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Sample records for babesia equi trophozoites

  1. Inhibition of the in vitro growth of babesia bigemina, babesia caballi and theileria equi parasites by trifluralin analogues

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    Background: Bovine and equine babesiosis caused by Babesia bovis, B. bigemina and B. caballi, and equine theileriosis caused by Theileria equi are global tick borne hemoprotozoan diseases characterized by fever, anemia, weight losses and abortions. A common feature of these diseases are transition f...

  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

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    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. Theileria equi and Babesia caballi infection of equids in Punjab, India: a serological and molecular survey.

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    Sumbria, Deepak; Das Singla, Lachhman; Sharma, Amrita

    2016-01-01

    A cross-sectional study was conducted in Submountain undulating, Undulating plain, Western and Western plain agro-climatic zones of Punjab province, India, to determine the prevalence, agreement between diagnostic tests and associated related risk factors of Theileria equi and Babesia caballi infection in equids (horses, donkey, mules). An overall prevalence of 14.14 and 0.0% of T. equi and B. caballi was recorded by multiplex polymerase chain reaction targeting 18S ribosomal RNA (rRNA) for both the parasites and 75 and 1.11% by competitive enzyme-linked immunosorbent assay in a representative sample of 180 animals. Only two animals with positive antibody titre from B. caballi and none with PCR indicated T. equi as the predominant haemoprotozoan responsible for equine piroplasmosis in the study area. Among the PCR-positive animals, presence of tick vectors in farm vicinity was the most influential associated with T. equi infection (P = 0.002; odds ratio (OR) 9.30; 95% confidence interval (CI) = 3.32-27.10). For animals with higher anti-T. equi antibody titres, strong association of sero-prevalence for T. equi was recorded with age, sex, usage, tick infestation and deworming/vaccination status of host animals and farm management strategies. The study has demonstrated the possible absence of B. caballi in both conducive and non-conducive areas of Punjab and demonstrated T. equi as the potential agent of equine piroplasmosis in Punjab.

  5. Babesia equi-induced anemia in a Quarter Horse and subsequent regulatory response.

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    Beard, Laurie A; Pelzel, Angela M; Rush, Bonnie R; Wright, Abra M; Galgut, Bradley I; Hennager, Steven G; King, Alison O; Traub-Dargatz, Josie L

    2013-04-01

    A 7-year-old Quarter Horse gelding used for unsanctioned racing was examined because of fever and anorexia. Physical examination revealed fever, tachycardia, and tachypnea. Results of a CBC indicated anemia and mild thrombocytopenia. Results of microscopic examination of a blood smear indicated piroplasms in erythrocytes, consistent with Babesia spp. Regulatory authorities were contacted, and results of serologic testing at the National Veterinary Services Laboratories confirmed acute Babesia equi infection. Equids on the home premises of the index horse were placed under quarantine. Those equids were tested for piroplasmosis, and 6 of 63 horses had positive results for B equi. Another horse that had previously been housed on the index premises also had positive results for B equi. Competent tick vectors for piroplasmosis organisms were not identified. All 8 horses with piroplasmosis were Quarter Horses that participated in unsanctioned racing and were trained by the same person. Two of the horses were illegally removed from the index premises; these 2 horses and the other horse with piroplasmosis that was previously housed on the index premises could not be found. The other 5 horses with piroplasmosis were euthanized. Investigators concluded that transmission of B equi among horses was most likely iatrogenic. The United States has been considered piroplasmosis free. However, veterinarians should consider piroplasmosis in horses with signalments and clinical signs similar to those of the index horse of this report. Regulatory authorities should be contacted regarding horses in which piroplasmosis is suspected.

  6. Microscopic and Molecular detection of Theileria (Babesia equi infection in equids of Kurdistan Province, Iran

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    Gholamreza HABIBI

    2016-03-01

    Full Text Available Background: 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.Methods: 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.Results: 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%.  Conclusion: 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.

  7. 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.

  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).

  9. Molecular detection and prevalence of Theileria equi and Babesia caballi in horses of central Balkan.

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    Davitkov, Darko; Vucicevic, Milos; Stevanovic, Jevrosima; Krstic, Vanja; Slijepcevic, Dajana; Glavinic, Uros; Stanimirovic, Zoran

    2016-03-01

    Equine piroplasmosis is significant tick-borne disease with wide distribution. The prevalence of equine piroplasmosis in Serbia, Montenegro and Bosnia and Herzegovina is unknown. In aim to obtain a first insight into the prevalence we performed molecular epidemiological study which included 142 horses, on seven locations in these three countries. We first performed PCR for the detection of a 450bp long section of the 18S rRNA of piroplasma-specific region. For all positive samples we have done multiplex PCR for the species detection. Species determination was further confirmed by sequencing PCR products of 10 randomly selected Theileria equi and all Babesia caballi samples. The overall prevalence rates in analysed region for T. equi and B. caballi were 22.5% and 2.1%, respectively. Possible risk factors (such as location, age, sex and activity) associated with PCR positivity were evaluated. Marked differences were found in prevalence between geographic areas. There was no significant association between positivity and age group. T. equi was more prevalent in females and farming horses. This is the first report on the molecular survey of T. equi and B. caballi in central Balkan. Further prevalence studies on definitive host and vectors in this region are necessary.

  10. Serodiagnosis of Babesia equi in horses submitted to exercise stress Diagnóstico sorológico de infestação por Babesia equi em eqüinos submetidos ao estresse do exercício

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    Cristiane D. Baldani

    2007-04-01

    Full Text Available A complement fixation test (CFT, performed in microtitre plates, based upon the use of crude antigenic preparation of Babesia equi was adapted for the detection of antibodies in serum of infected horses. The indirect fluorescent antibody test (IFAT and enzyme-linked immunosorbent assay (ELISA were also used for the immunodiagnosis of B. equi. Serum samples from 15 apparently healthy horses, previously conditioned to a high-speed equine treadmill, were taken before and after exercise. All the samples analyzed were positive for B. equi infection. There were no significant differences (PA reação de fixação do complemento (RFC, realizada em microplacas, utilizando-se antígeno bruto de Babesia equi, foi adaptada para a detecção de anticorpos em soros de eqüinos infectados. A reação de imunofluorescência indireta (RIFI e o ensaio imunoenzimático (ELISA também foram utilizados para o imunodiagnóstico de B. equi. Amostras de soro foram obtidas de 15 eqüinos aparentemente sadios, submetidos a treinamento físico em esteira rolante de alto desempenho, sendo as amostras colhidas antes e após os exercícios. Todas as amostras testadas foram positivas para B. equi. Não houve diferença significativa (P<0,01 entre estes 3 testes sorológicos, ou entre a condição de estresse e repouso. A combinação da RFC com a RIFI ou ELISA pode ser recomendada a fim de evitar a entrada de eqüinos portadores em áreas consideradas livres da doença.

  11. Effects of dihydroorotate dehydrogenase (DHODH) inhibitors on the growth of Theileria equi and Babesia caballi in vitro.

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    Kamyingkird, Ketsarin; Cao, Shinuo; Tuvshintulga, Bumduuren; Salama, Akram; Mousa, Ahmed Abdelmoniem; Efstratiou, Artemis; Nishikawa, Yoshifumi; Yokoyama, Naoaki; Igarashi, Ikuo; Xuan, Xuenan

    2017-05-01

    Theileria equi and Babesia caballi are the causative agents of equine piroplasmosis (EP), which affects equine production in various parts of the world. However, a safe and effective drug is not currently available for treatment of EP. Dihydroorotate dehydrogenase (DHODH) is the fourth enzyme in the de novo pyrimidine synthesis pathway and has been known as a novel drug target for several apicomplexan protozoan parasites. In this study, we evaluated four DHODH inhibitors; atovaquone (ATV), leflunomide (LFN), brequinar (Breq), and 7-hydroxy-5-[1,2,4] triazolo [1,5,a] pyrimidine (TAZ) on the growth of T. equi and B. caballi in vitro and compared them to diminacene aceturate (Di) as the control drug. The growth of T. equi and B. caballi was significantly hindered by all inhibitors except TAZ. The half maximal inhibitory concentration (IC50) of ATV, LFN, Breq and Di against T. equi was approximately 0.028, 109, 11 and 40 μM, respectively, whereas the IC50 of ATV, LFN, Breq and Di against B. caballi was approximately 0.128, 193, 5.2 and 16.2 μM, respectively. Using bioinformatics and Western blot analysis, we showed that TeDHODH was similar to other Babesia parasite DHODHs, and confirmed that targeting DHODHs could be useful for the development of novel chemotherapeutics for treatment of EP.

  12. 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.

  13. Molecular and serological detection of Theileria equi and Babesia caballi infection in horses and ixodid ticks in Iran.

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    Abedi, Vali; Razmi, Golamreza; Seifi, Hesam; Naghibi, Abolghasem

    2014-04-01

    Equine piroplasmosis is a hemoprotozoan tick-borne disease with worldwide distribution that is caused by Theileria equi and Babesia caballi. However, the geographical distribution of equine piroplasmosis in Iran is unknown. The aim of the current study was to determine the causative agents and vector ticks of equine piroplasmosis in horses in the North Khorasan Province. In the year 2011, 100 horses were randomly selected from 14 villages. Blood samples and ixodid ticks were collected and examined using microscopical, molecular, and serological methods. Theileria equi infection was microscopically detected in 5 (5%) of the blood smears with low parasitemia, while serum samples were tested by the indirect immunofluorescent antibody test (IFAT). Antibodies against T. equi, B. caballi, and a mixed infection were detected in 48 (48%), 2 (2%), and 3 (3%) of the serum samples, respectively. A multiplex PCR was used to detect T. equi and B. caballi DNA in blood samples. No B. caballi infections could be found, but Theileria equi DNA was detected in 45 (45%) of the blood samples, and a BLAST analysis of the sequenced samples indicated a 99% similarity with T. equi 18S rRNA gene sequences in GenBank. Both molecular and serological results did not identify any significant association between T. equi infection and risk factors. A comparision of the results of 3 diagnostic methods demonstrated a poor agreement between microscopical examination with IFAT and PCR and a moderate agreement between IFAT and PCR. Thirty-seven adult ticks (20 females and 17 males) were collected from 15 horses. The most common tick was Hyalomma marginatum marginatum (n=19), followed by Hyalomma anatolicum excavatum (n=10), Rhipicephalus bursa (n=4), Hyalomma marginatum turanicum (n=3), and Hyalomma anatolicum anatolicum (n=1). The salivary glands and ovaries were also examined using PCR. The genomic DNA samples of the salivary glands of 3 ticks, H. a. excavatum (n=2) and R. bursa (n=1), had a

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

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    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.

  15. Detection of Theileria equi and Babesia caballi infections in Venezuelan horses using Competitive-Inhibition ELISA and PCR.

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    Rosales, Romel; Rangel-Rivas, Ariadna; Escalona, América; Jordan, Luis Segundo; Gonzatti, Mary Isabel; Aso, Pedro Maria; Perrone, Trina; Silva-Iturriza, Adriana; Mijares, Alfredo

    2013-09-01

    The focus of this study was the detection of equine piroplasmosis in Distrito Capital, Miranda, Aragua, Guárico and Apure States from Venezuela, using two methods: Competitive-Inhibition ELISA and multiplex PCR and the analysis of the possible differences in occurrence in relation to the primary purpose of the horses, which is related to varied degrees of exposure to tick. Antibody levels to Babesia caballi and Theileria equi were assessed in 694 equine serum samples using Competitive-Inhibition ELISA, while PCR assays were performed in 136 horses, using two sets of oligonucleotides to establish the presence of T. equi, B. caballi or both. The overall seroprevalence of equine piroplasmosis was 50.2%, antibodies to B. caballi were found in 161 horses (23.2%), whereas 97 (14.0%) were seropositive to T. equi and 90 (13.0%) were positives to both parasites (mixed infections). PCR determinations (n=136) showed a prevalence of 66.2%, distributed in 84 (61.8% positives) for T. equi and, 6 (4.4%) were positive to both parasites. The cELISA showed higher levels of prevalence of B. caballi and mixed infections, as compared to the PCR method. This discrepancy can be explained by the different parameters that are evaluated by each technique, PCR detect the parasite itself, while cELISA detects antibodies to the parasite. By PCR, the highest prevalence was found in Apure state, where 92.3% of the samples were positive to T. equi infections. In this locality, free grazing animals are used for livestock management. This high prevalence may be linked to the tick species present in that area. More epidemiological studies will be necessary to assess the epidemiological status of equine piroplasmosis in Venezuela.

  16. Seroepidemiological survey of Theileria equi and Babesia caballi in horses from a rural and from urban areas of Paraná State, southern Brazil.

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    Vieira, Thállitha S W J; Vieira, Rafael F C; Finger, Mariane A P; Nascimento, Denise A G; Sicupira, Patrícia M L; Dutra, Leonardo H; Deconto, Ivan; Barros-Filho, Ivan R; Dornbusch, Peterson T; Biondo, Alexander W; Vidotto, Odilon

    2013-12-01

    The objective of this study was to evaluate the seroepidemiological data of Babesia caballi and Theileria equi in horses from a rural settlement and carthorses from urban areas of Paraná State, southern Brazil. A total of 198 horses, including 32 from the rural settlement and 166 carthorses from Colombo (n=48), Pinhais (n=76), Londrina (n=24), and Curitiba city (n=18) was sampled and tested using a commercial competitive inhibition ELISA (cELISA) test. Out of the 198 horses, 193 (97.5%) were seropositive for at least one piroplasm species. Antibodies to T. equi were detected in 155/198 horses (78.3%), antibodies to B. caballi were detected in 137/198 horses (69.2%), and antibodies to both were detected in 99/198 (50.0%) horses. Horses living in the rural settlement and Colombo were more likely to be seropositive to T. equi than those in Curitiba (p0.05). In conclusion, the high seroprevalences to B. caballi and T. equi observed in this study emphasize that active surveillance programs are critical for monitoring animal health status, particularly because carthorses may act as urban disseminators of these piroplasms.

  17. Babesia caballi and Theileria equi infections in horses in Central-Southern Italy: Sero-molecular survey and associated risk factors.

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    Bartolomé Del Pino, Leticia Elisa; Nardini, Roberto; Veneziano, Vincenzo; Iacoponi, Francesca; Cersini, Antonella; Autorino, Gian Luca; Buono, Francesco; Scicluna, Maria Teresa

    2016-04-01

    Babesia caballi and Theileria equi are tick-borne pathogens, etiological agents of equine piroplasmosis that affect different species of Equidae causing relevantly important direct and indirect losses. A field study was conducted to evaluate the distribution of the equine piroplasms in an area of Central-Southern Italy and to identify correlated risk factors. Serum samples of 673 asymptomatic horses were collected during spring-summer of 2013 to estimate the seroprevalence of the parasites within the study area using T. equi and B. caballi Antibody test kit (VMRD(®), Inc, Pullman, WA, USA). The 273 seropositive samples were subsequently tested by real time PCR to verify the presence of the genome of the piroplasms, indicative of the carrier status of the subjects. The variables chosen to identify which were the risk factors associated with the serological and PCR-positivity for each of the equine piroplasms were the following: gender, age, breed, access to pasture, altitude, land cover, climatic zone, soil type and province location (coastal/inland). The resulting overall seroprevalence for T. equi was 39.8% (268/673) and for B. caballi was 8.9% (60/673) while 70.3% of the PCR tested samples (185/263) were positive for T. equi and 10.3% (27/263) for B. caballi. The univariate and multiple logistic regression models were used to assess the association of the risk factors with the different outcomes. The risk factors found to be associated with T. equi seropositivity were gender, age, breed, access to pasture, land cover, soil type and province location, while those associated with PCR-positivity were age, soil type and province location. As the number of B. caballi seropositive subjects was limited, the multiple logistic regression model was performed only for the PCR-positive status, identifying climatic zone and soil type as the sole risk factors. In the study area, a major diffusion of T. equi, in terms of seroprevalence and PCR-positivity was present when

  18. 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.

  19. 马巴贝西虫EMA-1蛋白基因在杆状病毒系统中的表达%Expression of Babesia equi EMA-1 in insect cells by the baculovirus

    Institute of Scientific and Technical Information of China (English)

    张守发; 鞠玉琳; 贾洪林

    2004-01-01

    马巴贝西虫(Babesia equi)是一种蜱传播的血液寄生原虫。马巴贝西虫裂殖子抗原EMA-1,是B.equi的一种主要的表面蛋白,基因长度为816bp,由272个氨基酸组成,分子量为34kDa,具有较强的免疫原性和特异性,被认为具有广泛的开发前景。本文利用杆状病毒真核表达系统对B.equi的EMA-1基因进行了体外表达,

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

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    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. Comparative genomic analysis and phylogenetic position of Theileria equi

    Science.gov (United States)

    Transmission of arthropod-borne apicomplexan parasites that cause disease and result in death or persistent infection represents a major challenge to global human and animal health. First described in 1901 as Piroplasma equi, this re-emergent apicomplexan parasite was renamed Babesia equi and subseq...

  2. A Duplex PCR Method for Detection of Babesia caballi and Theileria equi%驽巴贝虫和马泰勒虫双重PCR检测方法的建立

    Institute of Scientific and Technical Information of China (English)

    张杨; 张玉婷; 王振宝; 波拉提; 李海; 巴音查汗

    2015-01-01

    目的 建立驽巴贝虫(Babesia caballi和马泰勒虫(Theileria equi)的双重PCR检测方法. 方法 根据GenBank发表的驽巴贝虫裂殖子mRNA BC48基因保守序列和马泰勒虫核糖体小亚基18 s rRNA基因保守序列,设计、合成2对特异性引物,经优化反应条件,建立双重PCR检测方法,并检测该方法的特异性和敏感性.用该双重PCR法对采集于伊犁地区马疑似病例全血样品进行检测,并与镜检法、常规PCR和荧光PCR的结果进行比较. 结果 建立的双重PCR法可特异性地扩增驽巴贝虫和马泰勒虫的相应目的条带,分别长约155 bp和280 bp,而对其他种属的双芽巴贝虫(B.bigemina)、环形泰勒虫(T.annulata)、瑟氏泰勒虫(T.sergenti、刚地弓形虫(Toxoplasma gondii)、犬新孢子虫(Neospora caninum)和伊氏锥虫(Trypanosoma evansi)进行扩增未见相应片段.该方法对驽巴贝虫和马泰勒虫的最低检出浓度分别为4.85×105拷贝/μl和4.85×104拷贝/μl.对采集的24份疑似血样进行双重PCR扩增,驽巴贝虫的阳性率为45.8% (11/24),马泰勒虫的阳性率为75.0% (18/24),镜检法、常规PCR和荧光PCR与双重PCR检测的符合率分别为91.7% (22/24)、95.8% (23/24)和95.8% (23/24). 结论 建立了可同时检测驽巴贝虫和马泰勒虫的双重PCR方法.

  3. 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.

  4. Comparative genomic analysis and phylogenetic position of Theileria equi

    Directory of Open Access Journals (Sweden)

    Kappmeyer Lowell S

    2012-11-01

    Full Text Available Abstract Background Transmission of arthropod-borne apicomplexan parasites that cause disease and result in death or persistent infection represents a major challenge to global human and animal health. First described in 1901 as Piroplasma equi, this re-emergent apicomplexan parasite was renamed Babesia equi and subsequently Theileria equi, reflecting an uncertain taxonomy. Understanding mechanisms by which apicomplexan parasites evade immune or chemotherapeutic elimination is required for development of effective vaccines or chemotherapeutics. The continued risk of transmission of T. equi from clinically silent, persistently infected equids impedes the goal of returning the U. S. to non-endemic status. Therefore comparative genomic analysis of T. equi was undertaken to: 1 identify genes contributing to immune evasion and persistence in equid hosts, 2 identify genes involved in PBMC infection biology and 3 define the phylogenetic position of T. equi relative to sequenced apicomplexan parasites. Results The known immunodominant proteins, EMA1, 2 and 3 were discovered to belong to a ten member gene family with a mean amino acid identity, in pairwise comparisons, of 39%. Importantly, the amino acid diversity of EMAs is distributed throughout the length of the proteins. Eight of the EMA genes were simultaneously transcribed. As the agents that cause bovine theileriosis infect and transform host cell PBMCs, we confirmed that T. equi infects equine PBMCs, however, there is no evidence of host cell transformation. Indeed, a number of genes identified as potential manipulators of the host cell phenotype are absent from the T. equi genome. Comparative genomic analysis of T. equi revealed the phylogenetic positioning relative to seven apicomplexan parasites using deduced amino acid sequences from 150 genes placed it as a sister taxon to Theileria spp. Conclusions The EMA family does not fit the paradigm for classical antigenic variation, and we propose a

  5. Seroepidemiological Detection of Babesia equi and Babesia caballi Infections in Qinghai Province%青海省马属小巴贝斯原虫和马巴贝斯原虫感染的血清流行病学检测

    Institute of Scientific and Technical Information of China (English)

    叶成玉; 王戈平; 陆艳; 蔡其刚; 牛小迎; 李晓卉; 马利青

    2011-01-01

    用重组蛋白作为ELISA抗原对青海地区马的上属小巴贝斯原虫[b.(Th.)equi]和马巴贝斯原虫(B.caballi)感染的流行情况进行了调查.经过对所收集的317份马属动物血清抗体的检测,共检出B.(Th.)equi阳性血清16份,B.caballi 阳性血清11份.结果初步说明我省的马属动物群中存在马巴贝斯原虫病的流行.

  6. The transmission of Babesia canis to the wild dog Lycaon pictus (Temminck) and black-backed jackal Canis mesomelas Schreber.

    Science.gov (United States)

    Van Heerden, J

    1980-06-01

    Babesia canis was successfully transmitted from the domestic dog to 3 wild dogs Lycaon pictus and 4 black-backed jackals Canis mesomelas. Both wild dogs and black-backed jackals showed no clinical signs or clinical pathological evidence of disease. Trophozoites of Babesia canis were found in peripheral blood smears from all experimental animals. The disease was also successfully transmitted from both black-backed jackals and wild dogs to the domestic dog.

  7. Rhodococcus equi foal pneumonia.

    Science.gov (United States)

    Cohen, Noah D

    2014-12-01

    Pneumonia caused by Rhodococcus equi is an important cause of disease and death in foals. This article reviews current knowledge of the epidemiology, clinical signs, diagnosis, treatment, prevention, and control of R equi pneumonia in foals. Copyright © 2014 Elsevier Inc. All rights reserved.

  8. Klossiella equi in a donkey--a first case report from Iran.

    Science.gov (United States)

    Rezaie, A; Bahrami, S; Ansari, M

    2013-09-01

    Klossiella equi is the only known and rarely reported coccidian parasite of the renal paranchyma of equids. An aged male donkey (Equus asinus asinus) was submitted to necropsy department of veterinary hospital. In histopathological study of renal sections different developmental stages of parasite were observed. These stages were as follow: Trophozoites, microgametes, macrogametes, sporont, budding sporont, sporoblasts, free sporoblasts, mature sporoblast and sporocyst. Parasitic infection with K. equi was encountered in the donkey. According to literature review this is the first report of donkey klossiellosis in Iran.

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

    Institute of Scientific and Technical Information of China (English)

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

    2016-01-01

    Background: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.Findings: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(R)) administration,the patient recovered from fever and tested negative for detection of Babesia-specific genes.Conclusion:Babesia sp.XXB/HangZhou is a novel Babesia species,which causes mild babesiosis in an immunocompetent patient.

  10. EquiMar

    DEFF Research Database (Denmark)

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

    2011-01-01

    / 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...... 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...

  11. Inhibitory effects of pepstatin A and mefloquine on the growth of Babesia parasites.

    Science.gov (United States)

    Munkhjargal, Tserendorj; AbouLaila, Mahmoud; Terkawi, Mohamad Alaa; Sivakumar, Thillaiampalam; Ichikawa, Madoka; Davaasuren, Batdorj; Nyamjargal, Tserendorj; Yokoyama, Naoaki; Igarashi, Ikuo

    2012-10-01

    We evaluated the inhibitory effects of pepstatin A and mefloquine on the in vitro and in vivo growths of Babesia parasites. The in vitro growth of Babesia bovis, B. bigemina, B. caballi, and B. equi was significantly inhibited (P mefloquine (50% inhibitory concentrations = 59.7, 56.7, 20.7, and 4 μM, respectively). Furthermore, both reagents either alone at a concentration of 5 mg/kg or in combinations (2.5/2.5 and 5/5 mg/kg) for 10 days significantly inhibited the in vivo growth of B. microti in mice. Mefloquine treatment was highly effective and the combination treatments were less effective than other treatments. Therefore, mefloquine may antagonize the actions of pepstatin A against babesiosis and aspartic proteases may play an important role in the asexual growth cycle of Babesia parasites.

  12. 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.

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

    Science.gov (United States)

    2012-01-01

    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. PMID:23151128

  14. 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.

  15. [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.

  16. Rhodococcus equi (Prescottella equi) vaccines; the future of vaccine development.

    Science.gov (United States)

    Giles, C; Vanniasinkam, T; Ndi, S; Barton, M D

    2015-09-01

    For decades researchers have been targeting prevention of Rhodococcus equi (Rhodococcus hoagui/Prescottella equi) by vaccination and the horse breeding industry has supported the ongoing efforts by researchers to develop a safe and cost effective vaccine to prevent disease in foals. Traditional vaccines including live, killed and attenuated (physical and chemical) vaccines have proved to be ineffective and more modern molecular-based vaccines including the DNA plasmid, genetically attenuated and subunit vaccines have provided inadequate protection of foals. Newer, bacterial vector vaccines have recently shown promise for R. equi in the mouse model. This article describes the findings of key research in R. equi vaccine development and looks at alternative methods that may potentially be utilised.

  17. Evaluation of in vitro inhibitory effect of enoxacin on Babesia and Theileria parasites.

    Science.gov (United States)

    Omar, Mosaab A; Salama, Akram; Elsify, Ahmed; Rizk, Mohamed Abdo; Al-Aboody, Mohammad Saleh; AbouLaila, Mahmoud; El-Sayed, Shimaa Abd El-Salam; Igarashi, Ikuo

    2016-02-01

    Enoxacin is a broad-spectrum 6-fluoronaphthyridinone antibacterial agent (fluoroquinolones) structurally related to nalidixic acid used mainly in the treatment of urinary tract infections and gonorrhea. Also it has been shown recently that it may have cancer inhibiting effect. The primary antibabesial effect of Enoxacin is due to inhibition of DNA gyrase subunit A, and DNA topoisomerase. In the present study, enoxacin was tested as a potent inhibitor against the in vitro growth of bovine and equine Piroplasms. The in vitro growth of five Babesia species that were tested was significantly inhibited (P Theileria equi, respectively). Enoxacin IC50 values for Babesia and Theileria parasites were satisfactory as the drug is potent antibacterial drug with minimum side effects. Therefore, enoxacin might be used for treatment of Babesiosis and Theileriosis especially in case of mixed infections with bacterial diseases or incase of animal sensitivity against diminazin toxicity.

  18. Genetic Susceptibility to Rhodococcus equi.

    Science.gov (United States)

    McQueen, C M; Dindot, S V; Foster, M J; Cohen, N D

    2015-01-01

    Rhodococcus equi pneumonia is a major cause of morbidity and mortality in neonatal foals. Much effort has been made to identify preventative measures and new treatments for R. equi with limited success. With a growing focus in the medical community on understanding the genetic basis of disease susceptibility, investigators have begun to evaluate the interaction of the genetics of the foal with R. equi. This review describes past efforts to understand the genetic basis underlying R. equi susceptibility and tolerance. It also highlights the genetic technology available to study horses and describes the use of this technology in investigating R. equi. This review provides readers with a foundational understanding of candidate gene approaches, single nucleotide polymorphism-based, and copy number variant-based genome-wide association studies, and next generation sequencing (both DNA and RNA). Copyright © 2015 The Authors. Journal of Veterinary Internal Medicine published by Wiley Periodicals, Inc. on behalf of the American College of Veterinary Internal Medicine.

  19. Rhodococcus equi Infections in Dogs.

    Science.gov (United States)

    Bryan, L K; Clark, S D; Díaz-Delgado, J; Lawhon, S D; Edwards, J F

    2017-01-01

    Five cases of Rhodococcus equi infection in dogs were identified from 2003 to 2014. Three of the dogs had severe, internal lesions attributable to R. equi that have not been previously described: endophthalmitis, endocarditis, and suppurative pleuropneumonia. Isolates from 4 of the dogs were analyzed by polymerase chain reaction for Rhodococcus virulence-associated plasmid (vap) genes. One isolate was vapA-positive, 2 lacked a virulence plasmid, and 1 carried the novel vapN-associated plasmid (pVAPN) recently characterized in bovine isolates. The pVAPN plasmid has not been described in isolates cultured from companion animals. Four of the dogs either were receiving immunosuppressive drugs or had endocrinopathies. R. equi has the potential to cause significant infections in dogs, and immunocompromised animals should be considered at risk for infection.

  20. Equi-Gaussian Curvature Folding

    Indian Academy of Sciences (India)

    E M El-Kholy; El-Said R Lashin; Salama N Daoud

    2007-08-01

    In this paper we introduce a new type of folding called equi-Gaussian curvature folding of connected Riemannian 2-manifolds. We prove that the composition and the cartesian product of such foldings is again an equi-Gaussian curvature folding. In case of equi-Gaussian curvature foldings, $f:M→ P_n$, of an orientable surface onto a polygon $P_n$ we prove that (i) $f\\in\\mathcal{F}_{EG}(S^2)\\Leftrightarrow n=3$ (ii) $f\\in\\mathcal{F}_{EG}(T^2)\\Rightarrow n=4$ (iii) $f\\in\\mathcal{F}_{EG}(\\# 2T^2)\\Rightarrow n=5, 6$ and we generalize (iii) for $\\# nT^2$.

  1. Development of EMA-2 recombinant antigen based enzyme-linked immunosorbent assay for seroprevalence studies of Theileria equi infection in Indian equine population.

    Science.gov (United States)

    Kumar, Sanjay; Kumar, Rajender; Gupta, Ashok K; Yadav, Suresh C; Goyal, Sachin K; Khurana, Sandip K; Singh, Raj K

    2013-11-15

    Equine piroplasmosis is a tick-transmitted protozoan disease caused by Theileria equi and/or Babesia caballi. In the present study, we expressed a 53kDa protein from the truncated EMA-2 gene of T. equi (Indian strain) and developed EMA-2ELISA using this expressed protein. This ELISA is able to detect T. equi-specific antibodies in experimentally infected animals as early as 9 days post-infection. The assay developed was validated with the OIE recommended competitive ELISA (cELISA) on 120 serum samples and significant agreement (kappa=0.93) was observed between results of both the ELISAs which indicates suitability of EMA-2ELISA for use in sero-diagnosis. Diagnostic sensitivity and specificity of EMA-2ELISA - as compared with cELISA - were 0.97 and 0.96, respectively. Analysis of 5651 equine serum samples - collected during 2007-2012 from 12 states of India representing eight agro-climatic zones - by EMA-2ELISA revealed 32.65% seroprevalence of T. equi in India. In conclusion, the EMA-2ELISA developed using the T. equi EMA-2 recombinant protein as antigen for detecting T. equi-specific antibodies has good diagnostic potential for sero-epidemiological surveys.

  2. Effect of synthetic antimicrobial peptides on Naegleria fowleri trophozoites.

    Science.gov (United States)

    Tiewcharoen, Supathra; Phurttikul, Watchara; Rabablert, Jundee; Auewarakul, Prasert; Roytrakul, Sittiruk; Chetanachan, Pruksawan; Atithep, Thassanant; Junnu, Virach

    2014-05-01

    We evaluated the effect of tritrpticin, lactoferrin, killer decapeptide and scrambled peptide in vitro against Naegleria fowleri trophozoites compared with amphotericin B. Tritrpticin (100 microg/ml) caused apoptosis of N. fowleri trophozoites (2x10(5) cells/ml), while lactoferrin, killer decapeptide and scrambled peptide did not. On Gormori trichrome staining, tritrpticin affected the elasticity of the surface membrane and reduced the size of the nuclei of N. fowleri trophozoites. The ultrastructure surface membrane and food cup formation of the trophozoites were 100% inhibited. These results are consistent with inhibition of the nfa1, Mp2CL5 of the treated trophozoite, which plays a role in food cup formation. Tritrpticin 100 microg/ml was not toxic against SK-N-MC cells. Our findings suggest tritrpticin has activity against the surface membrane and nfa1 and Mp2CL5 of N. fowleri trophozoites and could be developed as a potential therapeutic agent.

  3. ANTI-AMEBIC ACTIVITY OF DIOSGENIN ON NAEGLERIA FOWLERI TROPHOZOITES.

    Science.gov (United States)

    Rabablert, Jundee; Tiewcharoen, Supathra; Auewarakul, Prasert; Atithep, Thassanant; Lumlerdkij, Natchagorn; Vejaratpimol, Renu; Junnu, Virach

    2015-09-01

    The aim of this study was to investigate the activity of diosgenin against Naegleria fowleri trophozoites at the cellular and molecular levels. Diosgenin (100 μg/ml; 241.2 μM) had a 100% inhibitory effect on N. fowleri trophozoites (5 x 10(5) cell/ml). Scanning electron micrograph revealed diosgenin decreased the number of sucker-like apparatuses and food cup formation among N. fowleri trophozoites at 3 and 6 hours post-exposure, respectively. Diosgenin down-regulated the nf cysteine protease gene expression of N. fowleri trophozoites at 6 and 12 hours post-exposure. The toxicity to mammalian cells caused by diosgenin at therapeutic dose was less than amphotericin B, the current drug used to treat N. fowleri infections. Our findings suggest diosgenin has activity against the surface membrane and the nf cysteine pro tease of N. fowleri trophozoites. However, the other mechanisms of action of diosgenin against N. fowleri trophozoites require further exploration.

  4. Streptococcus equi meningoencephalomyelitis in a foal.

    Science.gov (United States)

    Finno, Carrie; Pusterla, Nicola; Aleman, Monica; Mohr, F Charles; Price, Teena; George, Jeanne; Holmberg, Tara

    2006-09-01

    A 4-month-old American Paint Horse colt was evaluated because of acute onset of ataxia, left-sided head tilt, and fever and a recently noticed heart murmur. Upper respiratory tract infection caused by Streptococcus equi subsp equi had been diagnosed at 3 months of age. Hematologic abnormalities included leukocytosis, mature neutrophilia, monocytosis, and mild anemia. Analysis of a CSF sample revealed high total protein concentration and total nucleated cell count; nucleated cells consisted mainly of degenerate neutrophils. Results of a real-time PCR assay were positive for S equi subsp equi, and a diagnosis of S equi subsp equi meningoencephalomyelitis was made. Treatment included administration of potassium penicillin and fluids, but the foal developed uroperitoneum and was subsequently euthanized. Postmortem examination revealed meningoencephalomyelitis, and S equi subsp equi was cultured from a brain aspirate. Additional findings included suppurative cystitis with rupture and neutrophilic myocarditis. Findings suggest that S equi subsp equi meningoencephalomyelitis should be considered in the differential diagnosis for foals with neurologic signs that have a history of strangles or exposure to affected horses.

  5. Treatment of Infections Caused by Rhodococcus equi.

    Science.gov (United States)

    Giguère, Steeve

    2017-04-01

    Pneumonia caused by Rhodococcus equi remains an important cause of disease and death in foals. The combination of a macrolide (erythromycin, azithromycin, or clarithromycin) with rifampin remains the recommended therapy for foals with clinical signs of infection caused by R equi. Most foals with small, subclinical ultrasonographic pulmonary lesions associated with R equi recover without therapy, and administration of antimicrobial agents to these subclinically affected foals does not hasten lesion resolution relative to administration of a placebo. Resistance to macrolides and rifampin in isolates of R equi is increasing. Copyright © 2016 Elsevier Inc. All rights reserved.

  6. Molecular characterization of virulence genes of Streptococcus equi subsp. equi and Streptococcus equi subsp. zooepidemicus in equines

    Science.gov (United States)

    Javed, R.; Taku, A. K.; Gangil, Rakhi; Sharma, R. K.

    2016-01-01

    Aim: The aim was to determine the occurrence of streptococci in equines in Jammu (R. S. Pura, Katra), characterization of Streptococci equi subsp. equi and Streptococcus equi subsp. zooepidemicus with respect to their virulence traits and to determine antibiotic sensitivity pattern of virulent Streptococcus isolates. Materials and Methods: A total of 96 samples were collected from both clinically affected animals (exhibiting signs of respiratory tract disease) and apparently healthy animals and were sent to laboratory. The organisms were isolated on Columbia nalidixic acid agar containing 5% sheep blood as well as on sheep blood agar and confirmed by cultural characteristics and biochemical tests. Molecular detection of Streptococcus was done directly from cultures using sodA and seM gene-based polymerase chain reaction (PCR). Antibiogram was performed against five antibiotics such as amoxicillin, penicillin G, streptomycin, rifampicin, and methicillin. Results: During this study, a total 40 streptococcal isolates were obtained out of which 2 isolates were of S. equi subsp. equi, 12 isolates were from S. equi subsp. zooepidemicus. In the PCR-based detection, we revealed amplicons of 235 bp and 679 bp for confirmation of sodA and seM gene, respectively. In antibiogram, two isolates of S. equi subsp. equi were found resistant to penicillin G, and all other isolates were found sensitive to amoxicillin and streptomycin. Conclusion: The majority of streptococcal infections was due to S. equi subsp. Zooepidemicus, and thus was recognized as a potential pathogen of diseases of equines besides S. equi subsp. equi. PMID:27651677

  7. Molecular characterization of virulence genes of Streptococcus equi subsp. equi and Streptococcus equi subsp. zooepidemicus in equines

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    R. Javed

    2016-08-01

    Full Text Available Aim: The aim was to determine the occurrence of streptococci in equines in Jammu (R. S. Pura, Katra, characterization of Streptococci equi subsp. equi and Streptococcus equi subsp. zooepidemicus with respect to their virulence traits and to determine antibiotic sensitivity pattern of virulent Streptococcus isolates. Materials and Methods: A total of 96 samples were collected from both clinically affected animals (exhibiting signs of respiratory tract disease and apparently healthy animals and were sent to laboratory. The organisms were isolated on Columbia nalidixic acid agar containing 5% sheep blood as well as on sheep blood agar and confirmed by cultural characteristics and biochemical tests. Molecular detection of Streptococcus was done directly from cultures using sodA and seM gene-based polymerase chain reaction (PCR. Antibiogram was performed against five antibiotics such as amoxicillin, penicillin G, streptomycin, rifampicin, and methicillin. Results: During this study, a total 40 streptococcal isolates were obtained out of which 2 isolates were of S. equi subsp. equi, 12 isolates were from S. equi subsp. zooepidemicus. In the PCR-based detection, we revealed amplicons of 235 bp and 679 bp for confirmation of sodA and seM gene, respectively. In antibiogram, two isolates of S. equi subsp. equi were found resistant to penicillin G, and all other isolates were found sensitive to amoxicillin and streptomycin. Conclusion: The majority of streptococcal infections was due to S. equi subsp. Zooepidemicus, and thus was recognized as a potential pathogen of diseases of equines besides S. equi subsp. equi.

  8. Antimicrobial resistance in Rhodococcus equi.

    Science.gov (United States)

    Cisek, Agata A; Rzewuska, Magdalena; Witkowski, Lucjan; Binek, Marian

    2014-01-01

    Rhodococcus equi is an important etiologic agent of respiratory- and non-respiratory tract infections, diseases of animals and humans. Therapy includes the use of various group of chemotherapeutic agents, however resistance acquirement is quite common. To date there is no preferred treatment protocol for infections caused by isolates resistant to macrolides and rifampicin. The resistance acquirement is a result of many molecular mechanisms, some of which include alterations in the cell envelope composition and structure, activity of the efflux pumps, enzymatic destruction or inactivation of antibiotics, and changes in the target site. This paper contains an overview of antimicrobial susceptibility of R. equi, and explains the possible molecular mechanisms responsible for antimicrobial resistance in this particular microorganism.

  9. Entamoeba histolytica trophozoites transfer lipophosphopeptidoglycans to enteric cell layers.

    Science.gov (United States)

    Lauwaet, Tineke; Oliveira, Maria José; De Bruyne, Georges; Bruchhaus, Iris; Duchêne, Michael; Mareel, Marc; Leroy, Ancy

    2004-04-01

    Transfer of antigens frequently follows adhesion of protozoan parasites to host cells. We were interested in such transfer from the Entamoeba surface to enterocytes following adhesion of trophozoites. Therefore, cocultures of enterocytes in vitro and ex vivo with Entamoeba histolytica (strain HM-1:IMSS) or Entamoeba dispar (strain SAW760) trophozoites were processed for immunocytochemistry. The EH5 monoclonal antibody against amoebic proteophosphoglycans marked a dotted pattern on the apical side of enterocytes in in vitro cocultures with HM-1:IMSS and SAW760 trophozoites. Basolateral staining was present in cocultures following dysfunction of tight junctions, or when trophozoites made direct contact with the basolateral side of enterocytes in in vitro and ex vivo cocultures. Based on the molecular mass in Western blot, the transferred proteophosphoglycan was identified as a lipophosphopeptidoglycan. In conclusion, trophozoites transfer LPPG to the apical side of enterocytes following adhesion and prior to dysfunction of tight junctions.

