrRNA gene copies or double infection with different B. canis isolates. None of the dogs had a recent travel history, but one had frequently travelled to Hungary and had suffered twice from clinical babesiosis 18 and 24 months prior to the outbreak in autumn 2011. Retrospective sequencing of a stored blood DNA sample of this dog revealed B. canis, with an identical sequence to the Babesia involved in the outbreaks. For the first time in Switzerland, the partial 18S rRNA gene of B. canis could be amplified from DNA isolated from 19 out of 23 adult Dermacentor reticulatus ticks flagged in the same area. The sequence was identical to that found in the dogs. Furthermore, one affected dog carried a female D. reticulatus tick harbouring B. canis DNA. Our findings illustrate that, under favourable biogeographic and climatic conditions, the life-cycle of B. canis can relatively rapidly establish itself in previously non-endemic areas. Canine babesiosis should therefore always be a differential diagnosis when dogs with typical clinical signs are presented, regardless of known endemic areas. Copyright © 2013 Elsevier GmbH. All rights reserved.

  8. Using msa-2b as a molecular marker for genotyping Mexican isolates of Babesia bovis.

    Science.gov (United States)

    Genis, Alma D; Perez, Jocelin; Mosqueda, Juan J; Alvarez, Antonio; Camacho, Minerva; Muñoz, Maria de Lourdes; Rojas, Carmen; Figueroa, Julio V

    2009-12-01

    Variable merozoite surface antigens of Babesia bovis are exposed glycoproteins having a role in erythrocyte invasion. Members of this gene family include msa-1 and msa-2 (msa-2c, msa-2a(1), msa-2a(2) and msa-2b). To determine the sequence variation among B. bovis Mexican isolates using msa-2b as a genetic marker, PCR amplicons corresponding to msa-2b were cloned and plasmids carrying the corresponding inserts were purified and sequenced. Comparative analysis of nucleotide and deduced amino acid sequences revealed distinct degrees of variability and identity among the coding gene sequences obtained from 16 geographically different Mexican B. bovis isolates and a reference strain. Clustal-W multiple alignments of the MSA-2b deduced amino acid sequences performed with the 17 B. bovis Mexican isolates, revealed the identification of three genotypes with a distinct set each of amino acid residues present at the variable region: Genotype I represented by the MO7 strain (in vitro culture-derived from the Mexico isolate) as well as RAD, Chiapas-1, Tabasco and Veracruz-3 isolates; Genotype II, represented by the Jalisco, Mexico and Veracruz-2 isolates; and Genotype III comprising the sequences from most of the isolates studied, Tamaulipas-1, Chiapas-2, Guerrero-1, Nayarit, Quintana Roo, Nuevo Leon, Tamaulipas-2, Yucatan and Guerrero-2. Moreover, these three genotypes could be discriminated against each other by using a PCR-RFLP approach. The results suggest that occurrence of indels within the variable region of msa-2b sequences can be useful markers for identifying a particular genotype present in field populations of B. bovis isolated from infected cattle in Mexico.

  9. Diversity of Babesia bovis merozoite surface antigen genes in the Philippines.

    Science.gov (United States)

    Tattiyapong, Muncharee; Sivakumar, Thillaiampalam; Ybanez, Adrian Patalinghug; Ybanez, Rochelle Haidee Daclan; Perez, Zandro Obligado; Guswanto, Azirwan; Igarashi, Ikuo; Yokoyama, Naoaki

    2014-02-01

    Babesia bovis is the causative agent of fatal babesiosis in cattle. In the present study, we investigated the genetic diversity of B. bovis among Philippine cattle, based on the genes that encode merozoite surface antigens (MSAs). Forty-one B. bovis-positive blood DNA samples from cattle were used to amplify the msa-1, msa-2b, and msa-2c genes. In phylogenetic analyses, the msa-1, msa-2b, and msa-2c gene sequences generated from Philippine B. bovis-positive DNA samples were found in six, three, and four different clades, respectively. All of the msa-1 and most of the msa-2b sequences were found in clades that were formed only by Philippine msa sequences in the respective phylograms. While all the msa-1 sequences from the Philippines showed similarity to those formed by Australian msa-1 sequences, the msa-2b sequences showed similarity to either Australian or Mexican msa-2b sequences. In contrast, msa-2c sequences from the Philippines were distributed across all the clades of the phylogram, although one clade was formed exclusively by Philippine msa-2c sequences. Similarities among the deduced amino acid sequences of MSA-1, MSA-2b, and MSA-2c from the Philippines were 62.2-100, 73.1-100, and 67.3-100%, respectively. The present findings demonstrate that B. bovis populations are genetically diverse in the Philippines. This information will provide a good foundation for the future design and implementation of improved immunological preventive methodologies against bovine babesiosis in the Philippines. The study has also generated a set of data that will be useful for futher understanding of the global genetic diversity of this important parasite. © 2013.

  10. Genotypic diversity of merozoite surface antigen 1 of Babesia bovis within an endemic population.

    Science.gov (United States)

    Lau, Audrey O T; Cereceres, Karla; Palmer, Guy H; Fretwell, Debbie L; Pedroni, Monica J; Mosqueda, Juan; McElwain, Terry F

    2010-08-01

    Multiple genetically distinct strains of a pathogen circulate and compete for dominance within populations of animal reservoir hosts. Understanding the basis for genotypic strain structure is critical for predicting how pathogens respond to selective pressures and how shifts in pathogen population structure can lead to disease outbreaks. Evidence from related Apicomplexans such as Plasmodium, Toxoplasma, Cryptosporidium and Theileria suggests that various patterns of population dynamics exist, including but not limited to clonal, oligoclonal, panmictic and epidemic genotypic strain structures. In Babesia bovis, genetic diversity of variable merozoite surface antigen (VMSA) genes has been associated with disease outbreaks, including in previously vaccinated animals. However, the extent of VMSA diversity within a defined population in an endemic area has not been examined. We analyzed genotypic diversity and temporal change of MSA-1, a member of the VMSA family, in individual infected animals within a reservoir host population. Twenty-eight distinct MSA-1 genotypes were identified within the herd. All genotypically distinct MSA-1 sequences clustered into three groups based on sequence similarity. Two thirds of the animals tested changed their dominant MSA-1 genotypes during a 6-month period. Five animals within the population contained multiple genotypes. Interestingly, the predominant genotypes within those five animals also changed over the 6-month sampling period, suggesting ongoing transmission or emergence of variant MSA-1 genotypes within the herd. This study demonstrated an unexpected level of diversity for a single copy gene in a haploid genome, and illustrates the dynamic genotype structure of B. bovis within an individual animal in an endemic region. Co-infection with multiple diverse MSA-1 genotypes provides a basis for more extensive genotypic shifts that characterizes outbreak strains.

  11. Genetic diversity and antigenicity variation of Babesia bovis merozoite surface antigen-1 (MSA-1) in Thailand.

    Science.gov (United States)

    Tattiyapong, Muncharee; Sivakumar, Thillaiampalam; Takemae, Hitoshi; Simking, Pacharathon; Jittapalapong, Sathaporn; Igarashi, Ikuo; Yokoyama, Naoaki

    2016-07-01

    Babesia bovis, an intraerythrocytic protozoan parasite, causes severe clinical disease in cattle worldwide. The genetic diversity of parasite antigens often results in different immune profiles in infected animals, hindering efforts to develop immune control methodologies against the B. bovis infection. In this study, we analyzed the genetic diversity of the merozoite surface antigen-1 (msa-1) gene using 162 B. bovis-positive blood DNA samples sourced from cattle populations reared in different geographical regions of Thailand. The identity scores shared among 93 msa-1 gene sequences isolated by PCR amplification were 43.5-100%, and the similarity values among the translated amino acid sequences were 42.8-100%. Of 23 total clades detected in our phylogenetic analysis, Thai msa-1 gene sequences occurred in 18 clades; seven among them were composed of sequences exclusively from Thailand. To investigate differential antigenicity of isolated MSA-1 proteins, we expressed and purified eight recombinant MSA-1 (rMSA-1) proteins, including an rMSA-1 from B. bovis Texas (T2Bo) strain and seven rMSA-1 proteins based on the Thai msa-1 sequences. When these antigens were analyzed in a western blot assay, anti-T2Bo cattle serum strongly reacted with the rMSA-1 from T2Bo, as well as with three other rMSA-1 proteins that shared 54.9-68.4% sequence similarity with T2Bo MSA-1. In contrast, no or weak reactivity was observed for the remaining rMSA-1 proteins, which shared low sequence similarity (35.0-39.7%) with T2Bo MSA-1. While demonstrating the high genetic diversity of the B. bovis msa-1 gene in Thailand, the present findings suggest that the genetic diversity results in antigenicity variations among the MSA-1 antigens of B. bovis in Thailand. Copyright © 2016 Elsevier B.V. All rights reserved.

  12. Riboflavin and ultraviolet light reduce the infectivity of Babesia microti in whole blood.

    Science.gov (United States)

    Tonnetti, Laura; Thorp, Aaron M; Reddy, Heather L; Keil, Shawn D; Goodrich, Raymond P; Leiby, David A

    2013-04-01

    Babesia microti is the parasite most frequently transmitted by blood transfusion in the United States. Previous work demonstrated the efficacy of riboflavin (RB) and ultraviolet (UV) light to inactivate B.microti in apheresis plasma and platelet units. In this study we investigated the effectiveness of RB and UV light to reduce the levels of B.microti in whole blood (WB). WB units were spiked with B. microti-infected hamster blood. Spearman-Karber methods were used to calculate infectivity of each sample in terms of hamster infectious dose 50% (HID50 ) value. After RB addition, the units were illuminated with 80 J/mLRBC UV light. Two samples were collected: one before illumination and one after illumination. The samples were serially diluted and dilutions injected into a group of five naive hamsters. Four weeks postinoculation (PI), blood was collected from the animals and evaluated by microscopic observation. One pilot study showed a good dose response in the animals and demonstrated that sample infectivity could be calculated in terms of an HID50 . Three additional replicates were performed in the same manner as the pilot study, but with fewer dilutions. Infectivity values were consistent between the experiments and were used to calculate log reduction. The posttreatment reduction of B. microti for all the experiments was more than 5 log. The data collected indicate that use of RB and UV is able to decrease the parasite load in WB units thus reducing the risk of transfusion-transmitted B. microti from blood components containing B. microti-infected RBCs. © 2012 American Association of Blood Banks.

  13. Development of a microtitre-based spectrophotometric method to monitor Babesia divergens growth in vitro.

    Science.gov (United States)

    Malandrin, Laurence; Marchand, Anne-Marie; Chauvin, Alain

    2004-09-01

    Babesia divergens multiplication cycle involves erythrocyte invasion, intracellular division, and erythrocyte lysis with the simultaneous liberation of hemoglobin. We have decided to set up a spectrophotometric protocol based on hemoglobin concentration in the culture supernatants to monitor B. divergens in vitro growth. After the selection of 405 nm as the most appropriate endpoint hemoglobin wavelength in our conditions (hemoglobin concentration in the supernatant), cultures were standardized [1 x 10(9) red blood cell (RBC)/ml, 1-2.5 x 10(5) infected red blood cell (iRBC)/ml] to allow their monitoring over 3 days. The protocol was then compared to the most commonly used growth measurement methods: parasitemia counting and [(3)H]hypoxanthine incorporation. An excellent correlation was demonstrated between A(405) of the culture supernatant and parasitemia of the iRBC, whatever the RBC concentration used in the medium. This correlation was also evidenced between A(405) and [(3)H]hypoxanthine incorporation for [(3)H]hypoxanthine concentrations lower than 4 microCi/ml. Our assays also highlighted the inhibitory effect of [(3)H]hypoxanthine on B. divergens growth even when used at low concentrations (0.8 microCi/ml) and for a short incorporation duration (24 h). This effect was confirmed by both A(405) and parasitemia counting. In conclusion, A(405) measurement of B. divergens culture supernatant represents a simple, rapid, safe, and reliable way to measure the in vitro growth of this parasite. Generation times of three different B. divergens strains were then determined by the protocol described here and varied between 8 h 36 min and 13 h 8 min.