  10. Production of recombinant EMA-1 protein and its application for the diagnosis of Theileria equi using an enzyme immunoassay in horses from São Paulo State, Brazil.

    Science.gov (United States)

    Baldani, Cristiane Divan; Hilario, Eduardo; Nakaghi, Andréa Cristina Higa; Bertolini, Maria Célia; Machado, Rosangela Zacarias

    2011-01-01

    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.

  11. Differential expression of three members of the multidomain adhesion CCp family in babesia bigemina, babesia bovis and theileria equi

    Science.gov (United States)

    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 tr...

  12. The identification of MacSe in Streptococcus equi ssp. equi

    Institute of Scientific and Technical Information of China (English)

    Jiande YANG; Yanfei LIU; Jun XU; Jifei MA

    2009-01-01

    Streptococcus equi subsp, equi (S. equi ssp. equi) causes equine strangles, a highly contagious and widespread purulent lymphadenitis of the head and neck. We have identified MacSe, a novel protein of S. equi, by screening a phage library of 3-8 kb random DNA fragments of S. equi CF32. MacSe shares 62% and 67.5% amino acid homology with Mac5005 and Mac8345 of S. pyogenes respectively. Expression during infection was shown by strong reactivity of the protein with convalescent sera and mucosal wash IgA of ponies infected by commingling exposure. Release into the culture medium was detected during the log phase of growth. Dose dependent anti-phagocytic activity for equine neutrophils involved interaction of MacSe with C3 and neutrophils.

  13. Babesia species in questing Ixodes ricinus, Sweden.

    Science.gov (United States)

    Karlsson, Maria E; Andersson, Martin O

    2016-02-01

    Babesiosis is an emerging tick-transmitted zoonosis in large parts of the world. In Sweden, the occurrence and diversity of Babesia species is largely unknown. In order to estimate the exposure to Babesia from infected ticks, we collected questing Ixodes ricinus from several sites across southern Sweden during two consecutive field seasons and investigated the occurrence of Babesia species. We report for the first time the occurrence of the zoonotic species Babesia venatorum in Swedish ticks, with a prevalence of 1%. We also detected B. microti (prevalence 3.2%) and B. divergens (prevalence 0.2%). The incidence of Babesia in questing ticks is substantially lower than that of several other tick-borne diseases in Sweden. Nevertheless, babesiosis should not be neglected as a possible diagnosis following tick bites in humans and animals in Sweden.

  14. Rhodococcus equi: A pathogen in immunocompetent patients

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    Meena Dias

    2013-01-01

    Full Text Available Rhodococcus equi is an uncommon human pathogen known to cause lung infections in immunocompromised patients. We report two cases of Rhodococcus infections in immunocompetent individuals, who were treated successfully.

  15. Detection of Theileria equi in spleen and blood of asymptomatic piroplasm carrier horses.

    Science.gov (United States)

    Ribeiro, Isabel B; Câmara, Antônio Carlos L; Bittencourt, Marta V; Marçola, Tatiana G; Paludo, Giane R; Soto-Blanco, Benito

    2013-06-01

    This study aimed to determine whether asymptomatic horses naturally infected with Theileria equi retain infected erythrocytes in the spleen and whether the presence of the hemoparasite in this organ is associated with parasitemia. We collected samples from 25 adult horses without clinical signs of any disease. From each animal, we collected whole blood samples from the jugular vein and a splenic puncture blood sample. All samples were submited to blood cell counts and detection of Theileria or Babesia. DNA extraction and PCR were performed in all samples for identification of piroplasm infection (T. equi and B. caballi). From the 25 horses evaluated for piroplasm detection by PCR, seven horses (28%) were positive in jugular vein blood but negative in splenic blood samples, five horses (20%) were positive in splenic blood samples but negative in jugular vein blood samples, and 13 horses (52%) were positive in both jugular vein and splenic blood samples. The hematological evaluation revealed anemia in 13 of 25 (52%) infected horses, lymphopenia in five (20%), neutrophilia in two (8%), neutropenia in one (4%), and thrombocytopenia in one (4%) infected horse. The present study demonstrated that several (20%) of the asymptomatic piroplasm carrier horses did not show parasitemia, but show infected erythrocytes in the spleen.

  16. Ex vivo laser confocal microscopy findings of cultured Acanthamoeba trophozoites

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    Yamazaki N

    2012-08-01

    Full Text Available Natsuko Yamazaki,1 Akira Kobayashi,1 Hideaki Yokogawa,1 Yasuhisa Ishibashi,2 Yosaburo Oikawa,3 Masaharu Tokoro,4 Kazuhisa Sugiyama11Department of Ophthalmology, Kanazawa University Graduate School of Medical Science, Kanazawa, Japan; 2Department of Ophthalmology, East Washinomiya Hospital, Kuki, Japan; 3Department of Medical Zoology, Kanazawa Medical University, Kahoku, Japan; 4Department of Parasitology, Kanazawa University Graduate School of Medical Science, Kanazawa, JapanPurpose: The purpose of the current study was to investigate ex vivo laser confocal microscopic findings of cultured Acanthamoeba trophozoites obtained from Acanthamoeba keratitis patients.Methods: Eight cultured samples of Acanthamoeba trophozoites from eight eyes of seven patients (mean age, 26.9 years; age range, 18–52 years were used. Seven samples were from corneal scrapings of Acanthamoeba keratitis patients and one sample was from the solution in a soft contact lens case. Ex vivo laser confocal microscopy was performed to qualitatively evaluate the shape and degree of light reflection of the living Acanthamoeba trophozoites.Results: Ex vivo laser confocal microscopy demonstrated highly reflective, high-contrast Acanthamoeba trophozoites with no walls (mean size, 25.4 µm; range, 17.1–58.5 µm. The shapes of the trophozoites were highly pleomorphic, and some showed characteristic acanthopodia by laser confocal microscopy.Conclusion: Ex vivo laser confocal microscopy was effective in demonstrating cultured Acanthamoeba trophozoites of various shapes and sizes. The observations of the current study may be helpful when similar structures are identified under in vivo conditions.Keywords: Acanthamoeba, trophozoite, laser confocal microscopy

  17. Quantitative aspects of fecal Rhodococcus (Corynebacterium) equi in foals.

    OpenAIRE

    Takai, S.; Ohkura, H; Watanabe, Y.; Tsubaki, S

    1986-01-01

    Quantitative aspects of fecal Rhodococcus (Corynebacterium) equi in newborn foals for 12 weeks after birth were investigated on two horse breeding farms. R. equi was found in the feces of foals during week 1 of life. The greatest numbers of R. equi were present in the feces of foals during the first 8 weeks of their lives, which coincides with the age when foals are most liable to be exposed to R. equi.

  18. COMPARABLE CYTOLOGICAL DIAGNOSTIC OF BLOOD SMEARS ON BABESIA INFECTION

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    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

  19. 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.

  20. Babesia canis and Babesia rossi co-infection in an untraveled Nigerian dog.

    Science.gov (United States)

    Kamani, Joshua; Sannusi, Abdulrahim; Dogo, A Goni; Tanko, James T; Egwu, Kinsley O; Tafarki, Agbadu E; Ogo, Isaac N; Kemza, Sarah; Onovoh, Emmanuel; Shamaki, David; Lombin, Lami H; Catto, Victoria; Birkenheuer, Adam J

    2010-10-29

    A sexually intact 6-month-old female Alsatian dog was presented to the Veterinary Clinic of the National Veterinary Research Institute, Vom, Plateau State, Nigeria, for the following complaints: anorexia, hemoglobinuria, fever, tick infestation and general malaise. Microscopy revealed piroplasms with a wide range of sizes (1-5 μm in length) in red blood cells, raising a suspicion of a co-infection with two or more Babesia species. Specific PCR assays for canine Babesia spp. and DNA sequencing revealed the presence of Babesia canis and Babesia rossi co-infection. This study constitutes the first report of co-infection with B. canis and B. rossi in the West African sub-region and the first report of autochthonous B. canis on the African continent. Practitioners should be aware of potential changes in the species/sub-species of Babesia causing canine babesiosis in this region.

  1. 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

  2. Study of lysozyme resistance in Rhodococcus equi.

    Science.gov (United States)

    Hébert, Laurent; Bidaud, Pauline; Goux, Didier; Benachour, Abdellah; Laugier, Claire; Petry, Sandrine

    2014-03-01

    Lysozyme is an important and widespread component of the innate immune response that constitutes the first line of defense against bacterial pathogens. The bactericidal effect of this enzyme relies on its capacity to hydrolyze the bacterial cell wall and also on a nonenzymatic mechanism involving its cationic antimicrobial peptide (CAMP) properties, which leads to membrane permeabilization. In this paper, we report our findings on the lysozyme resistance ability of Rhodococcus equi, a pulmonary pathogen of young foals and, more recently, of immunocompromised patients, whose pathogenic capacity is conferred by a large virulence plasmid. Our results show that (i) R. equi can be considered to be moderately resistant to lysozyme, (ii) the activity of lysozyme largely depends on its muramidase action rather than on its CAMP activity, and (iii) the virulence plasmid confers part of its lysozyme resistance capacity to R. equi. This study is the first one to demonstrate the influence of the virulence plasmid on the stress resistance capacity of R. equi and improves our understanding of the mechanisms enabling R. equi to resist the host defenses.

  3. Canine babesiosis in Romania due to Babesia canis and Babesia vogeli: a molecular approach.

    Science.gov (United States)

    Ionita, Mariana; Mitrea, Ioan Liviu; Pfister, Kurt; Hamel, Dietmar; Buzatu, Catalin Marius; Silaghi, Cornelia

    2012-05-01

    Canine babesiosis is a tick-borne disease caused by the protozoa Babesia spp. that affects dogs worldwide. In Romania, canine babesiosis has become quite frequent in the last few years, with a wide variety of clinical signs, ranging from mild, nonspecific illness to peracute collapse, and even death. Traditionally, a Babesia infection in dogs is diagnosed based on the morphologic appearance of the intraerythrocytic piroplasms observed in peripheral blood smears. To date, no data on genetic characterization of Babesia species in dogs has been documented for Romania. Therefore, a molecular survey on natural Babesia infections of dogs in Romania using polymerase chain reaction and genetic sequence analysis of a fragment of the ssRNA gene was performed. A total number of 16 blood samples were tested for the presence of Babesia DNA. Blood samples were collected from 11 dogs with symptoms of babesiosis and microscopically proven positive for Babesia and from a group of five asymptomatic dogs, not tested microscopically for Babesia, which were included in the study for comparative analysis. The piroplasm-specific PCR amplifying the partial 18S rRNA gene confirmed Babesia spp. infection in all 11 samples from dogs with clinical babesiosis, and in one of the clinically normal dogs. Sequence analysis revealed the presence of Babesia canis in all clinically affected dogs and Babesia vogeli in one clinically normal dog. This is the first molecular evidence of B. canis and B. vogeli in dogs from Romania. The results of the study provide basic information toward a better understanding of the epidemiology of canine babesiosis in Romania and will help to promote an effective control program.

  4. Proteomic profiling of the infective trophozoite stage of Acanthamoeba polyphaga.

    Science.gov (United States)

    Caumo, Karin Silva; Monteiro, Karina Mariante; Ott, Thiely Rodrigues; Maschio, Vinicius José; Wagner, Glauber; Ferreira, Henrique Bunselmeyer; Rott, Marilise Brittes

    2014-12-01

    Acanthamoeba polyphaga is a free-living protozoan pathogen, whose infective trophozoite form is capable of causing a blinding keratitis and fatal granulomatous encephalitis in humans. The damage caused by A. polyphaga trophozoites in human corneal or brain infections is the result of several different pathogenic mechanisms that have not yet been elucidated at the molecular level. We performed a comprehensive analysis of the proteins expressed by A. polyphaga trophozoites, based on complementary 2-DE MS/MS and gel-free LC-MS/MS approaches. Overall, 202 non-redundant proteins were identified. An A. polyphaga proteomic map in the pH range 3-10 was produced, with protein identification for 184 of 370 resolved spots, corresponding to 142 proteins. Additionally, 94 proteins were identified by gel-free LC-MS/MS. Functional classification revealed several proteins with potential importance for pathogen survival and infection of mammalian hosts, including surface proteins and proteins related to defense mechanisms. Our study provided the first comprehensive proteomic survey of the trophozoite infective stage of an Acanthamoeba species, and established foundations for prospective, comparative and functional studies of proteins involved in mechanisms of survival, development, and pathogenicity in A. polyphaga and other pathogenic amoebae.

  5. Bacteremia due to Rhodococcus equi in an immunocompetent infant

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    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.

  6. 基于Hsp70基因的马梨形虫分类学定位分析%Taxonomic Status Assays of Theileria equi Based on Hsp70 Gene

    Institute of Scientific and Technical Information of China (English)

    罗金; 刘光远; 田占成; 谢俊仁; 张萍; 沈辉; 党根生

    2011-01-01

    [Objective] The objective of the study is to determine the taxonomic status of Theileria equi (T. equi). [Method] According to the Hsp70 gene sequence (AB248743.1) of Babesia equi (B.equi) in GenBank, the primers TeHsp70F and TeHsp70R were designed. And a fragment with the length of 1 920 bp was obtained by PCR. The gene sequence was compared with other 23 amino acids sequences. [Result] The fragment encods 639 amino acids, 208 hydrophobic amino acids and 167 polar amino acid. Identity analysis showed that T. equi has a closest relationship with B.equi (BAF0262S.1), and has a little relationship with T. parva (XP764717.1) and T. annulata (AAA30130.1), while T. equi has a distant relationship with B. caballi (BAF02619.1), which infects equine. [ Conclusion ] It was a correct taxonomic status for B. equi to Theileria species. And the taxonomic status was error before, for T. equi to Babesia species. In this paper, a proof was provided for taxonomic status of Theileria equi.%[目的]为了进一步确定马泰勒虫的分类学定位.[方法]根据GenBank上登录的马巴贝斯虫Hsp70基因序列(AB248743.1),设计引物TeHsp70F,TeHsp70R,以马泰勒虫基因组DNA为模板进行扩增,获得全长为1920 bp的核酸片段.将该基因序列与GenBank中23种已知虫种的相应序列进行分析比较.[结果]该片段编码639个氨基酸,其疏水性氨基酸达到208个,极性氨基酸167个.同一性分析显示,该基因片段与报道的马巴贝斯虫Hsp70基因(B.equi BAF02625.1)亲缘关系最近,其次是小泰勒虫(T.parva XP764717.1)和环形泰勒虫(T.annulataAAA30130.1),而与感染马属动物的另一个虫种驽巴贝斯虫(B.caballi BAF02619.1)亲缘关系较远.[结论]马泰勒虫Hsp70基因的克隆及系统发育分析显示,之前被定名为马巴贝斯虫(B.equi)的虫种隶属于泰勒虫虫种.本试验为马巴贝斯虫正确更名为马泰勒虫(T.equi)的分类地位提供了又一佐证.

  7. Association of perinatal exposure to airborne Rhodococcus equi with risk of pneumonia caused by R equi in foals.

    Science.gov (United States)

    Cohen, Noah D; Chaffin, M Keith; Kuskie, Kyle R; Syndergaard, Melissa K; Blodgett, Glenn P; Takai, Shinji

    2013-01-01

    To determine whether the concentrations of airborne virulent Rhodococcus equi in stalls housing foals during the first 2 weeks after birth are associated with subsequent development of R equi pneumonia in those foals. Air samples collected from foaling stalls and holding pens in which foals were housed during the first 2 weeks after birth. At a breeding farm in Texas, air samples (500 L each) were collected (January through May 2011) from stalls and pens in which 121 foals were housed on day 1 and on days 4, 7, and 14 after birth. For each sample, the concentration of airborne virulent R equi was determined with an immunoblot technique. The association between development of pneumonia and airborne R equi concentration was evaluated via random-effects Poisson regression analysis. Some air samples were not available for analysis. Of the 471 air samples collected from stalls that housed 121 foals, 90 (19%) contained virulent R equi. Twenty-four of 121 (20%) foals developed R equi pneumonia. Concentrations of virulent R equi in air samples from stalls housing foals that developed R equi pneumonia were significantly higher than those in samples from stalls housing foals that did not develop pneumonia. Accounting for disease effects, air sample concentrations of virulent R equi did not differ significantly by day after birth or by month of birth. Exposure of foals to airborne virulent R equi during the first 2 weeks after birth was significantly (and likely causally) associated with development of R equi pneumonia.

  8. Transcriptional silencing of multiple genes in trophozoites of Entamoeba histolytica.

    Directory of Open Access Journals (Sweden)

    Rivka Bracha

    2006-05-01

    Full Text Available In a previous work we described the transcriptional silencing of the amoebapore A (AP-A gene (Ehap-a of Entamoeba histolytica strain HM-1:IMSS. The silencing occurred following transfection with a plasmid containing a 5' upstream region (473 bp of Ehap-a that included a truncated segment (140 bp of a short interspersed nuclear element (SINE1. Silencing remained in effect even after removal of the plasmid (clone G3. Neither short interfering RNA nor methylated DNA were detected, but the chromatin domain of Ehap-a in the gene-silenced trophozoites was modified. Two other similar genes (Ehap-b and one encoding a Saposin-like protein, SAPLIP 1 also became silenced. In the present work we demonstrate the silencing of a second gene of choice, one that encodes the light subunit of the Gal/GalNAc inhibitable lectin (Ehlgl1 and the other, the cysteine proteinase 5 (EhCP-5. This silencing occurred in G3 trophozoites transfected with a plasmid in which the 473 bp 5' upstream Ehap-a fragment was directly ligated to the second gene. Transcriptional silencing occurred in both the transgene and the chromosomal gene. SINE1 sequences were essential, as was a direct connection between the Ehap-a upstream region and the beginning of the open reading frame of the second gene. Gene silencing did not occur in strain HM-1:IMSS with any of these plasmid constructs. The trophozoites with two silenced genes were virulence-attenuated as were those of clone G3. In addition, trophozoites not expressing Lgl1 and AP-A proteins had a significantly reduced ability to cap the Gal/GalNAc-lectin to the uroid region when incubated with antibodies against the heavy (170 kDa subunit of the lectin. Lysates of trophozoites lacking cysteine proteinase 5 and AP-A proteins had 30% less cysteine proteinase activity than those of HM-1:IMSS strain or the G3 clone. Silencing of other genes in G3 amoebae could provide a model to study their various functions. In addition, double gene

  9. Transcriptional silencing of multiple genes in trophozoites of Entamoeba histolytica.

    Directory of Open Access Journals (Sweden)

    2006-05-01

    Full Text Available In a previous work we described the transcriptional silencing of the amoebapore A (AP-A gene (Ehap-a of Entamoeba histolytica strain HM-1:IMSS. The silencing occurred following transfection with a plasmid containing a 5' upstream region (473 bp of Ehap-a that included a truncated segment (140 bp of a short interspersed nuclear element (SINE1. Silencing remained in effect even after removal of the plasmid (clone G3. Neither short interfering RNA nor methylated DNA were detected, but the chromatin domain of Ehap-a in the gene-silenced trophozoites was modified. Two other similar genes (Ehap-b and one encoding a Saposin-like protein, SAPLIP 1 also became silenced. In the present work we demonstrate the silencing of a second gene of choice, one that encodes the light subunit of the Gal/GalNAc inhibitable lectin (Ehlgl1 and the other, the cysteine proteinase 5 (EhCP-5. This silencing occurred in G3 trophozoites transfected with a plasmid in which the 473 bp 5' upstream Ehap-a fragment was directly ligated to the second gene. Transcriptional silencing occurred in both the transgene and the chromosomal gene. SINE1 sequences were essential, as was a direct connection between the Ehap-a upstream region and the beginning of the open reading frame of the second gene. Gene silencing did not occur in strain HM-1:IMSS with any of these plasmid constructs. The trophozoites with two silenced genes were virulence-attenuated as were those of clone G3. In addition, trophozoites not expressing Lgl1 and AP-A proteins had a significantly reduced ability to cap the Gal/GalNAc-lectin to the uroid region when incubated with antibodies against the heavy (170 kDa subunit of the lectin. Lysates of trophozoites lacking cysteine proteinase 5 and AP-A proteins had 30% less cysteine proteinase activity than those of HM-1:IMSS strain or the G3 clone. Silencing of other genes in G3 amoebae could provide a model to study their various functions. In addition, double gene

  10. Entamoeba histolytica Trophozoites and Lipopeptidophosphoglycan Trigger Human Neutrophil Extracellular Traps.

    Science.gov (United States)

    Ávila, Eva E; Salaiza, Norma; Pulido, Julieta; Rodríguez, Mayra C; Díaz-Godínez, César; Laclette, Juan P; Becker, Ingeborg; Carrero, Julio C

    2016-01-01

    Neutrophil defense mechanisms include phagocytosis, degranulation and the formation of extracellular traps (NET). These networks of DNA are triggered by several immune and microbial factors, representing a defense strategy to prevent microbial spread by trapping/killing pathogens. This may be important against Entamoeba histolytica, since its large size hinders its phagocytosis. The aim of this study was to determine whether E. histolytica and their lipopeptidophosphoglycan (EhLPPG) induce the formation of NETs and the outcome of their interaction with the parasite. Our data show that live amoebae and EhLPPG, but not fixed trophozoites, induced NET formation in a time and dose dependent manner, starting at 5 min of co-incubation. Although immunofluorescence studies showed that the NETs contain cathelicidin LL-37 in close proximity to amoebae, the trophozoite growth was only affected when ethylene glycol tetra-acetic acid (EGTA) was present during contact with NETs, suggesting that the activity of enzymes requiring calcium, such as DNases, may be important for amoeba survival. In conclusion, E. histolytica trophozoites and EhLPPG induce in vitro formation of human NETs, which did not affect the parasite growth unless a chelating agent was present. These results suggest that NETs may be an important factor of the innate immune response during infection with E. histolytica.

  11. Entamoeba histolytica Trophozoites and Lipopeptidophosphoglycan Trigger Human Neutrophil Extracellular Traps

    Science.gov (United States)

    Ávila, Eva E.; Rodríguez, Mayra C.; Díaz-Godínez, César; Laclette, Juan P.; Becker, Ingeborg; Carrero, Julio C.

    2016-01-01

    Neutrophil defense mechanisms include phagocytosis, degranulation and the formation of extracellular traps (NET). These networks of DNA are triggered by several immune and microbial factors, representing a defense strategy to prevent microbial spread by trapping/killing pathogens. This may be important against Entamoeba histolytica, since its large size hinders its phagocytosis. The aim of this study was to determine whether E. histolytica and their lipopeptidophosphoglycan (EhLPPG) induce the formation of NETs and the outcome of their interaction with the parasite. Our data show that live amoebae and EhLPPG, but not fixed trophozoites, induced NET formation in a time and dose dependent manner, starting at 5 min of co-incubation. Although immunofluorescence studies showed that the NETs contain cathelicidin LL-37 in close proximity to amoebae, the trophozoite growth was only affected when ethylene glycol tetra-acetic acid (EGTA) was present during contact with NETs, suggesting that the activity of enzymes requiring calcium, such as DNases, may be important for amoeba survival. In conclusion, E. histolytica trophozoites and EhLPPG induce in vitro formation of human NETs, which did not affect the parasite growth unless a chelating agent was present. These results suggest that NETs may be an important factor of the innate immune response during infection with E. histolytica. PMID:27415627

  12. 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.

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

    Science.gov (United States)

    Erganis, Osman; Sayin, Zafer; Hadimli, Hasan Huseyin; Sakmanoglu, Asli; Pinarkara, Yasemin; Ozdemir, Ozgur; Maden, Mehmet

    2014-01-01

    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. PMID:24982958

  15. 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 ass

  16. 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

  17. 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 ass

  18. Rhodococcus equi isolation from sputum of patients with suspected tuberculosis

    Directory of Open Access Journals (Sweden)

    Paulo da Silva

    2010-03-01

    Full Text Available Rhodococcus equi has emerged as an opportunistic pathogen associated with pulmonary, invasive or systemic infections in immunocompromised patients. We report the identification of 51 R. equi isolates found in sputum samples of 546 individuals suspected to have pulmonary tuberculosis in two Public Health Hospital Units in Brazil. The epidemiology of R. equi infection as well as the phenotypic identification and drug susceptibility profile of isolates are described in this paper.

  19. Pangenome and Phylogenomic Analysis of the Pathogenic Actinobacterium Rhodococcus equi.

    Science.gov (United States)

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

    2016-10-23

    We report a comparative study of 29 representative genomes of the animal pathogen Rhodococcus equi The analyses showed that R. equi is genetically homogeneous and clonal, with a large core genome accounting for ≈80% of an isolates' gene content. An open pangenome, even distribution of accessory genes among the isolates, and absence of significant core-genome recombination, indicated that gene gain/loss is a main driver of R. equi genome evolution. Traits previously predicted to be important in R. equi physiology, virulence and niche adaptation were part of the core genome. This included the lack of a phosphoenolpyruvate:carbohydrate transport system (PTS), unique among the rhodococci except for the closely related Rhodococcus defluvii, reflecting selective PTS gene loss in the R. equi-R. defluvii sublineage. Thought to be asaccharolytic, rbsCB and glcP non-PTS sugar permease homologues were identified in the core genome and, albeit inefficiently, R. equi utilized their putative substrates, ribose and (irregularly) glucose. There was no correlation between R. equi whole-genome phylogeny and host or geographical source, with evidence of global spread of genomovars. The distribution of host-associated virulence plasmid types was consistent with the exchange of the plasmids (and corresponding host shifts) across the R. equi population, and human infection being zoonotically acquired. Phylogenomic analyses demonstrated that R. equi occupies a central position in the Rhodococcus phylogeny, not supporting the recently proposed transfer of the species to a new genus.

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

    DEFF Research Database (Denmark)

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

    1991-01-01

    Rhodococcus equi is an aerobic, gram-positive, non-motile pleomorphic bacillus infecting immunocompromised patients. Forty-nine cases of Rhodococcus equi infection have been reported, mainly in patients infected with the human immunodeficiency virus (HIV). A case in which Rhodococcus equi caused...... severe pulmonary infection, the most common presentation, is described. Clinically, patients have symptoms of pneumonia with hemoptysis as a prominent feature. X-ray will often show a cavitating upper-lobe infiltrate, resembling infection with mycobacteria. Rhodococcus equi is easily cultured from blood......-term antibiotic treatment with erythromycin plus rifampicin, or vancomycin in combination with either of these antibiotics....

  1. Caprine vertebral osteomyelitis caused by Rhodococcus equi : clinical communication

    Directory of Open Access Journals (Sweden)

    P.N. Kabongo

    2005-06-01

    Full Text Available There have been reports of Rhodococcus equi infections in goats in Australia, America and India. In this study, R. equi was isolated from an inflamed vertebra in a Boer goat in South Africa. At autopsy, there was a purulent inflammatory reaction in the 1st cervical vertebra. Histopathologically, a neutrophilic infiltration was encountered in the bone. Aerobic culture of swabs collected from the abscesses yielded R. equi in pure culture that was identified on biochemical tests. R. equi has become important as the cause of an opportunistic infection in people suffering from HIV.

  2. Scanning electron microscopic study of trophozoite and cyst stages of Naegleria fowleri.

    Science.gov (United States)

    Antonios, Sanaa N

    2010-04-01

    Whole trophozoites and cysts of axenically cultivated Naegleria fowleri were prepared for study of their surface morphology by scanning electron microscopy (SEM). Trophozoites and cyst stages were studied from Chang's culture media. Some trophozoites were examined after animal inoculation and brain isolation to compare the changes in surface features. Photomicrographs of freeze-dried and critical point-dried organisms fixed with glutaraldehyde were presented along with views of both isolates of trophozoites to compare the surface features. SEM revealed the surface of trophozoites to be undulating, wrinkled and covered at irregular intervals by protruding vesicles. There were also surface extensions which were long and thin in brain isolates which may help in the contact and cytolysis of host cells at some distance from the trophozoite. Some cysts appeared wrinkled while others smooth, and empty cysts were also seen with many pores on the surface.

  3. Cocktail of Theileria equi antigens for detecting infection in equines

    Institute of Scientific and Technical Information of China (English)

    Shimaa; Abd; El-Salam; El-Sayed; Mohamed; Abdo; Rizk; Mohamed; Alaa; Terkawi; Ahmed; Mousa; El; Said; El; Shirbini; El; Said; Gehad; Elsayed; Mohamed; Fouda; Naoaki; Yokoyama; Ikuo; Igarashi

    2015-01-01

    Objective:To use two diagnostic antigens belonging to the frequently associated in Theileria domain,Theileria equi(T.equi)protein 82(Te 82)and T.equi 104 k Da microneme-rhoptry antigen precursor(Te 43),to diagnose T.equi infection in horses as compared with equi merozoite antigen-2(EMA-2).Methods:In the current study,we applied a cocktail-ELISA containing two antigens(EMA-2+Te 82)to diagnose T.equi infection either in experimentally infected horses or in field infection.Results:Our findings have revealed that a cocktail formula of EMA-2+Te 82 provided a more practical and sensitive diagnostic candidate for diagnosing T.equi infection in horses as compared with Te 82 or Te 43 alone.Conclusions:The ELISA technique using a cocktail formula of EMA-2+Te 82 offers a practical and sensitive diagnostic tool for diagnosing T.equi infection in horses and using of this promising cocktail formula will be applicable for epidemiological surveys and will help control the infection in horses.

  4. Cocktail of Theileria equi antigens for detecting infection in equines

    Institute of Scientific and Technical Information of China (English)

    Shimaa Abd El-Salam El-Sayed; Mohamed Abdo Rizk; Mohamed Alaa Terkawi; Ahmed Mousa; El Said El Shirbini El Said; Gehad Elsayed; Mohamed Fouda; Naoaki Yokoyama; Ikuo Igarashi

    2015-01-01

    Objective: To use two diagnostic antigens belonging to the frequently associated in Theileria domain, Theileria equi (T. equi) protein 82 (Te 82) and T. equi 104 kDa microneme-rhoptry antigen precursor (Te 43), to diagnose T. equi infection in horses as compared with equi merozoite antigen-2 (EMA-2). Methods: In the current study, we applied a cocktail-ELISA containing two antigens (EMA-2+Te 82) to diagnose T. equi infection either in experimentally infected horses or in field infection. Results: Our findings have revealed that a cocktail formula of EMA-2+Te 82 provided a more practical and sensitive diagnostic candidate for diagnosing T. equi infection in horses as compared with Te 82 or Te 43 alone. Conclusions: The ELISA technique using a cocktail formula of EMA-2+Te 82 offers a practical and sensitive diagnostic tool for diagnosing T. equi infection in horses and using of this promising cocktail formula will be applicable for epidemiological surveys and will help control the infection in horses.

  5. Klossiella equi in the Kidneys of a Horse

    OpenAIRE

    Austin, R J; Dies, K H

    1981-01-01

    The protozoan, Klossiella equi was found in the kidneys of an aged Shetland mare raised in the Fredericton area of New Brunswick. This is the first published report of K. equi in a horse in Canada. The microscopic appearance of the parasite in the kidney is described. A brief discussion of other conditions seen in the horse is also presented.

  6. Klossiella equi in the kidneys of a horse.

    Science.gov (United States)

    Austin, R J; Dies, K H

    1981-05-01

    The protozoan, Klossiella equi was found in the kidneys of an aged Shetland mare raised in the Fredericton area of New Brunswick. This is the first published report of K. equi in a horse in Canada. The microscopic appearance of the parasite in the kidney is described. A brief discussion of other conditions seen in the horse is also presented.

  7. 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)

  8. Phenotypic and genotypic characterization of Rhodococcus equi isolated from sputum

    Directory of Open Access Journals (Sweden)

    Paulo da Silva

    Full Text Available INTRODUCTION: Rhodococcus equi is an opportunistic pathogen, causing rhodococcosis, a condition that can be confused with tuberculosis. Often, without identifying M. tuberculosis, physicians initiate empiric treatment for tuberculosis. R. equi and M. tuberculosis have different susceptibility to drugs. Identification of R. equi is based on a variety of phenotypic, chromatographic, and genotypic characteristics. OBJECTIVE: This study aimed to characterize bacterial isolates from sputum samples suggestive of R. equi. METHODS: The phenotypic identification included biochemical assays; thin-layer chromatography (TLC and polymerase chain reaction (PCR were used for genotypic identification. RESULTS: Among 78 Gram-positive and partially acid-fast bacilli isolated from the sputum of tuberculosis-suspected patients, 51 were phenotypically and genotypically characterized as R. equi based on literature data. Mycolic acid analysis showed that all suspected R. equi had compounds with a retention factor (Rf between 0.4-0.5. Genotypic characterization indicated the presence of the choE gene 959 bp fragments in 51 isolates CAMP test positive. Twenty-two CAMP test negative isolates were negative for the choE gene. Five isolates presumptively identified as R. equi, CAMP test positive, were choE gene negative, and probably belonged to other bacterial species. CONCLUSIONS: The phenotypic and molecular techniques used constitute a good methodological tool to identify R. equi.

  9. Phenotypic and genotypic characterization of Rhodococcus equi isolated from sputum.

    Science.gov (United States)

    Silva, Paulo da; Santos, Adolfo Carlos Barreto; Sato, Daisy Nakamura; Silva, Jaqueline Otero; Medeiros, Marta Inês Cazentini; Carneiro, Ana Maria Machado; Leite, Sergio Roberto de Andrade; Leite, Clarice Queico Fujimura

    2012-01-01

    Rhodococcus equi is an opportunistic pathogen, causing rhodococcosis, a condition that can be confused with tuberculosis. Often, without identifying M. tuberculosis, physicians initiate empiric treatment for tuberculosis. R. equi and M. tuberculosis have different susceptibility to drugs. Identification of R. equi is based on a variety of phenotypic, chromatographic, and genotypic characteristics. This study aimed to characterize bacterial isolates from sputum samples suggestive of R. equi. The phenotypic identification included biochemical assays; thin-layer chromatography (TLC) and polymerase chain reaction (PCR) were used for genotypic identification. Among 78 Gram-positive and partially acid-fast bacilli isolated from the sputum of tuberculosis-suspected patients, 51 were phenotypically and genotypically characterized as R. equi based on literature data. Mycolic acid analysis showed that all suspected R. equi had compounds with a retention factor (R(f)) between 0.4-0.5. Genotypic characterization indicated the presence of the choE gene 959bp fragments in 51 isolates CAMP test positive. Twenty-two CAMP test negative isolates were negative for the choE gene. Five isolates presumptively identified as R. equi, CAMP test positive, were choE gene negative, and probably belonged to other bacterial species. The phenotypic and molecular techniques used constitute a good methodological tool to identify R. equi. Copyright © 2012 Elsevier Editora Ltda. All rights reserved.

  10. Genotypic and phenotypic detection of efflux pump in Rhodococcus equi.

    Science.gov (United States)

    Gressler, Letícia Trevisan; de Vargas, Agueda Castagna; da Costa, Mateus Matiuzzi; Pötter, Luciana; da Silveira, Bibiana Petri; Sangioni, Luis Antônio; de Avila Botton, Sônia

    2014-01-01

    The req_39680 gene, associated to a putative efflux system, was detected in 60% (54/90) of R. equi isolates by PCR. The phenotypic expression of efflux mechanism was verified in 20% of the isolates using ethidium bromide. For the first time, the expression of efflux mechanism was demonstrated in R. equi.

  11. Genotypic and phenotypic detection of efflux pump in Rhodococcus equi

    Directory of Open Access Journals (Sweden)

    Letícia Trevisan Gressler

    2014-06-01

    Full Text Available The req_39680 gene, associated to a putative efflux system, was detected in 60% (54/90 of R. equi isolates by PCR. The phenotypic expression of efflux mechanism was verified in 20% of the isolates using ethidium bromide. For the first time, the expression of efflux mechanism was demonstrated in R. equi.

  12. Activity of 10 antimicrobial agents against intracellular Rhodococcus equi.

    Science.gov (United States)

    Giguère, Steeve; Berghaus, Londa J; Lee, Elise A

    2015-08-05

    Studies with facultative intracellular bacterial pathogens have shown that evaluation of the bactericidal activity of antimicrobial agents against intracellular bacteria is more closely associated with in vivo efficacy than traditional in vitro susceptibility testing. The objective of this study was to determine the relative activity of 10 antimicrobial agents against intracellular Rhodococcus equi. Equine monocyte-derived macrophages were infected with virulent R. equi and exposed to erythromycin, clarithromycin, azithromycin, rifampin, ceftiofur, gentamicin, enrofloxacin, vancomycin, imipenem, or doxycycline at concentrations achievable in plasma at clinically recommended dosages in foals. The number of intracellular R. equi was determined 48h after infection by counting colony forming units (CFUs). The number of R. equi CFUs in untreated control wells were significantly higher than those of monolayers treated with antimicrobial agents. Numbers of R. equi were significantly lower in monolayers treated with enrofloxacin followed by those treated with gentamicin, and vancomycin, when compared to monolayers treated with other antimicrobial agents. Numbers of R. equi in monolayers treated with doxycycline were significantly higher than those of monolayers treated with other antimicrobial agents. Differences in R. equi CFUs between monolayers treated with other antimicrobial agents were not statistically significant. Enrofloxacin, gentamicin, and vancomycin are the most active drugs in equine monocyte-derived macrophages infected with R. equi. Additional studies will be needed to determine if these findings correlate with in vivo efficacy. Copyright © 2015 Elsevier B.V. All rights reserved.

  13. 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.

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

    DEFF Research Database (Denmark)

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

    1991-01-01

    Rhodococcus equi is an aerobic, gram-positive, non-motile pleomorphic bacillus infecting immunocompromised patients. Forty-nine cases of Rhodococcus equi infection have been reported, mainly in patients infected with the human immunodeficiency virus (HIV). A case in which Rhodococcus equi caused...... severe pulmonary infection, the most common presentation, is described. Clinically, patients have symptoms of pneumonia with hemoptysis as a prominent feature. X-ray will often show a cavitating upper-lobe infiltrate, resembling infection with mycobacteria. Rhodococcus equi is easily cultured from blood...... 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...

  15. 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.

  16. Severe human Babesia divergens infection in Norway

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    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.

  17. Molecular detection and identification of Babesia bovis and Babesia bigemina in cattle in northern Thailand.

    Science.gov (United States)

    Cao, Shinuo; Aboge, Gabriel Oluga; Terkawi, Mohamad Alaa; Yu, Longzheng; Kamyingkird, Ketsarin; Luo, Yuzi; Li, Yan; Goo, Youn-Kyoung; Yamagishi, Junya; Nishikawa, Yoshifumi; Yokoyama, Naoaki; Suzuki, Hiroshi; Igarashi, Ikuo; Maeda, Ryuichiro; Inpankaew, Tawin; Jittapalapong, Sathaporn; Xuan, Xuenan

    2012-09-01

    Although Babesia bovis and Babesia bigemina infections cause economic losses in the cattle industry in northern Thailand, there is inadequate information on Babesia isolates present in the area. Therefore, to determine the prevalence and genetic relationship between Babesia isolates, we screened 200 blood samples of cattle from Chiang Rai, Chiang Mai, and Lumpang provinces of northern Thailand. A nested polymerase chain reaction using primers targeting B. bovis spherical body protein 2 (BboSBP2) and B. bigemina rhoptry-associated protein 1a (BbiRAP-1a) genes revealed a prevalence of 12 and 21 % for B. bovis and B. bigemina, respectively, while that of mixed infections was 6.5 % samples. The prevalences of B. bovis in Chiang Rai, Chiang Mai, and Lumpang were 9.5, 3.7, and 25.5 %, respectively. For B. bigemina, the prevalences were 15.8, 12.9, and 39.2 % in Chiang Rai, Chiang Mai, and Lumpang, respectively. Mixed infections with B. bovis and B. bigemina were 6.3 % in Chiang Rai, 1.9 % in Chiang Mai, and 13.7 % in Lumpang. The identical sequences of either BboSBP2 gene or BbiRAP-1a gene were shared among the Babesia isolates in the three provinces of northern Thailand. Further analysis using the internal transcribed spacer gene revealed at least four genotypes for B. bovis and five genotypes for B. bigemina in northern Thailand, while the sequences present great genetic diversities in the different isolates. Overall, we have demonstrated a high prevalence and polymorphism of Babesia parasites in northern Thailand calling for the need to design effective control programs for bovine babesiosis.

  18. Comparative genomic analyses reveal a lack of a substantial signature of host adaptation in Rhodococcus equi ('Prescottella equi').