  14. Delayed-type hypersensitivity to Babesia microti-infected erythrocytes in mice

    International Nuclear Information System (INIS)

    Ruebush, M.J.; Troutman, E.H.; Kennedy, D.A.

    1986-01-01

    Strong delayed-type hypersensitivity (DTH) to Babesia microti was elicited when intraerythrocytic parasites (IEP) were inoculated subcutaneously into the flank of normal mice 6 to 14 days before challenge in the ipsilateral footpad with 10(8) IEP. Intraperitoneal or intravenous administration of antigen did not sensitize mice for DTH. When challenge was given 21 days after immunization, the response was approximately half of the maximum and then rose again slowly over the next 3 weeks to levels that were not significantly different from those maximal values. The response was classified as a true DTH reaction on the basis of kinetics, histology, and the transfer of responsiveness with immune T lymphocytes of the Ly 1+ phenotype, but not with serum. The reaction was specific for IEP since control groups given two injections of red blood cells from uninfected syngeneic mice (NRBC) or one injection of NRBC or sheep red blood cells (SRBC) and one of IEP never developed significant footpad swelling. Freed parasites obtained by osmotic rupture, density gradient sedimentation, and lethally irradiated IEP were also effective for elicitation of DTH. Anti-IEP DTH was expressed in a dose-dependent fashion with 10(6), 10(7), or 10(8) parasites sufficing for immunizing inoculum as long as 10(8) parasites were used as the challenge dose. Mice immunized and challenged with 10(8) lethally irradiated IEP (60 krad, 60Co), were protected against subsequent intraperitoneal challenge with 10(8) viable IEP. If mice were infected intraperitoneally with 10(8) IEP at any time between 21 days before immunization to 2 hr after challenge, their ability to respond to immunization and challenge was profoundly depressed. Development of a strong anti-parasite DTH response can occur in parallel with resistance to infection, but is not a rapid sequela of bloodborne infection

  15. Molecular characterization of the Babesia caballi rap-1 gene and epidemiological survey in horses in Israel.

    Science.gov (United States)

    Rapoport, Adi; Aharonson-Raz, Karin; Berlin, Dalia; Tal, Saar; Gottlieb, Yuval; Klement, Eyal; Steinman, Amir

    2014-04-01

    Equine piroplasmosis imposes great concerns for the equine industry regarding international horse movement, and therefore requires reliable diagnostic tools. Recent studies from South Africa and Jordan, including a preliminary study in Israel, reported extremely low seroprevalence to Babesia caballi (B. caballi) (0-1%) using the acceptable rhoptry-associated protein-1 (RAP-1) cELISA. In accordance with the study from South Africa demonstrating a significant heterogeneity in the rap-1 gene sequence of South African B. caballi isolates, the objectives of this study were to phylogenetically characterize the rap-1 gene of the Israeli isolates and determine the prevalence of B. caballi in horses in Israel. Out of 273 horses tested using the RAP-1 cELISA, only one was sero-positive, while 9.3% were positive on PCR performed on the rap-1 gene. Phylogenetic analysis of the rap-1 gene grouped the Israeli isolates in a cluster together with the South African strains (99% nt identity), but in a separate cluster from the American/Caribbean strains (81-82% nt identity). These findings support the existence of heterogeneity in the RAP-1 amino-acid sequences of the Israeli and South African isolates as compared to that used in the cELISA commercial kit and raise doubts as to the ability of this assay to serve as a sole regulatory test for international horse movement. Risk factor analysis found management and age to significantly associate with prevalence of B. caballi, as higher prevalence was noted in horses held out on pasture and a negative association was recorded with age. In addition, B. caballi was not detected in horses in the steppe-arid and extreme-arid climatic regions as compared to the wetter regions. Findings of this study emphasize the need to combine several detection methods to ameliorate the control and spread of the disease. Copyright © 2014 Elsevier B.V. All rights reserved.

  16. Gliding motility of Babesia bovis merozoites visualized by time-lapse video microscopy.

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    Masahito Asada

    Full Text Available BACKGROUND: Babesia bovis is an apicomplexan intraerythrocytic protozoan parasite that induces babesiosis in cattle after transmission by ticks. During specific stages of the apicomplexan parasite lifecycle, such as the sporozoites of Plasmodium falciparum and tachyzoites of Toxoplasma gondii, host cells are targeted for invasion using a unique, active process termed "gliding motility". However, it is not thoroughly understood how the merozoites of B. bovis target and invade host red blood cells (RBCs, and gliding motility has so far not been observed in the parasite. METHODOLOGY/PRINCIPAL FINDINGS: Gliding motility of B. bovis merozoites was revealed by time-lapse video microscopy. The recorded images revealed that the process included egress of the merozoites from the infected RBC, gliding motility, and subsequent invasion into new RBCs. The gliding motility of B. bovis merozoites was similar to the helical gliding of Toxoplasma tachyzoites. The trails left by the merozoites were detected by indirect immunofluorescence assay using antiserum against B. bovis merozoite surface antigen 1. Inhibition of gliding motility by actin filament polymerization or depolymerization indicated that the gliding motility was driven by actomyosin dependent process. In addition, we revealed the timing of breakdown of the parasitophorous vacuole. Time-lapse image analysis of membrane-stained bovine RBCs showed formation and breakdown of the parasitophorous vacuole within ten minutes of invasion. CONCLUSIONS/SIGNIFICANCE: This is the first report of the gliding motility of B. bovis. Since merozoites of Plasmodium parasites do not glide on a substrate, the gliding motility of B. bovis merozoites is a notable finding.

  17. Multiple ionization of noble gases by 2.0 MeV proton impact: comparison with equi-velocity electron impact ionization

    International Nuclear Information System (INIS)

    Melo, W.S.; Santos, A.C.F.; Sant'Anna, M.M.; Sigaud, G.M.; Montenegro, E.C.

    2002-01-01

    Absolute single- and multiple-ionization cross sections of rare gases (He, Ne, Ar, Kr and Xe) have been measured for collisions with 2.0 MeV p + . A comparison is made with equi-velocity electron impact ionization cross sections as well as with the available proton impact data. For the light rare gases the single-ionization cross sections are essentially the same for both proton and electron impacts, but increasing differences appear for the heavier targets. (author). Letter-to-the-editor

  18. Infection rate of Babesia spp. sporokinetes in engorged Boophilus microplus from an area of enzootic stability in the State of Minas Gerais, Brazil

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    Quintão-Silva Maria G

    2003-01-01

    Full Text Available The infection rates of Babesia sporokinetes in engorged Boophilus microplus were evaluated during a 2-year period in a dairy farm located in an area of enzootic stability. Every 14 days engorged females were collected from calves and from adult animals. Ticks were incubated at 27 ± 0.5ºC and 80-90% relative humidity and Babesia infection rates were determined by microscopic examination of Giemsa-stained hemolymph smears. After 52 collections, 2105 ticks were obtained, from which 982 were collected from calves and 1123 from cows. The total Babesia infection rate was 10%, however the incidence was higher (p < 0.05 in ticks collected from calves (17.5% than in those collected from cows (3.6%. Females collected from cows showed the highest infection rates in January, March, and August, and absence of infection in April and May. Ticks feeding on calves were infected throughout the experimental period. The infection rates of engorged females collected from naturally infected calves that were artificially infested with Babesia-free-larvae of B. microplus gradually decreased until the calves were four months old. No differences were observed among infection rates of ticks collected from calves maintained under natural conditions.

  19. A study on the determination of risk factors associated with babesiosis and prevalence of Babesia sp., by PCR amplification, in small ruminants from Southern Punjab (Pakistan

    Directory of Open Access Journals (Sweden)

    Iqbal F.

    2011-08-01

    Full Text Available Babesiosis is a parasitic infection due to the multiplication of tick borne parasite, Babesia sp., in erythrocytes of host, which includes a wide variety of vertebrates including small ruminants causing decreased livestock output and hence economic losses. The objective of the present study was to establish a PCR based method for the detection of Babesia sp. in small ruminant population in Southern Punjab and to determine the risk factors involve in the spread of babesiosis. A total of 107 blood samples were collected from 40 sheep and 67 goats in seven districts of Southern Punjab from randomly selected herds. Data on the characteristics of the animals and the herd were collected through questionnaires. 36 blood samples (34% of total produced the DNA fragment specific for 18S rRNA gene of Babesia sp., by PCR amplification, of which 20 were sheep and 16 were goats. Samples from all seven district contained Babesia positive samples and prevalence varied between 18 to 68%. It was observed that male animals (P = 0.009 and young animals under one year of age (P = 0.01 were more prone to the parasite. It was observed that herds consist of more than 15 animals (P = 0.007, composed of mixed species of small ruminants (P = 0.022, associated with dogs (P = 0.003 and dogs having ticks on their bodies (P = 0.011 were among the major risk factors for the spread of babesiosis in small ruminants.

  20. A study on the determination of risk factors associated with babesiosis and prevalence of Babesia sp., by PCR amplification, in small ruminants from Southern Punjab (Pakistan).

    Science.gov (United States)

    Iqbal, F; Fatima, M; Shahnawaz, S; Naeem, M; Shaikh, Rs; Ali, M; Shaikh, As; Aktas, M; Ali, M

    2011-08-01

    Babesiosis is a parasitic infection due to the multiplication of tick borne parasite, Babesia sp., in erythrocytes of host, which includes a wide variety of vertebrates including small ruminants causing decreased livestock output and hence economic losses. The objective of the present study was to establish a PCR based method for the detection of Babesia sp. in small ruminant population in Southern Punjab and to determine the risk factors involve in the spread of babesiosis. A total of 107 blood samples were collected from 40 sheep and 67 goats in seven districts of Southern Punjab from randomly selected herds. Data on the characteristics of the animals and the herd were collected through questionnaires. 36 blood samples (34% of total) produced the DNA fragment specific for 18S rRNA gene of Babesia sp., by PCR amplification, of which 20 were sheep and 16 were goats. Samples from all seven district contained Babesia positive samples and prevalence varied between 18 to 68%. It was observed that male animals (P = 0.009) and young animals under one year of age (P = 0.01) were more prone to the parasite. It was observed that herds consist of more than 15 animals (P = 0.007), composed of mixed species of small ruminants (P = 0.022), associated with dogs (P = 0.003) and dogs having ticks on their bodies (P = 0.011) were among the major risk factors for the spread of babesiosis in small ruminants.

  1. A molecular survey of Theileria and Babesia parasites in cattle, with a note on the distribution of ticks in Tunisia.

    Science.gov (United States)

    M'ghirbi, Y; Hurtado, A; Barandika, J F; Brandika, J; Khlif, K; Ketata, Z; Bouattour, A

    2008-07-01

    Between October and November 2006, a total of 278 bovine blood samples were examined, and 104 (37.4%) were positive for piroplasms by microscopy. A reverse line blot hybridisation with polymerase chain reaction detected Theileria annulata, T. buffeli, Babesia bovis and B. bigemina in cattle accounting for 48.6% of positive samples. The most frequently found species was T. buffeli, which was present in 39.2% of the samples. T. annulata was found in 48 samples (17.3%). Babesia infections were less frequently detected: B. bovis was found in 6.8% of the samples and B. bigemina in 4.3%. Mixed infections were detected in 45 samples, accounting for seven different combinations of species. Seven Ixodid tick species (Boophilus annulatus, Ixodes ricinus, Hyalomma marginatum, Hyalomma excavatum, Hyalomma detritum, Haemaphysalis punctata and Haemaphysalis sulcata) were collected from examined cattle in the 23 visited farms. I. ricinus was the dominant species (36%), mainly collected in the humid zone, while it seemed to be very rare in the semi-arid zone (where only 15 specimens were collected), whereas B. annulatus was the most commonly collected species in the sub-humid area (68.5% of ticks collected in this zone).

  2. Natural Babesia bovis Infection in Water Buffaloes (Bubalus bubalis and Crossbred Cattle under Field Conditions in Egypt: a Preliminary Study.