    Science.gov (United States)

    Sangal, Vartul; Jones, Amanda L; Goodfellow, Michael; Sutcliffe, Iain C; Hoskisson, Paul A

    2014-08-01

    Rhodococcus equi ('Prescottella equi') is a pathogenic actinomycete primarily infecting horses but has emerged as an opportunistic human pathogen. We have sequenced the genome of the type strain of this species, R. equi strain C7(T) , and compared the genome with that of another foal isolate 103S and of a human isolate ATCC 33707. The R. equi strains are closely related to each other and yet distantly related to other rhodococci and Nocardia brasiliensis. The comparison of gene contents among R. equi strains revealed minor differences that could be associated with host adaptation from foals to humans, including the presence of a paa operon in the human isolate, which is potentially involved in pathogenesis. © 2013 Federation of European Microbiological Societies. Published by John Wiley & Sons Ltd. All rights reserved.

  19. Management of Rhodococcus equi pneumonia in foals

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    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. Immune response to Babesia bigemina infection in pregnant cows.

    Science.gov (United States)

    García, T D; Figueroa, M J V; Ramos, A J A; Rojas, M C; Cantó, A G J; Falcón, N A; Alvarez, M J A

    2004-10-01

    Babesiosis is a tick-borne disease of cattle caused by Babesia bigemina and Babesia bovis and is transmitted by the tick vector Boophilus microplus. In this study, we investigate B. bigemina infection regarding the clinical infection, T cell distribution, and cytokine profile during the acute phase of an experimental infection in pregnant cows.

  1. The third described case of transfusion-transmitted Babesia duncani

    NARCIS (Netherlands)

    Bloch, Evan M.; Herwaldt, Barbara L.; Leiby, David A.; Shaieb, Annette; Herron, Ross M.; Chervenak, Michael; Reed, William; Hunter, Robert; Ryals, Rosilyn; Hagar, Ward; Xayavong, Maniphet V.; Slemenda, Susan B.; Pieniazek, Norman J.; Wilkins, Patricia P.; Kjemtrup, Anne M.

    2012-01-01

    BACKGROUND: Almost all of the reported US tick-borne and transfusion-associated Babesia cases have been caused by Babesia microti, which is endemic in the Northeast and upper Midwest. We investigated a case caused by B. duncani (formerly, the WA1-type parasite), in a 59-year-old California resident

  2. Molecular identification of ovine Babesia spp. in north of Iran

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    Seyed Mousa Motavalli Haghi

    2013-02-01

    Conclusion: Our results showed ovine Babesiosis was common in the Iranian Northern provinces .Moreover, Babesia ovis is the main causative agent of ovine Babesiosis in this areas. The relatively high prevalence of Babesia infection in livestock indicates the epizootic stability status of babesiosis in the northern part of Iran.

  3. In vitro amoebicidal activities of Satureja cuneifolia and Melissa officinalis on Acanthamoeba castellanii cysts and trophozoites.

    Science.gov (United States)

    Malatyali, E; Tepe, B; Degerli, S; Berk, S

    2012-06-01

    Amoebic keratitis is difficult to treat without total efficacy in some patients because of cysts, which are less susceptible than trophozoites to the usual treatments. The aim of this study is to evaluate the in vitro amoebicidal activity of the methanolic extracts of Satureja cuneifolia and Melissa officinalis. In the presence of methanolic extracts (ranging from 1.0 to 32.0 mg/ml), numbers of the viable Acanthamoe castellanii trophozoites and cysts were decreased during the experimental process. Both extracts showed a time- and dose-dependent amoebicidal action on the trophozoites and cysts. Among the extracts tested, S. cuneifolia showed the strongest amoebicidal effect on the trophozoites and cysts. In the presence of 32 mg/ml extract, no viable trophozoites were observed within 24 h. At the same concentration value, the extract was found effective against the cysts at a rate of 46.3% within 72 h of the experimental process. At 16 mg/ml extract concentration, no viable trophozoites were also observed in the 24th hour of the experiment. At the end of the experimental process, 34.7% of the cysts were killed by the extract. M. officinalis showed moderate amoebicidal effect. At the concentration of 32 mg/ml, 44.3% and 30.0% of the trophozoites and cysts were killed by the extract, respectively. Results obtained from these concentration values were found statistically different in terms of their actions both on trophozoites and cysts (p<0.05).

  4. Disseminated Rhodococcus equi infection in dromedary camels (Camelus dromedarius).

    Science.gov (United States)

    Kinne, J; Madarame, H; Takai, S; Jose, S; Wernery, U

    2011-04-21

    Rhodococcus (R). equi, a recognized pathogen in horses, is emerging as a human opportunistic pathogen, especially in immunocompromized people. It affects also New World camelids, but there are no reports of R. equi infection in Old World camelids yet. Four cases of disseminated R. equi infection in adult breeding dromedaries occurred at one camel farm near Dubai within 16 months of each other. At necropsy the lungs were diffusely consolidated with large caseous areas. Histology revealed severe suppurative to necrotising pneumonia with multiple encapsulated abscesses. Immunohistochemistry enabled the detection of 15- to 17-kDa antigens (VapA) of R. equi in the lung sections. High numbers of R. equi were isolated from the lung lesions as well as from liver, spleen and mediastinal lymph nodes, indicative of septicaemia. The isolated strains were PCR-positive for the specific virulence plasmid (VapA-Gen) of R. equi, indicating virulent strains and containing an 85-kb type I plasmid. This is the first report of disseminated R. equi infection in Old World camelids. Since adult camels in general do not suffer from bacterial caused pneumonia (except tuberculosis), this is a new emerging disease for camels.

  5. Molecular characterization of Rhodococcus equi isolates in equines

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    Rabyia Javed

    2017-01-01

    Full Text Available Aim: The aim was to determine the occurrence of Rhodococcus equi in equines and their environment in Jammu (R.S. Pura, Katra, molecular characterization and to determine the antibiotic resistance pattern of R. equi. Materials and Methods: A total of 96 nasopharyngeal swab samples were collected from equines. The organism was isolated on Columbia nalidixic acid agar containing 5% sheep blood as well as on sheep blood agar and was later confirmed by cultural characteristics and biochemical tests. Molecular detection of R. equi isolates was done by 16S rRNA gene amplification followed by virulence associated protein A (Vap A gene amplification. Antibiogram was performed against five antibiotics, viz., amoxicillin, penicillin G, streptomycin, rifampicin, and methicillin. Results: During the study, 9 R. equi isolates were identified on the basis of cultural and biochemical tests. In the polymerase chain reaction based detection, 3 among the 9 rhodococcal isolates were positive for species-specific 16S rRNA gene and revealed amplicon of 450 bp for confirmation of 16S rRNA gene. None of the sample was found positive for Vap A gene. In antibiogram, R. equi isolates were found sensitive for amoxicillin, while some isolates were also found resistant to the most conventional antibiotic penicillin G. Conclusion: From this study, it was concluded that R. equi infection is prevalent in equines in Jammu region of India and the indiscriminate use of the antibiotics is leading toward the development of resistant strains of R. equi.

  6. Molecular characterization of Rhodococcus equi isolates in equines.

    Science.gov (United States)

    Javed, Rabyia; Taku, A K; Sharma, R K; Badroo, Gulzaar Ahmed

    2017-01-01

    The aim was to determine the occurrence of Rhodococcus equi in equines and their environment in Jammu (R.S. Pura, Katra), molecular characterization and to determine the antibiotic resistance pattern of R. equi. A total of 96 nasopharyngeal swab samples were collected from equines. The organism was isolated on Columbia nalidixic acid agar containing 5% sheep blood as well as on sheep blood agar and was later confirmed by cultural characteristics and biochemical tests. Molecular detection of R. equi isolates was done by 16S rRNA gene amplification followed by virulence associated protein A (Vap A) gene amplification. Antibiogram was performed against five antibiotics, viz., amoxicillin, penicillin G, streptomycin, rifampicin, and methicillin. During the study, 9 R. equi isolates were identified on the basis of cultural and biochemical tests. In the polymerase chain reaction based detection, 3 among the 9 rhodococcal isolates were positive for species-specific 16S rRNA gene and revealed amplicon of 450 bp for confirmation of 16S rRNA gene. None of the sample was found positive for Vap A gene. In antibiogram, R. equi isolates were found sensitive for amoxicillin, while some isolates were also found resistant to the most conventional antibiotic penicillin G. From this study, it was concluded that R. equi infection is prevalent in equines in Jammu region of India and the indiscriminate use of the antibiotics is leading toward the development of resistant strains of R. equi.

  7. Seroprevalence of Rhodococcus equi in horses in Israel

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    Sharon Tirosh-Levy

    2017-01-01

    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.

  8. 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.

  9. Molecular characterization of Rhodococcus equi isolates in equines

    Science.gov (United States)

    Javed, Rabyia; Taku, A. K.; Sharma, R. K.; Badroo, Gulzaar Ahmed

    2017-01-01

    Aim: The aim was to determine the occurrence of Rhodococcus equi in equines and their environment in Jammu (R.S. Pura, Katra), molecular characterization and to determine the antibiotic resistance pattern of R. equi. Materials and Methods: A total of 96 nasopharyngeal swab samples were collected from equines. The organism was isolated on Columbia nalidixic acid agar containing 5% sheep blood as well as on sheep blood agar and was later confirmed by cultural characteristics and biochemical tests. Molecular detection of R. equi isolates was done by 16S rRNA gene amplification followed by virulence associated protein A (Vap A) gene amplification. Antibiogram was performed against five antibiotics, viz., amoxicillin, penicillin G, streptomycin, rifampicin, and methicillin. Results: During the study, 9 R. equi isolates were identified on the basis of cultural and biochemical tests. In the polymerase chain reaction based detection, 3 among the 9 rhodococcal isolates were positive for species-specific 16S rRNA gene and revealed amplicon of 450 bp for confirmation of 16S rRNA gene. None of the sample was found positive for Vap A gene. In antibiogram, R. equi isolates were found sensitive for amoxicillin, while some isolates were also found resistant to the most conventional antibiotic penicillin G. Conclusion: From this study, it was concluded that R. equi infection is prevalent in equines in Jammu region of India and the indiscriminate use of the antibiotics is leading toward the development of resistant strains of R. equi. PMID:28246441

  10. Babesia canis canis, Babesia canis vogeli, Babesia canis rossi: differentiation of the three subspecies by a restriction fragment length polymorphism analysis on amplified small subunit ribosomal RNA genes.

    Science.gov (United States)

    Carret, C; Walas, F; Carcy, B; Grande, N; Précigout, E; Moubri, K; Schetters, T P; Gorenflot, A

    1999-01-01

    The parasites Babesia canis and Babesia gibsoni (phylum Apicomplexa) are responsible for canine babesiosis throughout the world. Babesia canis was previously described as a group of three biologically different subspecies, namely B. canis canis, B. canis vogeli, and B. canis rossi. We report partial sequences of small subunit ribosomal RNA gene (ssu-rDNA) of each subspecies amplified in vitro with primers derived from a semi-conserved region of the ssu-rDNA genes in other Babesia species. The polymerase chain reaction combined with a restriction fragment length polymorphism analysis, using HinfI and TaqI restriction enzymes, confirmed the separation of B. canis into three subspecies. These sequences were compared with previously published sequences of other Babesia species. A phylogenetic approach showed that the three subspecies of B. canis belong to the clade of Babesia species sensu stricto where B. canis canis clusters with B. canis rossi whereas B. canis vogeli might form a monophyletic group with the cluster B. divergens and B. odocoilei. Our results show that the three subspecies of B. canis can readily be differentiated at the molecular level and suggest that they might be considered as true species.

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

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    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.

  12. An Adenoviral Vector Based Vaccine for Rhodococcus equi.

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    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.

  13. [Periprosthetic joint infection caused by Rhodococcus equi. Case report].

    Science.gov (United States)

    Sallai, Imre; Péterfy, Nóra; Sanatkhani, Mohammad; Bejek, Zoltán; Antal, Imre; Prinz, Gyula; Kristóf, Katalin; Skaliczki, Gábor

    2017-07-01

    Rhodococcus equi is a rare pathogen in humans causing infections mostly in immunocompromised hosts. We present the first case of periprosthetic joint infection caused by Rhodococcus equi. An 88-year-old male patient was referred to our clinic with a history of fever and right hip pain. The patient had multiple hip surgeries including total joint arthroplasty and revision for aseptic loosening on the right side. He was immunocompetent, but his additional medical history was remarkable for diabetes mellitus, diabetic nephropathy and stroke with hemiplegia resulting in immobilization. Radiography showed stable components, joint aspirate yielded Rhodococcus equi. Irrigation and debridement was proposed, but the patient refused any surgical intervention. Therefore antibiotic therapy was administered. At the last follow-up the patient is free of complaints but the C-reactive protein level is still elevated. This case illustrates the possible role of Rhodococcus equi in medical device-associated infections. Orv Hetil. 2017; 158(27): 1071-1074.

  14. An Adenoviral Vector Based Vaccine for Rhodococcus equi.

    Science.gov (United States)

    Giles, Carla; Ndi, Olasumbo; Barton, Mary D; Vanniasinkam, Thiru

    2016-01-01

    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.

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

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    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.

  16. Persistent detection of Babesia EU1 and Babesia microti in Ixodes ricinus in The Netherlands during a 5-year surveillance: 2003-2007

    NARCIS (Netherlands)

    Wielinga, P.R.; Fonville, M.; Sprong, H.; Gaasenbeek, C.P.H.; Borgsteede, F.H.M.

    2009-01-01

    We report the finding of Babesia EU1 and Babesia microti in Ixodes ricinus ticks in the Netherlands. During 5 years of surveillance between 2003 and 2007, 1488 ticks were collected in a dune forest area near the North Sea and were screened for Babesia infections. In 17 ticks, DNA of the protozoan pa

  17. Susceptibility testing of Rhodococcus equi: An interlaboratory test.

    Science.gov (United States)

    Riesenberg, Anne; Kaspar, Heike; Feßler, Andrea T; Werckenthin, Christiane; Schwarz, Stefan

    2016-10-15

    Due to the importance of antimicrobial susceptibility testing (AST) for veterinary diagnostics, a standardised protocol for AST of Rhodococcus equi by broth microdilution has recently been developed and approved by the Clinical and Laboratory Standards Institute (CLSI). The aim of the present study was to test this protocol in an interlaboratory comparative study for its fitness for use in routine laboratory diagnostics. All of the 18 participating laboratories determined the minimum inhibitory concentrations (MIC) of two R. equi strains against 24 antimicrobial agents. The modal MIC values were determined and the acceptable ranges were set as the modal MIC ±1 dilution step. The R. equi field strain Rh110 showed a slightly better performance than the type strain R. equi ATCC(®) 25729. For the different antimicrobial agents tested, the percentage of MIC values within the acceptable ranges varied from 75.9 to 100% for R. equi ATCC(®) 25729, and from 85.2 to 100% for R. equi Rh110. The most homogeneous MIC results (i.e. modal MIC ±1 dilution step) were obtained for oxacillin and vancomycin, while the most divergent results were seen with cefotaxime and ceftiofur. Using a success rate of at least 80% of the strain-specific MICs being within the acceptable ranges as an arbitrary cut-off, only one of the participating laboratories failed to reach this cut-off value for one of the two R. equi strains. Thus, we consider the new protocol fit for use in routine AST of R. equi. Copyright © 2015 Elsevier B.V. All rights reserved.

  18. Streptococcus zooepidemicus and Streptococcus equi evolution: the role of CRISPRs.

    Science.gov (United States)

    Waller, Andrew S; Robinson, Carl

    2013-12-01

    The host-restricted bacterium Streptococcus equi is the causative agent of equine strangles, the most frequently diagnosed infectious disease of horses worldwide. The disease is characterized by abscessation of the lymph nodes of the head and neck, leading to significant welfare and economic cost. S. equi is believed to have evolved from an ancestral strain of Streptococcus zooepidemicus, an opportunistic pathogen of horses and other animals. Comparison of the genome of S. equi strain 4047 with those of S. zooepidemicus identified examples of gene loss due to mutation and deletion, and gene gain through the acquisition of mobile genetic elements that have probably shaped the pathogenic specialization of S. equi. In particular, deletion of the CRISPR (clustered regularly interspaced short palindromic repeats) locus in the ancestor of S. equi may have predisposed the bacterium to acquire and incorporate new genetic material into its genome. These include four prophages and a novel integrative conjugative element. The virulence cargo carried by these mobile genetic elements is believed to have shaped the ability of S. equi to cause strangles. Further sequencing of S. zooepidemicus has highlighted the diversity of this opportunistic pathogen. Again, CRISPRs are postulated to influence evolution, balancing the need for gene gain over genome stability. Analysis of spacer sequences suggest that these pathogens may be susceptible to a limited range of phages and provide further evidence of cross-species exchange of genetic material among Streptococcus pyogenes, Streptococcus agalactiae and Streptococcus dysgalactiae.

  19. Entamoeba histolytica: Adhesins and Lectins in the Trophozoite Surface

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    Magdalena Aguirre García

    2015-02-01

    Full Text Available Entamoeba histolytica is the causative agent of amebiasis in humans and is responsible for 100,000 deaths annually, making it the third leading cause of death due to a protozoan parasite. Pathogenesis appears to result from the potent cytotoxic activity of the parasite, which kills host cells within minutes. Although the mechanism is unknown, it is well established to be contact-dependent. The life cycle of the parasite alternates with two forms: the resistant cyst and the invasive trophozoite. The adhesive interactions between the parasite and surface glycoconjugates of host cells, as well as those lining the epithelia, are determinants for invasion of human tissues, for its cytotoxic activity, and finally for the outcome of the disease. In this review we present an overview of the information available on the amebic lectins and adhesins that are responsible of those adhesive interactions and we also refer to their effect on the host immune response. Finally, we present some concluding remarks and perspectives in the field.

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

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    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.

  1. Prevalence of Theileria and Babesia species in Tunisian sheep.

    Science.gov (United States)

    Rjeibi, Mohamed R; Darghouth, Mohamed A; Gharbi, Mohamed

    2016-05-24

    In this study, the prevalence of Theileria and Babesia species in sheep was assessed with Giemsastained blood smear examination and polymerase chain reaction to identify the different piroplasms in 270 sheep from three Tunisian bioclimatic zones (north, centre, and south). The overall infection prevalence by Babesia spp. and Theileria spp. in Giemsa-stained blood smears was 2.9% (8/270) and 4.8% (13/270) respectively. The molecular results showed that sheep were more often infected by Theileria ovis than Babesia ovis with an overall prevalence of 16.3% (44/270) and 7.8% (21/270) respectively (p = 0.01). The molecular prevalence by Babesia ovis was significantly higher in females than in males (p distribution.

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

    OpenAIRE

    2015-01-01

    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 ...

  3. Interaction of secretory immunoglobulin A antibodies with Naegleria fowleri trophozoites and collagen type I.

    Science.gov (United States)

    Shibayama, Mineko; Serrano-Luna, José de Jesús; Rojas-Hernández, Saúl; Campos-Rodríguez, Rafael; Tsutsumi, Victor

    2003-03-01

    In this work, we analyzed the in vitro interaction of human secretory immunoglobulin A (sIgA) antibodies with Naegleria fowleri trophozoites and the capacity of these antibodies to inhibit amoeba adherence to collagen type I. We also studied N. fowleri antigens that are recognized by sIgA, using immunoblot assays. Immunocytochemical analysis of the interaction showed a redistribution of antigens on the surface of trophozoites by sIgA antibodies. Ultrastructural analysis of antibody-amoeba interaction showed that besides the patching and cap formation, parasites were capable of eliminating the antigen-antibody complex produced on the surface. sIgA antibodies were capable of inhibiting the in vitro adhesion of trophozoites to collagen type I. We suggest that nonsymptomatic infections by N. fowleri may stimulate a local specific immunity that prevents trophozoite adhesion and invasion of nasal mucosa.

  4. Cavitary Rhodococcus equi pneumonia with endobronchial granulomas: report of an unusual case

    Energy Technology Data Exchange (ETDEWEB)

    Fidvi, Shabnam A.; Brudnicki, Adele R. [Department of Radiology, Westchester Medical Center, Valhalla, NY (United States); Chowdhury, M.Ishraq; Beneck, Debra [Department of Pathology, Westchester Medical Center, Valhalla, NY (United States)

    2003-02-01

    An unusual case of cavitary Rhodococcus equi pneumonia with endobronchial granulomas in congenital HIV infection is presented. The clinical features and radiological manifestations of pulmonary R. equi infection are discussed.

  5. In vitro amoebicidal activity of Origanum syriacum and Origanum laevigatum on Acanthamoeba castellanii cysts and trophozoites.

    Science.gov (United States)

    Degerli, Serpil; Tepe, Bektas; Celiksoz, Ali; Berk, Seyda; Malatyali, Erdogan

    2012-05-01

    In some patients, complete treatment of amoebic keratitis is difficult because of the resistance of cysts to therapeutic agents. The aim of this study was to evaluate the in vitro amoebicidal activity of methanolic extracts of Origanum syriacum and Origanum laevigatum. In the presence of methanolic extracts (ranging from 1.0 to 32.0mg/ml), numbers of the viable Acanthamoeba castellanii trophozoites and cysts were decreased. Both extracts showed a time and dose dependent amoebicidal action on the trophozoites and cysts. Of the extracts tested, O. syriacum showed the stronger amoebicidal effect on the trophozoites and cysts. In the presence of 32 mg/ml extract, no viable trophozoites were observed within third hour. The extract was also found effective against the cysts within 24th hour. In the case of O. laevigatum, no viable trophozoites were observed within 72nd hour at the concentrations of 16 and 32 mg/ml. As expected, cysts were found more resistant to the extracts than the trophozoites.

  6. PCR-based detection of the transovarial transmission of Uruguayan Babesia bovis and Babesia bigemina vaccine strains.

    Science.gov (United States)

    Gayo, V; Romito, M; Nel, L H; Solari, M A; Viljoen, G J

    2003-09-01

    Bovine babesiosis is responsible for serious economic losses in Uruguay. Haemovaccines play an important role in disease prevention, but concern has been raised about their use. It is feared that the attenuated Babesia bovis and Babesia bigemina vaccine strains may be transmitted by the local tick vector Boophilus microplus, and that reversion to virulence could occur. We therefore investigated the possibility that these strains could be transmitted via the transovarial route in ticks using a Babesia species-specific polymerase chain reaction (PCR) assay. DNA was extracted from the developmental stages of the tick vector that had fed on calves immunized with the haemovaccine. It was possible to detect Babesia DNA not only in adult ticks, but also in their eggs and larvae. In addition, it was shown that calves infested with larvae derived from eggs laid by ticks fed on acutely infected calves, were positive for Babesia using PCR. Caution should therefore be shown with the distribution of the haemovaccine in marginal areas. It is still advisable that suitable tick control measures be used to prevent transovarial transmission and the potential risk of attenuated Babesia reverting to virulence.

  7. 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.

  8. 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.

  9. Transcriptional changes are involved in phenotype switching in Streptococcus equi subspecies equi.

    Science.gov (United States)

    Steward, Karen F; Robinson, Carl; Waller, Andrew S

    2016-04-01

    Phenotypic heterogeneity within a population of bacteria, through genetic or transcriptional variation, enables survival and persistence in challenging and changing environments. We report here that a recent clinical isolate of S. equi, strain 1691 (Se1691), yielded a mixture of reduced capsule and mucoid colonies on primary isolation when grown on colistin-oxolinic acid blood agar (COBA) streptococcal selective plates. Passaging colonies of Se1691, with a reduced capsule phenotype maintained this mixed phenotype. In contrast, passaging mucoid colonies fixed the mucoid phenotype, suggesting adaptive genetic or transcriptional changes in response to growth on artificial media. However, despite obvious phenotypic and transcriptional differences, there were no apparent differences in the genome sequences of Se1691 recovered from colonies with a mucoid or reduced capsule phenotype. We identified 105 differentially transcribed genes in the transcriptomes of reduced capsule and mucoid colonies. The reduced capsule phenotype was associated with a significant reduction in transcription of the has locus (SEQ_0269 Q = 0.0015, SEQ_0270 Q = 0.0015, SEQ_0271 Q = 0.0285) and the amount of hyaluronic acid on the surface of S. equi recovered from non-mucoid colonies (P = 0.017). Significant differences in the transcription of 21 surface and secreted proteins were also observed. Our data show that changes in the bacterial transcriptome are linked to the mixed colony phenotype of Se1691.

  10. Disseminated Rhodococcus equi infection in a patient with Hodgkin lymphoma.

    Science.gov (United States)

    Mikić, Dragan; Djordjević, Zoran; Sekulović, Leposava; Kojić, Miroslav; Tomanović, Branka

    2014-03-01

    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. We reported the extremly rare case of a 43-year-old woman with Hodgkin 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 fa cial 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 x 10(12)/L, whyite blood cells (WBC) 2.8 x 10(9)/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. 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.

  11. Unusual Extrapulmonary Rhodococcus Equi Infection in a Kidney Transplant Patient.

    Science.gov (United States)

    Varotti, Giovanni; Barabani, Caterina; Dodi, Ferdinando; Bertocchi, Massimo; Mondello, Rosalia; Cupo, Pierpaolo; Santori, Gregorio; Palombo, Domenico; Fontana, Iris

    2016-12-01

    Rhodococcus equi is a well-recognized pathogen in veterinary medicine that can also affect immuno-compromised human subjects. The most common clinical features in humans include necrotizing pneumonia with subacute pulmonary disease, progressive cough, chest pain and fever. We report a case of a 49-year-old kidney transplant patient who developed a Rhodococcus equi infection characterized by multiple abscesses of the soft tissues and muscles without any respiratory manifestation. Combining specific antibiotic therapy and surgical management of the abscesses without immunosuppression discontinuation led to a complete recovery of both patient and graft.

  12. Clinicopathological findings in dogs naturally infected dogs with Babesia

    Directory of Open Access Journals (Sweden)

    Egon Andoni

    2013-05-01

    Full Text Available The aim of the study was to describe the clinicopathological findings in dogs naturally infected with Babesia. During the period of February 2011-February 2012 twenty-nine cases of Babesia Canis were diagnosed in the clinic of Small Animal at the Veterinary Faculty of Tirana. The animal was considered infected when pear shaped babesia was observed inside infected erythrocytes in Wright-Giemsa stained blood smears. The animal stay for one week in our clinic and during this period we evaluated the canine babesiosis cases by means of clinical history, physical examination, hematological and biochemical tests. Based in the clinical sing of babesia the animal was divided in two groups with uncomplicated and complicate babesia. The duration of clinical signs ranged from 1 to 5 days prior to the arrival at the clinic and all the animals were positive with B. c. canis infections. The main clinical signs were dehydration and anemia in (79%, apathy (74%, anorexia or decrease appetite (70% and fever (68%. The anemia was present in the dogs and classified severe (13% as mild (45%, and moderate (52% of all the cases. In conclusion, the main clinicopathological findings were a mild to moderate hemolytic anemia and mild to severe thrombocytopenia.

  13. 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.

  14. 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.

  15. Description of novel Babesia species and associated lesions from common murres (Uria aalge) from California.

    Science.gov (United States)

    Yabsley, Michael J; Greiner, Ellis; Tseng, Florina S; Garner, Michael M; Nordhausen, Robert W; Ziccardi, Michael H; Borjesson, Dori L; Zabolotzky, Shanon

    2009-10-01

    A novel Babesia species is described from blood collected from 3 common murres (also known as common guillemots) (Uria aalge) from California. This is the first report of a Babesia species from the Alcidae and the second from the Charadriiformes. Babesia uriae n. sp. is morphologically similar to several other avian Babesia species but can be differentiated from Babesia bennetti, the only other species from a host in the Charadriiformes, by size of round and amoeboid forms. Phylogenetic analysis of near-full length 18S rRNA, ITS-1, 5.8S rRNA, and ITS-2 sequences indicated that the Babesia sp. from the common murre is closely related to Babesia poelea, a parasite of brown boobies (Sula leucogaster).

  16. First detection of Babesia occultans in Hyalomma ticks from Tunisia.

    Science.gov (United States)

    Ros-García, A; M'Ghirbi, Y; Bouattour, A; Hurtado, A

    2011-04-01

    Descriptions of Babesia occultans have previously been restricted to sub-Saharan Africa. Here, we report the finding, for the first time, of this low or non-pathogenic bovine Babesia species in Tunisia, northern Africa. B. occultans DNA was detected by molecular methods in Hyalomma marginatum unfed ticks collected in 3 bioclimatic regions of Tunisia. The near-full-length 18S rRNA gene was sequenced and compared with related sequences retrieved from GenBank. Phylogenetic analysis indicated that other sequences deposited as Babesia sp. could also correspond to B. occultans, suggesting that this species may have a wide distribution in Mediterranean and Asiatic regions, and not only in sub-Saharan Africa as previously described. A B. occultans-specific Reverse Line Blot (RLB) oligonucleotide probe was designed for future epidemiological studies that would help to clarify this possibility.

  17. Hematologic Changes In Dogs Naturally Infected With Babesia

    Directory of Open Access Journals (Sweden)

    Egon Andoni

    2012-09-01

    Full Text Available Canine babesiosis caused by tick-borne organisms of the genus Babesia, is one of the most significant disease worldwide. The aims of this study were to determine the hematological changes in six dogs naturally infected with Babesia canis. The hematological evaluation included Hemoglobin (Hb, Red Blood Cell (RBC, Platelet, Packet Cell Volume (PCV, Mean Corpuscular Volume (MCV, Mean Corpuscular Hemoglobin Concentration (MCHC and Reticulocyte counts. They were estimated using an autonomic hematologic analyzer. The animal had hypocytic hypochromic anemia and 20% of the cases had a packed cell volume (PCV less of 24%. All the animal had thrombocytopenia and platelets counts was lower than 50x103 cell/ µl The results of this study show that Babesia infection in dogs caused anemia and thrombocytopenia.

  18. 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.

  19. Entamoeba histolytica calreticulin: an endoplasmic reticulum protein expressed by trophozoites into experimentally induced amoebic liver abscesses.

    Science.gov (United States)

    González, Enrique; de Leon, Maria del Carmen García; Meza, Isaura; Ocadiz-Delgado, Rodolfo; Gariglio, Patricio; Silva-Olivares, Angelica; Galindo-Gómez, Silvia; Shibayama, Mineko; Morán, Patricia; Valadez, Alicia; Limón, Angelica; Rojas, Liliana; Hernández, Eric G; Cerritos, René; Ximenez, Cecilia

    2011-02-01

    Entamoeba histolytica calreticulin (EhCRT) is remarkably immunogenic in humans (90-100% of invasive amoebiasis patients). Nevertheless, the study of calreticulin in this protozoan is still in its early stages. The exact location, biological functions, and its role in pathogenesis are yet to be fully understood. The aim of the present work is to determine the location of EhCRT in virulent trophozoites in vivo and the expression of the Ehcrt gene during the development of experimentally induced amoebic liver abscesses (ALA) in hamsters. Antibodies against recombinant EhCRT were used for the immunolocalization of EhCRT in trophozoites through confocal microscopy; immunohistochemical assays were also performed on tissue sections of ALAs at different times after intrahepatic inoculation. The expression of the Ehcrt gene during the development of ALA was estimated through both in situ RT-PCR and real-time RT-PCR. Confocal assays of virulent trophozoites showed a distribution of EhCRT in the cytoplasmic vesicles of different sizes. Apparently, EhCRT is not exported into the hepatic tissue. Real-time RT-PCR demonstrated an over-expression of the Ehcrt gene at 30 min after trophozoite inoculation, reaching a peak at 1-2 h; thereafter, the expression fell sharply to its original levels. These results demonstrate for the first time in an in vivo model of ALA, the expression of Ehcrt gene in E. histolytica trophozoites and add evidence that support CRT as a resident protein of the ER in E. histolytica species. The in vivo experiments suggest that CRT may play an important role during the early stages of the host-parasite relationship, when the parasite is adapting to a new environment, although the protein seems to be constitutively synthesized. Moreover, trophozoites apparently do not export EhCRT into the hepatic tissue in ALA.

  20. 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...

  1. In vitro activity of ponazuril against Theileria equi

    Science.gov (United States)

    The equid hemoprotozoan parasite Theileria equi is endemic in most regions worldwide. Infection of horses is a cause of significant economic loss due to costs associated with disease and restriction of trade with non-endemic nations. The ability of certain drugs such as imidocarb dipropionate to eli...

  2. Amblyomma cajennense is an intrastadial biological vector of theileria equi

    Science.gov (United States)

    Background: The apicomplexan hemoprotozoan parasite Theileria equi is one of the etiologic agents causing equine piroplasmosis (EP), a disease of horses and their relatives that are endemic throughout large parts of the world. Prior to 2009 the United States had been considered to be free of this pa...

  3. Cocktail of Theileria equi antigens for detecting infection in equines

    Directory of Open Access Journals (Sweden)

    Shimaa Abd El-Salam El-Sayed

    2015-12-01

    Conclusions: The ELISA technique using a cocktail formula of EMA-2 + Te 82 offers a practical and sensitive diagnostic tool for diagnosing T. equi infection in horses and using of this promising cocktail formula will be applicable for epidemiological surveys and will help control the infection in horses.

  4. Clinical and microbiological features of bacteremia with Streptococcus equi.

    Science.gov (United States)

    Trell, Kristina; Nilson, Bo; Petersson, Ann-Cathrine; Rasmussen, Magnus

    2017-02-01

    Streptococcus equi (SE) rarely causes human infections. We identified 18 SE isolates from blood cultures. The focus of infection was unknown (n = 5), arthritis (n = 3), catheter-related (n = 2), pneumonia (n = 2), or other (n = 6). There were no fatalities. Several patients had animal contacts but there were no indications of clonal outbreaks.

  5. Control of Bovicola Equi (Phthiraptera: Trichodectidae) with Dimilin and Permethrin

    Science.gov (United States)

    Lice are wingless ectoparasitic insects that can irritate and injure their hosts and transmit pathogens. Horses and ponies can be infested with a chewing louse, Bovicola equi (Denny) (Phthiraptera: Trichodectidae) that irritates the animals, creates skin lesions, causes hair loss, and generally redu...

  6. Proteomic Study of Entamoeba histolytica Trophozoites, Cysts, and Cyst-Like Structures.

    Science.gov (United States)

    Luna-Nácar, Milka; Navarrete-Perea, José; Moguel, Bárbara; Bobes, Raúl J; Laclette, Juan P; Carrero, Julio C

    2016-01-01

    The cyst stage of Entamoeba histolytica is a promising therapeutic target against human amoebiasis. Our research team previously reported the production in vitro of Cyst-Like Structures (CLS) sharing structural features with cysts, including rounded shape, size reduction, multinucleation, and the formation of a chitin wall coupled to the overexpression of glucosamine 6-phosphate isomerase, the rate-limiting enzyme of the chitin synthesis pathway. A proteomic study of E. histolytica trophozoites, cysts, and in vitro-produced CLS is reported herein to determine the nature of CLS, widen our knowledge on the cyst stage, and identify possible proteins and pathways involved in the encystment process. Total protein extracts were obtained from E. histolytica trophozoites, CLS, and partially purified cysts recovered from the feces of amoebic human patients; extracts were trypsin-digested and analyzed by LC-MS/MS. In total, 1029 proteins were identified in trophozoites, 550 in CLS, and 411 in cysts, with 539, 299, and 84 proteins unique to each sample, respectively, and only 74 proteins shared by all three stages. About 70% of CLS proteins were shared with trophozoites, even though differences were observed in the relative protein abundance. While trophozoites showed a greater abundance of proteins associated to a metabolically active cell, CLS showed higher expression of proteins related to proteolysis, redox homeostasis, and stress response. In addition, the expression of genes encoding for the cyst wall proteins Jessie and Jacob was detected by RT-PCR and the Jacob protein identified by Western blotting and immunofluorescence in CLS. However, the proteomic profile of cysts as determined by LC-MS/MS was very dissimilar to that of trophozoites and CLS, with almost 40% of hypothetical proteins. Our global results suggest that CLS are more alike to trophozoites than to cysts, and they could be generated as a rapid survival response of trophozoites to a stressful condition

  7. Proteomic Study of Entamoeba histolytica Trophozoites, Cysts, and Cyst-Like Structures.

    Directory of Open Access Journals (Sweden)

    Milka Luna-Nácar

    Full Text Available The cyst stage of Entamoeba histolytica is a promising therapeutic target against human amoebiasis. Our research team previously reported the production in vitro of Cyst-Like Structures (CLS sharing structural features with cysts, including rounded shape, size reduction, multinucleation, and the formation of a chitin wall coupled to the overexpression of glucosamine 6-phosphate isomerase, the rate-limiting enzyme of the chitin synthesis pathway. A proteomic study of E. histolytica trophozoites, cysts, and in vitro-produced CLS is reported herein to determine the nature of CLS, widen our knowledge on the cyst stage, and identify possible proteins and pathways involved in the encystment process. Total protein extracts were obtained from E. histolytica trophozoites, CLS, and partially purified cysts recovered from the feces of amoebic human patients; extracts were trypsin-digested and analyzed by LC-MS/MS. In total, 1029 proteins were identified in trophozoites, 550 in CLS, and 411 in cysts, with 539, 299, and 84 proteins unique to each sample, respectively, and only 74 proteins shared by all three stages. About 70% of CLS proteins were shared with trophozoites, even though differences were observed in the relative protein abundance. While trophozoites showed a greater abundance of proteins associated to a metabolically active cell, CLS showed higher expression of proteins related to proteolysis, redox homeostasis, and stress response. In addition, the expression of genes encoding for the cyst wall proteins Jessie and Jacob was detected by RT-PCR and the Jacob protein identified by Western blotting and immunofluorescence in CLS. However, the proteomic profile of cysts as determined by LC-MS/MS was very dissimilar to that of trophozoites and CLS, with almost 40% of hypothetical proteins. Our global results suggest that CLS are more alike to trophozoites than to cysts, and they could be generated as a rapid survival response of trophozoites to a

  8. 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.

  9. Oral Administration of Electron-Beam Inactivated Rhodococcus equi Failed to Protect Foals against Intrabronchial Infection with Live, Virulent R. equi.

    Directory of Open Access Journals (Sweden)

    Joana N Rocha

    Full Text Available There is currently no licensed vaccine that protects foals against Rhodococcus equi-induced pneumonia. Oral administration of live, virulent R. equi to neonatal foals has been demonstrated to protect against subsequent intrabronchial challenge with virulent R. equi. Electron beam (eBeam-inactivated R. equi are structurally intact and have been demonstrated to be immunogenic when administered orally to neonatal foals. Thus, we investigated whether eBeam inactivated R. equi could protect foals against developing pneumonia after experimental infection with live, virulent R. equi. Foals (n = 8 were vaccinated by gavaging with eBeam-inactivated R. equi at ages 2, 7, and 14 days, or gavaged with equal volume of saline solution (n = 4, and subsequently infected intrabronchially with live, virulent R. equi at age 21 days. The proportion of vaccinated foals that developed pneumonia following challenge was similar among the vaccinated (7/8; 88% and unvaccinated foals (3/4; 75%. This vaccination regimen did not appear to be strongly immunogenic in foals. Alternative dosing regimens or routes of administration need further investigation and may prove to be immunogenic and protective.

  10. Oral Administration of Electron-Beam Inactivated Rhodococcus equi Failed to Protect Foals against Intrabronchial Infection with Live, Virulent R. equi.

    Science.gov (United States)

    Rocha, Joana N; Cohen, Noah D; Bordin, Angela I; Brake, Courtney N; Giguère, Steeve; Coleman, Michelle C; Alaniz, Robert C; Lawhon, Sara D; Mwangi, Waithaka; Pillai, Suresh D

    2016-01-01

    There is currently no licensed vaccine that protects foals against Rhodococcus equi-induced pneumonia. Oral administration of live, virulent R. equi to neonatal foals has been demonstrated to protect against subsequent intrabronchial challenge with virulent R. equi. Electron beam (eBeam)-inactivated R. equi are structurally intact and have been demonstrated to be immunogenic when administered orally to neonatal foals. Thus, we investigated whether eBeam inactivated R. equi could protect foals against developing pneumonia after experimental infection with live, virulent R. equi. Foals (n = 8) were vaccinated by gavaging with eBeam-inactivated R. equi at ages 2, 7, and 14 days, or gavaged with equal volume of saline solution (n = 4), and subsequently infected intrabronchially with live, virulent R. equi at age 21 days. The proportion of vaccinated foals that developed pneumonia following challenge was similar among the vaccinated (7/8; 88%) and unvaccinated foals (3/4; 75%). This vaccination regimen did not appear to be strongly immunogenic in foals. Alternative dosing regimens or routes of administration need further investigation and may prove to be immunogenic and protective.

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

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    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.

  12. Prevalence and diversity of Babesia spp. in questing Ixodes ricinus ticks from Norway

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    Øines Øivind

    2012-08-01

    Full Text Available Abstract Background Ixodes ricinus ticks transmit Babesia species to vertebrate hosts. Using molecular tools we were able to detect the presence of this piroplasmid in its vector. The aims of this study were to investigate the prevalence and identity of Babesia species in questing ticks collected in various areas of Norway. Methods DNA from questing l. ricinus ticks were examined with a realtime PCR for the presence of Babesia. Positive samples of tick DNA were identified to species using PCR, and sequence analysis. Results From a total of 1908 questing l. ricinus ticks, 17 (0.9% indicated the presence of Babesia spp. after realtime-PCR screening. Ixodes ricinus harbouring Babesia spp. was detected in 9 out of 22 localities. Further molecular analyses of DNA from these positive ticks indicate the presence of Babesia venatorum, B. divergens, B. capreoli and a currently undescribed Babesia in Norwegian ticks. The most prevalent was B. venatorum found in 71% of the positive ticks. Conclusions A total of 17 out of 1908 (0.9% ticks were positive for Babesia. Our data confirm that there are several Babesia species in ticks in Norway. Babesia venatorum was the most prevalent. This species has a zoonotic potential and may cause human babesiosis following a tick bite.

  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. Contact-independent cell death of human microglial cells due to pathogenic Naegleria fowleri trophozoites.

    Science.gov (United States)

    Kim, Jong-Hyun; Kim, Daesik; Shin, Ho-Joon

    2008-12-01

    Free-living Naegleria fowleri leads to a fatal infection known as primary amebic meningoencephalitis in humans. Previously, the target cell death could be induced by phagocytic activity of N. fowleri as a contact-dependent mechanism. However, in this study we investigated the target cell death under a non-contact system using a tissue-culture insert. The human microglial cells, U87MG cells, co-cultured with N. fowleri trophozoites for 30 min in a non-contact system showed morphological changes such as the cell membrane destruction and a reduction in the number. By fluorescence-activated cell sorter (FACS) analysis, U87MG cells co-cultured with N. fowleri trophozoites in a non-contact system showed a significant increase of apoptotic cells (16%) in comparison with that of the control or N. fowleri lysate. When U87MG cells were co-cultured with N. fowleri trophozoites in a non-contact system for 30 min, 2 hr, and 4 hr, the cytotoxicity of amebae against target cells was 40.5, 44.2, and 45.6%, respectively. By contrast, the cytotoxicity of non-pathogenic N. gruberi trophozoites was 10.2, 12.4, and 13.2%, respectively. These results suggest that the molecules released from N. fowleri in a contact-independent manner as well as phagocytosis in a contact-dependent manner may induce the host cell death.

  15. High-speed microscopic imaging of flagella motility and swimming in Giardia lamblia trophozoites.

    Science.gov (United States)

    Lenaghan, Scott C; Davis, Corinne A; Henson, William R; Zhang, Zhili; Zhang, Mingjun

    2011-08-23

    We report, in this paper, several findings about the swimming and attachment mechanisms of Giardia lamblia trophozoites. These data were collected using a combination of a high-contrast CytoViva imaging system and a particle image velocimetry camera, which can capture images at speeds greater than 800 frames/s. Using this system, we discovered that, during rapid swimming of Giardia trophozoites, undulations of the caudal region contributed to forward propulsion combined with the beating of the flagella pairs. It was also discovered, in contrast to previous studies with 10 times slower image sampling technique, that the anterior and posterolateral flagella beat with a clearly defined power stroke and not symmetrical undulations. During the transition from free swimming to attachment, trophozoites modified their swimming behavior from a rapid rotating motion to a more stable planar swimming. While using this planar swimming motion, the trophozoites used the flagella for propulsion and directional control. In addition to examination of the posterolateral and anterior flagella, a model to describe the motion of the ventral flagella was derived, indicating that the ventral flagella beat in an expanding sine wave. In addition, the structure of the ventrocaudal groove creates boundary conditions that determine the form of beating of the ventral flagella. The results from this study indicate that Giardia is able to simultaneously generate both ciliary beating and typical eukaryotic flagellar beating using different pairs of flagella.

  16. Sensitivity of free-living amoeba trophozoites and cysts to water disinfectants.

    Science.gov (United States)

    Dupuy, Mathieu; Berne, Florence; Herbelin, Pascaline; Binet, Marie; Berthelot, Nelsie; Rodier, Marie-Hélène; Soreau, Sylvie; Héchard, Yann

    2014-03-01

    Free-living amoebae are naturally present in water. These protozoa could be pathogenic and could also shelter pathogenic bacteria. Thus, they are described as a potential hazard for health. Also, free-living amoebae have been described to be resistant to biocides, especially under their cyst resistant form. There are several studies on amoeba treatments but none of them compare sensitivity of trophozoites and cysts from different genus to various water disinfectants. In our study, we tested chlorine, monochloramine and chlorine dioxide on both cysts and trophozoites from three strains, belonging to the three main genera of free-living amoebae. The results show that, comparing cysts to trophozoites inactivation, only the Acanthamoeba cysts were highly more resistant to treatment than trophozoites. Comparison of the disinfectant efficiency led to conclude that chlorine dioxide was the most efficient treatment in our conditions and was particularly efficient against cysts. In conclusion, our results would help to adapt water treatments in order to target free-living amoebae in water networks.

  17. Interaction of Escherichia coli K1 and K5 with Acanthamoeba castellanii trophozoites and cysts.

    Science.gov (United States)

    Matin, Abdul; Jung, Suk-Yul

    2011-12-01

    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. coli K12 was also applied. The association ratio of E. coli K1 with A. castellanii was 4.3 cfu per amoeba for 1 hr but E. coli K5 with A. castellanii was 1 cfu per amoeba for 1 hr. By invasion and survival assays, E. coli K5 was recovered less than E. coli K1 but still alive inside A. castellanii. E. coli K1 and K5 survived and multiplied intracellularly in A. castellanii. The survival assay was performed under a favourable condition for 22 hr and 43 hr with the encystment of A. castellanii. Under the favourable condition for the transformation of trophozoites into cysts, E. coli K5 multiplied significantly. Moreover, the pathogenic potential of E. coli K1 from A. castellanii cysts exhibited no changes as compared with E. coli K1 from A. castellanii trophozoites. E. coli K5 was multiplied in A. castellanii trophozoites and survived in A. castellanii cysts. Therefore, this study suggests that E. coli K5 can use A. castellanii as a reservoir host or a vector for the bacterial transmission.

  18. Morphological Features and In Vitro Cytopathic Effect of Acanthamoeba griffini Trophozoites Isolated from a Clinical Case

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    Arturo González-Robles

    2014-01-01

    Full Text Available Light and transmission electron microscopy observations are reported on the structure and in vitro cytopathic effect of Acanthamoeba griffini trophozoites isolated from a clinical case. Live trophozoites were moderately active with a remarkable pleomorphism which changed from ovoid to quite elongated shapes. When moving, amoebae formed cytoplasmic projections such as wide lamellae and acanthopodia of diverse size and thickness which contain a significant amount of actin. Ultrastructurally, the cytoplasm showed the main organelles found in other free-living amoebae. Coincubation of trophozoites with MDCK cell monolayers resulted in a local damage to target cells after 24 h of interaction, suggesting that the cytopathic effect is contact-dependent. By transmission electron microscopy, amoebae appeared to engulf small portions of the MDCK cells; however, the cells that were not in contact with trophozoites had an unaltered morphology. When epithelial monolayers were incubated with conditioned medium for 24 h, small areas of cell injury were also observed. The phylogenetical analysis as well as the sequencing of the acquired amplified product for the DF3 region of the amoebae isolate confirmed that it belongs to genotype T3, which includes other pathogenic amoebae; besides the activity of two drugs currently used against Acanthamoeba was tested on A. griffini.

  19. Mitosomes in trophozoites and cysts of the reptilian parasite Entamoeba invadens.

    Science.gov (United States)

    Siegesmund, Maria A; Hehl, Adrian B; van der Giezen, Mark

    2011-11-01

    Heat shock protein genes led to the discovery of mitosomes in Entamoeba histolytica, but mitosomes have not been described for any other Entamoeba species, nor have they been identified in the cyst stage. Here, we show that the distantly related reptilian pathogen Entamoeba invadens contains mitosomes, in both trophozoites and cysts, suggesting all Entamoeba species contain these organelles.

  20. In Vitro Evaluation of Drug Susceptibilities of Babesia divergens Isolates

    OpenAIRE

    Brasseur, Philippe; Lecoublet, Sophie; Kapel, Nathalie; Favennec, Loic; Ballet, Jean J.

    1998-01-01

    The susceptibilities of three bovine and two human Babesia divergens isolates to antimicrobial agents were evaluated in vitro by a tritiated hypoxanthine incorporation assay. The MICs at which 50% of isolates are inhibited (MIC50s) for mefloquine (chlorhydrate), chloroquine (sulfate), quinine (chlorhydrate), clindamycin (phosphate), pentamidine (isethionate), phenamidine (isethionate) plus oxomemazine (chlorhydrate), lincomycin (chlorhydrate monohydrate), and imidocarb (dipropionate) were det...

  1. Babesia microti in rodents and raccoons from northeast Florida.

    Science.gov (United States)

    Clark, Kerry; Savick, Kyla; Butler, Joseph

    2012-12-01

    Human babesiosis in the United States is caused most commonly by the intraerythrocytic protozoan parasite, Babesia microti . Although a few reports have described evidence of Babesia species in animals in Florida, to date Babesia microti specifically has not been reported from Florida or most other southern states. To determine if the organism is present in vertebrates in the region, small mammals were trapped and sampled at 2 sites in northeastern Florida, and DNA extracts from blood samples were screened for B. microti DNA via PCR assays targeting portions of the nuclear small subunit rRNA (18S rDNA) and beta-tubulin genes. Amplified fragments from representative samples of PCR-positive hosts were sequenced and compared phylogenetically to reference strains of Babesia species. The B. microti strains found in cotton rats ( Sigmodon hispidus ) most closely resembles B. microti sensu stricto strains that are pathogenic to humans, and strains found in raccoons ( Procyon lotor ) most closely resembles previously described raccoon-related strains of B. microti sensu lato. The results of this study suggest that B. microti is prevalent among cotton rats and raccoons at some sites in northeast Florida and may pose a risk to humans in the region.

  2. First Report of Babesia microti-Caused Babesiosis in Spain.

    Science.gov (United States)

    Arsuaga, Marta; Gonzalez, Luis M; Lobo, Cheryl A; de la Calle, Fernando; Bautista, Jose M; Azcárate, Isabel G; Puente, Sabino; Montero, Estrella

    2016-10-01

    Babesiosis is an emerging zoonosis now found in several areas of the world. Using PCR and indirect immunofluorescence assay, we have diagnosed the first case of human babesiosis caused by Babesia microti in Spain. Diagnosis was delayed because of the nonspecific clinical symptoms that occurred in an immunocompetent patient.

  3. Trophozoites of Entamoeba histolytica epigenetically silenced in several genes are virulence-attenuated

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    Mirelman D.

    2008-09-01

    Full Text Available The human intestinal parasite Entamoeba histolytica causes amoebic colitis and amoebic liver abscesses. Three classes of amoebic molecules have been identified as the major virulence factors, the Gal/GalNAc inhibitable lectin that mediates adherence to mammalian cells, the amoebapores which cause the formation of membrane ion channels in the target cells and the cysteine proteinases which degrade the matrix proteins, the intestinal mucus and secretory IgA. Transcriptional silencing of the amoebapore (Ehap-a gene occurred after transfection of trophozoites with a plasmid containing a segment of the 5’ upstream region of the gene. Transcriptional silencing of the Ehap-a gene continued even after the removal of the plasmid and the cloned amoebae were termed G3. Transfection of G3 trophozoites with a plasmid construct containing the cysteine proteinase (EhCP-5 gene and the light subunit of the Gal- lectin (Ehlgl1 gene, each under the 5’ upstream sequences of the amoebapore gene, caused the simultaneous epigenetic silencing of expression of these two genes. The resulting trophozoites, termed RB-9, were cured from the plasmid and they do not express the three types of virulent genes. The RB-9 amoeba are virulence attenuated and are incapable of killing mammalian cells, they can not induce the formation of liver abscesses and they do not cause ulcerations in the cecum of experimental animals. The gene-silenced amoebae express the same surface antigens which are present in virulent strains and following intra peritoneal inoculation of live trophozoites into hamsters they evoked a protective immune response. Further studies are needed to find out if RB-9 trophozoites could be used for vaccination against amoebaisis.

  4. Genomic analyses confirm close relatedness between Rhodococcus defluvii and Rhodococcus equi (Rhodococcus hoagii)

    OpenAIRE

    Sangal, Vartul; Jones, Amanda; GOODFELLOW, Michael; Hoskisson, Paul; Kämpfer, Peter; Sutcliffe, Iain

    2015-01-01

    Rhodococcus defluvii strain Ca11T was isolated from a bioreactor involved in extensive phosphorus removal. We have sequenced the whole genome of this strain and our comparative genomic and phylogenetic analyses confirm its close relatedness with Rhodococcus equi (Rhodococcus hoagii) strains, which share >80% of the gene content. The R. equi virulence plasmid is absent though most of the chromosomal R. equi virulence-associated genes are present in R. defluvii Ca11T. These data suggest that al...

  5. Mouse lung infection model to assess Rhodococcus equi virulence and vaccine protection.

    Science.gov (United States)

    González-Iglesias, Patricia; Scortti, Mariela; MacArthur, Iain; Hapeshi, Alexia; Rodriguez, Héctor; Prescott, John F; Vazquez-Boland, José A

    2014-08-06

    The pathogenic actinomycete Rhodococcus equi causes severe purulent lung infections in foals and immunocompromised people. Although relatively unsusceptible to R. equi, mice are widely used for in vivo studies with this pathogen. The most commonly employed mouse model is based on systemic (intravenous) infection and determination of R. equi burdens in spleen and liver. Here, we investigated the murine lung for experimental infection studies with R. equi. Using a 10(7)CFU intranasal challenge in BALB/c mice, virulent R. equi consistently survived in quantifiable numbers up to 10 days in the lungs whereas virulence-deficient R. equi bacteria were rapidly cleared. An internally controlled virulence assay was developed in which the test R. equi strains are co-inoculated and monitored in the same mouse. Isogenic R. equi bacteria lacking either the plasmid vapA gene or the entire virulence plasmid were compared using this competitive assay. Both strains showed no significant differences in in vivo fitness in the lung, indicating that the single loss of the virulence factor VapA was sufficient to account for the full attenuation seen in the absence of the virulence plasmid. To test the adequacy of the lung infection model for monitoring R. equi vaccine efficacy, BALB/c mice were immunized with live R. equi and challenged intranasally. Vaccination conferred protection against acute pulmonary challenge with virulent R. equi. Our data indicate that the murine lung infection model provides a useful tool for both R. equi virulence and vaccine studies. Copyright © 2014 Elsevier B.V. All rights reserved.

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

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    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.

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

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    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.

  8. Rhodococcus equi Sepsis in a Renal Transplant Recipient: A Case Study

    Science.gov (United States)

    Macken, Eline; de Jonge, Hylke; Van Caesbroeck, Daniël; Verhaegen, Jan; Van Kerkhoven, Dana; Van Wijngaerden, Eric; Kuypers, Dirk

    2015-01-01

    Abstract Rhodococcus equi is an unusual cause of infection in humans, but has emerged as an opportunistic pathogen among immunocompromised patients. Primary pulmonary involvement is the most common clinical presentation, although the spectrum of disease is broad. Diagnosing R. equi infections remains challenging, both from clinical and microbiological view, and no standard treatment has been established. In this report, we present a detailed case of a 57-year-old male renal transplant recipient who developed R. equi bacteremia with a concomitant Pneumocystis jirovecii pneumonia. We describe the clinical features of R. equi infections, highlight the importance of an early diagnosis, and briefly review treatment options for this rare infection. PMID:27500216

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

    Science.gov (United States)

    Bordin, Angela I; Pillai, Suresh D; Brake, Courtney; Bagley, Kaytee B; Bourquin, Jessica R; Coleman, Michelle; Oliveira, Fabiano N; Mwangi, Waithaka; McMurray, David N; Love, Charles C; Felippe, Maria Julia B; Cohen, Noah D

    2014-01-01

    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.

  10. Rhodococcus equi infection after reduction mammaplasty in an immunocompetent patient

    Directory of Open Access Journals (Sweden)

    Uriel Sandkovsky

    2011-10-01

    Full Text Available The majority of infections caused by R. equi occur in hosts with some degree of cell-mediated immunodeficiency. Immunocompetent individuals are infrequently affected and usually present with localized disease. Infections of the skin or related structures are uncommon and are usually related to environmental contamination. The microbiology laboratory plays a key role in the identification of the organism since it may be mistaken for common skin flora. We describe a 31 year-old woman without medical problems who presented nine weeks after breast reduction with right breast cellulitis and purulent drainage from the surgical wound. She underwent incision and drainage, and cultures of the wound yielded Rhodococcus equi. The patient completed six weeks of antimicrobial therapy with moxifloxacin and rifampin with complete resolution.

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

    OpenAIRE

    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 D...

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

    OpenAIRE

    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 ca...

  13. Characterization of the arginine deiminase of Streptococcus equi subsp. zooepidemicus.

    Science.gov (United States)

    Hong, Kyongsu

    2006-09-01

    Streptococcus equi subsp. zooepidemicus is an important cause of infectious diseases in horses and rarely humans. Little is known about the virulence factors or protective antigens of S. equi subsp. zooepidemicus. In the present study, I designed original primers based on an alignment of the gene sagp(arcA) from Streptococcus pyogenes encoding streptococcal acid glycoprotein-arginine deiminase (SAGP/AD) to amplify the S. equi subsp. zooepidemicus counterpart sequence by polymerase chain reaction, and I analyzed the sagp(arcA) gene of the organism. Using chromosomal walking steps, I identified a contiguous eight-gene locus involved in SAGP/AD production. Their open reading frames were found to share significant homologies and to correspond closely in molecular mass to previously sequenced arc genes of S. pyogenes, thus they were designated ahrC.2 (arginine repressor), arcR (CRP/FNR transcription regulator), sagp(arcA) (streptococcal acid glycoprotein-arginine deiminase), putative acetyltransferase gene, arcB (ornithine carbamyl transferase), arcD (arginine-ornithine antiporter), arcT (Xaa-His peptidase), and arcC (carbamate kinase). The SAGP homologue of S. equi subsp. zooepidemicus (SzSAGP), encoded by arcA gene of the bacteria (arcA(SZ)), was successfully expressed in Escherichia coli and purified to homogeneity. When in vitro growth inhibitory activity of the recombinant SzSAGP was tested against MOLT-3 cells, it inhibited the growth of the cells during the 3 days of culture in a dose-dependent manner, accompanied by the induction of apoptotic cell death. The recombinant protein also possessed AD activity. By immunoblot analysis using both anti-SzSAGP-SfbI(H8) and anti-SfbI(H8) sera, I was able to demonstrate that the SzSAGP protein is expressed on the streptococcal surface.

  14. In vivo programmed cell death of Entamoeba histolytica trophozoites in a hamster model of amoebic liver abscess.

    Science.gov (United States)

    Villalba-Magdaleno, José D'Artagnan; Pérez-Ishiwara, Guillermo; Serrano-Luna, Jesús; Tsutsumi, Víctor; Shibayama, Mineko

    2011-05-01

    Entamoeba histolytica trophozoites can induce host cell apoptosis, which correlates with the virulence of the parasite. This phenomenon has been seen during the resolution of an inflammatory response and the survival of the parasites. Other studies have shown that E. histolytica trophozoites undergo programmed cell death (PCD) in vitro, but how this process occurs within the mammalian host cell remains unclear. Here, we studied the PCD of E. histolytica trophozoites as part of an in vivo event related to the inflammatory reaction and the host-parasite interaction. Morphological study of amoebic liver abscesses showed only a few E. histolytica trophozoites with peroxidase-positive nuclei identified by terminal deoxynucleotidyltransferase enzyme-mediated dUTP nick end labelling (TUNEL). To better understand PCD following the interaction between amoebae and inflammatory cells, we designed a novel in vivo model using a dialysis bag containing E. histolytica trophozoites, which was surgically placed inside the peritoneal cavity of a hamster and left to interact with the host's exudate components. Amoebae collected from bags were then examined by TUNEL assay, fluorescence-activated cell sorting (FACS) and transmission electron microscopy. Nuclear condensation and DNA fragmentation of E. histolytica trophozoites were observed after exposure to peritoneal exudates, which were mainly composed of neutrophils and macrophages. Our results suggest that production of nitric oxide by inflammatory cells could be involved in PCD of trophozoites. In this modified in vivo system, PCD appears to play a prominent role in the host-parasite interaction and parasite cell death.

  15. 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.

  16. A Study on Babesia Infection in Sheep of Pars Abad Moghan Region, Ardabil Province (2014

    Directory of Open Access Journals (Sweden)

    SHahram Dastouri Dastgir

    2016-07-01

    Full Text Available Background & objectives: Pars Abad Moghan city, due to the mild weather and numerous Pastures for animals feeding, is a suitable place for breeding sheep and goat. Babesiosis is one of the most important parasitic infection in animals, especially ruminants. Babesia is a protozoa living inside the red blood cells and transmitted to the host by hard Tick especially Rhipicephalus, Hyalomma, Dermacentor and Haemaphysalis. The aim of this study was to consider Babesia infection in sheep of Pars Abad Moghan city. Methods: In order to investigate the prevalence of babesia, 105 sheep in two different seasons of spring and summer were prepared and blood sample was taken and thin and thick smears were done. Thin blood smear was fixed with methanol and in order to determine the presence of the protozoan parasite, Giemsa with ratio of 1 to 14 in distilled water was used for staining. Results: The results showed that the number of 33 samples (31.42% were infected with Babesia. Among these, 18 samples (17.14% were infected with Babesia ovis, 9 (8.57% with babesia motasi and 6 samples (5.71% were infected simultaneously with both species of Babesia ovis and Babesia motasi. Conclusion: The rate of infection to Babesia among the sheep in Pars Abad Moghan city is remarkable. Early diagnosis and early treatment can prevent spread of the disease in this area.

  17. Assessment of human natural killer and lymphokine-activated killer cell cytotoxicity against Toxoplasma gondii trophozoites and brain cysts

    Energy Technology Data Exchange (ETDEWEB)

    Dannemann, B.R.; Morris, V.A.; Araujo, F.G.; Remington, J.S. (Palo Alto Medical Foundation, CA (USA))

    1989-10-15

    Because previous work has suggested that NK cells may be important in host resistance against the intracellular parasite Toxoplasma gondii we examined whether human NK cells and lymphokine-activated killer (LAK) cells have activity against trophozoites and cysts of this organism in vitro. A method to radiolabel Toxoplasma trophozoites with 51Cr was developed and direct cytotoxic activity was determined by using modifications of the standard 51Cr release assay. Viability of 51Cr-labeled trophozoites assessed by both methylene blue staining and trypan blue exclusion was greater than 90%. Significantly more 51Cr was released by anti-Toxoplasma antibody and C than by antibody in the absence of C. Incubation of trophozoites with freshly isolated human NK cells or NK cells activated with either rIL-2 or rIFN-alpha did not result in significant release of 51Cr (specific lysis was 0 to 2.3%). In contrast, the average specific lysis of radiolabeled trophozoites by LAK cells was significant. In a series of separate experiments, preincubation of radiolabeled trophozoites with heat-inactivated normal or Toxoplasma antibody-positive human serum increased the cytotoxicity of LAK cells from a mean specific lysis of 15% +/- 4.5 to 39% +/- 8.5, respectively, as assessed by 51Cr release. Because previous work has shown that radioisotope release from parasites may be nonspecific, separate experiments were performed to determine the cytotoxicity of LAK cells against antibody-coated trophozoites by using ethidium bromide-acridine orange staining to assess effector cell damage. LAK cells had a mean specific lysis of 51% against antibody-coated trophozoites by ethidium bromide-acridine orange staining. Preincubation with heat-inactivated Toxoplasma-antibody positive human serum did not increase activity of rIL-2-activated NK cells against 51CR-labeled trophozoites.

  18. Protective effects of passively transferred merozoite-specific antibodies against Theileria equi in horses with severe combined immunodeficiency

    Science.gov (United States)

    Theileria equi immune plasma was infused into young horses (foals) with severe combined immunodeficiency. Although all foals became infected following intravenous challenge with homologous T. equi merozoite stabilate, delayed time-to-peak parasitemia and enhanced survival occurred. Protective effect...

  19. The Steroid Catabolic Pathway of the Intracellular Pathogen Rhodococcus equi Is Important for Pathogenesis and a Target for Vaccine Development

    NARCIS (Netherlands)

    van der Geize, R.; Grommen, A. W. F.; Hessels, G. I.; Jacobs, A. A. C.; Dijkhuizen, L.

    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

  20. 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

  1. 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.

  2. Giardia duodenalis: effects of an ozonized sunflower oil product (Oleozon) on in vitro trophozoites.

    Science.gov (United States)

    Hernández, Frank; Hernández, Damian; Zamora, Zullyt; Díaz, Maritza; Ancheta, Odelsa; Rodriguez, Sandra; Torres, Dinorah

    2009-03-01

    The ozonized sunflower oil product (Oleozon) was investigated to explore its cytotoxic activity on Giardia duodenalis in vitro cultivated trophozites. Oleozon produced inactivation of Giardia trophozoites in a dose- and cell density-dependent manner. Thirty microliter of Oleozon with peroxide index value of 500 equivalent-mmol of activated oxygen per kilogram were used to achieve a 100% inhibition (density-dependent effect must be taken into account when prescription of this product for giardiasis treatment in humans.

  3. 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. ...

  4. Autochthonous canine babesiosis caused by Babesia canis canis in Latvia.

    Science.gov (United States)

    Berzina, Inese; Capligina, Valentina; Baumanis, Viesturs; Ranka, Renate; Cirule, Dina; Matise, Ilze

    2013-09-23

    This is the first report of confirmed canine babesiosis in Latvia supporting the observed geographical expansion of this disease. Between 2009 and 2011 three dogs which have not traveled outside of Latvia were diagnosed with babesiosis. Hematological analysis and serological tests for granulocytic anaplasmosis, ehrlichiosis and borreliosis were negative (Idexx SNAP 4Dx test). Peripheral blood erythrocytes of the three dogs contained large Babesia that were identified as Babesia canis canis by PCR. Sequences of partial 18S rRNA gene were 98-100% similar to the sequences of B. canis canis isolated from dogs in other European countries. We conclude that these are the first autochthonous canine babesiosis cases reported from Latvia. Copyright © 2013 Elsevier B.V. All rights reserved.

  5. Transmission of Babesia microti Parasites by Solid Organ Transplantation.

    Science.gov (United States)

    Brennan, Meghan B; Herwaldt, Barbara L; Kazmierczak, James J; Weiss, John W; Klein, Christina L; Leith, Catherine P; He, Rong; Oberley, Matthew J; Tonnetti, Laura; Wilkins, Patricia P; Gauthier, Gregory M

    2016-11-01

    Babesia microti, an intraerythrocytic parasite, is tickborne in nature. In contrast to transmission by blood transfusion, which has been well documented, transmission associated with solid organ transplantation has not been reported. We describe parasitologically confirmed cases of babesiosis diagnosed ≈8 weeks posttransplantation in 2 recipients of renal allografts from an organ donor who was multiply transfused on the day he died from traumatic injuries. The organ donor and recipients had no identified risk factors for tickborne infection. Antibodies against B. microti parasites were not detected by serologic testing of archived pretransplant specimens. However, 1 of the organ donor's blood donors was seropositive when tested postdonation and had risk factors for tick exposure. The organ donor probably served as a conduit of Babesia parasites from the seropositive blood donor to both kidney recipients. Babesiosis should be included in the differential diagnosis of unexplained fever and hemolytic anemia after blood transfusion or organ transplantation.

  6. Efficacy of imidocarb dipropionate in the elimination of Theileria equi in experimentally infected horses

    Science.gov (United States)

    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 United States. In an effort to exclude T. equi, the U.S. practices stringent serological screening of horses prior to entry. Current regulato...

  7. Neutrophils extracellular traps damage Naegleria fowleri trophozoites opsonized with human IgG.

    Science.gov (United States)

    Contis-Montes de Oca, A; Carrasco-Yépez, M; Campos-Rodríguez, R; Pacheco-Yépez, J; Bonilla-Lemus, P; Pérez-López, J; Rojas-Hernández, S

    2016-08-01

    Naegleria fowleri infects humans through the nasal mucosa causing a disease in the central nervous system known as primary amoebic meningoencephalitis (PAM). Polymorphonuclear cells (PMNs) play a critical role in the early phase of N. fowleri infection. Recently, a new biological defence mechanism called neutrophil extracellular traps (NETs) has been attracting attention. NETs are composed of nuclear DNA combined with histones and antibacterial proteins, and these structures are released from the cell to direct its antimicrobial attack. In this work, we evaluate the capacity of N. fowleri to induce the liberation of NETs by human PMN cells. Neutrophils were cocultured with unopsonized or IgG-opsonized N. fowleri trophozoites. DNA, histone, myeloperoxidase (MPO) and neutrophil elastase (NE) were stained, and the formation of NETs was evaluated by confocal microscopy and by quantifying the levels of extracellular DNA. Our results showed N. fowleri induce the liberation of NETs including release of MPO and NE by human PMN cells as exposure interaction time is increased, but N. fowleri trophozoites evaded killing. However, when trophozoites were opsonized, they were susceptible to the neutrophils activity. Therefore, our study suggests that antibody-mediated PMNs activation through NET formation may be crucial for antimicrobial responses against N. fowleri.

  8. In vitro effect of Acanthospermum australe (Asteraceae extracts on Acanthamoeba polyphaga trophozoites

    Directory of Open Access Journals (Sweden)

    LC Castro

    2013-01-01

    Full Text Available Acanthamoeba is a free-living protozoan widely distributed in the environment, occurring in vegetative trophozoite and resistance cyst stages during its life cycle. It constitutes an etiological agent of Acanthamoeba keratitis, a disease that may cause severe ocular inflammation and blindness. New drugs can be developed from molecules found in plants and thus help in its difficult treatment. Acanthospermum australe (Asteraceae, a plant used in folk medicine, had its effect tested on Acanthamoeba polyphaga. Aqueous and ethanolic extracts of A. austral were obtained from aerial parts for infusion and static maceration, respectively. Concentrations of 10, 5, 2.5, 1.25 and 0.625 mg/ml of the extract were tested against Acanthamoeba polyphaga trophozoites. The cytotoxic effect of the extracts was tested in mammalian cells using the 3-(4, 5-dimethylthiazol-2-yl-2, 5-diphenyltetrazolium bromide (MTT assay. RESULTS: The 10 mg/ml concentration of ethanolic extract was lethal to 100% of the A. polyphaga trophozoites in 24 h and both extracts presented cytotoxic effect against mammalian cells. These findings suggest that the A. austral ethanolic extract may have compounds with relevance to the development of new amoebicidal drugs.

  9. Microarray analysis of differentially expressed genes between cysts and trophozoites of Acanthamoeba castellanii.

    Science.gov (United States)

    Moon, Eun-Kyung; Xuan, Ying-Hua; Chung, Dong-Il; Hong, Yeonchul; Kong, Hyun-Hee

    2011-12-01

    Acanthamoeba infection is difficult to treat because of the resistance property of Acanthamoeba cyst against the host immune system, diverse antibiotics, and therapeutic agents. To identify encystation mediating factors of Acanthamoeba, we compared the transcription profile between cysts and trophozoites using microarray analysis. The DNA chip was composed of 12,544 genes based on expressed sequence tag (EST) from an Acanthamoeba ESTs database (DB) constructed in our laboratory, genetic information of Acanthamoeba from TBest DB, and all of Acanthamoeba related genes registered in the NCBI. Microarray analysis indicated that 701 genes showed higher expression than 2 folds in cysts than in trophozoites, and 859 genes were less expressed in cysts than in trophozoites. The results of real-time PCR analysis of randomly selected 9 genes of which expression was increased during cyst formation were coincided well with the microarray results. Eukaryotic orthologous groups (KOG) analysis showed an increment in T article (signal transduction mechanisms) and O article (posttranslational modification, protein turnover, and chaperones) whereas significant decrement of C article (energy production and conversion) during cyst formation. Especially, cystein proteinases showed high expression changes (282 folds) with significant increases in real-time PCR, suggesting a pivotal role of this proteinase in the cyst formation of Acanthamoeba. The present study provides important clues for the identification and characterization of encystation mediating factors of Acanthamoeba.

  10. 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.

  11. Elongation factor 1-alpha is released into the culture medium during growth of Giardia intestinalis trophozoites.

    Science.gov (United States)

    Skarin, Hanna; Ringqvist, Emma; Hellman, Ulf; Svärd, Staffan G

    2011-04-01

    The molecular pathogenesis of the intestinal parasite Giardia intestinalis is still not fully understood but excretory-secretory products have been suggested to be important during host-parasite interactions. Here we used SDS-PAGE gels and MALDI-TOF analysis to identify proteins released by Giardia trophozoites during in vitro growth. Serum proteins (mainly bovine serum albumin) in the growth medium, bind to the parasite surface and they are continuously released, which interfere with parasite secretome characterization. However, we identified two released Giardia proteins: elongation factor-1 alpha (EF-1α) and a 58 kDa protein, identified as arginine deiminase (ADI). This is the first description of EF-1α as a released/secreted Giardia protein, whereas ADI has been identified in an earlier secretome study. Two genes encoding EF-1α were detected in the Giardia WB genome 35 kbp apart with almost identical coding sequences but with different promoter and 3' regions. Promoter luciferase-fusions showed that both genes are transcribed in trophozoites. The EF-1α protein localizes to the nuclear region in trophozoites but it relocalizes to the cytoplasm during host-cell interaction. Recombinant EF-1α is recognized by serum from giardiasis patients. Our results suggest that released EF-1α protein can be important during Giardia infections.

  12. Entamoeba histolytica: inflammatory process during amoebic liver abscess formation involves cyclooxygenase-2 expression in macrophages and trophozoites.

    Science.gov (United States)

    Gutiérrez-Alarcón, A; Moguel-Torres, M; Mata-Leyva, O; Cuellar-Nevárez, G; Siqueiros-Cendón, T; Erosa, G; Ramos-Martínez, E; Talamás-Rohana, P; Sánchez-Ramírez, B

    2006-11-01

    It has been demonstrated that expression of cyclooxygenase-2 (COX-2) isoform is induced by Entamoeba histolytica in macrophages and polymorphonuclear cells during amoebic liver abscess (ALA) formation in hamsters. Trophozoites present in the lesion were also positive for COX-2 signal. However, no cross reactivity of the anti-COX-2 antibody with protein extract of cultivated trophozoites was found. To clarify if trophozoites are involved in PGE(2) production during ALA development, COX-2 expression was detected by in situ hybridization and RT-PCR in liver tissue from intrahepatically infected hamsters. COX-2 mRNA was in polymorphonuclear cells since 4h postinfection, and subsequently, local macrophages expressed COX-2 mRNA in a similar way. Additionally, a positive signal for COX-2 mRNA expression was detected in E. histolytica trophozoites, suggesting that, in vivo, parasite COX expression may be an important mechanism to promote inflammation.

  13. Prevalence of Theileria and Babesia species in Tunisian sheep

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    Mohamed R. Rjeibi

    2016-03-01

    Full Text Available In this study, the prevalence of Theileria and Babesia species in sheep was assessed with Giemsastained blood smear examination and polymerase chain reaction to identify the different piroplasms in 270 sheep from three Tunisian bioclimatic zones (north, centre, and south. The overall infection prevalence by Babesia spp. and Theileria spp. in Giemsa-stained blood smears was 2.9% (8/270 and 4.8% (13/270 respectively. The molecular results showed that sheep were more often infected by Theileria ovis than Babesia ovis with an overall prevalence of 16.3% (44/270 and 7.8% (21/270 respectively (p = 0.01. The molecular prevalence by Babesia ovis was significantly higher in females than in males (p < 0.05. According to localities B. ovis was found exclusively in sheep from the centre of Tunisia (Kairouan whereas Theileria ovis was found in all regions. Infections with T. ovis and B. ovis were confirmed by sequencing. The sequence of T. ovis in this study (accession numbers KM924442 falls into the same clade as T. ovis deposited in GenBank. The T. ovis amplicons (KM924442 showed 99%–100% identities with GenBank sequences. Moreover, comparison of the partial sequences of 18S rRNA gene of B. ovis described in this study (KP670199 revealed 99.4% similarity with B. ovis recently reported in northern Tunisia from sheep and goats. Three nucleotides were different at positions 73 (A/T, 417 (A/T, and 420 (G/T. It also had 99% identity with B. ovis from Spain, Turkey and Iraq. The results suggest a high T. ovis prevalence in Tunisia with a decreasing north-south gradient. This could be correlated to the vector tick distribution.

  14. Seroprevalence of Babesia infections of dairy cows in northern Thailand.

    Science.gov (United States)

    Iseki, Hiroshi; Zhou, Lijia; Kim, Chulmin; Inpankaew, Tawin; Sununta, Chainirun; Yokoyama, Naoaki; Xuan, Xuenan; Jittapalapong, Sathaporn; Igarashi, Ikuo

    2010-06-24

    The present study was conducted to demonstrate the epidemiological distribution of bovine babesiosis in the northern regions of Thailand. A total of 700 serum samples of dairy cows in the northern provinces (Chiang Rai, Chiang Mai, Lumpang, and Mae Hong Sorn) were tested for antibodies against Babesia bovis and B. bigemina. Species-specific enzyme-linked (rRAP-1/CTs) were performed. According to the results, 517 (73.8%) and 484 (69.1%) were positive for B. bovis and B. bigemina, respectively. In addition, 370 (52.9%) were positive for mixed infections by both ELISAs. On the other hand, all samples were also examined by the indirect fluorescent-antibody test (IFAT) with B. bovis- and B. bigemina-infected blood smears. According to the IFAT, 482 (68.8%) and 531 (75.8%) were positive for these infections, respectively. The overall concordances between the ELISA and IFAT techniques were 93.6% and 90.7% for B. bovis and B. bigemina infections, respectively. These results indicated that babesia infections are widespread in the northern parts of Thailand. To our knowledge, this is the first report describing the epidemiology of Babesia infections using rRAP-1/CT-based ELISAs in these areas.

  15. Molecular detection of Babesia spp. in ticks in northern Serbia

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    Mihaljica D.

    2012-01-01

    Full Text Available In order to evaluate the prevalence rate of Babesia spp. in ticks collected from vegetation at seven localities in northern Serbia, tick samples were subjected to molecular analysis. A total of 132 unfed adult ticks of five different species (Dermacentor marginatus, Dermacentor reticulatus, Ixodes ricinus, Haemaphysalis concinna and Haemaphysalis punctata, were examined by PCR for the presence of Babesia spp. Out of the analyzed ticks, 10.61% (14/132 were positive for babesial DNA. The presence of babesiae was found at the localities Pančevački Rit, Titov Gaj, Makiš, PKB and Kljajićevo. Prevalence in D. reticulatus ticks was 21.57% (11/51 and in H. concinna ticks, 8.57% (3/35. Sequencing and phylogenetic analysis showed a clustering of the obtained sequences with those of B. canis from the GenBank database. These results add to the knowledge of the distribution of babesial pathogens and their vectors in Serbia. [Projekat Ministarstva nauke Republike Srbije, br. ON 173006

  16. Molecular surveillance of Theileria equi and Anaplasma phagocytophilum infections in horses from Ukraine, Poland and Slovakia.

    Science.gov (United States)

    Slivinska, Kateryna; Víchová, Bronislava; Werszko, Joanna; Szewczyk, Tomasz; Wróblewski, Zbigniew; Peťko, Branislav; Ragač, Ondrej; Demeshkant, Vitaliy; Karbowiak, Grzegorz

    2016-01-15

    A survey was undertaken to assess the prevalence of Theileria equi and Anaplasma phagocytophilum in some regions of Ukraine, Poland and Slovakia. Using a specific PCR assays, blood samples from 215 horses were tested. The prevalence of T. equi and A. phagocytophilum infection was 13.95% and 1.4%, respectively. BLAST analysis showed the isolates closest to the T. equi 18S rRNA and A. phagocytophilum msp4 gene sequences in GenBank with a similarity of ≥99%. No significant association was found between the T. equi PCR positivity and the age or sex of the horses. There was a significant association between the origin of horses and T. equi-PCR positivity. No significant association was found between the A. phagocytophilum-PCR positivity and the age, sex or origin.

  17. Rhodococcus equi pericarditis in a patient living with HIV/AIDS.

    Science.gov (United States)

    Gundelly, Praveen; Thornton, Alice; Greenberg, Richard N; McCormick, Malkanthie; Myint, Thein

    2014-01-01

    Rhodococcus equi, previously called Corynebacterium equi, is known to cause pneumonia in foals and swine. Although it was known to cause infection rarely in humans, R equi infection in humans has increased with the advent of HIV and increased use of immunosuppressants. We report a case of a 48-year-old male patient with newly diagnosed HIV/AIDS presenting with recurrent R equi bacteremia, pericardial effusion, and pericardial cyst. The infection was treated with drainage of the pericardial effusion and cyst and 2 weeks of intravenous vancomycin and 6 months of oral azithromycin and levofloxacin. Rhodococcus equi causes pericarditis and pericardial effusion. It can be effectively treated with debridement, drainage, and a prolonged course of antibiotics. In vitro antibiotic susceptibility should be checked as resistance to antibiotics can develop, especially if drainage is inadequate.