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    Yasser Mahmmod

    2014-06-01

    Full Text Available There is a little or no data available on the natural Babesia bovis (B. bovis infection in water buffaloes (Bubalus bubalis comparing to the available one for cattle. This study was conducted to investigate the natural B. bovis infection in water buffaloes in comparison to crossbred cattle under field conditions in Egypt.A total of 35 buffaloes and cattle were clinically and laboratory investigated from March to June 2008. Twenty-nine buffaloes and cattle out of 35 were naturally infected with B. bovis and showed signs of bovine babesiosis. Three cows and three buffaloes showed no clinical signs and were free from external, internal, and blood parasites served as control group.Babesia bovis-infected cattle showed typical signs of bovine babesiosis while B. bovis-infected buffaloes showed a milder form (less severe of the clinical signs. Advanced cases of cattle showed dark brown to dark red (coffee-color urine, hemoglobinuria and nervous manifestations while these manifestations were not detected in the infected buffaloes. Hematological changes in both species however, these changes were less significant in buffaloes than those reported in cattle.This paper documents the first description of natural B. bovis infection in water buffaloes which were found to be more likely to be tolerant than cattle to the natural clinical infection with B. bovis and its subsequent haematological changes. Our finding may lead to a better understanding of the disease pattern of B. bovis infection under field conditions in buffaloes.

  3. Efecto de la Esplenectomia en la infección a Babesias de la ardilla Citellus Tridecemlineatus

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    Arístides Herrer

    1956-07-01

    Full Text Available Observaciones en el Citellus tridecemilineatus con infecciones latentes a babesias ponen de manifiesto que la esplenectomía desencadena una inusitada intensificación del parasitismo, marcada anemia y otras alteraciones que pueden conducir a la muerte del animal. De cifras inferiores a 1 por mil que usualmente se observa en las infecciones latentes, con la esplenectomía la parasitemia se intensifica en tal forma que en ciertas ocasiones suele comprender a algo más del 80 por ciento de los eritrocitos. De igual manera aumenta notablemente la intensidad con que son parasitados los hematies, verificándose después de la extirpación del bazo eritrocitos con 8, 10 y 12 parásitos. Los resultados de estos estudios señalan la enorme importancia que el bazo tendría en la defensa contra la infección latente de esta especie de babesia, de manera parecida a lo que sucede principalmente en las infecciones a bartonellas de ciertos animales.

  4. Genome-wide analysis of gene expression and protein secretion of Babesia canis during virulent infection identifies potential pathogenicity factors.

    Science.gov (United States)

    Eichenberger, Ramon M; Ramakrishnan, Chandra; Russo, Giancarlo; Deplazes, Peter; Hehl, Adrian B

    2017-06-13

    Infections of dogs with virulent strains of Babesia canis are characterized by rapid onset and high mortality, comparable to complicated human malaria. As in other apicomplexan parasites, most Babesia virulence factors responsible for survival and pathogenicity are secreted to the host cell surface and beyond where they remodel and biochemically modify the infected cell interacting with host proteins in a very specific manner. Here, we investigated factors secreted by B. canis during acute infections in dogs and report on in silico predictions and experimental analysis of the parasite's exportome. As a backdrop, we generated a fully annotated B. canis genome sequence of a virulent Hungarian field isolate (strain BcH-CHIPZ) underpinned by extensive genome-wide RNA-seq analysis. We find evidence for conserved factors in apicomplexan hemoparasites involved in immune-evasion (e.g. VESA-protein family), proteins secreted across the iRBC membrane into the host bloodstream (e.g. SA- and Bc28 protein families), potential moonlighting proteins (e.g. profilin and histones), and uncharacterized antigens present during acute crisis in dogs. The combined data provides a first predicted and partially validated set of potential virulence factors exported during fatal infections, which can be exploited for urgently needed innovative intervention strategies aimed at facilitating diagnosis and management of canine babesiosis.

  5. Detection and molecular characterization of Babesia, Theileria, and Hepatozoon species in hard ticks collected from Kagoshima, the southern region in Japan.

    Science.gov (United States)

    Masatani, Tatsunori; Hayashi, Kei; Andoh, Masako; Tateno, Morihiro; Endo, Yasuyuki; Asada, Masahito; Kusakisako, Kodai; Tanaka, Tetsuya; Gokuden, Mutsuyo; Hozumi, Nodoka; Nakadohzono, Fumiko; Matsuo, Tomohide

    2017-06-01

    To reveal the distribution of tick-borne parasites, we established a novel nested polymerase chain reaction (PCR) system to detect the most common agents of tick-borne parasitic diseases, namely Babesia, Theileria, and Hepatozoon parasites. We collected host-seeking or animal-feeding ticks in Kagoshima Prefecture, the southernmost region of Kyusyu Island in southwestern Japan. Twenty of the total of 776 tick samples displayed a specific band of the appropriate size (approximately 1.4-1.6kbp) for the 18S rRNA genes in the novel nested PCR (20/776: 2.58%). These PCR products have individual sequences of Babesia spp. (from 8 ticks), Theileria spp. (from 9 ticks: one tick sample including at least two Theileria spp. sequences), and Hepatozoon spp. (from 3 ticks). Phylogenetic analyses revealed that these sequences were close to those of undescribed Babesia spp. detected in feral raccoons in Japan (5 sequences; 3 sequences being identical), Babesia gibsoni-like parasites detected in pigs in China (3 sequences; all sequences being identical), Theileria spp. detected in sika deer in Japan and China (10 sequences; 2 sequences being identical), Hepatozoon canis (one sequence), and Hepatozoon spp. detected in Japanese martens in Japan (two sequences). In summary, we showed that various tick-borne parasites exist in Kagoshima, the southern region in Japan by using the novel nested PCR system. These including undescribed species such as Babesia gibsoni-like parasites previously detected in pigs in China. Importantly, our results revealed new combinations of ticks and protozoan parasites in southern Japan. The results of this study will aid in the recognition of potential parasitic animal diseases caused by tick-borne parasites. Copyright © 2017 Elsevier GmbH. All rights reserved.

  6. Survey of vector-borne agents in feral cats and first report of Babesia gibsoni in cats on St Kitts, West Indies.

    Science.gov (United States)

    Kelly, Patrick John; Köster, Liza; Li, Jing; Zhang, Jilei; Huang, Ke; Branford, Gillian Carmichael; Marchi, Silvia; Vandenplas, Michel; Wang, Chengming

    2017-11-13

    As there is little data on vector-borne diseases of cats in the Caribbean region and even around the world, we tested feral cats from St Kitts by PCR to detect infections with Babesia, Ehrlichia and spotted fever group Rickettsia (SFGR) and surveyed them for antibodies to Rickettsia rickettsii and Ehrlichia canis. Whole blood was collected from apparently healthy feral cats during spay/ neuter campaigns on St Kitts in 2011 (N = 68) and 2014 (N = 52). Sera from the 52 cats from 2014 were used to detect antibodies to Ehrlichia canis and Rickettsia rickettsii using indirect fluorescent antibody tests and DNA extracted from whole blood of a total of 119 cats (68 from 2011, and 51 from 2014) was used for PCRs for Babesia, Ehrlichia and Rickettsia. We could not amplify DNA of SFG Rickettsia in any of the samples but found DNA of E. canis in 5% (6/119), Babesia vogeli in 13% (15/119), Babesia gibsoni in 4% (5/119), mixed infections with B. gibsoni and B. vogeli in 3% (3/119), and a poorly characterized Babesia sp. in 1% (1/119). Overall, 10% of the 52 cats we tested by IFA for E. canis were positive while 42% we tested by indirect fluorescent antibody (IFA) for R. rickettsii antigens were positive. Our study provides the first evidence that cats can be infected with B. gibsoni and also indicates that cats in the Caribbean may be commonly exposed to other vector-borne agents including SFGR, E. canis and B. vogeli. Animal health workers should be alerted to the possibility of clinical infections in their patients while public health workers should be alerted to the possibility that zoonotic SFGR are likely circulating in the region.

  7. Babesia spp. and Anaplasma phagocytophilum in questing ticks, ticks parasitizing rodents and the parasitized rodents--analyzing the host-pathogen-vector interface in a metropolitan area.

    Science.gov (United States)

    Silaghi, Cornelia; Woll, Dietlinde; Hamel, Dietmar; Pfister, Kurt; Mahling, Monia; Pfeffer, Martin

    2012-09-05

    The aims of this study were to evaluate the host-tick-pathogen interface of Babesia spp. and Anaplasma phagocytophilum in restored areas in both questing and host-attached Ixodes ricinus and Dermacentor reticulatus and their small mammalian hosts. Questing ticks were collected from 5 sites within the city of Leipzig, Germany, in 2009. Small mammals were trapped at 3 of the 5 sites during 2010 and 2011. DNA extracts of questing and host-attached I. ricinus and D. reticulatus and of several tissue types of small mammals (the majority bank voles and yellow-necked mice), were investigated by PCR followed by sequencing for the occurrence of DNA of Babesia spp. and by real-time PCR for A. phagocytophilum. A selected number of samples positive for A. phagocytophilum were further investigated for variants of the partial 16S rRNA gene. Co-infection with Rickettsia spp. in the questing ticks was additionally investigated. 4.1% of questing I. ricinus ticks, but no D. reticulatus, were positive for Babesia sp. and 8.7% of I. ricinus for A. phagocytophilum. Sequencing revealed B. microti, B. capreoli and Babesia spp. EU1 in Leipzig and sequence analysis of the partial 16S RNA gene of A. phagocytophilum revealed variants either rarely reported in human cases or associated with cervid hosts. The statistical analysis revealed significantly less ticks infected with A. phagocytophilum in a city park in Leipzig as compared to the other sampling sites. A. phagocytophilum-DNA was detected in 2 bank voles, DNA of B. microti in 1 striped field-mouse and of Babesia sp. EU1 in the skin tissue of a mole. Co-infections were detected. Our results show the involvement of small mammals in the natural endemic cycles of tick-borne pathogens. A more thorough understanding of the interactions of ticks, pathogens and hosts is the essential basis for effective preventive control measures.

  8. Babesia spp. and Anaplasma phagocytophilum in questing ticks, ticks parasitizing rodents and the parasitized rodents – Analyzing the host-pathogen-vector interface in a metropolitan area

    Directory of Open Access Journals (Sweden)

    Silaghi Cornelia

    2012-09-01

    Full Text Available Abstract Background The aims of this study were to evaluate the host-tick-pathogen interface of Babesia spp. and Anaplasma phagocytophilum in restored areas in both questing and host-attached Ixodes ricinus and Dermacentor reticulatus and their small mammalian hosts. Methods Questing ticks were collected from 5 sites within the city of Leipzig, Germany, in 2009. Small mammals were trapped at 3 of the 5 sites during 2010 and 2011. DNA extracts of questing and host-attached I. ricinus and D. reticulatus and of several tissue types of small mammals (the majority bank voles and yellow-necked mice, were investigated by PCR followed by sequencing for the occurrence of DNA of Babesia spp. and by real-time PCR for A. phagocytophilum. A selected number of samples positive for A. phagocytophilum were further investigated for variants of the partial 16S rRNA gene. Co-infection with Rickettsia spp. in the questing ticks was additionally investigated. Results 4.1% of questing I. ricinus ticks, but no D. reticulatus, were positive for Babesia sp. and 8.7% of I. ricinus for A. phagocytophilum. Sequencing revealed B. microti, B. capreoli and Babesia spp. EU1 in Leipzig and sequence analysis of the partial 16S RNA gene of A. phagocytophilum revealed variants either rarely reported in human cases or associated with cervid hosts. The statistical analysis revealed significantly less ticks infected with A. phagocytophilum in a city park in Leipzig as compared to the other sampling sites. A. phagocytophilum-DNA was detected in 2 bank voles, DNA of B. microti in 1 striped field-mouse and of Babesia sp. EU1 in the skin tissue of a mole. Co-infections were detected. Conclusion Our results show the involvement of small mammals in the natural endemic cycles of tick-borne pathogens. A more thorough understanding of the interactions of ticks, pathogens and hosts is the essential basis for effective preventive control measures.