  18. Rhodococcus equi pneumonia in the foal--part 1: pathogenesis and epidemiology.

    Science.gov (United States)

    Muscatello, Gary

    2012-04-01

    Rhodococcus equi pneumonia is a worldwide infectious disease of major concern to the equine breeding industry. The disease typically manifests in foals as pyogranulomatous bronchopneumonia, resulting in significant morbidity and mortality. Inhalation of aerosolised virulent R. equi from the environment and intracellular replication within alveolar macrophages are essential components of the pathogenesis of R. equi pneumonia in the foal. Recently documented evidence of airborne transmission between foals indicates the potential for an alternative contagious route of disease transmission. In the first of this two-part review, the complexity of the host, pathogen and environmental interactions that underpin R. equi pneumonia will be discussed through an exploration of current understanding of the epidemiology and pathogenesis of R. equi pneumonia in the foal. Copyright © 2011 Elsevier Ltd. All rights reserved.

  19. Characterisation and expression analysis of trophozoite and cyst proteins of Acanthamoeba spp. isolated from Acanthamoeba keratitis (AK) patient.

    Science.gov (United States)

    Behera, Himansu Sekhar; Satpathy, Gita

    2016-01-01

    The study was carried out to characterise and analyze the expression pattern of proteins of infective trophozoite and cyst forms of Acanthamoeba spp. isolated from an amoebic keratitis patient. Protein was isolated from the trophozoites and cysts of Acanthamoeba spp. isolates and subjected to SDS PAGE, 2D PAGE analysis where a large number of protein bands and protein spots were observed. Four prominent protein spots i.e. 2 from trophozoites and 2 from cysts that appeared more intense compared to the corresponding spots in other corresponding gel were excised from the 2D PAGE gels and analysed by MALDI-TOF/TOF MS assay and Mascot search software. Protein spots from trophozoites were identified as "hypothetical protein ACA1" and "eukaryotic porin protein" and those from cysts were identified as "chaperone protein DnaK" and "chaperonin protein" respectively. Proteomic results of 4 proteins were further validated by reverse genomics using quantitative real time PCR assay which showed a 1388 fold and 4.35 fold increase in expression of "hypothetical protein ACA1" gene and "eukaryotic porin protein" gene respectively in trophozoites compared to cysts and a 15 fold and 12.36 fold increase in expression of "chaperone protein DnaK" gene and "chaperonin protein" gene respectively in cysts compared to trophozoites. "Hypothetical protein ACA1" of trophozoites, whose function is unknown might have some important role in the parasite division and pathogenicty of Acanthamoeba spp. which needs further study. As trophozoites are the active and feeding form of Acanthamoeba spp., "eukaryotic porin" proteins may have some important role in efflux of toxic metabolites and exudates from interior of cell to outside along with some role in pathogenicity. Similarly proteins such as "chaperone protein DnaK" and "chaperonin protein" which belongs to group of heat shock proteins may have a role in folding of cyst specific proteins in cyst which needs further study.

  20. Glycoproteins and Gal-GalNAc cause Cryptosporidium to switch from an invasive sporozoite to a replicative trophozoite.

    Science.gov (United States)

    Edwinson, Adam; Widmer, Giovanni; McEvoy, John

    2016-01-01

    The apicomplexan parasite Cryptosporidium causes cryptosporidiosis, a diarrheal disease that can become chronic and life threatening in immunocompromised and malnourished people. There is no effective drug treatment for those most at risk of severe cryptosporidiosis. The disease pathology is due to a repeated cycle of host cell invasion and parasite replication that amplifies parasite numbers and destroys the intestinal epithelium. This study aimed to better understand the Cryptosporidium replication cycle by identifying molecules that trigger the switch from invasive sporozoite to replicative trophozoite. Our approach was to treat sporozoites of Cryptosporidium parvum and Cryptosporidium hominis, the species causing most human cryptosporidiosis, with various media under axenic conditions and examine the parasites for rounding and nuclear division as markers of trophozoite development and replication, respectively. FBS had a concentration-dependent effect on trophozoite development in both species. Trophozoite development in C. parvum, but not C. hominis, was enhanced when RPMI supplemented with 10% FBS (RPMI-FBS) was conditioned by HCT-8 cells for 3h. The effect of non-conditioned and HCT-8 conditioned RPMI-FBS on trophozoite development was abrogated by proteinase K and sodium metaperiodate pretreatment, indicating a glycoprotein trigger. Cryptosporidium parvum and C. hominis trophozoite development also was triggered by Gal-GalNAc in a concentration-dependent manner. Cryptosporidium parvum replication was greatest following treatments with Gal-GalNAc, followed by conditioned RPMI-FBS and non-conditioned RPMI-FBS (PGalNAc (1mM).

  1. The first detection of species of Babesia Starcovici, 1893 in moose, Alces alces (Linnaeus), in Norway.

    Science.gov (United States)

    Puraite, Irma; Rosef, Olav; Radzijevskaja, Jana; Lipatova, Indre; Paulauskas, Algimantas

    2016-04-01

    Babesiosis is an emerging zoonotic disease and various wildlife species are reservoir hosts for zoonotic species of Babesia Starcovici, 1893. The objective of the present study was to investigate the presence and prevalence of Babesia spp. in moose Alces alces (Linnaeus) in two regions of Norway. A total of 99 spleen samples were collected from animals of various ages from an area with the occurrence of the tick Ixodes ricinus (Linnaeus, 1758), and from an area where the ticks are known to be absent. Infection was detected by the amplification of different regions of the 18S rRNA gene by using two different PCR primer sets specific of Babesia. Babesia spp. were found in the spleen samples of four moose. All Babesia-infected animals were from an area where ticks occur, with an infection rate of 6% (4 of 70). Babesia-positive samples were obtained from a five-month old moose calf and three adults. Two Babesia species, Babesia capreoli (Enigk et Friedhoff, 1962) and a B. odocoilei-like, were identified. Co-infection with Anaplasma phagocytophilum was obtained in two animals. This is the first report of the occurrence of B. capreoli and B. odocoilei-like species in moose.

  2. Sequence heterogeneity in the equi merozoite antigen gene (ema-1) of Theileria equi and development of an ema-1-specific TaqMan MGB assay for the detection of T. equi.

    Science.gov (United States)

    Bhoora, Raksha; Quan, Melvyn; Matjila, Paul T; Zweygarth, Erich; Guthrie, Alan J; Collins, Nicola E

    2010-08-27

    Although a quantitative real-time PCR assay (qPCR) assay for the detection of Theileria equi has been developed and evaluated, it is possible that additional, as yet undetected 18S rRNA gene sequence variants may exist. A qPCR assay targeting a different gene, used in conjunction with the T. equi 18S rRNA qPCR assay, could assist in the detection of all T. equi genotypes in field samples. A T. equi ema-1-specific qPCR (Ueti et al., 2003) was tested on 107 South African field samples, 90 of which tested positive for T. equi antibody using the immuno-fluorescent antibody test (IFAT). The qPCR assay performed poorly, as T. equi was detected in only 67 of the 90 IFAT-positive field samples at quantification cycle (C(q)) values ranging from 27 to 39.95. Furthermore, a high C(q) value of 36.18 was obtained from DNA extracted from a South African in vitro-cultured T. equi WL isolate [1.38% parasitized erythrocytes (PE)] when a low C(q) value (indicative of a high T. equi DNA concentration) was expected. Approximately 600 bp of the ema-1 gene from 38 South African samples were sequenced and BLASTN analysis confirmed all sequences to be merozoite surface protein genes, with an identity of 87.1-100% to previously published T. equi ema-1 gene sequences. Alignment of the sequences revealed extensive sequence variations in the target regions of the primers and probes (Ueti et al., 2003), explaining the poor performance of the qPCR assay. Based on these observations, we developed a new TaqMan minor-groove binder (MGB) probe-based qPCR assay, targeting a more conserved region of the ema-1 gene. This assay was shown to be efficient and specific, and the detection limit, defined as the concentration at which 95% of T. equi-positive samples are detected, was determined to be 1.4 x 10(-4)% PE. The two ema-1 assays were compared by testing 41 South African field samples in parallel. The results suggested that the new assay was more sensitive than the original assay, as T. equi was

  3. 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.

  4. 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

  5. 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.

  6. 驽巴贝斯虫二温式 PCR 检测方法的建立及初步应用%Establishment and Application of Two-temperature PCR for Babesia caballi Detection

    Institute of Scientific and Technical Information of China (English)

    张杨; 达伊力特; 刘进; 李佳; 巴音查汗

    2014-01-01

    为了建立一种能快速、特异、敏感地检测马驽巴贝斯虫的方法,根据 GenBank 中的驽巴贝斯虫(Babesia caballi )Bc-48基因序列设计了一对特异性引物,建立检测驽巴贝斯虫的二温式 PCR 方法。该方法能够特异地扩增155 bp 的片段,与双芽巴贝斯虫、环形泰勒虫、马泰勒虫、瑟氏泰勒虫均无交叉反应,能检测到的最低 DNA 拷贝数为5.17×102 copies/μL。应用所建立的方法检测了采集于和静县焉耆马匹样品,结果显示,马驽巴贝斯虫感染率为69.51%(57/82),与血液涂片检查结果37.8%(31/82)相比,其检出率更高。表明该二温式 PCR 检测方法具有较高的特异性和敏感性,可用于马驽巴贝斯虫病的早期诊断。%To establish a rapid,specific and sensitive two-temperature PCR assay for detection of Babesia caballi ,specific primers were designed according to Bc-48 gene of Babesia caballi published in GenBank.A gene fragment of 155 bp was amplified by this method,and there was no cross reaction with B.bigemina, T.annulata,T.equi,T.sergenti,and the lowest detection limit of Babesia caballi DNA was 5.17 × 10 2 copies/μL.Eighty-two blood samples collected from horses in Hejing were detected by the established method.The results showed that the positive rate was 69.51%(57/82),while that of blood smear test was 37.8%(31/82).This study indicated that established two-temperature PCR method could be used for early diagnosis of B.caballi infection with its high specificity and sensitivity.

  7. Epizootiology of Babesia bovis and Babesia bigemina in free-ranging white-tailed deer in northeastern Mexico.

    Science.gov (United States)

    Cantu-C, Antonio; Ortega-S, J Alfonso; García-Vázquez, Zeferino; Mosqueda, Juan; Henke, Scott E; George, John E

    2009-06-01

    Species of Rhipicephalus (Boophilus) ticks are the vectors of babesiosis (cattle fever tick), which are distributed worldwide. White-tailed deer (Odocoileus virginianus) are important secondary hosts for the cattle fever ticks, Rhipicephalus (B.) annulatus and Rhipicephalus (B.) microplus. White-tailed deer are capable of sustaining Boophilus spp. tick populations in the presence or absence of cattle. The objectives of this study were to determine the frequency of Babesia bovis and Babesia bigemina and the prevalence of antibodies to them and identify possible risk factors for bovine babesiosis in white-tailed deer in 3 northeastern states of México. Whole blood and serum samples (n = 457) were collected from white-tailed deer in the states of Coahuila, Nuevo Leon, and Tamaulipas during the spring of 2004. Samples were tested for B. bovis and B. bigemina by nested polymerase chain reaction (n-PCR) (the primers for B. bovis identified the gene Rap-1 and B. bigemina were specific primers) and by an indirect immunofluorescence antibody test (IFAT). A questionnaire was given to each ranch to obtain information about management practices. Logistic regression methods were used to test the association between management factors and the dependent variable of positive n-PCR or IFAT. Nineteen (4.2%) samples were positive to B. bigemina and 6 (1.7%) were positive to B. bovis by n-PCR. Serological testing showed 59.9% (n = 274) of deer sampled were positive to B. bovis and 5.4% (n = 25) were positive to B. bigemina antibodies. The logistic model varied with different dependent variables. With positive n-PCR and B. bigemina as the dependent variable, 3 factors were associated: habitat (presence of brush and exotic grasses; odds ratio (OR), 3.3; 95% confidence interval (CI), 1.3-8.5), grazing system (continuous grazing OR 4.0; CI, 1.3-12.2), and tick treatment frequency (3-4 mo; OR 7.0, CI 1.4-34.3; 5-6 mo; OR, 11.0; CI, 1.9-62.7; > 6 mo; OR, 4.6; CI, 0.9-23.3). These findings

  8. Nested PCR Detection and Phylogenetic Analysis of Babesia bovis and Babesia bigemina in Cattle from Peri-Urban Localities in Gauteng Province, South Africa

    OpenAIRE

    2013-01-01

    ABSTRACT 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....

  9. 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.

  10. Biofilm formation by Rhodococcus equi and putative association with macrolide resistance

    Directory of Open Access Journals (Sweden)

    Letícia T. Gressler

    2015-10-01

    Full Text Available Abstract: Rhodococcus equi is a facultative intracellular pathogen, which cause severe pyogranulomatous pneumonia in foals and tuberculosis-like lesions in humans. Its ability to form biofilm was described in strains isolated from chronic diseases associated to treatment failures in humans. This study aimed to verify the biofilm formation by 113 R. equi isolated from equine samples (clinical and fecal using two different methods (biofilm-culturing with and without additional glucose and epifluorescence microscopy. We also aimed to determine the efficacy of azithromycin, clarithromycin and erythromycin on R. equi in established biofilm. We found 80.5% (26/41 and 63% (58/72 biofilm-positive isolates, in fecal and clinical samples, respectively. The additional glucose increased the biofilm formation by R. equi fecal samples, but not by clinical samples. The antimicrobials tested herein were not able to eradicate R. equi in biofilm even at higher concentrations. This is the first study showing the biofilm formation by R. equi isolated from equine samples. Our findings indicate that R. equi biofilm-producers may be more resistant to the antimicrobials evaluated. Further studies are warranted to test this hypothesis.

  11. Phenotypic mutants of the intracellular actinomycete Rhodococcus equi created by in vivo Himar1 transposon mutagenesis.

    Science.gov (United States)

    Ashour, Joseph; Hondalus, Mary K

    2003-04-01

    Rhodococcus equi is a facultative intracellular opportunistic pathogen of immunocompromised people and a major cause of pneumonia in young horses. An effective live attenuated vaccine would be extremely useful in the prevention of R. equi disease in horses. Toward that end, we have developed an efficient transposon mutagenesis system that makes use of a Himar1 minitransposon delivered by a conditionally replicating plasmid for construction of R. equi mutants. We show that Himar1 transposition in R. equi is random and needs no apparent consensus sequence beyond the required TA dinucleotide. The diversity of the transposon library was demonstrated by the ease with which we were able to screen for auxotrophs and mutants with pigmentation and capsular phenotypes. One of the pigmentation mutants contained an insertion in a gene encoding phytoene desaturase, an enzyme of carotenoid biosynthesis, the pathway necessary for production of the characteristic salmon color of R. equi. We identified an auxotrophic mutant with a transposon insertion in the gene encoding a putative dual-functioning GTP cyclohydrolase II-3,4-dihydroxy-2-butanone-4-phosphate synthase, an enzyme essential for riboflavin biosynthesis. This mutant cannot grow in minimal medium in the absence of riboflavin supplementation. Experimental murine infection studies showed that, in contrast to wild-type R. equi, the riboflavin-requiring mutant is attenuated because it is unable to replicate in vivo. The mutagenesis methodology we have developed will allow the characterization of R. equi virulence mechanisms and the creation of other attenuated strains with vaccine potential.

  12. Chloroquine inhibits Rhodococcus equi replication in murine and foal alveolar macrophages by iron-starvation.

    Science.gov (United States)

    Gressler, Leticia T; Bordin, Angela I; McQueen, Cole M; Cohen, Noah D; de Vargas, Agueda Castagna

    2016-05-30

    Rhodococcus equi preferentially infects macrophages causing pyogranulomatous pneumonia in young foals. Both the vapA and rhbC genes are up-regulated in an iron (Fe)-deprived environment, such as that found within macrophages. Chloroquine (CQ) is a drug widely used against malaria that suppresses the intracellular availability of Fe in eukaryotic cells. The main objective of this study was to evaluate the ability of CQ to inhibit replication of virulent R. equi within murine (J774A.1) and foal alveolar macrophages (AMs) and to verify whether the mechanism of inhibition could be Fe-deprivation-dependent. CQ effect on R. equi extracellular survival and toxicity to J774A.1 were evaluated. R. equi survival within J774A.1 and foal AMs was evaluated under CQ (10 and 20μM), bovine saturated transferrin (bHTF), and bovine unsaturated transferrin (bATF) exposure. To explore the action mechanism of CQ, the superoxide anion production, the lysozyme activity, as well as the relative mRNA expression of vapA and rhbC were examined. CQ at≤20μM had no effect on R. equi extracellular multiplication and J774A.1 viability. Exposure to CQ significantly and markedly reduced survival of R. equi within J774A.1 and foal AMs. Treatment with bHTF did not reverse CQ effect on R. equi. Exposure to CQ did not affected superoxide anion production or lysozyme activity, however vapA and rhbC expression was significantly increased. Our results reinforce the hypothesis that intracellular availability of Fe is required for R. equi survival, and our initial hypothesis that CQ can limit replication of R. equi in J774A.1 and foal AMs, most likely by Fe starvation. Copyright © 2016 Elsevier B.V. All rights reserved.

  13. Arthrobacter equi sp. nov., isolated from veterinary clinical material.

    Science.gov (United States)

    Yassin, A F; Spröer, C; Siering, C; Hupfer, H; Schumann, P

    2011-09-01

    A Gram-positive-staining, catalase-positive, non-spore-forming, rod-shaped bacterium, strain IMMIB L-1606(T), isolated from genital swabs of a horse, was characterized using a polyphasic approach. Comparative 16S rRNA gene sequence analysis showed that the organism was related to members of the genus Arthrobacter, displaying sequence similarities of 93.5-99.1 % with the type strains of recognized species of the genus. Cell-wall analysis revealed peptidoglycan type A3α L-Lys-L-Ser-L-Thr-L-Ala. DNA-DNA hybridization data and biochemical characterization of strain IMMIB L-1606(T) enabled the isolate to be differentiated genotypically and phenotypically from phylogenetically closely related species of the genus Arthrobacter. Therefore, it is concluded that strain IMMIB L-1606(T) represents a novel species of the genus Arthrobacter, for which the name Arthrobacter equi sp. nov. is proposed. The type strain of Arthrobacter equi sp. nov. is IMMIB L-1606(T) ( = DSM 23395(T) = CCUG 59597(T)).

  14. Acute phase proteins in Andalusian horses infected with Theileria equi.

    Science.gov (United States)

    Rodríguez, Rocío; Cerón, José J; Riber, Cristina; Castejón, Francisco; Gómez-Díez, Manuel; Serrano-Rodríguez, Juan M; Muñoz, Ana

    2014-10-01

    Clinical and laboratory findings were determined in 23 Andalusian horses in southern Spain that were positive for Theileria equi by PCR, including 16 mares at pasture (group A1) and seven stabled stallions (group B1). Five healthy mares at pasture (group A2) and five stabled stallions (group B2), all of which were negative for T. equi in Giemsa stained blood smears and by PCR, were used as controls. The most frequent clinical signs were anorexia, anaemia, depression and icterus (group A1), along with loss of performance or failure to train and depression (group B1). Thrombocytopoenia was evident in 5/7 horses in group B1. Lower serum iron concentrations were observed in both diseased groups compared with their respective control groups. There were no significant differences in APP concentrations between diseased and control groups; all affected horses had APP concentrations within reference limits. Serum haptoglobin, serum amyloid A and plasma fibrinogen concentrations were higher than the reference limits in 5/23, 3/23 and 1/23 diseased horses, respectively. It was concluded that horses with theileriosis exhibited only a mild systemic inflammatory response.

  15. Experimental amoebic liver abscess in hamsters caused by trophozoites of a Brazilian strain of Entamoeba dispar.

    Science.gov (United States)

    Guzmán-Silva, Maria Angélica; Santos, Helena Lúcia Carneiro; Peralta, Regina Saramago; Peralta, José Mauro; de Macedo, Heloisa Werneck

    2013-05-01

    It has been claimed that amoebic molecules such as amoebapore, galactose/N-acetyl galactosamine inhibitable lectin, and cysteine proteases are responsible for host tissue destruction and are present in both pathogenic Entamoeba histolytica and non-pathogenic Entamoeba dispar. Some reports have provided evidence that after infection with E. dispar, pathological changes may occur in some humans. The aim of this study was to evaluate E. dispar pathogenicity by comparing it to the pathogenicity of E. histolytica through liver abscesses induced in hamsters. Syrian golden hamsters were challenged by intrahepatic inoculation with the 03C E. dispar strain or with two strains of E. histolytica (HM1:IMSS and EGG) to compare their virulence grades. As control groups, we used bacterial flora and Pavlova's modified medium. Lesions were verified at 1, 3 and 6 days after inoculation. Multiplex Polymerase Chain Reaction was performed to characterize each strain using EdP1/EdP2 and EhP1/EhP2 primers. The EGG and HM1:IMSS E. histolytica strains and 03C E. dispar were able to cause liver lesions. The EGG strain caused extensive hepatic abscesses, and trophozoites were found in the lesions throughout the three periods of study. The HM1:IMSS strain caused smaller abscesses when compared to EGG lesions; however, trophozoites were observed at 1 and 3 days after inoculation. The 03C E. dispar strain caused intermediate abscesses when compared to the others; trophozoites were observed in all periods analyzed. The EGG strain caused progressive evolution of the injury, which differed from the HM1:IMSS and 03C strains. These results strongly suggest that the 03C E. dispar strain is pathogenic in the experimental hamster model. Additional studies are necessary to identify potential factors that regulate the manifestation of virulence of this strain and others. Copyright © 2013 Elsevier Inc. All rights reserved.

  16. Use and endocytosis of iron-containing proteins by Entamoeba histolytica trophozoites.

    Science.gov (United States)

    López-Soto, Fernando; León-Sicairos, Nidia; Reyes-López, Magda; Serrano-Luna, Jesús; Ordaz-Pichardo, Cynthia; Piña-Vázquez, Carolina; Ortiz-Estrada, Guillermo; de la Garza, Mireya

    2009-12-01

    Iron is essential for nearly all organisms; in mammals, it is part of proteins such as haemoglobin, and it is captured by transferrin and lactoferrin. Transferrin is present in serum, and lactoferrin is secreted by the mucosa and by neutrophils at infection sites, as a host iron-withholding response, sequestering iron away from invading microorganisms. Additionally, all cells contain ferritin, which sequesters iron when its intracellular levels are increased, detoxifying and preventing damage. Liver ferritin contains 50% of iron corporal reserves. During evolution, pathogens have evolved diverse strategies to obtain iron from their hosts in order to survive. The protozoan Entamoeba histolytica invades the intestinal mucosa, causing dysentery, and the trophozoites often travel to the liver producing hepatic abscesses; thus, intestine and liver proteins could be important iron supplies for E. histolytica. We found that E. histolytica trophozoites can grow in both ferrous and ferric iron, and that they can use haemoglobin, holo-transferrin, holo-lactoferrin, and ferritin as in vitro iron sources. These proteins supported the amoeba growth throughout consecutive passages, similarly to ferric citrate. By confocal microscopy and immunoblotting, iron-binding proteins were observed specifically bound to the amoeba surface, and they were endocytosed, trafficked through the endosomal/lysosomal route, and degraded by neutral and acidic cysteine-proteases. Transferrin and ferritin were mainly internalized through clathrin-coated vesicles, and holo-lactoferrin was mainly internalized by caveola-like structures. In contrast, apo-lactoferrin bound to membrane lipids and cholesterol, inducing cell death. The results suggest that in vivo trophozoites secrete products that can destroy enterocytes, erythrocytes, and hepatocytes, releasing transferrin, haemoglobin, ferritin, and other iron-containing proteins, which, together with lactoferrin derived from neutrophils and acinar cells

  17. Efficacy of liposomal gentamicin against Rhodococcus equi in a mouse infection model and colocalization with R. equi in equine alveolar macrophages.

    Science.gov (United States)

    Burton, Alexandra J; Giguère, Steeve; Berghaus, Londa J; Hondalus, Mary K; Arnold, Robert D

    2015-04-17

    Rhodococcus equi, a facultative intracellular pathogen and an important cause of pneumonia in foals, is highly susceptible to killing by gentamicin in vitro. However, gentamicin is not effective in vivo, due to its poor cellular penetration. Encapsulation of drugs in liposomes enhances cellular uptake. The objectives of this study were to compare liposomal gentamicin and free gentamicin with respect to their uptake by equine macrophages and intracellular colocalization with R. equi and to compare the efficacies of liposomal gentamicin, free gentamicin and clarithromycin with rifampin for the reduction of R. equi CFU in a mouse model of infection. After ex vivo exposure, a significantly higher mean (±SD) percentage of equine alveolar macrophages contained liposomal gentamicin (91.9±7.6%) as opposed to free gentamicin (16.8±12.5%). Intracellular colocalization of drug and R. equi, as assessed by confocal microscopy, occurred in a significantly higher proportion of cells exposed to liposomal gentamicin (81.2±17.8%) compared to those exposed to free gentamicin (10.4±8.7%). The number of R. equi CFU in the spleen was significantly lower in mice treated with liposomal gentamicin compared to that of mice treated with free gentamicin or to untreated control mice. Treatment with liposomal gentamicin also resulted in a significantly greater reduction in the number of R. equi CFU in the liver compared to treatment with clarithromycin in combination with rifampin. These results support further investigation of liposomal gentamicin as a new treatment for infections caused by R. equi. Copyright © 2015 Elsevier B.V. All rights reserved.

  18. Albendazole and its derivative JVG9 induce encystation on Giardia intestinalis trophozoites.

    Science.gov (United States)

    Pérez-Rangel, Armando; Hernández, José Manuel; Castillo-Romero, Araceli; Yépez-Mulia, Lilián; Castillo, Rafael; Hernández-Luis, Francisco; Nogueda-Torres, Benjamín; Luna-Arias, Juan Pedro; Radilla, Gerardo; León-Avila, Gloria

    2013-09-01

    In the present study, we evaluated the effect of an albendazole (ABZ) derivative JVG9 on cultured Giardia intestinalis. To assess the JVG9 effects, we evaluated the tubulin cytoskeleton by confocal microscopy, and we found that the characteristic staining was modified. The scanning electron microscopy images revealed extremely damaged trophozoites and cyst-like cells. The confocal images revealed that this drug triggered the expression of cyst wall protein 1 and encystation. We also found that at low doses, AL triggered the encystation process too.

  19. Activity of clarithromycin or rifampin alone or in combination against experimental Rhodococcus equi infection in mice.

    Science.gov (United States)

    Burton, Alexandra J; Giguère, Steeve; Berghaus, Londa J; Hondalus, Mary K

    2015-01-01

    Treatment of mice with the combination of clarithromycin with rifampin resulted in a significantly lower number of Rhodococcus equi CFU in the organs of mice than treatment with either drug alone or placebo. There was no significant difference in the number of R. equi CFU between mice treated with clarithromycin monotherapy, rifampin monotherapy, or placebo. The combination of clarithromycin with rifampin conferred a clear advantage over either drug as monotherapy in this model of chronic R. equi infection. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  20. Identification of Rhodococcus equi lipids recognized by host cytotoxic T lymphocytes

    Science.gov (United States)

    Harris, Seth P.; Fujiwara, Nagatoshi; Mealey, Robert H.; Alperin, Debra C.; Naka, Takashi; Goda, Reina

    2010-01-01

    Immune adult horses have CD8+ cytotoxic T lymphocytes (CTLs) that recognize and lyse Rhodococcus equi-infected cells in an equine lymphocyte alloantigen (ELA)-A [classical major histocompatibility complex (MHC) class I]-unrestricted fashion. As protein antigens are MHC class I-restricted, the lack of restriction suggests that the bacterial antigens being recognized by the host are not proteins. The goals of this study were to test the hypothesis that these CTLs recognize unique R. equi cell-wall lipids related to mycobacterial lipids. Initial experiments showed that treatment of soluble R. equi antigen with broadly reactive proteases did not significantly diminish the ability of the antigen to stimulate R. equi-specific CTLs. R. equi-specific CTLs were also shown to lyse target cells (equine macrophages) pulsed with an R. equi lipid extract. Analysis of the R. equi lipid by TLC and MS (MALDI-TOF and ES) indicated that the extracted antigen consisted of three primary fractions: trehalose monomycolate (TMM), trehalose dimycolate (TDM) and cardiolipin (CL). ELA-A-mismatched cells pulsed with purified TMM and CL, but not the TDM fraction, were recognized and lysed by R. equi-specific CTLs. Because of their role in immune clearance and pathogenesis, transcription of the cytokines gamma interferon (IFN-γ) and interleukin-4 (IL-4) was also measured in response to R. equi lipids by using real-time PCR; elevated IFN-γ, but not IL-4, was associated with host clearance of the bacteria. The whole-cell R. equi lipid and all three R. equi lipid fractions resulted in marked increases in IFN-γ transcription, but no increase in IL-4 transcription. Together, these data support the hypothesis that immune recognition of unique lipids in the bacterial cell wall is an important component of the protective immune response to R. equi. The results also identify potential lipid antigens not previously shown to be recognized by CTLs in an important, naturally occurring actinomycete

  1. Genomic analyses confirm close relatedness between Rhodococcus defluvii and Rhodococcus equi (Rhodococcus hoagii).

    Science.gov (United States)

    Sangal, Vartul; Jones, Amanda L; Goodfellow, Michael; Hoskisson, Paul A; Kämpfer, Peter; Sutcliffe, Iain C

    2015-01-01

    Rhodococcus defluvii strain Ca11(T) was isolated from a bioreactor involved in extensive phosphorus removal. We have sequenced the whole genome of this strain, and our comparative genomic and phylogenetic analyses confirm its close relatedness with Rhodococcus equi (Rhodococcus hoagii) strains, which share >80 % of the gene content. The R. equi virulence plasmid is absent though most of the chromosomal R. equi virulence-associated genes are present in R. defluvii Ca11(T). These data suggest that although R. defluvii is an environmental organism, it has the potential to colonize animal hosts.

  2. Eustachian tube diverticulum chondroids and neck abscessation in a case of Streptococcus equi subsp. equi : clinical communication

    Directory of Open Access Journals (Sweden)

    C. Furniss

    2007-06-01

    Full Text Available A 12-year-old, grey, crossbred pony mare was presented with a swelling in the neck over the area of the 2nd cervical vertebra (C2, which was found to be painful on palpation. The neck was held stiffly. Radiography of the cervical region showed a focal area of increased radio-opacity over the dorsal, caudal and lateral aspect of the dorsal spinous process of C2. Ultrasound confirmed the presence of a hypoechoic area approximately 15 cm in diameter superimposed over the dorsal spinous process of C2. An aspirate was taken of the mass, which revealed purulent exudate confirming the diagnosis of an abscess. The abscess was lanced with a scalpel blade and samples of the purulent material revealed a pure culture of Streptococcus equi subsp. equi. The guttural pouches (Eustachian tube diverticulae (ETD were then evaluated endoscopically and multiple chondroids were seen filling most of the right ETD. Surgery was subsequently performed and 189 chondroids were removed via a right-sided hyovertebrotomy. The ETDs were flushed and penicillin installed into both ETDs on 3 different occasions via a catheter introduced using a fibre optic scope. This procedure was repeated until a negative culture status was achieved in order to eliminate the carrier status.

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

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    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.

  4. Comparisons of serological tests for Babesia in British cattle.

    Science.gov (United States)

    Bidwell, D E; Turp, P; Joyner, L P; Payne, R C; Purnell, R E

    1978-11-11

    A comparison was made between the microplate enzyme linked immunosorbent assay (ELISA) and the indirect fluorescent antibody (IFA) and complement fixation (CF) tests for the detection of antibodies in the serum of cattle experimentally infected with Babesia divergens and B major. Antibodies were detected using all three tests but they were detected earlier using the CF test. However CF titres were consistently lower than those obtained using the other tests. Although there was little to choose between the IFA and ELISA tests, it was suggested that the ELISA may be preferable since it is less subject to operator error and operator stress, and can be adapted readily to field use.

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

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    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.

  6. Protease activity in Giardia duodenalis trophozoites of axenic strains isolated from symptomatic and asymptomatic patients

    Directory of Open Access Journals (Sweden)

    Guimarães Semíramis

    2003-01-01

    Full Text Available We have examined by gelatin-SDS-PAGE the protease activity in cell lysates of Giardia duodenalis trophozoites of two axenic strains isolated in Brazil from a symptomatic patient (BTU-11 and an asymptomatic carrier (BTU-10, and the reference strain Portland 1 (P1. The proteolysis band patterns showed differences among strains isolated from asymptomatic and symptomatic individuals. The lysate of the strain BTU-10, showed only five hydrolysis bands, while a greater number of bands (10-11 bands was seen in strains BTU-11 and P1. The protease activity in all lysates was inhibited by cysteine (E-64 and iodoacetamide and serine proteases (TPCK and TLCK inhibitors, but not by PMSF and EDTA. In general, the results revealed protease activities in G. duodenalis trophozoites of Brazilian axenic strains and the predominance of cysteine proteinases. It should be stressed the inter-strain difference in hydrolysis band patterns observed between strains isolated from symptomatic patients and the strain obtained from an asymptomatic carrier.

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

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    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.

  8. Temperature limitation may explain the containment of the trophozoites in the cornea during Acanthamoeba castellanii keratitis.

    Science.gov (United States)

    Nielsen, Mattias Kiel; Nielsen, Kim; Hjortdal, Jesper; Sørensen, Uffe B Skov

    2014-12-01

    Acanthamoeba keratitis is a serious sight-threatening disease. The relatively low temperature of the cornea may explain why amoebic infections usually are localized in this tissue and rarely spread to other parts of the eye. In this study, the growth rate of the amoeba Acanthamoeba castellanii was examined at different temperatures. The aim was to establish the optimal growth temperature for A. castellanii and to examine the growth within the vicinity of the core body temperature. The growth rates of four clinical and two environmental strains of A. castellanii were estimated at different temperatures, and temperature limitations for the trophozoite stage was established. Movements influenced by temperature gradients were monitored for two clinical strains of A. castellanii. The highest growth rate for each of the six amoebic strains tested was found to be close to 32 °C. The growth of the trophozoites of all examined strains was greatly reduced or completely halted at temperatures above 36 °C and encysted at the elevated temperature. Thus, the optimal growth temperature for the four strains of A. castellanii is close to the surface temperature of the human cornea, while the higher body core-temperature induced encysting of the amoebae. This may explain why most amoebic eye infections are confined to the cornea.

  9. Proteinase activities in total extracts and in medium conditioned by Acanthamoeba polyphaga trophozoites.

    Science.gov (United States)

    Alfieri, S C; Correia, C E; Motegi, S A; Pral, E M

    2000-04-01

    Acanthamoeba species can cause granulomatous encephalitis and keratitis in man. The mechanisms that underlie tissue damage and invasion by the amoebae are poorly understood, but involvement of as yet uncharacterized proteinases has been suggested. Here, we employed gelatin-containing gels and azocasein assays to examine proteinase activities in cell lysates and in medium conditioned by Acanthamoeba polyphaga trophozoites. Azocasein hydrolysis by cell lysates was optimally detected at pH 4.0-5.0 and was predominantly associated with the activity of cysteine proteinases. Compatible with enzyme activation during secretion, culture supernatants additionally contained a prominent azocasein hydrolyzing activity attributable to serine proteinases; these enzymes were better detected at pH 6.0 and above, and resolved at 47, 60, 75, 100, and >110 kDa in overlay gelatin gels. Although a similar banding profile was observed in gels of trophozoite lysates, intracellular serine proteinases were shown to be activated during electrophoresis and to split the substrate during migration in sodium dodecyl sulfate gels. Blockage of serine proteinases with phenylmethylsulfonylfluoride prior to electrophoresis permitted the detection of 43-, 59-, 70-, and 100-130-kDa acidic cysteine proteinases in cell lysates, and of 3 (43, 70, and 130 kDa) apparently equivalent enzymes in culture supernatants. Under the conditions employed, no band associated with a metalloproteinase activity could be depicted in substrate gels, although the discrete inhibition of supernatants' azocaseinolytic activity by 1,10-phenanthroline suggested secretion of some metalloproteinase.

  10. Immunolocalization of α18- and α12-giardin in Giardia lamblia trophozoites.

    Science.gov (United States)

    Wu, Sheng; Pan, Weida; Shi, Xianli; Abdullahi, Auwalu Yusuf; Wang, Zhen; Yu, Xingang; Jiang, Biao; Li, Kangxin; Xu, Chang; Li, Guoqing

    2016-11-01

    To study subcellular localization of α18- and α12-giardin in Giardia lamblia trophozoites, the α18- and α12-giardin genes were amplified from G. lamblia assemblage A, respectively. The PCR products were cloned into the prokaryotic expression vector pET-28a(+), and the positive recombinant plasmids were transformed into E. coli Rosetta (DE3) strain for the expression, and expressed α18- and α12-giardin fusion protein were purified by Ni-Agarose resin, respectively. Mice were immunized with purified fusion proteins for preparation of polyclonal antibody, and then the subcellular localization of α18- and α12-giardin was determined by fluorescence immunoassay. Results showed that the concentrations of purified α18- and α12-giardin fusion proteins were 1.20 and 0.86 mg/ml, respectively. The titers of anti-α18- and anti-α12-giardin polyclonal antibody were both as high as 1:25600 dilutions. Immunofluorescent analysis showed that α18- and α12-giardin proteins were mainly localized at four pairs of flagella and the cytoplasm of G. lamblia trophozoites, suggesting that α18- and α12-giardin are the flagella and cytoplasm-associated proteins, respectively. The above information would lay the foundation for research about the crystal structure and biological function of α18- and α12-giardin.

  11. The effects of saturated fatty acids on Giardia duodenalis trophozoites in vitro.

    Science.gov (United States)

    Rayan, Paran; Stenzel, Deborah; McDonnell, Pauline Ann

    2005-10-01

    Giardia duodenalis is a protozoal, intestinal parasite that is a common aetiological agent of infectious diarrhoea in humans worldwide. Chemotherapeutic intervention presently offers a limited range of drugs and these are usually only employed after clinical diagnosis. Moreover, these drugs are ineffective against the infectious cysts, can produce unpleasant side effects, and are expensive with limited availability in developing countries. Frequent reports of drug toxicity, treatment failure and parasite drug resistance have, in some instances, also resulted in the increasing reluctance to over-prescribe synthetic anti-microbials. Alternatively, there is now mounting evidence to suggest that some of the naturally derived, medium-chain, saturated fatty acids (MCSFAs) possess anti-microbial and anti-parasitic properties. We have therefore examined the effects of four different fatty acids on G. duodenalis trophozoites in vitro. Cytotoxicity was determined using fluorescence, scanning and transmission electron microscopic techniques and standard cytotoxicity assays. Our studies have confirmed that the MCSFA, dodecanoic acid (C: 12) (common name: lauric acid), is anti-giardial, with an LD50 concentration comparable to that of metronidazole, the drug of choice in the treatment of giardiasis. Dodecanoic acid appeared to induce trophozoite death by accumulating within the parasite cytoplasm resulting in rupture of the cell membrane. This study has opened fresh avenues for development of natural drug therapy in which food supplementation may augment, or even replace, some of the standard chemotherapeutic agents presently employed in the treatment of giardiasis and possibly other infectious intestinal diseases.

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

    Science.gov (United States)

    Aguilar-Díaz, Hugo; Díaz-Gallardo, Martha; Laclette, Juan P; Carrero, Julio C

    2010-02-16

    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.

  13. Effects of the extract and glycoalkaloids of Solanum lycocarpum St. Hill on Giardia lamblia trophozoites

    Directory of Open Access Journals (Sweden)

    Gilmarcio Z Martins

    2015-01-01

    Full Text Available Background: Solanum lycocarpum has great importance for food and medicinal traditional use. Recently, it was also evidenced that extracts of S. lycocarpum St. Hill (Solanaceae and its glycoalkaloids, solamargine (Sg and solasonine (Sn, are active against flagellated protozoa. Objective: The aim was to assess the effects of the extract of S. lycocarpum and its glycoalkaloids, Sn, and Sg, on Giardia lamblia trophozoites. Materials and Methods: A crude extract (96% ethanol (EB of fruits of S. lycocarpum was prepared and fractionated by partition with 40% ethanol and n-hexane: Ethyl acetate. Glycoalkaloids, Sn, and Sg were recognized in the ethanol fraction (EF and further isolated by column chromatography. EB, EF, the isolated Sn and Sg and a mixture (1:1 of both glycoalkaloids were tested on cultures of G. lamblia trophozoites and macrophages. Results: EB, EF and glycoalkaloids of S. lycocarpum showed activity against Giardia (95.0 < Inhibitory concentration 50 [IC 50 ] ≤120.3 μg/mL. The mixture of glycoalkaloids (1:1 was more active (IC 50 = 13.23 μg/mL than each one individually, suggesting a synergic effect. Moreover, the mixture is nontoxic to macrophage cells. Conclusion: Results are optimistic concerning the anti-Giardia potential of the mixture Sn + Sg. Further studies, in vitro and in vivo, will be required to consolidate the usefulness of the mixture of Sn + Sg in view of a new therapeutic strategy for giardiasis.