  9. Babesia spp. and Anaplasma phagocytophilum in questing ticks, ticks parasitizing rodents and the parasitized rodents – Analyzing the host-pathogen-vector interface in a metropolitan area

    Science.gov (United States)

    2012-01-01

    Background The aims of this study were to evaluate the host-tick-pathogen interface of Babesia spp. and Anaplasma phagocytophilum in restored areas in both questing and host-attached Ixodes ricinus and Dermacentor reticulatus and their small mammalian hosts. Methods Questing ticks were collected from 5 sites within the city of Leipzig, Germany, in 2009. Small mammals were trapped at 3 of the 5 sites during 2010 and 2011. DNA extracts of questing and host-attached I. ricinus and D. reticulatus and of several tissue types of small mammals (the majority bank voles and yellow-necked mice), were investigated by PCR followed by sequencing for the occurrence of DNA of Babesia spp. and by real-time PCR for A. phagocytophilum. A selected number of samples positive for A. phagocytophilum were further investigated for variants of the partial 16S rRNA gene. Co-infection with Rickettsia spp. in the questing ticks was additionally investigated. Results 4.1% of questing I. ricinus ticks, but no D. reticulatus, were positive for Babesia sp. and 8.7% of I. ricinus for A. phagocytophilum. Sequencing revealed B. microti, B. capreoli and Babesia spp. EU1 in Leipzig and sequence analysis of the partial 16S RNA gene of A. phagocytophilum revealed variants either rarely reported in human cases or associated with cervid hosts. The statistical analysis revealed significantly less ticks infected with A. phagocytophilum in a city park in Leipzig as compared to the other sampling sites. A. phagocytophilum-DNA was detected in 2 bank voles, DNA of B. microti in 1 striped field-mouse and of Babesia sp. EU1 in the skin tissue of a mole. Co-infections were detected. Conclusion Our results show the involvement of small mammals in the natural endemic cycles of tick-borne pathogens. A more thorough understanding of the interactions of ticks, pathogens and hosts is the essential basis for effective preventive control measures. PMID:22950642

  10. Structure of Rhodococcus equi virulence-associated protein B (VapB) reveals an eight-stranded antiparallel β-barrel consisting of two Greek-key motifs

    International Nuclear Information System (INIS)

    Geerds, Christina; Wohlmann, Jens; Haas, Albert; Niemann, Hartmut H.

    2014-01-01

    The structure of VapB, a member of the Vap protein family that is involved in virulence of the bacterial pathogen R. equi, was determined by SAD phasing and reveals an eight-stranded antiparallel β-barrel similar to avidin, suggestive of a binding function. Made up of two Greek-key motifs, the topology of VapB is unusual or even unique. Members of the virulence-associated protein (Vap) family from the pathogen Rhodococcus equi regulate virulence in an unknown manner. They do not share recognizable sequence homology with any protein of known structure. VapB and VapA are normally associated with isolates from pigs and horses, respectively. To contribute to a molecular understanding of Vap function, the crystal structure of a protease-resistant VapB fragment was determined at 1.4 Å resolution. The structure was solved by SAD phasing employing the anomalous signal of one endogenous S atom and two bound Co ions with low occupancy. VapB is an eight-stranded antiparallel β-barrel with a single helix. Structural similarity to avidins suggests a potential binding function. Unlike other eight- or ten-stranded β-barrels found in avidins, bacterial outer membrane proteins, fatty-acid-binding proteins and lysozyme inhibitors, Vaps do not have a next-neighbour arrangement but consist of two Greek-key motifs with strand order 41238567, suggesting an unusual or even unique topology

  11. Structure of Rhodococcus equi virulence-associated protein B (VapB) reveals an eight-stranded antiparallel β-barrel consisting of two Greek-key motifs

    Energy Technology Data Exchange (ETDEWEB)

    Geerds, Christina [Bielefeld University, Universitaetsstrasse 25, 33615 Bielefeld (Germany); Wohlmann, Jens; Haas, Albert [University of Bonn, Ulrich-Haberland Strasse 61a, 53121 Bonn (Germany); Niemann, Hartmut H., E-mail: hartmut.niemann@uni-bielefeld.de [Bielefeld University, Universitaetsstrasse 25, 33615 Bielefeld (Germany)

    2014-06-18

    The structure of VapB, a member of the Vap protein family that is involved in virulence of the bacterial pathogen R. equi, was determined by SAD phasing and reveals an eight-stranded antiparallel β-barrel similar to avidin, suggestive of a binding function. Made up of two Greek-key motifs, the topology of VapB is unusual or even unique. Members of the virulence-associated protein (Vap) family from the pathogen Rhodococcus equi regulate virulence in an unknown manner. They do not share recognizable sequence homology with any protein of known structure. VapB and VapA are normally associated with isolates from pigs and horses, respectively. To contribute to a molecular understanding of Vap function, the crystal structure of a protease-resistant VapB fragment was determined at 1.4 Å resolution. The structure was solved by SAD phasing employing the anomalous signal of one endogenous S atom and two bound Co ions with low occupancy. VapB is an eight-stranded antiparallel β-barrel with a single helix. Structural similarity to avidins suggests a potential binding function. Unlike other eight- or ten-stranded β-barrels found in avidins, bacterial outer membrane proteins, fatty-acid-binding proteins and lysozyme inhibitors, Vaps do not have a next-neighbour arrangement but consist of two Greek-key motifs with strand order 41238567, suggesting an unusual or even unique topology.

  12. Molecular detection of Hepatozoon canis and Babesia canis vogeli in domestic dogs from Cuiabá, Brazil Detecção molecular de Hepatozoon canis e Babesia canis vogeli em cães domésticos de Cuiabá, Brasil

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    Mariana Granziera Spolidorio

    2011-09-01

    Full Text Available The objective of this study was to report for the first time infection by Hepatozoon spp. and Babesia spp. in 10 dogs from the city of Cuiabá, State of Mato Grosso, central-western Brazil. A pair of primers that amplifies a 574 bp fragment of the 18S rRNA of Hepatozoon spp., and a pair of primers that amplifies a 551 bp fragment of the gene 18S rRNA for Babesia spp. were used. Six dogs were positive for Babesia spp., and 9 were positive for Hepatozoon spp. Co-infection of Babesia spp. and Hepatozoon spp. was seen in 5 dogs. Sequenced samples revealed 100% identity with B. canis vogeli, and H. canis. This is the first molecular detection of H. canis in domestic dogs from Cuiabá. Additionally, it is described for the first time the presence of B. canis vogeli circulating among dogs in Cuiabá.O objetivo deste estudo foi relatar pela primeira vez a infecção por Hepatozoon spp. e Babesia spp. em cães domésticos provenientes da cidade de Cuiabá, estado de Mato Grosso. Foram utilizados pares de primers que amplificam um fragmento de 574 pb do gene 18S rRNA de Hepatozoon spp., e 551 pb do gene 18S rRNA para Babesia spp. Dos 10 cães amostrados, 6 apresentaram-se positivos para Babesia spp., e 9 foram positivos para Hepatozoon spp. pela PCR. Co-infecção entre Babesia spp. e Hepatozoon spp. ocorreu em 5 cães. As amostras revelaram 100% de identidade com B. canis vogeli, e as amostras que foram positivas para Hepatozoon spp. foram 100% idênticas a H. canis. Esta é a primeira identificação molecular de H. canis em cães domésticos em Cuiabá. Adicionalmente, descrevemos pela primeira vez a presença de B. canis vogeli circulando entre cães em Cuiabá.

  13. Transfected Babesia bovis Expressing a Tick GST as a Live Vector Vaccine

    Science.gov (United States)

    Oldiges, Daiane P.; Laughery, Jacob M.; Tagliari, Nelson Junior; Leite Filho, Ronaldo Viana; Davis, William C.; da Silva Vaz, Itabajara; Termignoni, Carlos; Knowles, Donald P.; Suarez, Carlos E.

    2016-01-01

    The Rhipicephalus microplus tick is a notorious blood-feeding ectoparasite of livestock, especially cattle, responsible for massive losses in animal production. It is the main vector for transmission of pathogenic bacteria and parasites, including Babesia bovis, an intraerythrocytic apicomplexan protozoan parasite responsible for bovine Babesiosis. This study describes the development and testing of a live B. bovis vaccine expressing the protective tick antigen glutathione-S-transferase from Haemaphysalis longicornis (HlGST). The B. bovis S74-T3B parasites were electroporated with a plasmid containing the bidirectional Ef-1α (elongation factor 1 alpha) promoter of B. bovis controlling expression of two independent genes, the selectable marker GFP-BSD (green fluorescent protein–blasticidin deaminase), and HlGST fused to the MSA-1 (merozoite surface antigen 1) signal peptide from B. bovis. Electroporation followed by blasticidin selection resulted in the emergence of a mixed B. bovis transfected line (termed HlGST) in in vitro cultures, containing parasites with distinct patterns of insertion of both exogenous genes, either in or outside the Ef-1α locus. A B. bovis clonal line termed HlGST-Cln expressing intracellular GFP and HlGST in the surface of merozoites was then derived from the mixed parasite line HlGST using a fluorescent activated cell sorter. Two independent calf immunization trials were performed via intravenous inoculation of the HlGST-Cln and a previously described control consisting of an irrelevant transfected clonal line of B. bovis designated GFP-Cln. The control GFP-Cln line contains a copy of the GFP-BSD gene inserted into the Ef-1α locus of B. bovis in an identical fashion as the HIGST-Cln parasites. All animals inoculated with the HlGST-Cln and GFP-Cln transfected parasites developed mild babesiosis. Tick egg fertility and fully engorged female tick weight was reduced significantly in R. microplus feeding on HlGST-Cln-immunized calves

  14. Molecular characterization of a new Babesia bovis thrombospondin-related anonymous protein (BbTRAP2.

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    Mohamad Alaa Terkawi

    Full Text Available A gene encoding a Babesia bovis protein that shares significant degree of similarity to other apicomplexan thrombospondin-related anonymous proteins (TRAPs was found in the genomic database and designated as BbTRAP2. Recombinant protein containing a conserved region of BbTRAP2 was produced in E. coli. A high antigenicity of recombinant BbTRAP2 (rBbTRAP2 was observed with field B. bovis-infected bovine sera collected from geographically different regions of the world. Moreover, antiserum against rBbTRAP2 specifically reacted with the authentic protein by Western blot analysis and an indirect fluorescent antibody test. Three bands corresponding to 104-, 76-, and 44-kDa proteins were identified in the parasite lysates and two bands of 76- and 44-kDa proteins were detected in the supernatant of cultivated parasites, indicating that BbTRAP2 was proteolytically processed and shed into the culture. Apical and surface localizations of BbTRAP2 were observed in the intracellular and extracellular parasites, respectively, by confocal laser microscopic examination. Moreover, native BbTRAP2 was precipitated by bovine erythrocytes, suggesting its role in the attachment to erythrocytes. Furthermore, the specific antibody to rBbTRAP2 inhibited the growth of B. bovis in a concentration-dependent manner. Consistently, pre-incubation of the free merozoites with the antibody to rBbTRAP2 resulted in an inhibition of the parasite invasion into host erythrocytes. Interestingly, the antibody to rBbTRAP2 was the most inhibitive for the parasite's growth as compared to those of a set of antisera produced against different recombinant proteins, including merozoite surface antigen 2c (BbMSA-2c, rhoptry-associated protein 1 C-terminal (BbRAP-1CT, and spherical body protein 1 (BbSBP-1. These results suggest that BbTRAP2 might be a potential candidate for development of a subunit vaccine against B. bovis infection.

  15. Transfected Babesia bovis Expressing a Tick GST as a Live Vector Vaccine.

    Directory of Open Access Journals (Sweden)

    Daiane P Oldiges

    2016-12-01

    Full Text Available The Rhipicephalus microplus tick is a notorious blood-feeding ectoparasite of livestock, especially cattle, responsible for massive losses in animal production. It is the main vector for transmission of pathogenic bacteria and parasites, including Babesia bovis, an intraerythrocytic apicomplexan protozoan parasite responsible for bovine Babesiosis. This study describes the development and testing of a live B. bovis vaccine expressing the protective tick antigen glutathione-S-transferase from Haemaphysalis longicornis (HlGST. The B. bovis S74-T3B parasites were electroporated with a plasmid containing the bidirectional Ef-1α (elongation factor 1 alpha promoter of B. bovis controlling expression of two independent genes, the selectable marker GFP-BSD (green fluorescent protein-blasticidin deaminase, and HlGST fused to the MSA-1 (merozoite surface antigen 1 signal peptide from B. bovis. Electroporation followed by blasticidin selection resulted in the emergence of a mixed B. bovis transfected line (termed HlGST in in vitro cultures, containing parasites with distinct patterns of insertion of both exogenous genes, either in or outside the Ef-1α locus. A B. bovis clonal line termed HlGST-Cln expressing intracellular GFP and HlGST in the surface of merozoites was then derived from the mixed parasite line HlGST using a fluorescent activated cell sorter. Two independent calf immunization trials were performed via intravenous inoculation of the HlGST-Cln and a previously described control consisting of an irrelevant transfected clonal line of B. bovis designated GFP-Cln. The control GFP-Cln line contains a copy of the GFP-BSD gene inserted into the Ef-1α locus of B. bovis in an identical fashion as the HIGST-Cln parasites. All animals inoculated with the HlGST-Cln and GFP-Cln transfected parasites developed mild babesiosis. Tick egg fertility and fully engorged female tick weight was reduced significantly in R. microplus feeding on Hl

  16. Genetic diversity of merozoite surface antigens in Babesia bovis detected from Sri Lankan cattle.

    Science.gov (United States)

    Sivakumar, Thillaiampalam; Okubo, Kazuhiro; Igarashi, Ikuo; de Silva, Weligodage Kumarawansa; Kothalawala, Hemal; Silva, Seekkuge Susil Priyantha; Vimalakumar, Singarayar Caniciyas; Meewewa, Asela Sanjeewa; Yokoyama, Naoaki

    2013-10-01

    Babesia bovis, the causative agent of severe bovine babesiosis, is endemic in Sri Lanka. The live attenuated vaccine (K-strain), which was introduced in the early 1990s, has been used to immunize cattle populations in endemic areas of the country. The present study was undertaken to determine the genetic diversity of merozoite surface antigens (MSAs) in B. bovis isolates from Sri Lankan cattle, and to compare the gene sequences obtained from such isolates against those of the K-strain. Forty-four bovine blood samples isolated from different geographical regions of Sri Lanka and judged to be B. bovis-positive by PCR screening were used to amplify MSAs (MSA-1, MSA-2c, MSA-2a1, MSA-2a2, and MSA-2b), AMA-1, and 12D3 genes from parasite DNA. Although the AMA-1 and 12D3 gene sequences were highly conserved among the Sri Lankan isolates, the MSA gene sequences from the same isolates were highly diverse. Sri Lankan MSA-1, MSA-2c, MSA-2a1, MSA-2a2, and MSA-2b sequences clustered within 5, 2, 4, 1, and 9 different clades in the gene phylograms, respectively, while the minimum similarity values among the deduced amino acid sequences of these genes were 36.8%, 68.7%, 80.3%, 100%, and 68.3%, respectively. In the phylograms, none of the Sri Lankan sequences fell within clades containing the respective K-strain sequences. Additionally, the similarity values for MSA-1 and MSA-2c were 40-61.8% and 90.9-93.2% between the Sri Lankan isolates and the K-strain, respectively, while the K-strain MSA-2a/b sequence shared 64.5-69.8%, 69.3%, and 70.5-80.3% similarities with the Sri Lankan MSA-2a1, MSA-2a2, and MSA-2b sequences, respectively. The present study has shown that genetic diversity among MSAs of Sri Lankan B. bovis isolates is very high, and that the sequences of field isolates diverged genetically from the K-strain. Copyright © 2013 Elsevier B.V. All rights reserved.

  17. Molecular detection and genetic diversity of bovine Babesia spp., Theileria orientalis, and Anaplasma marginale in beef cattle in Thailand.

    Science.gov (United States)

    Jirapattharasate, Charoonluk; Adjou Moumouni, Paul Franck; Cao, Shinuo; Iguchi, Aiko; Liu, Mingming; Wang, Guanbo; Zhou, Mo; Vudriko, Patrick; Efstratiou, Artemis; Changbunjong, Tanasak; Sungpradit, Sivapong; Ratanakorn, Parntep; Moonarmart, Walasinee; Sedwisai, Poonyapat; Weluwanarak, Thekhawet; Wongsawang, Witsanu; Suzuki, Hiroshi; Xuan, Xuenan

    2017-02-01

    Babesia spp., Theileria orientalis, and Anaplasma marginale are significant tick-borne pathogens that affect the health and productivity of cattle in tropical and subtropical areas. In this study, we used PCR to detect the presence of Babesia bovis, Babesia bigemina, and T. orientalis in 279 beef cattle from Western Thailand and A. marginale in 608 beef cattle from the north, northeastern, and western regions. The PCRs were performed using species-specific primers based on the B. bovis spherical body protein 2 (BboSBP2), B. bigemina rhoptry-associated protein 1a (BbiRAP-1a), T. orientalis major piroplasm surface protein (ToMPSP), and A. marginale major surface protein 4 (AmMSP4) genes. To determine the genetic diversity of the above parasites, amplicons of B. bovis and B. bigemina ITS1-5.8s rRNA gene-ITS2 regions (B. bovis ITS, B. bigemina ITS), ToMPSP, and AmMSP4 genes were sequenced for phylogenetic analysis. PCR results revealed that the prevalence of B. bovis, B. bigemina, T. orientalis, and A. marginale in the Western region was 11.1, 12.5, 7.8, and 39.1 %, respectively. Coinfections of two or three parasites were observed in 17.9 % of the animals sampled. The study revealed that the prevalence of A. marginale in the western region was higher than in the north and northeastern regions (7 %). Sequence analysis showed the BboSBP2 gene to be more conserved than B. bovis ITS in the different isolates and, similarly, the BbiRAP-1a was more conserved than B. bigemina ITS. In the phylogenetic analysis, T. orientalis MPSP sequences were classified into types 3, 5, and 7 as previously reported. A. marginale MSP4 gene sequences shared high identity and similarity with each other and clustered with isolates from other countries. This study provides information on the prevalence and genetic diversity of tick-borne pathogens in beef cattle and highlights the need for effective strategies to control these pathogens in Thailand.

  18. Analysis of Babesia bovis infection-induced gene expression changes in larvae from the cattle tick, Rhipicephalus (Boophilus microplus

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    Heekin Andrew M

    2012-08-01

    Full Text Available Abstract Background Cattle babesiosis is a tick-borne disease of cattle that has severe economic impact on cattle producers throughout the world’s tropical and subtropical countries. The most severe form of the disease is caused by the apicomplexan, Babesia bovis, and transmitted to cattle through the bite of infected cattle ticks of the genus Rhipicephalus, with the most prevalent species being Rhipicephalus (Boophilus microplus. We studied the reaction of the R. microplus larval transcriptome in response to infection by B. bovis. Methods Total RNA was isolated for both uninfected and Babesia bovis-infected larval samples. Subtracted libraries were prepared by subtracting the B. bovis-infected material with the uninfected material, thus enriching for expressed genes in the B. bovis-infected sample. Expressed sequence tags from the subtracted library were generated, assembled, and sequenced. To complement the subtracted library method, differential transcript expression between samples was also measured using custom high-density microarrays. The microarray probes were fabricated using oligonucleotides derived from the Bmi Gene Index database (Version 2. Array results were verified for three target genes by real-time PCR. Results Ticks were allowed to feed on a B. bovis-infected splenectomized calf and on an uninfected control calf. RNA was purified in duplicate from whole larvae and subtracted cDNA libraries were synthesized from Babesia-infected larval RNA, subtracting with the corresponding uninfected larval RNA. One thousand ESTs were sequenced from the larval library and the transcripts were annotated. We used a R. microplus microarray designed from a R. microplus gene index, BmiGI Version 2, to look for changes in gene expression that were associated with infection of R. microplus larvae. We found 24 transcripts were expressed at a statistically significant higher level in ticks feeding upon a B. bovis-infected calf contrasted to ticks

  19. Identification of candidate vaccine antigens of bovine hemoparasites Theileria parva and Babesia bovis by use of helper T cell clones.

    Science.gov (United States)

    Brown, W C; Zhao, S; Logan, K S; Grab, D J; Rice-Ficht, A C

    1995-03-01

    Current vaccines for bovine hemoparasites utilize live attenuated organisms or virulent organisms administered concurrently with antiparasitic drugs. Although such vaccines can be effective, for most hemoparasites the mechanisms of acquired resistance to challenge infection with heterologous parasite isolates have not been clearly defined. Selection of potentially protective antigens has traditionally made use of antibodies to identify immunodominant proteins. However, numerous studies have indicated that induction of high antibody titers neither predicts the ability of an antigen to confer protective immunity nor correlates with protection. Because successful parasites have evolved antibody evasion tactics, alternative strategies to identify protective immunogens should be used. Through the elaboration of cytokines, T helper 1-(Th1)-like T cells and macrophages mediate protective immunity against many intracellular parasites, and therefore most likely play an important role in protective immunity against bovine hemoparasites. CD4+ T cell clones specific for soluble or membrane antigens of either Theileria parva schizonts or Babesia bovis merozoites were therefore employed to identify parasite antigens that elicit strong Th cell responses in vitro. Soluble cytosolic parasite antigen was fractionated by gel filtration, anion exchange chromatography or hydroxylapatite chromatography, or a combination thereof, and fractions were tested for the ability to induce proliferation of Th cell clones. This procedure enabled the identification of stimulatory fractions containing T. parva proteins of approximately 10 and 24 kDa. Antisera raised against the purified 24 kDa band reacted with a native schizont protein of approximately 30 kDa. Babesia bovis-specific Th cell clones tested against fractionated soluble Babesia bovis merozoite antigen revealed the presence of at least five distinct antigenic epitopes. Proteins separated by gel filtration revealed four patterns of

  20. Perfil de suscetibilidade antimicrobiana e presença do gene vapA em Rhodococcus equi de origem humana, ambiental e equina

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    Lilian Kolling Girardini

    2013-06-01

    Full Text Available Rhodococcus equi é um micro-organismo intracelular facultativo, agente etiológico da rodococose, uma importante enfermidade que acomete principalmente potros com menos de seis meses de idade, causando a morte geralmente em decorrência de lesões pulmonares. Este agente também tem potencial zoonótico e emergiu como um patógeno oportunista no mundo, acometendo humanos imunocomprometidos, especialmente os transplantados e infectados pelo vírus da imunodeficiência humana (HIV. Entretanto, infecções por R. equi em hospedeiros hígidos tem sido relatadas, principalmente em crianças e idosos. Estudos tem mostrado um nível crescente na resistência de isolados de R. equi em relação aos antimicrobianos comumente utilizados no tratamento de animais e seres humanos infectados por este agente. A virulência deste pode estar associada a fatores como a cápsula de polissacarídeo, fosfolipase C e à enzima colesterol oxidase (fator equi. No entanto, uma proteína localizada em um plasmídeo, designada vapA, é essencial para a sobrevivência e replicação do agente em macrófagos. Com isso, os objetivos deste estudo foram avaliar o perfil de suscetibilidade de isolados de R. equi de diferentes fontes em relação aos antimicrobianos mais comumente utilizados na terapêutica animal e humana, bem como verificar a associação entre a presença do gene vapA e o índice de resistência múltipla aos antimicrobianos (IRMA. Neste estudo, 67 isolados brasileiros de R. qui de diferentes fontes foram analisados: 30 provenientes de amostras clínicas de equinos, sete de humanos e 30 ambientais (seis do solo e 24 de fezes de equinos. Para avaliar o perfil de suscetibilidade dos isolados utilizou-se o método de disco difusão, sendo testadas 16 drogas de diferentes classes de antimicrobianos. As amostras clínicas de equinos apresentaram as maiores taxas de resistência à penicilina (86,7% e lincomicina (30%. Além disso, foram também resistentes a

  1. The infection of questing Dermacentor reticulatus ticks with Babesia canis and Anaplasma phagocytophilum in the Chernobyl exclusion zone.