  14. A real-time impedance based method to assess Rhodococcus equi virulence.

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    Aleksandra A Miranda-CasoLuengo

    Full Text Available Rhodococcus equi is a facultative intracellular pathogen of macrophages and the causative agent of foal pneumonia. R. equi virulence is usually assessed by analyzing intracellular growth in macrophages by enumeration of bacteria following cell lysis, which is time consuming and does not allow for a high throughput analysis. This paper describes the use of an impedance based real-time method to characterize proliferation of R. equi in macrophages, using virulent and attenuated strains lacking the vapA gene or virulence plasmid. Image analysis suggested that the time-dependent cell response profile (TCRP is governed by cell size and roundness as well as cytoxicity of infecting R. equi strains. The amplitude and inflection point of the resulting TCRP were dependent on the multiplicity of infection as well as virulence of the infecting strain, thus distinguishing between virulent and attenuated strains.

  15. Immune Reconstitution Syndrome secondary to Rhodococcus equi infection in a patient with HIV and Burkitt's lymphoma.

    Science.gov (United States)

    Darraj, Majid; Fainstein, Rachel; Kasper, Ken; Keynan, Yoav

    Immune Reconstitution Syndrome (IRIS) has been associated with a variety of infections in patients with human immunodeficiency virus (HIV). However, we are reporting the first case of IRIS secondary to Rhodococcus equi (R. equi) in a patient with HIV. We report the case of a 48-year-old male found to have HIV infection in the setting of Burkitt's lymphoma. While on anti-retroviral therapy and chemotherapy, he had developed IRIS secondary to R. equi that manifested as a cavitating pneumonia. This report outlines the successful management of the R. equi infection with the use of a combination of antibiotics, radiographic follow up and suppressive antibiotic while on chemotherapy. Copyright © 2016 King Saud Bin Abdulaziz University for Health Sciences. Published by Elsevier Ltd. All rights reserved.

  16. The correct name of the taxon that contains the type strain of Rhodococcus equi.

    Science.gov (United States)

    Tindall, B J

    2014-01-01

    Based on a nomenclatural point of view, the name Rhodococcus equi is associated, as required by the Bacteriological Code, with a defined position, rank and circumscription. A search of the literature indicates that the name Rhodococcus equi (Magnusson 1923) Goodfellow and Alderson 1977 has also been shown to be a synonym of Corynebacterium equi Magnusson 1923, Corynebacterium hoagii (Morse 1912) Eberson 1918 and Nocardia restricta (Turfitt 1944) McClung 1974. Application of the rules of the Bacteriological Code together with the currently inferred taxonomic concept associated with the species bearing the name Rhodococcus equi indicates that this is not the correct name of this taxon and the use of that name in the context of a circumscription that includes the type strain of the species Corynebacterium hoagii is contrary to the Rules of the Code.

  17. A real-time impedance based method to assess Rhodococcus equi virulence.

    Science.gov (United States)

    Miranda-CasoLuengo, Aleksandra A; Miranda-CasoLuengo, Raúl; Lieggi, Nora T; Luo, Haixia; Simpson, Jeremy C; Meijer, Wim G

    2013-01-01

    Rhodococcus equi is a facultative intracellular pathogen of macrophages and the causative agent of foal pneumonia. R. equi virulence is usually assessed by analyzing intracellular growth in macrophages by enumeration of bacteria following cell lysis, which is time consuming and does not allow for a high throughput analysis. This paper describes the use of an impedance based real-time method to characterize proliferation of R. equi in macrophages, using virulent and attenuated strains lacking the vapA gene or virulence plasmid. Image analysis suggested that the time-dependent cell response profile (TCRP) is governed by cell size and roundness as well as cytoxicity of infecting R. equi strains. The amplitude and inflection point of the resulting TCRP were dependent on the multiplicity of infection as well as virulence of the infecting strain, thus distinguishing between virulent and attenuated strains.

  18. The effect of bacterial dose and foal age at challenge on Rhodococcus equi infection.

    Science.gov (United States)

    Sanz, M; Loynachan, A; Sun, L; Oliveira, A; Breheny, P; Horohov, D W

    2013-12-27

    While Rhodococcus equi remains the most common cause of subacute or chronic granulomatous bronchopneumonia in foals, development of a relevant model to study R. equi infection has proven difficult. The objective of this study was to identify a challenge dose of R. equi that resulted in slow progressive disease, spontaneous regression of lung lesions and age-dependent susceptibility. Foals less than one-week of age were challenged intratracheally using either 10(6), 10(5), 10(4), 10(3) or 10(2) cfu of R. equi. Two doses (10(3) cfu and 10(5) cfu) were used to challenge 2 and 3-week-old, and 3 and 6-week-old foals, respectively. Physical examination, thoracic ultrasound and blood work were performed. Foals were euthanized at the end of the study or when clinical signs of pneumonia developed. All foals were necropsied and their lung lesions scored. Foals challenged with low concentrations of R. equi developed slow progressive pneumonia and approximately 50% of the foals recovered spontaneously. Likewise, macroscopic (>1cm diameter) pyogranulomatous lesions were only observed when low doses of R. equi were used. Clinical pneumonia was not seen after low dose challenge in the 3-week-old foals or in the 6-week-old foals. This study demonstrates that the use of low doses of R. equi to challenge neonatal foals provides an improved model for studying this disease. Furthermore, susceptibility to R. equi infection was shown to diminish early in the foal's life, as has been reported in the field. Copyright © 2013 Elsevier B.V. All rights reserved.

  19. Molecular epidemiology of Rhodococcus equi in slaughtered swine, cattle and horses in Poland.

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    Witkowski, Lucjan; Rzewuska, Magdalena; Takai, Shinji; Kizerwetter-Świda, Magdalena; Kita, Jerzy

    2016-05-27

    Rhodococcus equi is an emerging zoonotic presumably foodborne pathogen. Since the data on the worldwide prevalence of R. equi in meat animals are scarce, the present study aimed to investigate the molecular epidemiology of R. equi in swine, cattle and horse carcasses intended for human consumption in Poland. Totally 1028 lymph node samples were examined. R. equi was isolated from 26.6 % (105/395) swine and 1.3 % (3/234) bovine healthy submaxillary lymph nodes. In horses, R. equi was isolated only from 0.5 % (1/198) samples of middle tracheo-branchiales lymph node while no lymphocentrum retropharyngeum sample was positive (0/198). The purulent lesions were observed only in 0.8 % swine submaxillary lymph nodes samples (3/398) and in two of them R. equi was detected. All bovine and most of swine isolates (98.1 %) were vapB-positive. 87.9 % of swine isolates carried 95-kb type 5 plasmid, 3.7 % type 1 and plasmid types: 4, 7, 10, 11, 21, 31 were carried by a single isolate (0.9 %). All bovine isolates carried VAPB type 26. Single horse isolate was vapA-positive and carried plasmid VAPA 85-kb type I. The prevalence of vapB-positive R. equi in investigated healthy swine intended for human consumption was very high. Not only swine, but also even apparently healthy cattle or horse carcasses should be considered as a potential source of R. equi for humans, especially in countries where undercooked or raw beef or horsemeat is traditionally consumed.

  20. Use of Liposomal Gentamicin for Treatment of 5 Foals with Experimentally Induced Rhodococcus equi Pneumonia.

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    Cohen, N D; Giguère, S; Burton, A J; Rocha, J N; Berghaus, L J; Brake, C N; Bordin, A I; Coleman, M C

    2016-01-01

    Adverse effects of, and bacterial resistance to, macrolides used to treat Rhodococcus equi infections have prompted search for clinically effective alternative antimicrobials. Liposomal gentamicin (LG) is effective against R. equi in vitro and decreases tissue concentrations of R. equi in experimentally infected mice. Effectiveness of LG treatment of foals with R. equi pneumonia, however, has not been described. Liposomal gentamicin is safe and effective for treating foals with R. equi pneumonia. Ten foals with experimentally induced R. equi pneumonia. Pilot treatment trial. Foals with pneumonia induced by intrabronchial instillation of R. equi were randomly allocated to receive either clarithromycin combined with rifampin (CLR + RIF) PO or LG IV, and followed by daily physical examinations and weekly thoracic ultrasonography and serum creatinine concentration determinations until the resolution of clinical signs. Treatment success was defined as the resolution of clinical signs and ultrasonographically identified pulmonary abscesses. All 10 foals were successfully treated. Two of 5 foals treated with LG developed azotemia within 1 week; LG was discontinued and treatment switched to CLR + RIF for these foals. None of the CLR + RIF treated foals developed azotemia. Liposomal gentamicin IV can be effective for treatment of R. equi pneumonia, but nephrotoxicity indicates that an alternative dosing interval or route (such as nebulization) will be needed before LG is adequately safe for clinical use. Larger comparative trials will be needed to evaluate the relative efficacy of a safer LG dosage regimen. Copyright © 2015 The Authors. Journal of Veterinary Internal Medicine published by Wiley Periodicals, Inc. on behalf of the American College of Veterinary Internal Medicine.

  1. Effects of opsonization of Rhodococcus equi on bacterial viability and phagocyte activation.

    Science.gov (United States)

    Dawson, Dominic R; Nydam, Daryl V; Price, Christopher T; Graham, James E; Cynamon, Michael H; Divers, Thomas J; Felippe, Maria Julia B

    2011-11-01

    To investigate the effect of opsonization of Rhodococcus equi with R. equi-specific antibodies in plasma on bacterial viability and phagocyte activation in a cell culture model of infection. Neutrophils and monocyte-derived macrophages from 6 healthy 1-week-old foals and 1 adult horse. Foal and adult horse phagocytes were incubated with either opsonized or nonopsonized bacteria. Opsonization was achieved by use of plasma containing high or low concentrations of R. equi-specific antibodies. Phagocyte oxidative burst activity was measured by use of flow cytometry, and macrophage tumor necrosis factor (TNF)-α production was measured via an ELISA. Extracellular and intracellular bacterial viability was measured with a novel R. equi-luciferase construct that used a luminometer. Opsonized bacteria increased oxidative burst activity in adult horse phagocytes, and neutrophil activity was dependent on the concentration of specific antibody. Secretion of TNF-α was higher in macrophages infected with opsonized bacteria. Opsonization had no significant effect on bacterial viability in macrophages; however, extracellular bacterial viability was decreased in broth containing plasma with R. equi-specific antibodies, compared with viability in broth alone. The use of plasma enriched with specific antibodies for the opsonization of R. equi increased the activation of phagocytes and decreased bacterial viability in the extracellular space. Although opsonized R. equi increased TNF-α secretion and oxidative burst in macrophages, additional factors may be necessary for effective intracellular bacterial killing. These data have suggested a possible role of plasma antibody in protection of foals from R. equi pneumonia.

  2. In vitro synergy, pharmacodynamics, and postantibiotic effect of 11 antimicrobial agents against Rhodococcus equi.

    Science.gov (United States)

    Giguère, Steeve; Lee, Elise A; Guldbech, Kristen M; Berghaus, Londa J

    2012-11-09

    There are no studies investigating interactions between clarithromycin or azithromycin and rifampin or other commonly used antimicrobial agents against virulent isolates of Rhodococcus equi. In addition, there is no published data on the postantibiotic effects (PAEs) and pharmacodynamics properties of antimicrobial agents against R. equi. The objectives were to assess in vitro interactions, pharmacodynamics, and PAEs of 11 antimicrobial agents belonging to various antimicrobial classes against R. equi. Antimicrobial agents investigated (erythromycin, clarithromycin, azithromycin, rifampin, amikacin, gentamicin, enrofloxacin, vancomycin, imipenem, ceftiofur, and doxycycline) were selected based on in vitro activity against large numbers of isolates of R. equi and frequency of use in foals or humans infected with R. equi. Three virulent strains of R. equi were evaluated by time-kill curves and checkerboard assays, and the postantibiotic effect was measured at 5×MIC. Only amikacin, gentamicin, enrofloxacin, and vancomycin were bactericidal against R. equi. Combinations including a macrolide (erythromycin, clarithromycin, azithromycin) and either rifampin or doxycycline, and the combination doxycycline-rifampin were synergistic. Combinations containing amikacin and erythromycin, clarithromycin, azithromycin, or rifampin and the combination gentamicin-rifampin were antagonistic. The PAEs of rifampin, erythromycin, clarithromycin, vancomycin, and doxycycline were relatively long with median values ranging between 4.5 and 6.5h. Azithromycin, gentamicin, and imipenem had intermediate PAEs ranging between 3.3 and 3.5h. Amikacin, enrofloxacin, and ceftiofur had shorter PAEs ranging between 1.3 and 2.1h. Gentamicin, amikacin, enrofloxacin, and doxycycline exhibited concentration-dependent activity whereas erythromycin, clarithromycin, azithromycin, rifampin, ceftiofur, imipenem, and vancomycin exhibited time-dependent activity against R. equi. Copyright © 2012 Elsevier B

  3. Novel transferable erm(46) determinant responsible for emerging macrolide resistance in Rhodococcus equi.

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    Anastasi, Elisa; Giguère, Steeve; Berghaus, Londa J; Hondalus, Mary K; Willingham-Lane, Jennifer M; MacArthur, Iain; Cohen, Noah D; Roberts, Marilyn C; Vazquez-Boland, Jose A

    2015-12-01

    The objective of this study was to identify the molecular mechanism of macrolide resistance in the actinomycete Rhodococcus equi, a major equine pathogen and zoonotic agent causing opportunistic infections in people. Macrolide-resistant (n = 62) and macrolide-susceptible (n = 62) clinical isolates of R. equi from foals in the USA were studied. WGS of 18 macrolide-resistant and 6 macrolide-susceptible R. equi was performed. Representative sequences of all known macrolide resistance genes identified to date were used to search the genome assemblies for putative homologues. PCR was used to screen for the presence of the identified resistance determinant in the rest of the isolates. Mating experiments were performed to verify mobility of the gene. A novel erm gene, erm(46), was identified in all sequenced resistant isolates, but not in susceptible isolates. There was complete association between macrolide resistance and the presence of erm(46) as detected by PCR screening of all 124 clinical isolates of R. equi. Expression of erm(46) in a macrolide-susceptible strain of R. equi induced high-level resistance to macrolides, lincosamides and streptogramins B, but not to other classes of antimicrobial agents. Transfer of erm(46) to macrolide-susceptible R. equi was confirmed. The transfer frequency ranged from 3 × 10(-3) to 1 × 10(-2). This is the first molecular characterization of resistance to macrolides, lincosamides and streptogramins B in R. equi. Resistance was due to the presence of a novel erm(46) gene mobilizable likely by conjugation, which has spread among equine isolates of R. equi in the USA. © The Author 2015. Published by Oxford University Press on behalf of the British Society for Antimicrobial Chemotherapy. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.

  4. Relationship of mixed bacterial infection to prognosis in foals with pneumonia caused by Rhodococcus equi.

    Science.gov (United States)

    Giguère, S; Jordan, L M I; Glass, K; Cohen, N D

    2012-01-01

    Isolation of multiple bacterial species is common in foals with Rhodococcus equi pneumonia. There is no association between isolation of other microorganisms and outcome. 155 foals with pneumonia caused by R. equi. Case records of foals diagnosed with R. equi pneumonia based on culture of the respiratory tract were reviewed at 2 referral hospitals (University of Florida [UF] and Texas A&M University [TAMU]). R. equi was cultured from a tracheobronchial aspirate (TBA) in 115 foals and from lung tissue in 38 foals. Survival was significantly higher at UF (71%; 70/99) than at TAMU (50%; 28/56). R. equi was significantly more likely to grow in pure cultures from samples obtained from foals at UF (55%; 54/99) than from foals at TAMU (23%; 13/56). Microorganisms cultured with R. equi included Gram-positive bacteria in 40, Gram-negative bacteria in 41, and fungi in 23 foals. The most common bacteria isolated were beta-hemolytic streptococci (n = 26) and Escherichia coli (n = 18). Mixed infections were significantly more likely to be encountered in TBA than in lung tissue. Only foals from which R. equi was cultured from a TBA were included in the analysis for association between mixed infection and outcome. After adjusting for the effect of hospital using multivariate logistic regression, mixed culture, mixed bacterial culture, Gram-positive bacteria, beta-hemolytic streptococci, Gram-negative bacteria, enteric Gram-negative bacteria, nonenteric Gram-negative bacteria, and fungi were not significantly associated with outcome. Isolation of multiple bacteria or fungi from a TBA along with R. equi does not negatively impact prognosis. Copyright © 2012 by the American College of Veterinary Internal Medicine.

  5. Pleural malakoplakia caused by Rhodoccocus equi infection in a patient after stem cell transplantation

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    Behnes Carl

    2012-02-01

    Full Text Available Abstract Malakoplakia is a disease especially of the urinary tract with typical plaques most frequently observed in the urinary bladder's mucosa. In the context of immunosuppression malakoplakia can also occur in other organs. Some of these extravesical malakoplakias are associated with an infection by Rhodococcus equi, a rare human pathogen well known from veterinary medicine. Here we present the first case of a pleural malakoplakia without lung involvement caused by a proved Rhodococcus equi infection.

  6. Unusual outbreak of clinical mastitis in dairy sheep caused by Streptococcus equi subsp. zooepidemicus.

    Science.gov (United States)

    Las Heras, Alfonso; Vela, Ana I; Fernández, Elena; Legaz, Emilio; Domínguez, Lucas; Fernández-Garayzábal, Jose F

    2002-03-01

    This work describes an outbreak of clinical mastitis affecting 13 of 58 lactating ewes due to Streptococcus equi subsp. zooepidemicus. S. equi subsp. zooepidemicus was isolated in pure culture from all milk samples. All the clinical isolates had identical biochemical profiles and antimicrobial susceptibility patterns and also exhibited indistinguishable macrorestriction patterns by pulsed-field gel electrophoresis, indicating that all cases of mastitis were produced by a single strain.

  7. Babesia in a Nonsplenectomized Patient Requiring Exchange Transfusion

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    Dikshya Sharma

    2015-01-01

    Full Text Available Babesiosis is a tick born zoonosis caused by red blood cell parasites of the genus Babesia. It is caused predominantly by B. microti and B. divergens, microti being more common in the US. The parasites are transmitted by Ixodes tick to their host but infection can also spread by blood transfusion and perinatally. Clinical manifestations vary from subclinical infection to fulminating disease depending upon the immune status of the patient. About half of patients, hospitalized with babesiosis, develop complication with fatality rates of 6 to 9% which increase up to 21% among those with immunosuppression. A case of 58-year-old previously healthy man, infected by B. microti, was reported on 2000 who presented with severe disease characterized by severe anemia, DIC, and renal and respiratory failure. First case of overwhelming septic shock without respiratory involvement due to babesiosis in a healthy patient with an intact spleen was published in a case report on 2011. Since our patient here is an immunocompetent healthy male with intact spleen presenting with severe babesiosis requiring exchange transfusion, this presentation of Babesia is rare and warrants further study.

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

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    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.

  9. Diagnosis and prevalence of Theileria equi horses in western Mexico by nested PCR.

    Science.gov (United States)

    Ayala-Valdovinos, Miguel Angel; Lemus-Flores, Clemente; Galindo-García, Jorge; Bañuelos-Pineda, Jacinto; Rodríguez-Carpena, Javier Germán; Sánchez-Chiprés, David; Duifhuis-Rivera, Theodor

    2017-02-01

    Theileria equi infection prevalence was calculated from 1000 blood samples obtained from apparently healthy horses in western Mexico. Samples were sent to the Animal Biotechnology Laboratory of the University of Guadalajara (Mexico) for T. equi diagnosis. Nested polymerase chain reaction (nPCR) was used as a diagnostic method to detect pathogen DNA. Using primers for the merozoite antigen-1 (EMA-1) gene, 19.70±2.47% of the horses (95% CI, 17.23-22.17%) tested positive for T. equi. There was no significant association between gender and T. equi infection. However, prevalence was higher among stabled horses (25.81%) than that among grazing horses (15.02%). The positivity rate was also higher among Quarter Horse (24.70%), Lusitano (35.90%), and Costa Rican Saddle Horse (47.37%) breeds than that among the other seven breeds investigated in this study. The percentage of T. equi infection was higher among adult horses (≥ 4years old, 25.05%) than that among colts and fillies (2-4years old, 15.48%), yearlings (1-2years old, 10.49%), and foals (horses in Mexico by nPCR . The results indicate that the equine piroplasmosis (EP) caused by T. equi is enzootic in western Mexico. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

  10. Nitric oxide-mediated intracellular growth restriction of pathogenic Rhodococcus equi can be prevented by iron.

    Science.gov (United States)

    von Bargen, Kristine; Wohlmann, Jens; Taylor, Gregory Alan; Utermöhlen, Olaf; Haas, Albert

    2011-05-01

    Rhodococcus equi is an intracellular pathogen which causes pneumonia in young horses and in immunocompromised humans. R. equi arrests phagosome maturation in macrophages at a prephagolysosome stage and grows inside a privileged compartment. Here, we show that, in murine macrophages activated with gamma interferon and lipopolysaccharide, R. equi does not multiply but stays viable for at least 24 h. Whereas infection control of other intracellular pathogens by activated macrophages is executed by enhanced phagosome acidification or phagolysosome formation, by autophagy or by the interferon-inducible GTPase Irgm1, none of these mechanisms seems to control R. equi infection. Growth control by macrophage activation is fully mimicked by treatment of resting macrophages with nitric oxide donors, and inhibition of bacterial multiplication by either activation or nitric oxide donors is annihilated by cotreatment of infected macrophages with ferrous sulfate. Transcriptional analysis of the R. equi iron-regulated gene iupT demonstrates that intracellular R. equi encounters iron stress in activated, but not in resting, macrophages and that this stress is relieved by extracellular addition of ferrous sulfate. Our results suggest that nitric oxide is central to the restriction of bacterial access to iron in activated macrophages.

  11. Resistance studies of erythromycin and rifampin for Rhodococcus equi over a 10-year period

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    Buckley T

    2007-12-01

    Full Text Available Abstract This study sought to determine whether an increase in resistance of Rhodococcus equi to the antibiotics rifampin and erythromycin occurred over a 10-year period. This was carried out by the use of E test strips for rifampin and erythromycin to determine the MIC (minimum inhibitory concentration values of Rhodococcus equi to this combination of antibiotics. The findings of this study indicated that there was an increase in resistance of Rhodococcus equi to rifampin and erythromycin over the 10-year period. The MIC for rifampin increased from 0.081 μg/ml in 1996 to 0.187 μg/ml in 2006 and from 0.258 μg/ml for erythromycin during the years prior to 2000 to 0.583 μg/ml in 2006. This finding suggests that there may be a problem in the treatment of Rhodococcus equi infections in foals in the future, particularly as the number of drugs available for treatment of Rhodococcus equi infection is limited because of the intracellular capabilities of this bacterium. Antibiotics used in its treatment have to be able to penetrate the polysaccharide cell wall of Rhodococcus equi as well as the alveolar macrophages in which the bacterium is capable of surviving.

  12. Induction of proinflammatory cytokines in human lung epithelial cells during Rhodococcus equi infection.

    Science.gov (United States)

    Remuzgo-Martínez, Sara; Pilares-Ortega, Lilian; Alvarez-Rodríguez, Lorena; Aranzamendi-Zaldunbide, Maitane; Padilla, Daniel; Icardo, Jose Manuel; Ramos-Vivas, Jose

    2013-08-01

    Rhodococcus equi is an opportunistic human pathogen associated with immunosuppressed people. While the interaction of R. equi with macrophages has been comprehensively studied, little is known about its interactions with non-phagocytic cells. Here, we characterized the entry process of this bacterium into human lung epithelial cells. The invasion is inhibited by nocodazole and wortmannin, suggesting that the phosphatidylinositol 3-kinase pathway and microtubule cytoskeleton are important for invasion. Pre-incubation of R. equi with a rabbit anti-R. equi polyclonal antiserum resulted in a dramatic reduction in invasion. Also, the invasion process as studied by immunofluorescence and scanning electron microscopy indicates that R. equi make initial contact with the microvilli of the A549 cells, and at the structural level, the entry process was observed to occur via a zipper-like mechanism. Infected lung epithelial cells upregulate the expression of cytokines IL-8 and IL-6 upon infection. The production of these pro-inflammatory cytokines was significantly enhanced in culture supernatants from cells infected with non-mucoid plasmid-less strains when compared with cells infected with mucoid strains. These results demonstrate that human airway epithelial cells produce pro-inflammatory mediators against R. equi isolates.

  13. Nitric Oxide-Mediated Intracellular Growth Restriction of Pathogenic Rhodococcus equi Can Be Prevented by Iron▿

    Science.gov (United States)

    von Bargen, Kristine; Wohlmann, Jens; Taylor, Gregory Alan; Utermöhlen, Olaf; Haas, Albert

    2011-01-01

    Rhodococcus equi is an intracellular pathogen which causes pneumonia in young horses and in immunocompromised humans. R. equi arrests phagosome maturation in macrophages at a prephagolysosome stage and grows inside a privileged compartment. Here, we show that, in murine macrophages activated with gamma interferon and lipopolysaccharide, R. equi does not multiply but stays viable for at least 24 h. Whereas infection control of other intracellular pathogens by activated macrophages is executed by enhanced phagosome acidification or phagolysosome formation, by autophagy or by the interferon-inducible GTPase Irgm1, none of these mechanisms seems to control R. equi infection. Growth control by macrophage activation is fully mimicked by treatment of resting macrophages with nitric oxide donors, and inhibition of bacterial multiplication by either activation or nitric oxide donors is annihilated by cotreatment of infected macrophages with ferrous sulfate. Transcriptional analysis of the R. equi iron-regulated gene iupT demonstrates that intracellular R. equi encounters iron stress in activated, but not in resting, macrophages and that this stress is relieved by extracellular addition of ferrous sulfate. Our results suggest that nitric oxide is central to the restriction of bacterial access to iron in activated macrophages. PMID:21383050

  14. EquiFACS: The Equine Facial Action Coding System.

    Science.gov (United States)

    Wathan, Jen; Burrows, Anne M; Waller, Bridget M; McComb, Karen

    2015-01-01

    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.

  15. 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.

  16. 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

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    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.

  17. Use of Serial Quantitative PCR of the vapA Gene of Rhodococcus equi in Feces for Early Detection of R. equi Pneumonia in Foals.

    Science.gov (United States)

    Madrigal, R G; Shaw, S D; Witkowski, L A; Sisson, B E; Blodgett, G P; Chaffin, M K; Cohen, N D

    2016-01-01

    Current screening tests for Rhodococcus equi pneumonia in foals lack adequate accuracy for clinical use. Real-time, quantitative PCR (qPCR) for virulent R. equi in feces has not been systematically evaluated as a screening test. The objective of this study was to evaluate the accuracy of qPCR for vapA in serially collected fecal samples as a screening test for R. equi pneumonia in foals. One hundred and twenty-five foals born in 2011 at a ranch in Texas. Fecal samples were collected concurrently with thoracic ultrasonography (TUS) screening examinations at ages 3, 5, and 7 weeks. Affected (pneumonic) foals (n = 25) were matched by age and date-of-birth to unaffected (n = 25) and subclinical (ie, having thoracic TUS lesions but no clinical signs of pneumonia) foals (n = 75). DNA was extracted from feces using commercial kits and concentration of virulent R. equi in feces was determined by qPCR. Subsequently affected foals had significantly greater concentrations of vapA in feces than foals that did not develop pneumonia (unaffected and subclinical foals) at 5 and 7 weeks of age. Accuracy of fecal qPCR, however, was poor as a screening test to differentiate foals that would develop clinical signs of pneumonia from those that would remain free of clinical signs (including foals with subclinical pulmonary lesions attributed to R. equi) using receiver operating characteristic (ROC) methods. In the population studied, serial qPCR on feces lacked adequate accuracy as a screening test for clinical R. equi foal pneumonia. Copyright © 2016 The Authors. Journal of Veterinary Internal Medicine published by Wiley Periodicals, Inc. on behalf of the American College of Veterinary Internal Medicine.

  18. Imidocarb dipropionate clears persistent Babesia caballi infection with elimination of transmission potential

    Science.gov (United States)

    Antimicrobial treatment of persistent infection to eliminate transmission risk represents a specific challenge requiring compelling evidence of complete pathogen clearance. The limited repertoire of antimicrobial agents targeted at protozoal parasites magnifies this challenge. Using Babesia cabal...

  19. Phylogenetic analysis based on 28S rRNA of Babesia spp. in ruminants in China.

    Science.gov (United States)

    Gou, Huitian; Guan, Guiquan; Ma, Miling; Liu, Aihong; Liu, Zhijie; Ren, Qiaoyun; Li, Youquan; Yang, Jifei; Chen, Ze; Yin, Hong; Luo, Jianxun

    2013-04-01

    Molecular phylogenetic analyses are mainly based on the small ribosomal RNA subunit (18S rRNA), internal transcribed spacer regions, and other molecular markers. We compared the phylogenetic relationships of Babesia spp. using large subunit ribosomal RNA, i.e., 28S rRNA, and the united 28S + 18S rRNA sequence fragments from 11 isolates of Babesia spp. collected in China. Due to sequence length and variability, the 28S rRNA gene contained more information than the 18S rRNA gene and could be used to elucidate the phlyogenetic relationships of B. motasi, B. major, and B. bovis. Thus, 28S rRNA is another candidate marker that can be used for the phylogenetic analysis of Babesia spp. However, the united fragment (28S + 18S) analysis provided better supported phylogenetic relationships than single genes for Babesia spp. in China.

  20. Molecular and serological prevalence of Babesia bovis and Babesia bigemina in water buffaloes in the northeast region of Thailand.

    Science.gov (United States)

    Terkawi, Mohamad Alaa; Huyen, Nguyen Xuan; Shinuo, Cao; Inpankaew, Tawin; Maklon, Khuanwalai; Aboulaila, Mahmoud; Ueno, Akio; Goo, Youn-Kyoung; Yokoyama, Naoaki; Jittapalapong, Sathaporn; Xuan, Xuenan; Igarashi, Ikuo

    2011-06-10

    Bovine babesiosis is a tick-transmitted hemoprotozoan disease that is mainly caused by Babesia bovis and Babesia bigemina and is characterized by significant morbidity and mortality worldwide. The disease is widespread in the northeastern region of Thailand, where an increasingly large part of the livestock is composed of water buffaloes. The present study was therefore conducted to investigate the epidemiological distribution of B. bovis and B. bigemina in water buffaloes in the northeastern region of Thailand. A total of 305 buffalo blood samples were randomly collected from five provinces and simultaneously analyzed by the nested PCR (nPCR) assay, ELISA, and IFAT techniques. The overall prevalence of B. bovis and B. bigemina was 11.2% and 3.6% by nPCR, 14.7% and 5.9% by ELISA, and 16.8% and 5.6% by IFAT, respectively. The high concordance between the molecular and the serological detection tests revealed the specificity and sensitivity of the diagnostic assays used for the detection of infection as well as the endemic stability status of the parasites in the surveyed areas. Statistically significant differences in the prevalence of the two infections were observed on the basis of age and location but not gender. Our data provide valuable information regarding the epidemiology of B. bovis and B. bigemina infection in water buffaloes in the northeastern region of Thailand which will likely be very beneficial for management and control programs of this disease.

  1. Molecular and serological detection of Babesia Bovis and Babesia Bigemina in cattle in the Rio de Janeiro, Brazil

    Directory of Open Access Journals (Sweden)

    Jenevaldo Barbosa da Silva

    2014-12-01

    Full Text Available Bovine babesiosis is an important disease of cattle where Rhipicephalus microplus acts as a vector for the two causal organisms Babesia bovis and Babesia bigemina. A total of 22 calves were randomly monitored during three years by semi-nested PCR assay and ELISA test to determine prevalence of B. bovis and B. bigemina. The overall prevalence of B. bovis and B. bigemina was 30% and 35% by nested PCR (nPCR, and 70% and 75% by ELISA, respectively. Statistical analysis of the characteristics of animals showed that age and tick infestations (p<0.05 might play an important role in the spread of babesiosis as animal less than 6 months old. A high correlation (Kappa index of 0.70 for B. bovis and 0.65 for B. bigemina, respectively between serological and molecular tests suggests that the combination of the utilized techniques in the present study is suitable for babesiosis diagnosis in an endemic unstable area.

  2. Detection of Babesia microti parasites by highly sensitive 18S rRNA reverse transcription PCR.

    Science.gov (United States)

    Hanron, Amelia E; Billman, Zachary P; Seilie, Annette M; Chang, Ming; Murphy, Sean C

    2017-03-01

    Babesia are increasingly appreciated as a cause of transfusion-transmitted infection. Sensitive methods are needed to screen blood products. We report herein that B. microti 18S rRNA is over 1,000-fold more abundant than its coding genes, making reverse transcription PCR (RT-PCR) much more sensitive than PCR. Babesia 18S rRNA may be useful for screening the blood supply.

  3. Bronchopneumonia in wild boar (Sus scrofa) caused by Rhodococcus equi carrying the VapB type 8 plasmid.

    Science.gov (United States)

    de Vargas, Agueda Castagna; Monego, Fernanda; Gressler, Letícia Trevisan; de Avila Botton, Sônia; Lazzari, Andrea Maria; da Costa, Mateus Matiuzzi; Ecco, Roselene; Ribeiro, Márcio Garcia; Lara, Gustavo Henrique Batista; Takai, Shinji

    2013-03-25

    Rhodococcus equi is associated with pyogranulomatous infections, especially in foals, and this bacterium has also emerged as a pathogen for humans, particularly immunocompromised patients. R. equi infections in pigs, wild boar (Sus scrofa) and humans are mainly due to strains carrying the intermediate virulence (VapB) plasmid. In Brazil, R. equi carrying the VapB type 8 plasmid is the most common type recovered from humans co-infected with the human immunodeficiency virus (HIV). R. equi infection in pigs and wild boar is restricted predominantly to the lymphatic system, without any reports of pulmonary manifestations. This report describes the microbiological and histopathological findings, and molecular characterization of R. equi in two bronchopneumonia cases in wild boar using PCR and plasmid profile analysis by digestion with restriction endonucleases. The histological findings were suggestive of pyogranulomatous infection, and the plasmid profile of both R. equi isolates enabled the characterization of the strains as VapB type 8. This is the first report of bronchopneumonia in wild boar due to R. equi. The detection of the VapB type 8 plasmid in R. equi isolates emphasize that wild boar may be a potential source of pathogenic R. equi strains for humans.

  4. Detection of Babesia venatorum, Anaplasma phagocytophilum and Candidatus Neoehrlichia mikurensis in Ixodes persulcatus ticks from Mongolia.

    Science.gov (United States)

    Karnath, Carolin; Obiegala, Anna; Speck, Stephanie; Essbauer, Sandra; Derschum, Henri; Scholz, Holger; Kiefer, Daniel; Tserennorov, Damdindorj; Dashdavaa, Otgonbataar; Tsogbadrakh, Nyamdorj; Jigjav, Battsetseg; Pfeffer, Martin

    2016-03-01

    Information about the prevalence and geographical distribution of tick-borne pathogens Anaplasma phagocytophilum, Candidatus Neoehrlichia mikurensis, and Babesia spp. is still rare in Mongolia. We tested 275 Ixodes persulcatus ticks for A. phagocytophilum, Cand. N. mikurensis and Babesia spp. and 125 Dermacentor nuttalli ticks especially for Babesia spp. using different PCR methods. Ticks were collected from three provinces (Selenge, Arkhangai, Khentii) in Mongolia. DNA of A. phagocytophilum, Cand. N. mikurensis and Babesia spp. were found with a prevalence of 6.2%, 1.5% and 3.3% in each case in I. persulcatus ticks. This is the first time Cand. N. mikurensis was found in ticks from Mongolia. Sequence analysis of Babesia spp.-positive amplicons showed exclusively B. venatorum, which had also not been mentioned in Mongolia before. On the contrary, all D. nuttalli ticks tested negatively for Babesia spp. This study demonstrates that all three zoonotic pathogens are present in I. persulcatus ticks in Mongolia, and justify the need for further investigations of a more detailed genetic characterization of these pathogens.

  5. 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.

  6. Balamuthia mandrillaris: Further morphological observations of trophozoites by light, scanning and transmission electron microscopy.

    Science.gov (United States)

    González-Robles, Arturo; Lares-Villa, Fernando; Lares-Jiménez, Luis Fernando; Omaña-Molina, Maritza; Salazar-Villatoro, Lizbeth; Martínez-Palomo, Adolfo

    2015-10-01

    Additional morphological features of Balamuthia mandrillaris observed by light and electron microscopy are reported. Trophozoites were extremely pleomorphic: their cell shapes ranged from rounded to elongated and sometimes they appeared exceptionally stretched out and branched. By transmission electron microscopy it was possible to observe two different cytoplasmic areas, the ectoplasm and the endoplasm and often sections of rough endoplasmic reticulum were found in the transition zone. The cytoplasm was very fibrogranular and most of the organelles typically found in eukaryotic cells were observed. A particular finding was the presence of numerous mitochondria with a different structure from those of other free-living amoebae. The observations reported here may reinforce the morphological knowledge of this amoeba and provide a background for further analyses.

  7. Evaluation of the activity of new cationic carbosilane dendrimers on trophozoites and cysts of Acanthamoeba polyphaga.

    Science.gov (United States)

    Heredero-Bermejo, Irene; Copa-Patiño, Jose Luis; Soliveri, Juan; Fuentes-Paniagua, Elena; de la Mata, Francisco Javier; Gomez, Rafael; Perez-Serrano, Jorge

    2015-02-01

    Dendrimers are repetitively branched molecules with a broad spectrum of applications, mainly for their antimicrobial properties and as nanocarriers for other molecules. Recently, our research group have synthesized and studied their activity against Acanthamoeba sp., causative agent of a severe ocular disease in humans: Acanthamoeba keratitis. New cationic carbosilane dendrimers were tested against the protozoa forms at different concentrations and for different incubation times. Trophozoite viability was determined by manual counting and cyst viability by observing excystment in microplates with fresh culture medium. Cytotoxicity was checked on HeLa cells using the microculture tetrazolium assay. Alterations were observed by optical microscopy and by flow cytometry staining with propidium iodide. Six out of the 18 dendrimers tested were non-cytotoxic and effective against the trophozoite form, having one of them (dendrimer 14 with an IC50 of 2.4 + 0.1 mg/L) a similar activity to chlorhexidine digluconate (IC50 1.7 + 0.1 mg/L). This dendrimer has a polyphenoxo core and a sulphur atom close to the six -NH3+ terminal groups. On the other hand, only two dendrimers showed some effect against cysts (dendrimers 14 and 17). However, their minimum cysticidal concentrations were cytotoxic and less effective than the control drug. The alterations on the amoeba morphology produced by the treatment with dendrimers were size reduction, increased complexity, loss of acanthopodia and cell membrane disruption. In conclusion, these results suggest that some dendrimers may be studied in animal models to test their effect and that new dendrimers with similar features should be synthesized.

  8. Amebicidal activity of the essential oils of Lippia spp. (Verbenaceae) against Acanthamoeba polyphaga trophozoites.

    Science.gov (United States)

    Santos, Israel Gomes de Amorim; Scher, Ricardo; Rott, Marilise Brittes; Menezes, Leociley Rocha; Costa, Emmanoel Vilaça; Cavalcanti, Sócrates Cabral de Holanda; Blank, Arie Fitzgerald; Aguiar, Jaciana dos Santos; da Silva, Teresinha Gonçalves; Dolabella, Silvio Santana

    2016-02-01

    Amoebic keratitis and granulomatous amoebic encephalitis are caused by some strains of free-living amoebae of the genus Acanthamoeba. In the case of keratitis, one of the greatest problems is the disease recurrence due to the resistance of parasites, especially the cystic forms, to the drugs that are currently used. Some essential oils of plants have been used as potential active agents against this protist. Thus, the aim of this study was to determine the amebicidal activity of essential oils from plants of the genus Lippia against Acanthamoeba polyphaga trophozoites. To that end, 8 × 10(4) trophozoites were exposed for 24 h to increasing concentrations of essential oils from Lippia sidoides, Lippia gracilis, Lippia alba, and Lippia pedunculosa and to their major compounds rotundifolone, carvone, and carvacrol. Nearly all concentrations of oils and compounds showed amebicidal activity. The IC50 values for L. sidoides, L. gracilis L. alba, and L. pedunculosa were found to be 18.19, 10.08, 31.79, and 71.47 μg/mL, respectively. Rotundifolone, carvacrol, and carvone were determined as the major compounds showing IC50 of 18.98, 24.74, and 43.62 μg/mL, respectively. With the exception of oil from L. alba, the other oils evaluated showed low cytotoxicity in the NCI-H292 cell line. Given these results, the oils investigated here are promising sources of compounds for the development of complementary therapy against amoebic keratitis and granulomatous amoebic encephalitis and can also be incorporated into cleaning solutions to increase their amebicidal efficiency.

  9. Giardia canis: ultrastructural analysis of G. canis trophozoites transfected with full length G. canis virus cDNA transcripts

    Science.gov (United States)

    Giardia canis virus (GCV) is a double-stranded RNA (dsRNA) virus of the family Totiviridae. In this study, the full-length cDNA of the G. canis virus was constructed in pPoly2/sfinot vector and RNA was transcribed in vitro. Virus-free G. canis trophozoites were transfected with in vitro transcribed ...

  10. [Babesia choloepi n. sp. (Apicomplexa, Piroplasmida), a parasite of the two-toed sloth, Choloepus didactylus (Linnaeus, 1758) (Xenarthra, Bradypodidae), in French Guiana].

    Science.gov (United States)

    Dedet, J P; Veilly, M; Robin, Y; Bonnevie, O; Landau, I

    1988-01-01

    A two-toed sloth (Choloepus didactylus) from French Guiana showed an erythrocytic parasite morphologically related to the Babesiidae family, and described as Babesia choloepi. It is the first Babesia species described in the Edentata order.

  11. Air sampling in the breathing zone of neonatal foals for prediction of subclinical Rhodococcus equi infection.

    Science.gov (United States)

    Chicken, C; Muscatello, G; Freestone, J; Anderson, G A; Browning, G F; Gilkerson, J R

    2012-03-01

    Disease caused by Rhodococcus equi is a significant burden to the horse breeding industry worldwide. Early detection of rhodococcal pneumonia, albeit important to minimise treatment costs, is difficult because of the insidious nature of the disease and the lack of definitive diagnostic tests. To investigate air sampling from the breathing zone of neonatal foals as a predictor of subsequent rhodococcal pneumonia. Air samples were collected from the breathing zone of 53 neonatal foals (age ≤10 days) and again at the time of routine ultrasonographic screening for R. equi pneumonia (age 1-2 months). Pneumonia was diagnosed ultrasonographically in 23% of foals. Virulent R. equi was detected in air from the breathing zone of 19% of neonatal foals and 45% of foals at age 1-2 months. There was no association between virulent R. equi in the breathing zone of foals and the subsequent ultrasonographic diagnosis of rhodococcal pneumonia. The median concentration of virulent R. equi in the breathing zone of both neonates (0 [range 0-4] colony-forming units [cfu]/250 l) and older foals (0 [range 0-3] cfu/250 l) was not significantly different from that in background air samples (0 [range 0-6] cfu/250 l). There was no difference in the concentration of virulent R. equi in the breathing zone of older foals that were diagnosed with rhodococcal pneumonia or clinically normal foals. Detection of virulent R. equi in air from the breathing zone was not a positive predictor of rhodococcal pneumonia in foals up to age ≤2 months. Selective culture of air samples from the breathing zone of young foals is not better at diagnosing rhodococcal pneumonia than early ultrasonographic screening. However, culture of air samples from the breathing zone of older foals remains a useful herd-based epidemiological tool. © 2012 EVJ Ltd.

  12. In vitro evaluation of drug susceptibilities of Babesia divergens isolates.

    Science.gov (United States)

    Brasseur, P; Lecoublet, S; Kapel, N; Favennec, L; Ballet, J J

    1998-04-01

    The susceptibilities of three bovine and two human Babesia divergens isolates to antimicrobial agents were evaluated in vitro by a tritiated hypoxanthine incorporation assay. The MICs at which 50% of isolates are inhibited (MIC50s) for mefloquine (chlorhydrate), chloroquine (sulfate), quinine (chlorhydrate), clindamycin (phosphate), pentamidine (isethionate), phenamidine (isethionate) plus oxomemazine (chlorhydrate), lincomycin (chlorhydrate monohydrate), and imidocarb (dipropionate) were determined. Except for imidocarb, the MIC50s observed for the different isolates were close. Imidocarb and the combination of phenamidine plus oxomemazine exhibited the highest in vitro activity, while antimalarial agents such as mefloquine, choroquine, and quinine were inactive. Other drugs had intermediate activities. The data support further in vitro evaluation of antimicrobial agents active against B. divergens for the improvement of therapeutic strategies.

  13. 马巴贝西虫EMA-1基因的克隆与表达%Cloning and Expression of EMA-1 Gene of Babesia equi

    Institute of Scientific and Technical Information of China (English)

    张守发; 鞠玉林; 玄学南

    2003-01-01

    通过聚合酶链式反应扩增出体外培养的马巴西虫USDA株基因EMA-1,将该片段连接到克隆质粒载体pUC19的Bam HI酶切位点上,并对其序列进行测定.结果表明该基因长度为836bp,编码的表面蛋白由272个氨基酸残基组成,与佛罗里达(Florida)株的EMA-1的氨基酸序列有95%左右的同源性.再将EMA-1基因片段插入到表达质粒载体pGE-MEX-2的BamHI位点上,并导入大肠杆菌JM109(DE3)中.经SDSPAGE分析和Western blot检测的结果表明,马巴贝西虫USDA株基因EMA-1,在大肠杆菌内获得表达,融合蛋白的分子量为65kDa.将其免疫给小鼠可产生特异性的抗马巴贝西虫抗体,且同驽巴贝西虫无交叉反应.

  14. Blocking Babesia bovis vaccine reactions of dairy cattle in milk

    Directory of Open Access Journals (Sweden)

    Michael P. Combrink

    2012-12-01

    Full Text Available The use of 1.16 mg/kg (one third of the recommended dose of diminazene aceturate, administered indiscriminately to cattle on day seven of the unfrozen Babesia bovis and Babesia bigemina bivalent live blood vaccine reaction, was an infection and block treatment method of immunisation used successfully with no known adverse effect on the parasites or the development of protective immunity. Continuing with this practice after replacement of the unfrozen vaccine with deep-frozen monovalent B. bovis and B. bigemina live blood vaccines resulted in reports of vaccine failure. Laboratory investigation indicated the harmful effect of block treatment in preventing the development of durable immunity against B. bigemina as opposed to the much lesser effect it had on B. bovis. Consequently the practice was no longer recommended. A B. bovis vaccination attempt aimed at controlling the disease of dairy cows in milk (n = 30 resulted in 20% fatalities during the expected vaccine reaction period. The practice of block treating B. bovis was therefore reinvestigated, this time in a field trial using dairy cattle in milk (n = 11. Using 0.88 mg/kg (one quarter of the recommended dose of diminazene administered on day 12 of the B. bovis vaccine reaction resulted in only two animals (n = 5 testing ≥ 1/80 positive with the indirect fluorescent antibody test (IFAT although parasites could be demonstrated in three. In the untreated control group, by contrast, five of the vaccinated animals (n = 6 tested ≥ 1/80 positive with IFAT and parasites could be demonstrated in all. The unsatisfactory outcome obtained in this study, combined with that of the earlier investigation, indicated that there are more factors that influence successful vaccination than previously considered. It is therefore concluded that block treatment of the live frozen South African cattle babesiosis vaccines reactions is not recommended.

  15. Molecular and serological detection of Babesia bovis- and Babesiabigemina-infection in bovines and water buffaloes raised jointly in anendemic field

    Science.gov (United States)

    tBabesia bovis and Babesia bigemina are causative agents of bovine babesiosis, a tick-borne disease of cattlein tropical and subtropical regions. Babesia spp. infection adversely affects cattle health and can be fatalresulting in considerable economic loss worldwide. Under endemic stability conditio...

  16. Influence of Plasmid Type on the Replication of Rhodococcus equi in Host Macrophages.

    Science.gov (United States)

    Willingham-Lane, Jennifer M; Berghaus, Londa J; Giguère, Steeve; Hondalus, Mary K

    2016-01-01

    The soil-dwelling, saprophytic actinomycete Rhodococcus equi is a multihost, facultative intracellular pathogen of macrophages. When inhaled by susceptible foals, it causes severe bronchopneumonia. It is also a pathogen of pigs, which may develop submaxillary lymphadenitis upon exposure. R. equi isolates obtained from foals and pigs possess conjugative plasmids housing a pathogenicity island (PAI) containing a novel family of genes of unknown function called the virulence-associated protein or vap family. The PAI regions of the equine and swine plasmids differ in vap gene composition, with equine isolates possessing six vap genes, including the major virulence determinant vapA, while the PAIs of swine isolates house vapB and five other unique vap genes. Possession of the pVAPA-type virulence plasmid by equine isolates bestows the capacity for intramacrophage replication essential for disease development in vivo. Swine isolates of R. equi are largely unstudied. Here, we show that R. equi isolates from pigs, carrying pVAPB-type plasmids, are able to replicate in a plasmid-dependent manner in macrophages obtained from a variety of species (murine, swine, and equine) and anatomical locations. Similarly, equine isolates carrying pVAPA-type plasmids are capable of replication in swine macrophages. Plasmid swapping between equine and swine strains through conjugation did not alter the intracellular replication capacity of the parental strain, indicating that coevolution of the plasmid and chromosome is not crucial for this attribute. These results demonstrate that while distinct plasmid types exist among R. equi isolates obtained from equine and swine sources, this tropism is not determined by host species-specific intramacrophage replication capabilities. IMPORTANCE This work greatly advances our understanding of the opportunistic pathogen Rhodococcus equi, a disease agent of animals and immunocompromised people. Clinical isolates from diseased foals carry a

  17. Aspectos epidemiológicos do Rhodococcus equi em eqüinos do Município de Bagé, RS, Brasil Epidemiological aspects of Rhodococcus equi in horses from Bagé county, RS, Brazil

    Directory of Open Access Journals (Sweden)

    Andrea Lazzari

    1997-08-01

    Full Text Available Este trabalho foi realizado com o intuito de obter informações epidemiológicas sobre o Rhodococcus equi (R. equi em seis haras do município de Bagé, estado do Rio Grande do Sul. Foram coletadas 36 amostras de solo superficial e 143 de fezes de eqüinos, sendo semeadas em meio seletivo para o R. equi (NANAT e incubadas em aerobiose a 37°C por até 72 horas. Também foi coletada amostra de solo para determinação de pH e matéria orgânica (MO de cada haras. A identificação do R. equi baseou-se na pesquisa do "fator equi" e características morfológicas, tintoriais e bioquímicas. Isolou-se o microrganismo de 75% (27/36 das amostras de solo superficial e 66,43% (95/143 das amostras de fezes de eqüinos. O isolamento do R. equi em 100% dos haras analisados comprova a disseminação desta bactéria na região estudada. Os dados obtidos na análise de pH e MO do solo não demonstraram correlação com a taxa de isolamento do R. equi do solo superficial dos haras.Epidemiological aspects about Rhodococcus equi (R. equi at six horse-breeding farms from Bagé county, in the state of Rio Grande do Sul, Brazil, were analysed. Soil samples (36 and equine feces (143 were collected. The objective was to verify the presence of R. equi in those samples. Six soil samples were also used for pH and organic matter (OM determination. R. equi identification was based on equi factor search and morfological, tintorial and biochemistry behavior. All the horse-breeding farms had the R. equi present in soil and feces samples. Soil yielded 75.00% (27/36 positive samples and 66.43% (95/143 of equine feces were positive. The results obtained for pH and OM from the soil had no correlation with the isolation of R. equi from soil samples of the different horse-breeding farms.

  18. Oxidative stress and DNA damage in horses naturally infected with Theileria equi.

    Science.gov (United States)

    Radakovic, M; Davitkov, D; Borozan, S; Stojanovic, S; Stevanovic, J; Krstic, V; Stanimirovic, Z

    2016-11-01

    The aim of this study was to determine the concentrations of oxidative stress parameters and DNA damage in horses infected by Theileria equi. Initial screening of 110 horses with duplex PCR enabled the selection of 30 infected horses with T. equi and 30 free of infection (control). Specimens from the 60 horses were further analysed by determining the following oxidative stress parameters: extent of haemolysis (EH), plasma free haemoglobin (PHb), catalase (CAT), Cu,Zn superoxide dismutase (SOD1), paraoxonase (PON1), nitrite (NO2(-)), total nitrate and nitrite (NOx), malondialdehyde (MDA) and free thiol groups (-SH). In addition, relative distribution of lactate dehydrogenase (LDH1-LDH5) activity and the DNA-damaging effects of T. equi infection were evaluated. Compared to control horses, horses infected with T. equi had significantly higher SOD1 activities (P horses had significantly increased LDH5 isoenzyme activities (P horses, which resulted in changes in osmotic fragility, damage to lipids, proteins and DNA, haemolysis and hepatocellular damage. Oxidative stress in horses naturally infected with T. equi could contribute to the pathogenesis of the infection.

  19. 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.

  20. 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.

  1. Conjugal transfer of a virulence plasmid in the opportunistic intracellular actinomycete Rhodococcus equi.

    Science.gov (United States)

    Tripathi, V N; Harding, W C; Willingham-Lane, J M; Hondalus, M K

    2012-12-01

    Rhodococcus equi is a facultative intracellular, Gram-positive, soilborne actinomycete which can cause severe pyogranulomatous pneumonia with abscessation in young horses (foals) and in immunocompromised people, such as persons with AIDS. All strains of R. equi isolated from foals and approximately a third isolated from humans contain a large, ~81-kb plasmid which is essential for the intramacrophage growth of the organism and for virulence in foals and murine in vivo model systems. We found that the entire virulence plasmid could be transferred from plasmid-containing strains of R. equi (donor) to plasmid-free R. equi strains (recipient) at a high frequency and that plasmid transmission reestablished the capacity for intracellular growth in macrophages. Plasmid transfer required living cells and cell-to-cell contact and was unaffected by the presence of DNase, factors pointing to conjugation as the major means of genetic transfer. Deletion of a putative relaxase-encoding gene, traA, located in the proposed conjugative region of the plasmid, abolished plasmid transfer. Reversion of the traA mutation restored plasmid transmissibility. Finally, plasmid transmission to other Rhodococcus species and some additional related organisms was demonstrated. This is the first study showing a virulence plasmid transfer in R. equi, and it establishes a mechanism by which the virulence plasmid can move among bacteria in the soil.

  2. Elimination of Babesia microti Is Dependent on Intraerythrocytic Killing and CD4(+) T Cells.

    Science.gov (United States)

    Skariah, Sini; Arnaboldi, Paul; Dattwyler, Raymond J; Sultan, Ali A; Gaylets, Corey; Walwyn, Odaelys; Mulhall, Hannah; Wu, Xia; Dargham, Soha R; Mordue, Dana G

    2017-07-15

    Babesiosis is a tick-borne zoonosis caused by protozoans of the genus Babesia, apicomplexan parasites that replicate within erythrocytes. However, unlike related Plasmodium species, the pathogenesis of Babesia infection remains poorly understood. The primary etiological agent of babesiosis in the United States is B. microti. In healthy individuals, tick-transmitted infection with Babesia causes no specific clinical manifestations, with many having no symptoms at all. However, even in asymptomatic people, a Babesia carriage state can be established that can last up to a year or more. Current blood bank screening methods do not identify infected donors, and Babesia parasites survive blood-banking procedures and storage. Thus, Babesia can also be transmitted by infected blood, and it is currently the number one cause of reportable transfusion-transmitted infection in the United States. Despite a significant impact on human health, B. microti remains understudied. In this study, we evaluated the course of Babesia infection in three strains of mice, C57BL/6J, BALB/cJ, and C3H-HeJ, and examined the contribution of multiple immune parameters, including TLRs, B cells, CD4(+) cells, IFN-γ, and NO, on the level of parasitemia and parasite clearance during acute babesiosis. We found that B. microti reaches high parasitemia levels during the first week of infection in all three mice strains before resolving spontaneously. Our results indicate that resolution of babesiosis requires CD4 T cells and a novel mechanism of parasite killing within infected erythrocytes. Copyright © 2017 by The American Association of Immunologists, Inc.

  3. The Steroid Catabolic Pathway of the Intracellular Pathogen Rhodococcus equi Is Important for Pathogenesis and a Target for Vaccine Development

    NARCIS (Netherlands)

    van der Geize, R.; Grommen, A. W. F.; Hessels, G. I.; Jacobs, A. A. C.; Dijkhuizen, L.

    2011-01-01

    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 pu

  4. Sonoran propolis and some of its chemical constituents inhibit in vitro growth of Giardia lamblia trophozoites.

    Science.gov (United States)

    Alday-Provencio, Samuel; Diaz, Gabriela; Rascon, Lucila; Quintero, Jael; Alday, Efrain; Robles-Zepeda, Ramón; Garibay-Escobar, Adriana; Astiazaran, Humberto; Hernandez, Javier; Velazquez, Carlos

    2015-06-01

    Propolis is a cereus resin with a complex chemical composition that possesses a wide range of biological activities. The aim of this study was to evaluate the in vitro anti-Giardia lamblia activity of Sonoran propolis collected from three different areas of Sonoran Desert in northwestern Mexico (Caborca, Pueblo de Alamos, and Ures) and some of its chemical constituents. Additionally, we also analyzed the seasonal effect on the anti-G. lamblia activity of propolis. G. lamblia trophozoite cultures were treated with different concentrations of Sonoran propolis or chemical compounds during 48 h cell proliferation and cell viability were determined. Ures propolis showed the highest inhibitory activity against G. lamblia (IC50 63.8 ± 7.1 µg/mL) in a dose-dependent manner (Ures > Pueblo de Alamos > Caborca). Season had a significant effect on the in vitro anti-G. lamblia activity of Ures propolis. Summer propolis showed the highest inhibitory effect on the G. lamblia trophozoite growth (IC50 23.8 ± 2.3 µg/mL), followed by propolis collected during winter (IC50 59.2 ± 34.7 µg/mL), spring (IC50 102.5 ± 15.3 µg/mL), and autumn (IC50 125.0 ± 3.1 µg/mL). Caffeic acid phenethyl ester, an Ures propolis exclusive constituent, had the highest growth-inhibitory activity towards G. lamblia [IC50 63.1 ± 0.9 µg/mL (222.1 ± 3.2 µM)]. To our knowledge, this is the first study showing that caffeic acid phenethyl ester possesses antiparasitic activity against G. lamblia. Naringenin [IC50 125.7 ± 20.7 µg/mL (461.8 ± 76.3 µM)], hesperetin [IC50 149.6 ± 24.8 µg/mL (494.9 ± 82.2 µM)], and pinocembrin [IC50 174.4 ± 26.0 µg/mL (680.6 ± 101.7 µM)] showed weak anti-G. lamblia activity. On the other hand, chrysin and rutin did not show significant antiparasitic activity. In conclusion, our results suggest that Sonoran propolis and some of its chemical constituents had inhibitory effects on the

  5. First survey for Babesia bovis and Babesia bigemina infection in cattle from Central and Southern regions of Portugal using serological and DNA detection methods

    Science.gov (United States)

    Incidence of bovine babesiosis in Portugal is currently unknown. In this study, a first survey of Babesia bovis and B. bigemina infection in cattle was carried out using blood samples from 406 clinically healthy individuals from different districts from Central and Southern regions of Portugal and a...

  6. Redescription of Babesia capreoli (Enigk and Friedhoff, 1962) from roe deer (Capreolus capreolus): isolation, cultivation, host specificity, molecular characterisation and differentiation from Babesia divergens.

    Science.gov (United States)

    Malandrin, Laurence; Jouglin, Maggy; Sun, Yi; Brisseau, Nadine; Chauvin, Alain

    2010-03-01

    The recent use of the sole molecular identification of Babesia infecting European cervids has led to confusion between the closely related Babesia divergens and Babesia capreoli, and to their grouping together as "B. divergens-like". In order to clarify this taxonomic confusion, Babesia from roe deer, cattle and human blood were isolated, cultured and their biological as well as molecular characteristics compared. On this basis, we conclude that: (i) the parasites isolated from roe deer blood are B. capreoli; (ii) there are no intraspecific variations in the 18S rDNA within B. capreoli and B. divergens spp.; (iii) these two species are closely related as demonstrated by their morphology, serological cross-reactions and 99.83% identity in their 18S rDNA; (iv) these two species are distinct as demonstrated by their different abilities to grow in vitro in cattle, human and sheep erythrocytes, by their infectivity for gerbils, and by a conserved three bases difference at positions 631, 663 and 1637 of their 18S rDNA; (v) B. capreoli does not pose a threat to either humans or livestock. An integrated description is given of the host range, geographical distribution, biological and molecular characterisation of B. capreoli, and reference materials have been deposited at the Museum d'Histoire Naturelle de Paris.

  7. 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.

  8. Equid herpesvirus 1 and rhodococcus equi coinfection in a foal with bronchointerstitial pneumonia.

    Science.gov (United States)

    Perez-Ecija, Alejandro; Mendoza, Francisco Javier; Estepa, José Carlos; Bautista, María José; Pérez, José

    2016-10-01

    A 2-month-old foal with septic shock and severe respiratory distress was referred to the Veterinary Teaching Hospital. Due to poor prognosis, the foal was euthanized. Histopathology showed lesions suggestive of Rhodococcus equi infection associated with a diffuse interstitial infiltrate of foamy macrophages and syncytial cells presenting large acidophilic intranuclear inclusion bodies, fibrin exudates and hyaline membranes. Bacteriological examination from lung and respiratory exudates confirmed R. equi infection, whilst immunohistochemistry and PCR yielded a positive result for Equid herpesvirus type 1 (EHV-1). Several etiologies have been proposed for bronchointerstitial pneumonia in foals, although a multifactorial origin for this lesional pattern could be possible. This work is the first one describing a combined EHV-1 and R. equi infection in a foal affected with bronchointerstitial pneumonia.

  9. Mutant prevention concentration and mutant selection window for 10 antimicrobial agents against Rhodococcus equi.

    Science.gov (United States)

    Berghaus, Londa J; Giguère, Steeve; Guldbech, Kristen

    2013-10-25

    The objectives of this study were to determine the mutant prevention concentration (MPC), time above the MPC and mutant selection window for 10 antimicrobial agents against Rhodococcus equi and to determine if the combination of a macrolide with rifampin would decrease emergence of resistant mutants. Antimicrobial agents investigated (erythromycin, clarithromycin, azithromycin, rifampin, amikacin, gentamicin, enrofloxacin, vancomycin, imipenem, and doxycycline) were selected based on in vitro activity and frequency of use in foals or people infected with R. equi. Each antimicrobial agent or combination of agents was evaluated against four virulent strains of R. equi. MPC were determined using an agar plate assay. Pharmacodynamic parameters were calculated using published plasma and pulmonary pharmacokinetic variables. There was a significant (Pequi. Copyright © 2013 Elsevier B.V. All rights reserved.

  10. Susceptibility profile of Brazilian Rhodococcus equi isolates against azithromycin, clarithromycin and erythromycin

    Directory of Open Access Journals (Sweden)

    Letícia Trevisan Gressler

    2015-04-01

    Full Text Available Rhodococcus equi infection treatment is usually a macrolide (azithromycin - AZM, clarithromycin - CLR and erythromycin - ERY and rifampicin combination. However, resistance cases have been reported, especially for ERY. In view of the need of a study about Brazilian isolates susceptibility profile, this study aimed to characterize the minimum inhibitory concentration (MIC of the macrolides - AZM, CLR and ERY - against 44 R. equi isolates. It was found all isolates CLR and AZM sensitive; however, for ERY, 27% (12/44 were classified as intermediate sensitivity. R. equi Brazilian isolates used here showed a large susceptibility profile, except against ERY, for which it was observed some resistance evidence. In order to avoid failures in the equine rhodococcosis therapy it was highlighted the importance of microbiological culture and antimicrobial susceptibility testing in vitro before beginning treatment

  11. Rhodococcus equi human clinical isolates enter and survive within human alveolar epithelial cells.

    Science.gov (United States)

    Ramos-Vivas, J; Pilares-Ortega, L; Remuzgo-Martínez, S; Padilla, D; Gutiérrez-Díaz, J L; Navas-Méndez, J

    2011-05-01

    Rhodococcus equi is an emerging opportunistic human pathogen associated with immunosuppressed people, especially those infected with the human immunodeficiency virus (HIV). This pathogen resides primarily within lung macrophages of infected patients, which may explain in part its ability to escape normal pulmonary defense mechanisms. Despite numerous studies as a pulmonary pathogen in foals, where a plasmid seems to play an important role in virulence, information on the pathogenesis of this pathogen in humans is still scarce. In this study, fluorescence microscopy and vancomycin protection assays were used to investigate the ability of R. equi human isolates to adhere to and to invade the human alveolar epithelial cell line A549. Our findings indicate that some R. equi clinical strains are capable of adhering, entering and surviving within the alveolar cell line, which may contribute to the pathogen persistence in lung tissues. Copyright © 2011 Institut Pasteur. Published by Elsevier SAS. All rights reserved.

  12. 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.

  13. Multiplex PCR for molecular screening of Borrelia burgdorferi sensu lato, Anaplasma spp. and Babesia spp.

    Science.gov (United States)

    Rodríguez, Islay; Burri, Caroline; Noda, Angel A; Douet, Véronique; Gern, Lise

    2015-01-01

    Ticks transmit a great variety of pathogenic microorganisms to humans and animals. The detection of tick-borne pathogens (TBP) is mainly by molecular techniques based on polymerase chain reactions (PCR). To design and evaluate a multiplex PCR for the molecular screening of zoonotic TBP for exploratory studies. Control DNA from reference strains, DNA from experimentally-infected biological specimens, and from Rhipicephalus sanguineus ticks collected from domestic and homeless dogs were used. A multiplex PCR assay to detect the presence of Borrelia burgdorferi sensu lato, Anaplasma spp. and Babesia spp. was designed and optimized using primers previously reported for B. burgdorferi sensu lato and Anaplasma spp., while for Babesia spp. they were designed in silico. The multiplex PCR was evaluated on the DNA from biological samples. A new set of specific primers for Babesia spp. was designed. Adjustment of the master mix reactive concentrations and amplification conditions for the multiplex PCR allowed the successful amplification of the specific amplicons for each microbial group from the control DNA and experimentally-infected biological specimens. The efficiency of the multiplex PCR amplifying three DNA targets was confirmed. Individual and co-infection of Anaplasma spp. and Babesia spp. were detected in the R. sanguineus ticks from dogs. A multiplex PCR assay for the screening of three TBP is available. By using it, B. burgdorferi sensu lato, Anaplasma spp. and Babesia spp. can be detected accurately in one PCR reaction.

  14. In vitro growth inhibitory efficacy of some target specific novel drug molecules against Theileria equi.

    Science.gov (United States)

    Gopalakrishnan, A; Maji, C; Dahiya, R K; Suthar, A; Kumar, R; Gupta, A K; Dimri, U; Kumar, S

    2016-02-15

    The in vitro growth inhibitory efficacies of five drug molecules against Theileria equi were evaluated in in vitro cultured parasites. A continuous microaerophilic stationary-phase culture (MASP) system was established for propagation of T. equi parasites. This in vitro culture system was used to assess the growth inhibitory effect of harmaline hydrochloride dihydrate (HHD), hexadecyltrimethylammonium bromide (HDTAB), hesparidin methyl chalcone (HMC), andrographolide and imidocarb dipropionate against T. equi. The 50% inhibitory concentration value of HHD, HDTAB, HMC, and imidocarb dipropionate for T. equi growth were 17.42 μM, 14.00 μM, 246.34 μM and 0.279 μM (equivalent to 0.139 μg/ml), respectively (P<0.05). The andrographolide was not effective in inhibiting in vitro growth of T. equi in the present study. Furthermore, the in vitro cytotoxicity of these five drugs was evaluated on horse PBMC. At 2000 μM concentration of HHD, HDTAB, HMC, andrographolide and imidocarb dipropionate were 8.34, 46.44, 58.53, 31.06, 15.14% cytotoxic on PBMC, respectively. Out of our four tested drug molecules, HHD was having low IC50 value along with least cytotoxicity, as compared to reference drug imidocarb dipropionate. The difference in IC50 value of HDTAB and HHD was significant, but HDTAB was moderately more cytotoxic on PBMC cell lines. HHD and HDTAB are selective inhibitor for heat shock protein 90 (Hsp90) and choline kinase pathway. It can be concluded that HHD and HDTAB are potential drug molecules against T. equi parasite by acting on Hsp90 and choline kinase pathway.

  15. 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.

  16. 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.

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

    Science.gov (United States)

    McQueen, Cole M; Doan, Ryan; Dindot, Scott V; Bourquin, Jessica R; Zlatev, Zlatomir Z; Chaffin, M Keith; Blodgett, Glenn P; Ivanov, Ivan; Cohen, Noah D

    2014-01-01

    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.

  18. The hydroxamate siderophore rhequichelin is required for virulence of the pathogenic actinomycete Rhodococcus equi.

    Science.gov (United States)

    Miranda-Casoluengo, Raúl; Coulson, Garry B; Miranda-Casoluengo, Aleksandra; Vázquez-Boland, José A; Hondalus, Mary K; Meijer, Wim G

    2012-12-01

    We previously showed that the facultative intracellular pathogen Rhodococcus equi produces a nondiffusible and catecholate-containing siderophore (rhequibactin) involved in iron acquisition during saprophytic growth. Here, we provide evidence that the rhbABCDE cluster directs the biosynthesis of a hydroxamate siderophore, rhequichelin, that plays a key role in virulence. The rhbC gene encodes a nonribosomal peptide synthetase that is predicted to produce a tetrapeptide consisting of N(5)-formyl-N(5)-hydroxyornithine, serine, N(5)-hydroxyornithine, and N(5)-acyl-N(5)-hydroxyornithine. The other rhb genes encode putative tailoring enzymes mediating modification of ornithine residues incorporated into the hydroxamate product of RhbC. Transcription of rhbC was upregulated during growth in iron-depleted medium, suggesting that it plays a role in iron acquisition. This was confirmed by deletion of rhbCD, rendering the resulting strain R. equi SID2 unable to grow in the presence of the iron chelator 2,2-dipyridyl. Supernatant of the wild-type strain rescued the phenotype of R. equi SID2. The importance of rhequichelin in virulence was highlighted by the rapid increase in transcription levels of rhbC following infection and the inability of R. equi SID2 to grow within macrophages. Unlike the wild-type strain, R. equi SID2 was unable to replicate in vivo and was rapidly cleared from the lungs of infected mice. Rhequichelin is thus a key virulence-associated factor, although nonpathogenic Rhodococcus species also appear to produce rhequichelin or a structurally closely related compound. Rhequichelin biosynthesis may therefore be considered an example of cooption of a core actinobacterial trait in the evolution of R. equi virulence.

  19. Plasmid Profiles of Virulent Rhodococcus equi Strains Isolated from Infected Foals in Poland.

    Science.gov (United States)

    Kalinowski, Marcin; Grądzki, Zbigniew; Jarosz, Łukasz; Kato, Kiyoko; Hieda, Yu; Kakuda, Tsutomu; Takai, Shinji

    2016-01-01

    Rhodococcus equi is an important bacterial pathogen in foals up to 6 months old, widespread in horse farms all over the world. It was found that only virulent R. equi strains expressing 15-17 kDa virulence-associated protein (VapA) and having large virulence plasmid of 85-90 kb containing vapA gene are pathogenic for horses. To date, 12 plasmid types have been reported in VapA positive strains from horses. There are no data concerning plasmid types of Polish field R. equi strains isolated from horses and horse farm environment. The aim of the study is to determine plasmid profiles of virulent R. equi strains isolated in Poland from dead foals as well as from soil samples taken from horse breeding farms. Plasmid profiles of 10 clinical strains derived from 8 farms and 11 environmental strains from 3 farms, confirmed as virulent by PCR, were compared with 12 reference strains containing the known plasmid size and type. Plasmid DNAs were analysed by digestion with the restriction endonucleases BamHI, EcoRI, EcoT22I, and HindIII for detailed comparison and estimation of plasmid sizes. The results of RFLP analysis revealed that all except one isolates used in the study are classified as VapA 85 kb type I plasmid. One strain harboured VapA 87 kb type I plasmid. This is the first report of plasmid types of Polish field R. equi strains. The results of our preliminary investigations on horse farms located in central and eastern Poland indicate that the virulent R. equi strains thus far isolated from diseased foals and horse farms environment represent a highly uniform plasmid pattern.

  20. Multiplex PCR for detection of Trypanosoma evansi and Theileria equi in equids of Punjab, India.

    Science.gov (United States)

    Sumbria, Deepak; Singla, L D; Sharma, Amrita; Bal, M S; Kumar, Sanjay

    2015-07-30

    Multiplex PCR for simultaneous detection of Trypanosoma evansi and Theileria equi in single-step reaction was optimized and employed on 108 equids (99 horses and 9 donkeys/mules) blood samples collected from two agro-climatic zones (Sub-mountain undulating zone and Undulating plain zone) of Punjab to evaluate the status of concurrent infection and associated risk factors. The amplification products of 257 and 709 bp targeting repetitive nucleotide sequence of variable surface glycoproteins of T. evansi and 18S rRNA gene of T. equi, respectively expressed high fidelity of the primer pairs with sequence homology to neighboring geographic isolates. The overall prevalence of T. evansi and T. equi was 3.7 and 1.85%, with Undulating plain zone at higher infection risk for T. equi (OR=3.24, 95% CI=0.28-83.65); and Sub-mountain undulating zone (OR=∞, 95% CI=0.25-∞) for T. evansi. Multiplex PCR revealed higher risk of infection of both T. equi (OR=6.75, 95% CI=0.58-175.38) and T. evansi (OR=2.11, 95% CI=0.05-80.36) in the farms with inappropriate management system. The risk factor associated with the type of host species had an odds ratio of 12.35 (95% CI=0.29-508.37) for donkeys/mules versus horses for T. evansi infection. This group was also at higher risk of infection with Odds ratio (OR) of 4 (95% CI=0.14-53.99) for T. equi. The current investigation brings out various commodities at risk of infection pertaining to equid trypanosomosis and theileriosis evaluated by a rapid and sensitive multiplex PCR assay.

  1. 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...... 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....

  2. 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.

  3. In vitro efficacy of corifungin against Acanthamoeba castellanii trophozoites and cysts.

    Science.gov (United States)

    Debnath, Anjan; Tunac, Josefino B; Silva-Olivares, Angélica; Galindo-Gómez, Silvia; Shibayama, Mineko; McKerrow, James H

    2014-01-01

    Painful blinding keratitis and fatal granulomatous amebic encephalitis are caused by the free-living amebae Acanthamoeba spp. Several prescription eye medications are used to treat Acanthamoeba keratitis, but the infection can be difficult to control because of recurrence of infection. For the treatment of encephalitis, no single drug was found useful, and in spite of the use of a combination of multiple drugs, the mortality rate remains high. Therefore, efficient, novel drugs are urgently needed for the treatment of amebic keratitis and granulomatous amebic encephalitis. In this study, we identified corifungin, a water-soluble polyene macrolide, as amebicidal. In vitro, it was effective against both the trophozoites and the cysts. Transmission electron microscopy of Acanthamoeba castellanii incubated with corifungin showed the presence of swollen mitochondria, electron-dense granules, degeneration of cytoplasm architecture, and loss of nuclear chromatin structure. These changes were followed by lysis of amebae. Corifungin also induced the encystment process of A. castellanii. There were alterations in the cyst cell wall followed by lysis of the cysts. Corifungin is a promising therapeutic option for keratitis and granulomatous amebic encephalitis.

  4. Yersinia pseudotuberculosis IP32953 survives and replicates in trophozoites and persists in cysts of Acanthamoeba castellanii.

    Science.gov (United States)

    Santos-Montañez, Jennifer; Benavides-Montaño, Javier A; Hinz, Angela K; Vadyvaloo, Viveka

    2015-07-01

    Yersinia pseudotuberculosis is a foodborne enteric pathogen that causes a mild self-limiting diarrhea in humans. Yersinia pseudotuberculosis is able to persist in soil and water and in association with fresh produce, but the mechanism by which it persists is unknown. It has been shown that Y. pseudotuberculosis co-occurs with protozoans in these environments; therefore, this study investigates if bacterivorous free-living amoeba (FLA) are able to support persistence of Y. pseudotuberculosis. Coculture studies of Y. pseudotuberculosis and the prototype FLA, Acanthamoeba castellanii revealed that bacteria had an enhanced capacity to survive in association with amoeba and in the absence of any cytotoxic effects. Yersinia pseudotuberculosis is able to survive and replicate in trophozoites specifically localized within vacuoles, and persists within cysts over a period of at least a week. These data present the first evidence that Y. pseudotuberculosis is able to resist the bacterivorous nature of FLA and instead exhibits an enhanced ability to replicate and persist in coculture with amoeba. This study sheds light on the potential role of FLA in the ecology of Y. pseudotuberculosis which may have implications for food safety.

  5. Characterization of cyst and trophozoite proteins of environmental isolates of Acanthamoeba castellanii by two-dimensional gel electrophoresis.

    Science.gov (United States)

    Dao, Yap Ray Ming; Chooi, Lim Patricia Kim; Li, Chan Li; Fung, Wong Shew; Wah, Mak Joon

    2014-03-01

    Acanthamoeba castellanii has been known to possess pathogenic properties, such as acanthamoebic keratitis and granulomatous amoebic encephalitis. The role of proteases and proteins in the pathogenesis of these infections is still poorly understood. As Acanthamoeba sp is a ubiquitous protozoon found in the natural environment they can potentially cause human infections. This study characterized cyst and trophozoite proteins of 3 environmental A. castellanii isolates in comparison with a clinical isolate, ATCC 50492. The latter and environmental IMU1 isolate had 100% genotype identity with A. castellanii and demonstrated protein spots with higher molecular weights (> 95 kDa) at relatively higher isoelectric values (> pI 7.00) compared to the two other environmental isolates (IMU4 and IMU5) that had 99% genotype identity to A. castellanii based on 16 S rDNA sequence. Thus such trophozoite proteins may be involved with the parasite's ability to cause acanthamoebic keratitis.

  6. Transmissão transovariana de Babesia bovis em Boophilus microplus: obtenção de cepa de carrapato livre de Babesia spp. Babesia bovis transovarian transmission in Boophilus microplus: obtention of a Babesia free tick strain

    Directory of Open Access Journals (Sweden)

    Tânia Regina Bettin dos Santos

    2000-06-01

    Full Text Available O presente trabalho objetivou o estudo de parte do ciclo da Babesia bovis no seu hospedeiro invertebrado, o carrapato Boophilus microplus. Analisou-se a capacidade de infeccção e transmissão transovariana de B. bovis em partenóginas de B. microplus, alimentadas em bovinos portadores e enfermos por esse protozoário. No 18º dia após a infestação, coletaram-se partenóginas diretamente do corpo dos bovinos e teleóginas após o desprendimento natural, a partir do 21º dia. Todos os grupos foram incubados a 27ºC e umidade relativa superior a 70%. No 5º dia após o início da postura, realizou-se o exame de hemolinfa a fim de diagnosticar a infecção dos ínstares por B. bovis. A ausência de infecção detectada no exame de hemolinfa foi confirmada posteriormente com o teste biológico, revelando que partenóginas não transmitem B. bovis transovarianamente. Esses resultados oferecem uma técnica simplificada para a obtenção de cepas de carrapatos livres de B. bovis.In this experiment part of the life cycle of Babesia bovis in its invertebrate host, the tick Boophilus microplus was studied. In order to evaluate the capacity of infection and transmission of B. bovis were collected semi-engorged females of B. microplus fed on carrier and ill bovines. In the 18th day after infestation, semi-engorged females were collected directly from bovine bodies and after 21st day engorged females dropped on the ground. All the collected groups were incubated at 27°C and relative humidity greater than 70%. At the 5th day of oviposition the diagnosis was made by direct examination of haemolymph smears. The biological test reveled that B. bovis transovarial transmission doesn't happer in semi-engorged females. The results offer a simple techique to obtain strains of ticks free of B. bovis.

  7. The principal conductance in Giardia lamblia trophozoites possesses functional properties similar to the mammalian ClC-2 current.

    Science.gov (United States)

    Moreno-Galindo, Eloy G; Rodríguez-Elías, Julio C; Ramírez-Herrera, Mario A; Sánchez-Chapula, José A; Navarro-Polanco, Ricardo A

    2014-05-01

    The human intestinal pathogen Giardia lamblia is a flagellated unicellular protozoan with pronounced medical and biological relevance. However, the basic physiology of Giardia trophozoites has been sparsely studied, especially the electrical and ionic properties of their cellular membrane which are virtually unknown. In this study, we were able to record and characterize the macroscopic ionic currents of Giardia trophozoite membrane by electrophysiological methods of the patch clamp technique. Giardia trophozoites showed a high current density (∼600 pA/pF at -140 mV) that was activated upon hyperpolarization. This current was carried by a chloride-selective channel (I Cl-G) and it was the most important determinant of the membrane potential in Giardia trophozoites. Moreover, this conductance was able to carry other halide anions and the sequence of permeability was Br(-) > Cl(-) ≈ I(-) ≫ F(-). Besides the voltage-dependent inward-rectifying nature of I Cl-G, its activation and deactivation kinetics were comparable to those observed in ClC-2 channels. Extracellular pH modified the voltage-dependent properties of I Cl-G, shifting the activation curve from a V 1/2 = -79 ± 1 mV (pH 7.4) to -93 ± 2 mV (pH 8.4) and -112 ± 2 mV (pH 5.4). Furthermore, the maximal amplitude of I Cl-G measured at -100 mV showed dependence to external pH in a bell-shaped fashion reported only for ClC-2 channels. Therefore, our results suggest that I Cl-G possesses several functional properties similar to the mammalian ClC-2 channels.