    Science.gov (United States)

    Karbowiak, Grzegorz; Vichová, Bronislavá; Slivinska, Kateryna; Werszko, Joanna; Didyk, Julia; Peťko, Branislav; Stanko, Michal; Akimov, Igor

    2014-08-29

    Tick occurrence was studied in the Chernobyl exclusion zone (CEZ) during the August-October 2009-2012. Dermacentor reticulatus ticks were collected using the flagging method and then screened for infection with Anaplasma phagocytophilum and Babesia canis by a PCR method incorporating specific primers and sequence analysis. The prevalence of infection with B. canis canis and A. phagocytophilum was found to be 3.41% and 25.36%, respectively. The results present the first evidence of B. canis canis and A. phagocytophilum in questing D. reticulatus ticks from the Chernobyl exclusion zone. They also reveal the presence of tick-borne disease foci in areas with no human activity, and confirm that they can be maintained in areas after a nuclear disaster with radioactive contamination. Copyright © 2014 Elsevier B.V. All rights reserved.

  2. Babesia microti real-time polymerase chain reaction testing of Connecticut blood donors: potential implications for screening algorithms.

    Science.gov (United States)

    Johnson, Stephanie T; Van Tassell, Eric R; Tonnetti, Laura; Cable, Ritchard G; Berardi, Victor P; Leiby, David A

    2013-11-01

    Babesia microti, an intraerythrocytic parasite, has been implicated in transfusion transmission. B. microti seroprevalence in Connecticut (CT) blood donors is approximately 1%; however, it is not known what percentage of donors is parasitemic and poses a risk for transmitting infection. Therefore, we determined the prevalence of demonstrable B. microti DNA in donors from a highly endemic area of CT and compared observed rates with concurrent immunofluorescence assay (IFA) testing results. Blood samples from consenting donors in southeastern CT were collected from mid-August through early October 2009 and tested by IFA for immunoglobulin G antibodies and real-time polymerase chain reaction (PCR) for B. microti DNA. IFA specificity was determined using blood donor samples collected in northwestern Vermont (VT), an area nonendemic for Babesia. Of 1002 CT donors, 25 (2.5%) were IFA positive and three (0.3%) were real-time PCR positive. Among the three real-time PCR-positive donors, two were also IFA positive, while one was IFA negative and may represent a window period infection. The two IFA- and real-time PCR-positive donors appeared to subsequently clear infection. The other real-time PCR-positive donor did not provide follow-up samples. Of 1015 VT donors tested by IFA, only one (0.1%) was positive, but may have acquired infection during travel to an endemic area. We prospectively identified several real-time PCR-positive blood donors, including an IFA-negative real-time PCR-positive donor, in an area highly endemic for B. microti. These results suggest the need to include nucleic acid testing in planned mitigation strategies for B. microti. © 2013 American Association of Blood Banks.

  3. Spatial distribution, risk factors and haemato-biochemical alterations associated with Theileria equi infected equids of Punjab (India) diagnosed by indirect ELISA and nested PCR.

    Science.gov (United States)

    Sumbria, Deepak; Singla, L D; Kumar, Sanjay; Sharma, Amrita; Dahiya, Rajesh K; Setia, Raj

    2016-03-01

    Equine piroplasmosis is a febrile, tick-borne disease of equids predominately caused by obligatory intra-erythrocytic protozoa Theileria equi in the Indian sub-continent. A cross-sectional study was carried out on 464 equids (426 horses and 38 donkeys/mules) in Punjab, India to assess the level of exposure to equine piroplasmosis by 18S rRNA gene nested polymerase chain reaction (nPCR) and equine merozoite antigen-2 (EMA2) indirect-ELISA (enzyme linked immunosorbent assay), to investigate risk factors and haemato-biochemical alterations associated with the infection. The endemicity of the disease was confirmed by positive PCR amplification in 21.77% and positive antibody titers in 49.78% equid samples. There was a fair agreement between these two diagnostic techniques (Kappa coefficient=0.326). The spatial distribution analysis revealed an increasing trend of T. equi prevalence from north-eastern to south-western region of Punjab by both the techniques correspondingly, which proffered a direct relation with temperature and inverse with humidity variables. The relatively prominent risk factor associated with sero-positivity was the presence of other domestic animals in the herd, while the propensity of finding a positive PCR amplification was higher in donkeys/mules, animal kept at unorganised farm or those used for commercial purposes as compared to their counterparts. There was a significant increase in globulins, gamma glutamyl-transferase, total bilirubin, direct bilirubin, indirect bilirubin, glucose levels and decrease in total erythrocyte count, haemoglobin, packed cell volume by animals, which were revealed positive by nPCR (may or may not positive by indirect-ELISA) and increase in creatinine, total bilirubin, direct bilirubin, glucose and decrease in total erythrocytes count by animals, which were revealed positive by indirect-ELISA (alone). To our knowledge, this study, for the first time, brings out a comprehensive report on the status on spatial

  4. Comparison between conventional and molecular methods for diagnosis of bovine babesiosis (Babesia bovis infection) in tick infested cattle in upper Egypt.

    Science.gov (United States)

    Al-Hosary, Amira A T

    2017-03-01

    Ticks and tick-borne diseases are the main problems affecting the livestock production in Egypt. Bovine babesiosis has adverse effects on the animal health and production. A comparison of Giemsa stained blood smears, polymerase chain reaction (PCR) and nested PCR (nPCR) assays for detection of Babesia bovis infection in Egyptian Baladi cattle ( Bos taurus ) in reference to reverse line blot was carried out. The sensitivity of PCR and nested PCR (nPCR) assays were 65 and 100 % respectively. Giemsa stained blood smears showed the lowest sensitivity (30 %). According to these results using of PCR and nPCR target for B. bovis , [BBOV-IV005650 (BV5650)] gene are suitable for diagnosis of B. bovis infection. The 18Ss rRNA partial sequence confirmed that all the positive samples were Babesia bovis and all of them were deposited in the GenBank databases (Accession No: KM455548, KM455549 and KM455550).

  5. Validate or falsify: Lessons learned from a microscopy method claimed to be useful for detecting Borrelia and Babesia organisms in human blood

    Czech Academy of Sciences Publication Activity Database

    Aase, A.; Hajdušek, Ondřej; Øines, Ø.; Quarsten, H.; Wilhelmsson, P.; Herstad, T.K.; Kjelland, V.; Šíma, Radek; Jalovecká, Marie; Lindgren, P-E.; Aaberge, I.S.

    2016-01-01

    Roč. 48, č. 6 (2016), s. 411-419 ISSN 2374-4235 R&D Projects: GA ČR GP13-27630P; GA ČR GP13-12816P EU Projects: European Commission(XE) 316304 - MODBIOLIN Institutional support: RVO:60077344 Keywords : Lyme disease * Borrelia burgdorferi sensu lato * babesiosis * Babesia spp. * Lyme borreliosis * PCR * microscopy Subject RIV: EC - Immunology Impact factor: 1.119, year: 2016

  6. Infections and risk factors for livestock with species of Anaplasma, Babesia and Brucella under semi-nomadic rearing in Karamoja Region, Uganda.

    Science.gov (United States)

    Lolli, Chiara; Marenzoni, Maria Luisa; Strona, Paolo; Lappo, Pier Giorgio; Etiang, Patrick; Diverio, Silvana

    2016-03-01

    A survey was conducted to estimate the prevalence of Anaplasma, Babesia and Brucella spp. infections in cattle, goats and sheep in the Karamoja Region of Uganda and to identify possible risk factors existing in this semi-nomadic and pastoral area. Low cost laboratory tests were used to diagnose infections (Rose Bengal test for Brucella spp. antibodies and direct microscopic examination for Anaplasma and Babesia spp.). Multivariable logistic regression models were applied to identify possible risk factors linked to gender, animal species, age (only for cattle) and districts. A total of 3935 cattle, 729 goats and 306 sheep of five districts of the Karamoja Region were tested. Seroprevalence for Brucella was 9.2 % (CI, 95 %: 8.4-10), for Anaplasma 19.5 % (CI 95 %: 18.4-20.6) and for Babesia 16 % (CI 95 %: 15-17.1). Significant differences in infections prevalence were observed against risk factors associated with districts and species. Cattle were the species with higher risk of the infections. Female gender was identified as at risk only for Brucella spp. infection. Cattle more than one year old had greater likelihood to be Brucella seropositive. Co-infections of Anaplasma and Babesia spp. were statistically associated, especially in goats and sheep. Further studies to identify risk factors related to host species and geographical districts are needed. The influence on the semi-nomadic agro-pastoral system in Karamoja of animal raids and animal mixing should be further investigated. Findings were important to sensitize Karamojong undertaking measures on infection control, especially on cattle, which are their main source of food.

  7. Detecção molecular de Ehrlichia canis e Babesia canis vogeli em Rhipicephalus sanguineus sensu lato de carrapatos em Cuba

    Directory of Open Access Journals (Sweden)

    Maylin Gonzalez Navarrete

    2016-12-01

    Full Text Available Os carrapatos (Acari: Ixodidae são de importância médica e veterinária relevantes em todo o mundo por causa da variedade de agentes patogênicos que podem transmitir. No presente trabalho, foi realizada uma pesquisa para identificar Babesia spp. e Ehrlichia spp. em carrapatos coletados de cães de Cuba. Foram coletados 431 carrapatos de 378 cães, tendo sido identificados como pertencentes às espécies de Ripicephalus sanguineus sensu lato (s. 1. O DNA genômico foi extraído com protocolo usando fenol/clorofórmio. Os carrapatos foram organizados em “pools” e o DNA extraído foi testado pela reação em cadeia da polimerase (nPCR para amplificar 398 pares de bases (pb do DNA ribossômico 16S (rDNA de Ehrlichia canis e PCR para amplificar aproximadamente 560 pb do DNA ribossômico 18S (rDNA. Dos 49 pools testados, 8,16% (n = 4/49 foram positivos para o E. canis por nPCR visando o gene do 16S rDNA e apenas um pool (n = 1/49; 2,04% foi positivo para o gene 18S rDNA para Babesia canis. As quatro sequências obtidas para o fragmento de 16S rDNA foram idênticas entre si e resultaram em 100% de identidade com E. canis de diferentes países. A sequência obtida do gene do 18S rDNA para Babesia spp. apresentou semelhança de 100% com Babesia canis vogeli quando comparada às sequências depositadas no Genbank. Esta foi a primeira detecção molecular desses agentes no carrapato R. sanguineus s. l. em Cuba.

  8. A retrospective study of Babesia macropus associated with morbidity and mortality in eastern grey kangaroos (Macropus giganteus and agile wallabies (Macropus agilis

    Directory of Open Access Journals (Sweden)

    Shannon L. Donahoe

    2015-08-01

    Full Text Available This is a retrospective study of 38 cases of infection by Babesia macropus, associated with a syndrome of anaemia and debility in hand-reared or free-ranging juvenile eastern grey kangaroos (Macropus giganteus from coastal New South Wales and south-eastern Queensland between 1995 and 2013. Infection with B. macropus is recorded for the first time in agile wallabies (Macropus agilis from far north Queensland. Animals in which B. macropus infection was considered to be the primary cause of morbidity had marked anaemia, lethargy and neurological signs, and often died. In these cases, parasitised erythrocytes were few or undetectable in peripheral blood samples but were sequestered in large numbers within small vessels of visceral organs, particularly in the kidney and brain, associated with distinctive clusters of extraerythrocytic organisms. Initial identification of this piroplasm in peripheral blood smears and in tissue impression smears and histological sections was confirmed using transmission electron microscopy and molecular analysis. Samples of kidney, brain or blood were tested using PCR and DNA sequencing of the 18S ribosomal RNA and heat shock protein 70 gene using primers specific for piroplasms. The piroplasm detected in these samples had 100% sequence identity in the 18S rRNA region with the recently described Babesia macropus in two eastern grey kangaroos from New South Wales and Queensland, and a high degree of similarity to an unnamed Babesia sp. recently detected in three woylies (Bettongia penicillata ogilbyi in Western Australia.

  9. Soroepidemiologia de Rhodococcus equi em equinos da região de Bagé, RS, pelo teste de inibição da hemólise sinérgica

    Directory of Open Access Journals (Sweden)

    Lazzari Andrea

    1997-01-01

    Full Text Available Com o objetivo de avaliar a situação soroepidemiológica da infecção por Rhodococcus equi na região de Bagé, RS, foram testadas 290 amostras de soro sanguíneo de éguas e potros aparentemente sadios, obtidos de 6 haras com diferentes histórias de rodococose. Para relacionar o resultado sorológico com a presença deste agente bacteriano no trato intestinal destes animais, foram coletadas 123 amostras de fezes. O teste sorológico utilizado foi a inibição da hemólise sinérgica (IHS que detecta anticorpos neutralizantes contra o "fator equi". Um percentual de 87,93% (255/290 dos animais amostrados apresentaram estes anticorpos. O título médio geométrico (GMT destes anticorpos foi mais elevado nos potros do que nas éguas. A soropositividade destes equinos ao teste sorológico teve correlação com o isolamento do R. equi nas fezes dos respectivos animais. A maior taxa de isolamento de R. equi das fezes dos equinos e o maior GMT, ocorreu no único haras com casos clínicos recentes de enfermidade causada por esta bactéria. No entanto, todos os animais deste e dos demais haras, encontravam-se aparentemente sadios, sendo necessário, estabelecer em trabalho futuro, a possível relação entre títulos de anticorpos e sua importância na detecção da enfermidade.

  10. Evaluation of an enzyme linked immunosorbent assay kit for the detection of Babesia bovis antibodies in cattle in Argentina

    Energy Technology Data Exchange (ETDEWEB)

    Echaide, S; Echaide, I E; Mangold, A J; Lugaresi, C I; Guglielmone, A A [Estacion Experimental Agropecuaria, Instituto Nacional de Tecnologia Agropecuaria, Rafaela, Santa Fe (Argentina); Gaido, A B [Estacion Experimental Agropecuaria Salta, Salta (Argentina)

    1998-11-01

    An enzyme linked immunosorbent assay (ELISA) for the detection of antibodies to Babesia bovis was evaluated by using sera of 874 cattle carrying B. bovis antibodies, 700 sera of uninfected cattle, and 357 sera from calves from 16 herds subjected to different B. bovis inoculation rates. The seropositive/ seronegative cut-off point set as double the mean percent positivity of negative cattle sera (= 16%). The sensitivity of the ELISA (four trials) ranged from 97.1% to 100% and the specificity (three trials) varied from 92.0% to 97.0%. The agreement between ELISA and immunofluorescent antibody test was {>=} 90.0% in 18 of 23 evaluations and it ranged from 86.0% to 88.0% in the remainder. The correlation coefficient between percentage of sera positive to ELISA and IFA test in 16 herds was 0.9958 (P<0.001). The ELISA has the advantages of a high sensitivity, objectivity and capacity to test large number of samples in short period of time and could replace the IFA test specially for epidemiological studies. (author) 11 refs, 1 fig., 2 tabs

  11. Molecular detection of Dirofilaria immitis, Hepatozoon canis, Babesia spp., Anaplasma platys and Ehrlichia canis in dogs on Costa Rica.

    Science.gov (United States)

    Wei, Lanjing; Kelly, Patrick; Ackerson, Kate; El-Mahallawy, Heba S; Kaltenboeck, Bernhard; Wang, Chengming

    2014-03-01

    Although vector-borne diseases are important causes of morbidity and mortality in dogs in tropical areas, there is little information on these conditions in Costa Rica. In PCRs of blood from dogs in Costa Rica, we did not detect DNAs of Rickettsia (R.) felis and Coxiella (C.) burnetii but we did find evidence of infection with Dirofilaria (D.) immitis (9/40, 22.5%), Hepatozoon (H.) canis (15/40, 37.5%), Babesia spp. (10/40, 25%; 2 with B. gibsoni and 8 with B. vogeli), Anaplasma (A.) platys (3/40, 7.5%) and Ehrlichia (E.) canis (20/40, 50%). Nine dogs (22.5%) were free of any vector-borne pathogens while 14 (35%) were infected with a single pathogen, 11 (27.5%) with two, 4 (10%) with three, 1 (2.5%) with four, and 1 (2.5%) with five pathogens. Dogs in Costa Rica are commonly infected with vector-borne agents.

  12. Molecular detection of Ehrlichia canis, Hepatozoon canis and Babesia canis vogeli in stray dogs in Mahasarakham province, Thailand.

    Science.gov (United States)

    Piratae, Supawadee; Pimpjong, Kiattisak; Vaisusuk, Kotchaphon; Chatan, Wasupon

    2015-01-01

    Canine tick borne diseases showing distribution worldwide have caused morbidity and mortality in dogs. This study observed the mainly tick borne pathogens described for dogs in Thailand, Ehrlichia canis, Hepatozoon canis and Babesia canis vogeli. From May to July 2014, blood samples were collected from 79 stray dogs from 7 districts of Mahasarakham province to molecular surveyed for 16s rRNA gene of E. canis and 18s rRNA gene of H. canis and B. canis vogeli. Twenty eight (35.44%) of stray dogs showed the infection with tick borne pathogens. The prevalence of E. canis infection was the highest with 21.5% (17/79). DNA of H. canis and B. canis vogeli were detected at the prevalence of 10.1% (8/79) and 6.3% (5/79), respectively. Co-infection between E. canis and B. canis vogeli were identified in 2 (2.5%) dogs. The results indicated that a wide range of tick borne pathogens are circulation in the canine population in Mahasarakham province. This study is the first report on prevalence of E. canis, H. canis and B. canis vogeli in stray dogs in Mahasarakham, a province in northern part of Thailand. This data providing is important to understand the prevalence of E. canis, H. canis and B. canis vogeli infection in stray dogs in this region, which will assist in the management of these blood parasite.

  13. Development of an enzyme-linked immunosorbent assay for detection of IgM antibodies to Babesia bigemina in cattle

    Directory of Open Access Journals (Sweden)

    Ruiz Patrícia M Gonçalves

    2001-01-01

    Full Text Available A crude antigenic preparation of Babesia bigemina was used to develop an ELISA for the detection of IgM antibodies. Optimal dilutions of the antigen, using positive and negative reference sera, were determined by checkerboard titrations. Negative sera from cattle imported from tick-free areas, serum samples collected from infected B. bigemina cattle were used to validate the test. The specificity was 94% and sensitivity of the Elisa 87.5%. Sera from 385 cattle deriving from areas free from tick-borne diseases, which were submitted to a preimmunization process, were screened by this technique. The Elisa detected seroconversion on the 14th day post-inoculation in animals either infested with Boophilus microplus ticks (infected with B. bigemina, or inoculated with B. bigemina infected blood. Antibody titers decreased after day 33; however, all animals remained positive until the end of the experiment (124 days. The ELISA described may prove to be an appropriate serological test for the detection of IgM antibodies against B. bigemina.

  14. Evaluation of an enzyme linked immunosorbent assay kit for the detection of Babesia bovis antibodies in cattle in Argentina

    International Nuclear Information System (INIS)

    Echaide, S.; Echaide, I.E.; Mangold, A.J.; Lugaresi, C.I.; Guglielmone, A.A.; Gaido, A.B.

    1998-01-01

    An enzyme linked immunosorbent assay (ELISA) for the detection of antibodies to Babesia bovis was evaluated by using sera of 874 cattle carrying B. bovis antibodies, 700 sera of uninfected cattle, and 357 sera from calves from 16 herds subjected to different B. bovis inoculation rates. The seropositive/ seronegative cut-off point set as double the mean percent positivity of negative cattle sera (= 16%). The sensitivity of the ELISA (four trials) ranged from 97.1% to 100% and the specificity (three trials) varied from 92.0% to 97.0%. The agreement between ELISA and immunofluorescent antibody test was ≥ 90.0% in 18 of 23 evaluations and it ranged from 86.0% to 88.0% in the remainder. The correlation coefficient between percentage of sera positive to ELISA and IFA test in 16 herds was 0.9958 (P<0.001). The ELISA has the advantages of a high sensitivity, objectivity and capacity to test large number of samples in short period of time and could replace the IFA test specially for epidemiological studies. (author)

  15. Molecular epidemiology of Anaplasma platys, Ehrlichia canis and Babesia vogeli in stray dogs in Paraná, Brazil

    Directory of Open Access Journals (Sweden)

    Claudia M. Ribeiro

    Full Text Available ABSTRACT: Hemoparasitic infections are tick-borne diseases, which affect animals and humans. Considering the importance of canine hemoparasitic infections in veterinary clinics, this study aimed to determine the occurrence of Anaplasma platys, Ehrlichia canis and Babesia vogeli in blood samples from 182 dogs not domiciled in the city of Pato Branco, southwestern region of Paraná State, Brazil, using polymerase chain reaction (PCR. The prevalence of A. platys and B. vogeli was 32.9% and 10.9% respectively, and A. platys infection prevailed (p<0.001. The number of dogs positive for A. platys was larger in Winter (p<0.05. All blood samples were negative for E. canis. In the dogs, infestation by Amblyomma cajennense predominated over that by Rhipicephalus sanguineus (p<0.001; but there was no significant association between PCR and the variables presence of ticks, sex and age. Dogs infected by A. platys and B. vogeli showed thrombocytopenia, lymphopenia and leukocytosis; but there was no correlation between such hematological changes and infection by hemoparasites. This appears to be the first molecular study that demonstrates the existence of A. platys and B. vogeli in dogs from the southwestern region of Paraná.

  16. The Babesia bovis hap2 gene is not required for blood stage replication, but expressed upon in vitro sexual stage induction.

    Directory of Open Access Journals (Sweden)

    Hala E Hussein

    2017-10-01

    Full Text Available Babesia bovis, is a tick borne apicomplexan parasite responsible for important cattle losses globally. Babesia parasites have a complex life cycle including asexual replication in the mammalian host and sexual reproduction in the tick vector. Novel control strategies aimed at limiting transmission of the parasite are needed, but transmission blocking vaccine candidates remain undefined. Expression of HAP2 has been recognized as critical for the fertilization of parasites in the Babesia-related Plasmodium, and is a leading candidate for a transmission blocking vaccine against malaria. Hereby we identified the B. bovis hap2 gene and demonstrated that it is widely conserved and differentially transcribed during development within the tick midgut, but not by blood stage parasites. The hap2 gene was disrupted by transfecting B. bovis with a plasmid containing the flanking regions of the hap2 gene and the GPF-BSD gene under the control of the ef-1α-B promoter. Comparison of in vitro growth between a hap2-KO B. bovis clonal line and its parental wild type strain showed that HAP2 is not required for the development of B. bovis in erythrocytes. However, xanthurenic acid-in vitro induction experiments of sexual stages of parasites recovered after tick transmission resulted in surface expression of HAP2 exclusively in sexual stage induced parasites. In addition, hap2-KO parasites were not able to develop such sexual stages as defined both by morphology and by expression of the B. bovis sexual marker genes 6-Cys A and B. Together, the data strongly suggests that tick midgut stage differential expression of hap2 is associated with the development of B. bovis sexual forms. Overall these studies are consistent with a role of HAP2 in tick stages of the parasite and suggest that HAP2 is a potential candidate for a transmission blocking vaccine against bovine babesiosis.

  17. The Babesia bovis hap2 gene is not required for blood stage replication, but expressed upon in vitro sexual stage induction

    Science.gov (United States)

    Hussein, Hala E.; Bastos, Reginaldo G.; Schneider, David A.; Johnson, Wendell C.; Adham, Fatma K.; Davis, William C.; Laughery, Jacob M.; Herndon, David R.; Alzan, Heba F.

    2017-01-01

    Babesia bovis, is a tick borne apicomplexan parasite responsible for important cattle losses globally. Babesia parasites have a complex life cycle including asexual replication in the mammalian host and sexual reproduction in the tick vector. Novel control strategies aimed at limiting transmission of the parasite are needed, but transmission blocking vaccine candidates remain undefined. Expression of HAP2 has been recognized as critical for the fertilization of parasites in the Babesia-related Plasmodium, and is a leading candidate for a transmission blocking vaccine against malaria. Hereby we identified the B. bovis hap2 gene and demonstrated that it is widely conserved and differentially transcribed during development within the tick midgut, but not by blood stage parasites. The hap2 gene was disrupted by transfecting B. bovis with a plasmid containing the flanking regions of the hap2 gene and the GPF-BSD gene under the control of the ef-1α-B promoter. Comparison of in vitro growth between a hap2-KO B. bovis clonal line and its parental wild type strain showed that HAP2 is not required for the development of B. bovis in erythrocytes. However, xanthurenic acid-in vitro induction experiments of sexual stages of parasites recovered after tick transmission resulted in surface expression of HAP2 exclusively in sexual stage induced parasites. In addition, hap2-KO parasites were not able to develop such sexual stages as defined both by morphology and by expression of the B. bovis sexual marker genes 6-Cys A and B. Together, the data strongly suggests that tick midgut stage differential expression of hap2 is associated with the development of B. bovis sexual forms. Overall these studies are consistent with a role of HAP2 in tick stages of the parasite and suggest that HAP2 is a potential candidate for a transmission blocking vaccine against bovine babesiosis. PMID:28985216

  18. Determination of Prevalence and Risk Factors of Infection with Babesia ovis in Small Ruminants from West Azerbaijan Province, Iran by Polymerase Chain Reaction

    Directory of Open Access Journals (Sweden)

    Bijan Esmaeilnejad

    2015-10-01

    Full Text Available Background: Small ruminants’ babesiosis caused by Babesia ovis, is transmitted during blood feeding by infected ticks and is the most economically important tick-borne disease in tropical and subtropical areas. This study was carried out to to estimate the infection rate of B. ovis in sheep and goats by PCR. We have analysed risk factors that might influence infection of sheep and goats with B. ovis.Methods: A total 402 blood samples were examined microscopically for the presence of Babesia infection. All samples were tested by PCR. During sampling, whole body of each animal and farm dogs was examined for the presence of ticks.Results: Forty-two animals (10.4% were positive for Babesia spp. upon microscopic examination, whereas 67 animals (16.7% yielded the specific DNA for B. ovis of which 52 animals were sheep and 15 animals were goats.Twenty-nine farms (72.5% were found positive for B. ovis. The percentage of positive animals in each location varied from 13 % to 20 %. The relative risk of the presence of ticks in sheep and goats (P< 0.01 and farm dogs (P< 0.01 for PCRpositive results forB. ovis in sheep and goats was found 3.8 and 2.9, respectively. A total of 747 ticks identified as Rhipicephalus bursa, R. sanguineus and R. turanicus on the basis of morphological features.Conclusion: Other animal species besides dogs may also be risk factors for babesiosis in sheep and goats. Also, R. bursa may play an important role as a vector of the parasite in Iran.

  19. Forward prediction of tunnel face by use of equi-traveltime planes; Tosojimen no riyo ni yoru tunnel kiriha zenpo tansa

    Energy Technology Data Exchange (ETDEWEB)

    Ashida, Y [Kyoto University, Kyoto (Japan). Faculty of Engineering; Hirano, T [Nishimatsu Construction Co. Ltd., Tokyo (Japan)

    1996-05-01

    The result is reported herein of a depth transform algorithm application utilizing equi-traveltime planes in reflection seismic exploration for the purpose of knowing the structure configuration in front of the tunnel face in a tunnel excavation process. A vibration producing hole was provided 0.5m above the ground surface while a vibration receiving hole was provided at 1.5m above the ground surface, and the excavated lengths were 1.5m and 0.5m, respectively. There were 24 recording channels, from which inline offset records were taken. The on-site records were subjected to 100-1000Hz band pass filtering. The elastic wave propagation rate in the rock used for depth conversion was set at 400m/s in consideration of the distance and direct wave travel time from the vibration source to the vibration receiving point. As the result of study, weak layers were found in the neighborhood of 230-240m, 260-270m, and 280-290m. This method requires less core memory for calculation and completes calculation in a shorter period of time, which enables a conclusion that it will be effective in grasping the 3-dimensional structure in front of the tunnel face in the working scene. 4 refs., 7 figs.

  20. Natural Babesia bovis infection in water buffaloes (Bubalus bubalis) and crossbred cattle under field conditions in Egypt

    DEFF Research Database (Denmark)

    Mahmmod, Yasser

    2014-01-01

    showed no clinical signs and were free from external, internal, and blood parasites served as control group. Results: Babesia bovis-infected cattle showed typical signs of bovine babesiosis while B. bovis-infected buffaloes showed a milder form (less severe) of the clinical signs. Advanced cases...... cattle under field conditions in Egypt. Methods: A total of 35 buffaloes and cattle were clinically and laboratory investigated from March to June 2008. Twenty-nine buffaloes and cattle out of 35 were naturally infected with B. bovis and showed signs of bovine babesiosis. Three cows and three buffaloes...

  1. Serum antibodies from a subset of horses positive for babesia caballi by competitive enzyme-linked immunosorbent assay demonstrate a protein recognition pattern that is not consistent with infection

    Science.gov (United States)

    Tick-borne pathogens that cause persistent infection are of major concern to the livestock industry because of transmission risk from persistently infected animals and the potential economic losses they pose. The recent re-emergence of Theileria equi in the U.S. prompted widespread national surveill...

  2. Screening of bat faeces for arthropod-borne apicomplexan protozoa: Babesia canis and Besnoitia besnoiti-like sequences from Chiroptera.

    Science.gov (United States)

    Hornok, Sándor; Estók, Péter; Kováts, Dávid; Flaisz, Barbara; Takács, Nóra; Szőke, Krisztina; Krawczyk, Aleksandra; Kontschán, Jenő; Gyuranecz, Miklós; Fedák, András; Farkas, Róbert; Haarsma, Anne-Jifke; Sprong, Hein

    2015-08-28

    Bats are among the most eco-epidemiologically important mammals, owing to their presence in human settlements and animal keeping facilities. Roosting of bats in buildings may bring pathogens of veterinary-medical importance into the environment of domestic animals and humans. In this context bats have long been studied as carriers of various pathogen groups. However, despite their close association with arthropods (both in their food and as their ectoparasites), only a few molecular surveys have been published on their role as carriers of vector-borne protozoa. The aim of the present study was to compensate for this scarcity of information. Altogether 221 (mostly individual) bat faecal samples were collected in Hungary and the Netherlands. The DNA was extracted, and analysed with PCR and sequencing for the presence of arthropod-borne apicomplexan protozoa. Babesia canis canis (with 99-100% homology) was identified in five samples, all from Hungary. Because it was excluded with an Ixodidae-specific PCR that the relevant bats consumed ticks, these sequences derive either from insect carriers of Ba. canis, or from the infection of bats. In one bat faecal sample from the Netherlands a sequence having the highest (99%) homology to Besnoitia besnoiti was amplified. These findings suggest that some aspects of the epidemiology of canine babesiosis are underestimated or unknown, i.e. the potential role of insect-borne mechanical transmission and/or the susceptibility of bats to Ba. canis. In addition, bats need to be added to future studies in the quest for the final host of Be. besnoiti.

  3. Equine Piroplasmosis

    Science.gov (United States)

    Equine piroplasmosis is an infectious, tick-borne disease caused by the hemoprotozoan parasites Theileria (previously Babesia) equi and Babesia caballi. Piroplasmosis affects all wild and domestic equid species and causes signs related to intravascular hemolysis and associated systemic illness. Infe...

  4. BQP35 is a novel member of the intrinsically unstructured protein (IUP) family which is a potential antigen for the sero-diagnosis of Babesia sp. BQ1 (Lintan) infection.

    Science.gov (United States)

    Guan, Guiquan; Moreau, Emmanuelle; Liu, Junlong; Ma, Miling; Rogniaux, Hélène; Liu, Aihong; Niu, Qingli; Li, Youquan; Ren, Qiaoyun; Luo, Jianxun; Chauvin, Alain; Yin, Hong

    2012-07-06

    A new gene of Babesia sp. BQ1 (Lintan) (BQP35) was cloned by screening a merozoite cDNA expression library with infected sheep serum and using rapid amplification of cDNA ends (RACE). The nucleotide sequence of the cDNA was 1140bp with an open reading frame (ORF) of 936bp encoding a 35-kDa predicted polypeptide with 311 amino acid residues. Comparison of BQP35 cDNA and genomic DNA sequences showed that BQP35 does not possess an intron. Recombinant BQP35 (rBQP35), expressed in a prokaryotic expression system, showed abnormally slow migration on SDS-PAGE. Gel shifting, amino acid sequence and in silico disorder region prediction indicated that BQP35 protein has characteristics of intrinsically unstructured proteins (IUPs). This is the first description of such proteins in the Babesia genus. BQP35 induced antibodies production as early as one week after Babesia sp. BQ1 (Lintan) infection in sheep. No cross-reaction was observed with sera from sheep infected with other ovine piroplasms dominant in China, except with Babesia sp. Tianzhu. The interest of BQP35 as a diagnostic antigen is discussed. Copyright © 2012 Elsevier B.V. All rights reserved.

  5. The application of the HyPer fluorescent sensor in the real-time detection of H2O2 in Babesia bovis merozoites in vitro.

    Science.gov (United States)

    Asada, Masahito; Hakimi, Hassan; Kawazu, Shin-Ichiro

    2018-05-15

    In recent years, genetically encoded fluorescent probes have allowed a dramatic advancement in time-lapse imaging, enabling this imaging modality to be used to investigate intracellular events in several apicomplexan parasite species. In this study, we constructed a plasmid vector to stably express a genetically encoded H 2 O 2 sensor probe called HyPer in Babesia bovis. The HyPer-transfected parasite population was successfully developed and subjected to a time-lapse imaging analysis under in vitro culture conditions. HyPer was capable of sensing an increasing H 2 O 2 concentration in the parasite cells which was induced by the administration of paraquat as a superoxide donor. HyPer fluorescence co-staining with MitoTracker Red indicated the mitochondria as the major source of reactive oxygen species (ROS) in parasite cells. The fluctuating ROS dynamics in the parasite gliding toward, attaching to, and invading the target red blood cell was visualized and monitored in real time with the HyPer expressing parasite population. This is the first report to describe the application of the HyPer probe in an imaging analysis involving Babesia parasites. Hyper-expressing parasites can be widely utilized in studies to investigate the mechanisms of emergence and the reduction of oxidative stress, as well as the signal transduction in the parasite cells during host invasion and intercellular development. Copyright © 2018 Elsevier B.V. All rights reserved.

  6. High throughput pyrosequencing technology for molecular differential detection of Babesia vogeli, Hepatozoon canis, Ehrlichia canis and Anaplasma platys in canine blood samples.

    Science.gov (United States)

    Kaewkong, Worasak; Intapan, Pewpan M; Sanpool, Oranuch; Janwan, Penchom; Thanchomnang, Tongjit; Kongklieng, Amornmas; Tantrawatpan, Chairat; Boonmars, Thidarut; Lulitanond, Viraphong; Taweethavonsawat, Piyanan; Chungpivat, Sudchit; Maleewong, Wanchai

    2014-06-01

    Canine babesiosis, hepatozoonosis, ehrlichiosis, and anaplasmosis are tick-borne diseases caused by different he