  8. Inhibitory effects of (-)-epigallocatechin-3-gallate from green tea on the growth of Babesia parasites.

    Science.gov (United States)

    Aboulaila, M; Yokoyama, N; Igarashi, I

    2010-04-01

    (-)-Epigallocatechin-3-gallate (EGCG) is the major tea catechin and accounts for 50-80% of the total catechin in green tea. (-)-Epigallocatechin-3-gallate has antioxidant, anti-inflammatory, anti-microbial, anti-cancer, and anti-trypanocidal activities. This report describes the inhibitory effect of (-)-Epigallocatechin-3-gallate on the in vitro growth of bovine Babesia parasites and the in vivo growth of the mouse-adapted rodent babesia B. microti. The in vitro growth of the Babesia species was significantly (Pparasites showed no re-growth at 25 microM for B. bovis and B. bigemina in the subsequent viability test. The drug significantly (Pparasites completely cleared on day 14 and 16 post-inoculation in the 5 and 10 mg/kg treated groups, respectively. These findings highlight the potentiality of (-)-Epigallocatechin-3-gallate as a chemotherapeutic drug for the treatment of babesiosis.

  9. 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

    2015-10-01

    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.

  10. Naegleria fowleri glycoconjugates with residues of α-D-mannose are involved in adherence of trophozoites to mouse nasal mucosa.

    Science.gov (United States)

    Carrasco-Yepez, Maricela; Campos-Rodriguez, Rafael; Godinez-Victoria, Marycarmen; Rodriguez-Monroy, Marco Aurelio; Jarillo-Luna, Adriana; Bonilla-Lemus, Patricia; De Oca, Arturo Contis-Montes; Rojas-Hernandez, Saul

    2013-10-01

    We analyzed the possible role of glycoconjugates containing α-D-mannose and α-D-glucose residues in adherence of trophozoites to mouse nasal epithelium. Trophozoites incubated with 20 μg of one of three different lectins which preferentially recognized these residues were inoculated intranasally in Balb/c mice. Mouse survival was 40% with Pisum sativum and Canavalia ensiformis and 20% with Galanthus nivalis amebic pretreatment, compared with 0% survival for control animals administered trophozoites without pretreatment. Possibly some of the glycoproteins found in Naegleria fowleri represent an adherence factor. Differences in the saccharide sequences of the Naegleria species, even on the same glycoconjugate structure, could explain the different results corresponding to the distinct pretreatments (C. ensiformis, G. nivalis, and P. sativum). We found a higher expression of glycoconjugates recognized by P. sativum in Naegleria lovaniensis than N. fowleri, probably due to the higher number of oligosaccharides containing an α-1,6-linked fucose moiety expressed on the former species.

  11. The EhADH112 recombinant polypeptide inhibits cell destruction and liver abscess formation by Entamoeba histolytica trophozoites.

    Science.gov (United States)

    Martínez-López, Carolina; Orozco, Esther; Sánchez, Tomás; García-Pérez, Rosa María; Hernández-Hernández, Fidel; Rodríguez, Mario A

    2004-04-01

    The Entamoeba histolytica EhCPADH complex, formed by a cysteine proteinase (EhCP112) and an adhesin (EhADH112), is involved in adherence, phagocytosis and cytolysis. This makes this complex an attractive candidate as a vaccine against amoebiasis. Here, we produced the recombinant polypeptide EhADH243, which includes the adherence epitope detected by a monoclonal antibody against the EhCPADH complex. EhADH243 was purified, and the effect of the polypeptide on in vitro and in vivo virulence was studied. Antibodies against EhADH243 reacted with the EhCPADH complex and with the recombinant polypeptide. EhADH243 and antibodies against this polypeptide inhibited adherence, phagocytosis and destruction of cell monolayers by live trophozoites, but had little effect on cell monolayer destruction by trophozoite extracts. EhADH243 recognized a 97 kDa protein in the MDCK membrane fraction that could be a putative receptor for E. histolytica trophozoites. Hamsters immunized with EhADH243 developed humoral response against EhCPADH, and animals were partially protected from amoebic liver abscess.

  12. 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

  13. 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

  14. Transfer of the virulence-associated protein a-bearing plasmid between field strains of virulent and avirulent Rhodococcus equi.

    Science.gov (United States)

    Stoughton, W; Poole, T; Kuskie, K; Liu, M; Bishop, K; Morrissey, A; Takai, S; Cohen, N

    2013-01-01

    Virulent and avirulent isolates of Rhodococcus equi coexist in equine feces and the environment and are a source of infection for foals. The extent to which plasmid transfer occurs among field strains is ill-defined and this information is important for understanding the epidemiology of R. equi infections of foals. To estimate the frequency of transfer of the virulence plasmid between virulent and avirulent strains of R. equi derived from foals and their environment. None. In vitro study; 5 rifampin-susceptible, virulent R. equi isolates obtained from clinically affected foals or air samples from a farm with a history of recurrent R. equi foal pneumonia were each mixed with 5 rifampin-resistant, avirulent isolates derived from soil samples, using solid medium, at a ratio of 10 donor cells (virulent) per recipient cell. Presumed transconjugates were detected by plating on media with rifampin and colony immunoblotting to detect the presence of the virulence-associated protein A. Three presumed transconjugates were detected among 2,037 recipient colonies, indicating an overall estimated transfer frequency of 0.15% (95% CI, 0.03–0.43%). All 3 transconjugates were associated with a single donor and 2 recipient strains. Genotyping and multiplex PCR of presumed transconjugates demonstrated transfer of the virulence-associated protein A-bearing plasmid between virulent and avirulent R. equi. Transfer of the virulence plasmid occurs with relatively high frequency. These findings could impact strategies to control or prevent R. equi through environmental management.

  15. First report of Babesia divergens infection in an HIV patient

    Directory of Open Access Journals (Sweden)

    Luis M. González

    2015-04-01

    Full Text Available Human babesiosis is a zoonosis primarily transmitted through Ixodes ticks and alternatively by routes such as blood transfusions from asymptomatic donors. We report the first case of human babesiosis caused by Babesia divergens in a patient with HIV. This study also focuses on elucidating the possible transmission route of infection in this patient, who received numerous blood transfusions but showed patent symptoms only after splenectomy. A battery of detection tools along with a novel Western-Blot Assay and Enzyme Linked Immunosorbent Assay using the major surface protein of B. divergens (Bd37 as a target were used to evaluate the presence of B. divergens or antibodies against the parasite in samples from the patient and the blood donors involved in this case. A retrospective study of the humoral status against the parasite revealed B. divergens IgG antibodies in one of the implicated donors, but also showed that the patient had been already exposed to the parasite before any transfusion. Thus, this analysis of natural and transfusion transmission routes suggests a pre-existing subclinical babesiosis in the patient.

  16. Tick passage results in enhanced attenuation of Babesia bovis.

    Science.gov (United States)

    Sondgeroth, Kerry S; McElwain, Terry F; Ueti, Massaro W; Scoles, Glen A; Reif, Kathryn E; Lau, Audrey O T

    2014-10-01

    Serial blood passage of virulent Babesia bovis in splenectomized cattle results in attenuated derivatives that do not cause neurologic disease. Tick transmissibility can be lost with attenuation, but when retained, attenuated B. bovis can revert to virulence following tick passage. This study provides data showing that tick passage of the partially attenuated B. bovis T2Bo derivative strain further decreased virulence compared with intravenous inoculation of the same strain in infected animals. Ticks that acquired virulent or attenuated parasites by feeding on infected cattle were transmission fed on naive, splenectomized animals. While there was no significant difference between groups in the number of parasites in the midgut, hemolymph, or eggs of replete female ticks after acquisition feeding, animals infected with the attenuated parasites after tick transmission showed no clinical signs of babesiosis, unlike those receiving intravenous challenge with the same attenuated strain prior to tick passage. Additionally, there were significantly fewer parasites in blood and tissues of animals infected with tick-passaged attenuated parasites. Sequencing analysis of select B. bovis genes before and after tick passage showed significant differences in parasite genotypes in both peripheral blood and cerebral samples. These results provide evidence that not only is tick transmissibility retained by the attenuated T2Bo strain, but also it results in enhanced attenuation and is accompanied by expansion of parasite subpopulations during tick passage that may be associated with the change in disease phenotype. Copyright © 2014, American Society for Microbiology. All Rights Reserved.

  17. New methods for the diagnosis of Babesia bigemina infection

    Directory of Open Access Journals (Sweden)

    S. Morzaria

    1992-01-01

    Full Text Available Accurate diagnosis of Babesia bigemina infection, an economically important tick-transmitted protozoan parasite of cattle, is essential in the management of disease control and in epidemiological studies. The currentlyused methods of diagnosis are blood smear examination and serological tests which include agglutination and immunofluorescence tests. These testes have been used the fild but because they lack sensitivity and specificity, never and improved methods of diagnosis are being developed. The quantitative buffy coat (OBC method, using microhaematocrit tubes and acridine orange staining allows rapid and quicker diagnosis of B. bigemina and other blood parasites compared to light microscopic examination of stained smears. Parasite specific monoclonal antibodies have been used in antigen/antibody capture enzymelinked immunosorbent assays with grater sensitivity and specificity than previously described serological tests. Similary, DNA probes, derived from a repetitive sequence of the B. bigemina genome, offer a method of detecting very small numbers of parasites which are undetectable by conventional microscopy. An extrachromosomal DNA element, present in all the tick-borne protozoan parasites so far tested, provides an accurate means of diferentiating mixed parasite populations in infected animals. These improved methods will greatly facilitate epidemiological studies.

  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...... increased and may play a role in the lack of a bleeding phenotype, despite severe thrombocytopenia, in canine babesiosis....

  19. Effects of administration of live or inactivated virulent Rhodococccus equi and age on the fecal microbiome of neonatal foals.

    Directory of Open Access Journals (Sweden)

    Angela I Bordin

    Full Text Available BACKGROUND: Rhodococcus equi is an important pathogen of foals. Enteral administration of live, virulent R. equi during early life has been documented to protect against subsequent intrabronchial challenge with R. equi, indicating that enteral mucosal immunization may be protective. Evidence exists that mucosal immune responses develop against both live and inactivated micro-organisms. The extent to which live or inactivated R. equi might alter the intestinal microbiome of foals is unknown. This is an important question because the intestinal microbiome of neonates of other species is known to change over time and to influence host development. To our knowledge, changes in the intestinal microbiome of foals during early life have not been reported. Thus, the purpose of this study was to determine whether age (during the first month of life or administration of either live virulent R. equi (at a dose reported to protect foals against subsequent intrabronchial challenge, viz., 1×10(10 colony forming units [CFU] or inactivated virulent R. equi (at higher doses, viz., 2×10(10 and 1×10(11 [CFU] altered the fecal microbiome of foals. METHODOLOGY/PRINCIPAL FINDINGS: Fecal swab samples from 42 healthy foals after vaccination with low-dose inactivated R. equi (n = 9, high-dose inactivated R. equi (n = 10, live R. equi (n = 6, control with cholera toxin B (CTB, n = 9, and control without CTB (n = 8 were evaluated by 454-pyrosequencing of the 16S rRNA gene and by qPCR. No impact of treatment was observed among vaccinated foals; however, marked and significant differences in microbial communities and diversity were observed between foals at 30 days of age relative to 2 days of age. CONCLUSIONS: The results suggest age-related changes in the fecal microbial population of healthy foals do occur, however, mucosal vaccination does not result in major changes of the fecal microbiome in foals.

  20. 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

  1. Rhodococcus equi infection in HIV-positive subjects: a retrospective analysis of 24 cases.

    Science.gov (United States)

    Arlotti, M; Zoboli, G; Moscatelli, G L; Magnani, G; Maserati, R; Borghi, V; Andreoni, M; Libanore, M; Bonazzi, L; Piscina, A; Ciammarughi, R

    1996-01-01

    Rhodococcus equi causes a rare infection in immunocompromised hosts. We describe 24 cases of infection in patients with AIDS-related complex (ARC)/acquired immunodeficiency syndrome (AIDS). Pneumonia was always the first manifestation of R. equi infection, but extrapulmonary involvement was also observed. The main sources of bacteria were sputum, bronchial washings and blood. The strains isolated were mainly susceptible to erythromycin, vancomycin, teicoplanin, rifampicin, imipenem and aminoglycosides. Initial treatment should involve an intravenously administered antibiotic combination therapy including imipenem or vancomycin or teicoplanin, followed by orally administered maintenance combination therapy. Drug combinations should be investigated for serum bactericidal activity in vitro. Surgery does not increase survival time and should only be performed in cases that do not respond to antibiotic treatment. Presumptive risks of infection (contact with horses or farm dust, or cohabiting with people affected by R. equi infection) were present in more than 50% of patients. This finding, and the frequency of bacteria in the sputum, are not sufficient proof of transmission between humans, but do suggest the need for respiratory isolation of patients affected by R. equi pneumonia.

  2. Development of loop-mediated isothermal amplification (LAMP) for detection of Theileria equi.

    Science.gov (United States)

    Xie, Junren; Liu, Guangyuan; Tian, Zhancheng; Luo, Jin

    2013-09-01

    Several approaches have been developed for diagnosis of Theileria equi infection in horses and donkeys but all of them have limitations in practice. Due to numerous strengths including easy operation, cheapness and high sensitivity and specificity, LAMP has been already extensively used for surveillance of a number of diseases. We here set up a LAMP assay based on 18S rRNA gene for T. equi diagnosis. The approach was specific enough to differentiate T. equi from other evolutionary-related protozoa. Moreover, it was sensitive enough that LAMP was capable of detecting as much low as 10 copy target gene and 1 pg/μl blood genomic DNA. It was further demonstrated that LAMP was much more sensitive than canonical blood smear and comparable to PCR using test and field samples. The present results support an idea that LAMP developed in this study is reliable, reproducible and highly sensitive and specific, being a potential to be globally used for surveillance of T. equi infection in the field.

  3. 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. equ

  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 in AIDS patients: retrospective analysis of 13 patients in Argentina].

    Science.gov (United States)

    Corti, Marcelo; Solari, Rubén; De Carolis, Luis; Palmieri, Omar; Rollet, Raquel; Shah, Haroun N

    2014-08-01

    Rhodococcus equi is a gram positive coccoid rod that causes pulmonary infections in immunosuppressed patients. We retrospectively analyzed epidemiological, clinical, microbiological, radiological, and immunological features as well as the outcomes of 13 AIDS patients with R. equi infection. Between January 1994 and December 2012, 13 patients attending the AIDS department of the Infectious Diseases reference hospital in Buenos Aires were diagnosed with R. equi infection. All were men, the median age was 27 years. At the time of diagnosis, the median of CD4+ T cell counts was 11 cells/μl Twelve patients presented pulmonary disease with isolation of the microorganism from sputum or bronchoalveolar lavage; in the other patient the diagnosis was postmortem with positive culture of cerebrospinal fluid. The most frequent clinical manifestations were fever, haemoptysis, and weight loss. The predominant radiological finding was lobe consolidation with cavitation. Nine patients died after a median survival of 5.5 months. In all of them, cultures persisted positive until the last admission. The other 4 patients did continue clinical follow-ups. The insidious course of R. equi disease and the difficulties in the isolation of the microorganism contribute to the delay in the diagnosis and to the high mortality rate of this opportunistic infection.

  6. 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.

  7. Studies on a new candidate vaccine for Rhodococcus equi infections in foals

    NARCIS (Netherlands)

    Jacobs, Anton A. C.; Grommen, Ries; Hessels, Gerda; Dijkhuizen, Lubbert; van der Geize, Robert

    2012-01-01

    Rhodococcus (R.) equi is a facultative intracellular pathogen that causes severe pyogranulomatous pneumonia in foals up to 5 months of age. Despite the great need for a prophylactic measure against this devastating disease, no commercial vaccine is available. Today only long-term and cumbersome

  8. Development of a loop-mediated isothermal amplification method for detecting virulent Rhodococcus equi.

    Science.gov (United States)

    Kinoshita, Yuta; Niwa, Hidekazu; Higuchi, Tohru; Katayama, Yoshinari

    2016-09-01

    Rhodococcus equi is the most important causative bacterium of severe pneumonia in foals. We report herein the development of a specific loop-mediated isothermal amplification (LAMP) assay, which targets a gene encoding vapA for detecting virulent R. equi The detection limit of the LAMP assay was 10(4) colony forming units (CFU)/mL, which was equal to 10 CFU/reaction. The clinical efficacy of the LAMP assay was compared with those of 2 published PCR-based methods: nested PCR and quantitative real-time (q)PCR. Agreements between bacterial culture, which is the gold standard for detection of R. equi, and each of the 3 molecular tests were measured by calculating a kappa coefficient. The kappa coefficients of the LAMP (0.760), nested PCR (0.583), and qPCR (0.888) indicated substantial agreement, moderate agreement, and almost perfect agreement, respectively. Although the clinical efficacy of LAMP was not the best among the 3 methods tested, LAMP could be more easily introduced into less well-equipped clinics because it does not require special equipment (such as a thermocycler) for gene amplification. Veterinary practitioners could diagnose R. equi pneumonia more quickly by using LAMP and could use the results to select an appropriate initial treatment. © 2016 The Author(s).

  9. Influence of Rhodococcus equi on the respiratory burst of resident alveolar macrophages from horses

    Energy Technology Data Exchange (ETDEWEB)

    Brumbaugh, G.W.

    1986-01-01

    Rhodococcus equi is the etiologic agent of a devastating pneumonia of sporadic incidence in foals. The purpose of this study was to evaluate the influence of R. equi on the superoxide anion production, measured spectrophotometrically as the reduction of cytochrome C, and hexose monophosphate shunt activity, measured by /sup 14/CO/sub 2/ liberation from /sup 14/C-1-D-glucose, of alveolar macrophages from horses. Alveolar macrophages were harvested from 6 anesthetized, healthy, light-breed, adult horses by bronchoalveolar lavage. Following a randomized complete block design, the suspension of cells was divided into aliquots of 10/sup 6/ viable alveolar macrophages which were exposed to 1, 10 or 100 g. of opsonized R. equi or opsonized zymosan A at 37 C for 2 hours. In this study the respiratory burst of equine alveolar macrophages was only evidenced by the hexose monophosphate shunt activity and superoxide anion was not coincidentally produced. Rhodococcus equi did not adversely affect that response. The insignificant superoxide anion production by the alveolar macrophages suggests that this may not be a significant oxygen metabolite in those cells.

  10. 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

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

    Science.gov (United States)

    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

    2014-08-01

    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.

  12. 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.

  13. Imaging and pathological findings of AIDS complicated by pulmonary Rhodococcus equi infection

    Institute of Scientific and Technical Information of China (English)

    LI Hong-jun; CHENG Jing-liang

    2011-01-01

    Background Rhodococcus equi (R.equi) infection commonly occurs in grazing areas,especially in patients with AIDS or with T-lymphocyte immuno-deficiencies. Literature reviews revealed that cases radiologically and pathologically diagnosed of AIDS complicated by R. equi infection are rare. This study aimed to investigate the imaging features and pathological basis of AIDS complicated by pulmonary R. equi infection.Methods A total of 13 cases of AIDS complicated by pulmonary R. equi infection were retrospectively analyzed based on their imaging,bacterial culture and pathological data,including 10 cases by chest CT scanning and X-ray radiology and 3 cases by only X-ray radiology. All 13 cases were definitely diagnosed by bacterial culture,including one by CT-guided pulmonary puncture with following H&E staining and periodic acid-Schiff (PAS) staining for diagnostic biopsy and another one by bronchial biopsy with following H&E staining and PAS staining for pathological diagnosis. The imaging findings and the pathological findings of AIDS complicated by pulmonary R. equi infection were compared and evaluated.Results Totally 9 subjects (70%) had radiological demonstrations of central ball liked high density shadows in unilateral pulmonary hilus areas;10 (77%),cavities and liquefied levels;3 (23%),pleural effussion. The foci were found in pulmonary inner zone in 10 subjects (77%) and in pulmonary outer zone in one subject (7%). The pathological findings included intra-alveolar hemorrhage,lymphocyte infiltration and granulation tissue proliferation,which were in line with the pathological process of necrotic pneumonia. After 8-month follow-up of anti-R. equi therapy of these 13 cases,9 cases had obviously decreased or shrunk pulmonary cavities,one died,one missed follow-up,one completely absorbed foci and one did not receive reexaminations.Conclusions The radiological demonstrations of AIDS complicated by pulmonary R. equiinfection are central ball liked high density areas

  14. 马泰勒虫EMA1基因的克隆与生物学特性分析%Cloning and phylogenetic analysis of EMA1 gene of Theileria equi

    Institute of Scientific and Technical Information of China (English)

    罗金; 刘光远; 田占成; 谢俊仁; 沈辉; 田美媛

    2012-01-01

    根据GenBank上登录的马泰勒虫美国株(AB043618)裂殖子表面蛋白抗原1(EMA1)基因,设计了2对引物,采用PCR技术首次扩增出我国马泰勒虫肇源株EMA1基因。将该基因克隆到pMD19-T载体后,进行序列测定及分析。将鉴定为阳性的克隆重组到原核表达载体pET-30a后进行表达,并纯化表达的重组蛋白,进行Western-blotting试验,以鉴定其生物学活性。结果,EMA1基因长819bp,编码272个氨基酸,将该基因编码蛋白的氨基酸序列与GenBank中登录的11种已知梨形虫的相应氨基酸序列进行比较分析,马泰勒虫肇源分离株与已报道的马泰勒虫(马巴贝斯虫)亲缘关系最近,其次是环形泰勒虫和斑羚泰勒虫,而与东方泰勒虫、瑟氏泰勒虫的亲缘关系较远。表达的重组蛋白是分子质量约为38ku的融合蛋白,且表达产物能被马泰勒虫标准阳性血清识别。结果表明,马泰勒虫EMA1作为一种良好的抗原分子,在虫株的分类学研究、流行病学调查以及候选疫苗的研制中有着重要的意义。%To clarify the genetic background and molecular epidemiology of Theileria equi(T.equi),meanwhile providing the foundation for prevention and diagnosis of equine theileriosis in this study,primers for merozoite surface protein antigen 1(EMA1) gene of T.equi based on the USDA strain sequence in the GenBank(AB043618) were designed to amplified the EMA1 gene of Zhaoyuan strain.Then the gene was cloned into the pMD19-T vector,sequenced and analyzed.A recombinant expression plasmid pET-30a-EMA1 was constructed by subcloning the cloned EMA1 gene into the linearized pET-30a vector.Then the plasmid pET-30a-EMA1 was expressed in Escherichia coli(BL21).In result,the gene was 819 bp in size and encoded 272 amino acids.Comparison with 11 known amino acid sequence in the GenBank showed that the gene had the closest relationship with Zhaoyuan T.equi,followed by T.equi(Babesia equi

  15. 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.

  16. Assessing the Genotypic Differences between Strains of Corynebacterium pseudotuberculosis biovar equi through Comparative Genomics.

    Science.gov (United States)

    Baraúna, Rafael A; Ramos, Rommel T J; Veras, Adonney A O; Pinheiro, Kenny C; Benevides, Leandro J; Viana, Marcus V C; Guimarães, Luís C; Edman, Judy M; Spier, Sharon J; Azevedo, Vasco; Silva, Artur

    2017-01-01

    Seven genomes of Corynebacterium pseudotuberculosis biovar equi were sequenced on the Ion Torrent PGM platform, generating high-quality scaffolds over 2.35 Mbp. This bacterium is the causative agent of disease known as "pigeon fever" which commonly affects horses worldwide. The pangenome of biovar equi was calculated and two phylogenomic approaches were used to identify clustering patterns within Corynebacterium genus. Furthermore, other comparative analyses were performed including the prediction of genomic islands and prophages, and SNP-based phylogeny. In the phylogenomic tree, C. pseudotuberculosis was divided into two distinct clades, one formed by nitrate non-reducing species (biovar ovis) and another formed by nitrate-reducing species (biovar equi). In the latter group, the strains isolated from California were more related to each other, while the strains CIP 52.97 and 1/06-A formed the outermost clade of the biovar equi. A total of 1,355 core genes were identified, corresponding to 42.5% of the pangenome. This pangenome has one of the smallest core genomes described in the literature, suggesting a high genetic variability of biovar equi of C. pseudotuberculosis. The analysis of the similarity between the resistance islands identified a higher proximity between the strains that caused more severe infectious conditions (infection in the internal organs). Pathogenicity islands were largely conserved between strains. Several genes that modulate the pathogenicity of C. pseudotuberculosis were described including peptidases, recombination enzymes, micoside synthesis enzymes, bacteriocins with antimicrobial activity and several others. Finally, no genotypic differences were observed between the strains that caused the three different types of infection (external abscess formation, infection with abscess formation in the internal organs, and ulcerative lymphangitis). Instead, it was noted that there is a higher phenetic correlation between strains isolated at

  17. Assessing the Genotypic Differences between Strains of Corynebacterium pseudotuberculosis biovar equi through Comparative Genomics

    Science.gov (United States)

    Ramos, Rommel T. J.; Veras, Adonney A. O.; Pinheiro, Kenny C.; Benevides, Leandro J.; Edman, Judy M.; Spier, Sharon J.; Azevedo, Vasco; Silva, Artur

    2017-01-01

    Seven genomes of Corynebacterium pseudotuberculosis biovar equi were sequenced on the Ion Torrent PGM platform, generating high-quality scaffolds over 2.35 Mbp. This bacterium is the causative agent of disease known as “pigeon fever” which commonly affects horses worldwide. The pangenome of biovar equi was calculated and two phylogenomic approaches were used to identify clustering patterns within Corynebacterium genus. Furthermore, other comparative analyses were performed including the prediction of genomic islands and prophages, and SNP-based phylogeny. In the phylogenomic tree, C. pseudotuberculosis was divided into two distinct clades, one formed by nitrate non-reducing species (biovar ovis) and another formed by nitrate-reducing species (biovar equi). In the latter group, the strains isolated from California were more related to each other, while the strains CIP 52.97 and 1/06-A formed the outermost clade of the biovar equi. A total of 1,355 core genes were identified, corresponding to 42.5% of the pangenome. This pangenome has one of the smallest core genomes described in the literature, suggesting a high genetic variability of biovar equi of C. pseudotuberculosis. The analysis of the similarity between the resistance islands identified a higher proximity between the strains that caused more severe infectious conditions (infection in the internal organs). Pathogenicity islands were largely conserved between strains. Several genes that modulate the pathogenicity of C. pseudotuberculosis were described including peptidases, recombination enzymes, micoside synthesis enzymes, bacteriocins with antimicrobial activity and several others. Finally, no genotypic differences were observed between the strains that caused the three different types of infection (external abscess formation, infection with abscess formation in the internal organs, and ulcerative lymphangitis). Instead, it was noted that there is a higher phenetic correlation between strains isolated at

  18. 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.

  19. Minimum inhibitory concentrations of erythromycin and rifampin for Rhodococcus equi during the years 2007-2014.

    Science.gov (United States)

    Fenton, Caitriona S; Buckley, Thomas C

    2015-01-01

    Rhodococcus equi is a gram positive, intracellular pathogen of foals worldwide. The aim of this study was to determine whether there was an increasing resistance occurring in Rhodococcus equi towards the antibiotics rifampin and erythromycin over a seven year period. The investigation was carried out with the use of E test strips (epsilometers) for rifampin and erythromycin in order to determine the Minimum Inhibitory Concentrations (MIC) values of Rhodococcus equi to these antibiotics. The main results of this study found that the mean MICs were higher for erythromycin than for rifampin for every year analysed apart from 2008. The results highlight that 75 % (6/8) of the mean MICs for erythromycin were above the threshold of susceptibility of 0.5 μg/ml and one of the yearly mean MICs for rifampin (2008) was above the level of ≤ 1 μg/ml. Two soil samples analysed had high MIC values of 2 μg/ml and 3 μg/ml for rifampin and erythromycin respectively. These samples can be said to have acquired resistance as they are above 1 μg/ml. The significance of these findings is that R. equi is already a problematic pathogen to treat and if the bacteria keeps gaining resistance to these antibiotics at rate that has been shown over the last decade, then a new form of treatment will have to be introduced. Further research into the genomics of Rhodococcus equi will, in time, shed more light on possible alternatives such as vaccines or new, more effective antimicrobials.

  20. Serological detection of Anaplasma marginale, Babesia bovis and Babesia bigemina in beef cattle of the northern and central-western regions of Brazil

    Directory of Open Access Journals (Sweden)

    Jenevaldo Barbosa da Silva

    2015-06-01

    Full Text Available The objective of this study was to assess the occurrence of antibodies against Anaplasma marginale, Babesia bovis and Babesia bigemina in beef cattle of the northern and central-western regions of Brazil. A total of 1703 blood samples were collected from cattle from 100 farms distributed among 37 municipalities in the states of Mato Grosso, Pará and Tocantins. The search for antibodies was conducted using an indirect enzyme-linked immunosorbent assay (ELISA, and antibodies were observed for B. bovis, B. bigemina and A. marginale in cattle from Mato Grosso, Tocantins and Pará at rates of 99%, 90% and 41%; 99%, 70% and 52%; and 97%, 97% and 75%, respectively. The results show that the analyzed regions exhibit enzootic stability for infection with B. bovis and B. bigemina, whereas the same result was not observed with A. marginale.

  1. 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.

  2. 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.

  3. PCR detection of Babesia ovata from questing ticks in Japan.

    Science.gov (United States)

    Sivakumar, Thillaiampalam; Tattiyapong, Muncharee; Okubo, Kazuhiro; Suganuma, Keisuke; Hayashida, Kyoko; Igarashi, Ikuo; Zakimi, Satoshi; Matsumoto, Kotaro; Inokuma, Hisashi; Yokoyama, Naoaki

    2014-04-01

    Babesia ovata is a tick-transmitted hemoprotozoan parasite of cattle. In the present study, we analyzed tick DNA samples (n=1459) prepared from questing ticks collected from various cattle pastures in Hokkaido (Shibecha, Taiki, Otofuke, Memuro, and Shin-Hidaka districts) and Okinawa (Yonaguni Island) prefectures of Japan for B. ovata. When all the tick DNA samples were screened by a previously described B. ovata-specific apical membrane antigen-1 (AMA-1) gene-based polymerase chain reaction (PCR) assay, none of the DNA samples was positive. Therefore, we developed a PCR assay based on the protozoan beta-tubulin (β-tubulin) gene to detect B. ovata from ticks in Japan. In the specificity test, the PCR assay amplified the expected 444-bp target gene fragment from B. ovata DNA. No PCR products were amplified from DNA samples from other blood pathogens, bovine blood, or ticks. In addition, the PCR assay detected 100 fg of B. ovata-genomic DNA extracted from an in vitro culture of the parasites. Subsequently, when all the tick DNA samples were screened by this new PCR assay, 18 were positive for B. ovata. Positive samples were found only in the Yonaguni and Memuro areas. In Okinawa, where all the ticks were identified as Haemaphysalis longicornis, 9.7% of the samples were PCR-positive, while a single tick (Ixodes ovatus) from Memuro was infected with B. ovata. When the nucleotide sequences of the PCR amplicons were phylogenetically analyzed, they formed a separate clade containing a previously described β-tubulin gene sequence from B. ovata (Miyake strain), confirming that the PCR assay had detected only B. ovata from the tick DNA samples. This is the first report that describes the PCR detection of B. ovata in ticks. The findings warrant transmission experiments to evaluate I. ovatus as a potential vector of B. ovata.

  4. 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.

  5. 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

    2015-08-01

    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.

  6. Detection of Babesia divergens in southern Norway by using an immunofluorescence antibody test in cow sera

    OpenAIRE

    2010-01-01

    Background The incidence of bovine babesiosis, caused by Babesia divergens (Apicomplexa: Piroplasmida) has decreased markedly since the 1930 s, but may re-emerge as a consequence of climate change and changes in legislation and pasturing practices. This is a potentially serious disease, with both economical and animal welfare consequences. Therefore, there is a need to survey the distribution of B. divergens. Methods ...

  7. Molecular identification, genetic diversity and distribution of Theileria and Babesia species infecting small ruminants.

    Science.gov (United States)

    Altay, Kursat; Dumanli, Nazir; Aktas, Munir

    2007-06-20

    Detection and identification of Theileria and Babesia species in 920 apparently healthy small ruminants in eastern Turkey, as well as parasite genetic diversity, was investigated using a specifically designed reverse line blot (RLB) assay. The hypervariable V4 region of the 18S ribosomal RNA (rRNA) gene was amplified and hybridized to a membrane onto which catchall and species-specific oligonucleotide probes were covalently linked. Three Theileria and one Babesia genotype were identified. Comparison of the Theileria genotypes revealed 93.6-96.2% similarity among their 18S rRNA genes. Two Theileria shared 100% and 99.7% similarity with the previously described sequences of T. ovis and Theileria sp. OT3, respectively. A third Theileria genotype was found to be clearly different from previously described Theileria species. The genotype was provisionally designated as Theileria sp. MK. The Babesia genotype shared 100% similarity with Babesia ovis. The survey indicated a high prevalence of piroplasm infections in small ruminants (38.36%). Theileria spp. prevalence was 36.08%. Prevalence of B. ovis was 5.43%. The most abundant Theileria species identified was T. ovis (34.56%) followed by Theileia sp. MK (1.30%) and Theileria sp. OT3 (0.43%).

  8. Molecular detection of Babesia spp. and other haemoparasitic infections of cattle in Maputo Province, Mozambique.

    Science.gov (United States)

    Martins, Tiago M; Neves, Luís; Pedro, Olívia C; Fafetine, José M; Do Rosário, Virgílio E; Domingos, Ana

    2010-05-01

    Molecular detection of Babesia species in apparently healthy cattle within an endemic region was carried out in order to determine the prevalence of carriers and the geographical distribution of Babesia bigemina and Babesia bovis in Maputo Province, Mozambique. Samples from 477 animals at 5 localities were analysed using 2 techniques, the semi-nested hot-start PCR and the reverse line blot (RLB) assay. With the semi-nested hot-start PCR, detection of B. bigemina ranged between 30% and 89%, and of B. bovis between 27% and 83%. The RLB assay was comparatively less sensitive in this study and detection of B. bovis ranged from 0% to 17%, and B. bigemina was not detected at all by this technique. Analysis of new sequences of the 18S rRNA gene revealed that the current B. bigemina RLB probe is not specific for the identification of isolates in Mozambique. The RLB assay, however, resulted in the detection of 8 other haemoparasite species belonging to the genera Babesia, Theileria, Anaplasma and Ehrlichia. 18S rRNA gene sequences from the Theileria spp. were identified, and a phylogenic tree constructed with these sequences yielded a heterogeneous T. mutans-like group. In conclusion, infection with B. bigemina and B. bovis is endemic in Maputo Province, but rates of transmission vary. Furthermore, mixed infections with the haemoparasites responsible for several tick-borne diseases in cattle are common in Mozambique.

  9. 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...

  10. Analysis of Babesia bovis-induced gene expression changes in the cattle tick, Rhipcephalus (Boophilus) microplus.

    Science.gov (United States)

    Boophilus ticks are vectors of Babesia bovis, the protozoan causative agent of cattle fever, a disease which is responsible for significant production losses to cattle producers in much of Africa, Central and South America and Australia. We utilized subtractive cDNA library synthesis techniques to o...

  11. Clinical, hemato-biochemical alterations and oxidant–antioxidant biomarkers in Babesia-infected calves

    Directory of Open Access Journals (Sweden)

    Noha Y. Salem

    2016-06-01

    Full Text Available Babesia is one of the main causes of anemia in cattle, a lot of elucidations have been suggested to explain its pathogenesis. This study was designed to investigate clinical, hemato-biochemical and oxidant/antioxidant status and its relation with the resultant anemia in Babesia-infected calves. Seventeen (17 native breed calves were involved in this study, clinical signs and microscopic findings were recorded, also blood samples were taken to investigate hematologic changes, serum biochemical variations and oxidative stress biomarkers. The most commonly observed clinical signs were fever, emaciation, depression, icterus and hemoglobinuria. Significant reduction in PCV, HB, RBCs, MCHC, Total protein, and albumin along with significant increase in MCV, WBCs, monocytes and BUN were the most consistent hemato-biochemical changes. Oxidant/antioxidant and trace mineral assessment showed significant reduction in Superoxide dismutase “SOD”, Glutathione peroxidase “GPx”, Zn, Cu along with significant increase in malondialdehyde (MDA activities. In the current investigation, oxidant/antioxidant imbalance along with the synchronized alterations in antioxidant trace minerals was detected in Babesia-infected calves. These findings support notion that Babesia infection associated with oxidative stress and this process may be linked to the resultant anemia.

  12. Transfected babesia bovis expressing a tick GST as a live vector vaccine

    Science.gov (United States)

    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 pro...

  13. Expression of 6-Cys gene superfamily defines babesia bovis sexual stage development within rhipicephalus microplus

    Science.gov (United States)

    Babesia bovis, an intra-erythrocytic tick-borne apicomplexan protozoan, is one of the agents of bovine babesiosis. Its life cycle includes sexual reproduction within cattle fever ticks, Rhipicephalus spp. Six B. bovis 6-Cys gene superfamily members were previously identified (A, B, C, D, E, F) and t...

  14. Development of a tandem repeat-based multilocus typing system distinguishing Babesia bovis geographic isolates

    Science.gov (United States)

    Mini and microsatellite sequences have proven to be excellent tools for the differentiation of strains and populations in several protozoan parasites due to their high variability. In the present work we have searched the genome of the tick-transmitted bovine hemoprotozoon Babesia bovis for tandem r...

  15. 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.

  16. Infecção pulmonar pelo Rhodococcus equi na síndrome da imunodeficiência adquirida: Aspectos na tomografia computadorizada Rhodococcus equi infection in acquired immunodeficiency syndrome: Computed tomography aspects

    Directory of Open Access Journals (Sweden)

    Edson Marchiori

    2006-10-01

    Full Text Available OBJETIVO: Apresentar os aspectos na tomografia computadorizada da pneumonia pelo Rhodococcus equi em sete pacientes com síndrome da imunodeficiência adquirida. MÉTODOS: Estudo retrospectivo das tomografias de sete pacientes com síndrome da imunodeficiência adquirida e infecção pelo Rhodococcus equi. RESULTADOS: Os achados mais freqüentes foram: consolidação (n = 7 com escavação (n = 6, opacidades em vidro fosco (n = 6, nódulos do espaço aéreo (n = 4 e nódulos centrolobulares com árvore em brotamento (n = 3. CONCLUSÃO: Os achados mais comuns na infecção pulmonar pelo Rhodococcus equi em pacientes com síndrome da imunodeficiência adquirida foram as consolidações escavadas.OBJECTIVE: To present the computed tomography aspects of Rhodococcus equi pneumonia in seven patients with acquired immunodeficiency syndrome. METHODS: A retrospective study of the computed tomography scans of seven patients with acquired immunodeficiency syndrome and Rhodococcus equi infection. RESULTS: The most common findings were consolidation (n = 7, consolidation with cavitation (n = 6, ground glass opacities (n = 6, peribronchial nodules (n = 4 and centrilobular nodules presenting a "tree-in-bud" pattern (n = 3. CONCLUSION: The most common finding in patients with Rhodococcus equi pulmonary infection and acquired immunodeficiency syndrome was consolidation with cavitation.

  17. Experimental verification of the identity of variant-specific surface proteins in Giardia lamblia trophozoites.

    Science.gov (United States)

    Li, Wei; Saraiya, Ashesh A; Wang, Ching C

    2013-05-21

    The cell membrane of a Giardia lamblia trophozoite is covered with a single species of variant-specific surface protein (VSP) that is replaced by another VSP every 6 to 13 generations of cell growth, possibly for an evasion of host immunity. Experimentally, only six VSP species have been verified to localize to the cell membrane thus far. By assuming that VSP contains multiple CXXC motifs, 219 vsp genes were annotated in GiardiaDB of the WB isolate. By further assuming that VSP possesses both CXXC motifs and a CRGKA tail at the C terminus, Adam et al. (BMC Genomics 11:424, 2010) identified a total of 303 potential vsp genes in Giardia WB. The discrepancies between these two assumed VSP identities have caused some confusion. Here, we used experimental approaches to further verify what is required of the structures of a VSP to localize to the surface of cell membrane. The data led to the following conclusions. (i) The C-terminal CRGKA sequence is not essential for localizing VSPs to the cell membrane. (ii) A "motif 1" of 45 residues, consisting of two CXXCs separated by 12 to 15 amino acid residues, located close to the C terminus and a hydrophobic "motif 2" of 38 residues at the C terminus are both essential and sufficient for localizing the protein to the cell membrane. (ii) An N-terminal sequence upstream from motif 1 is not required for targeting VSPs to the cell membrane. By these criteria, we are able to identify 73 open reading frames as the putative vsp genes in Giardia. IMPORTANCE The intestinal pathogen Giardia lamblia expresses only one variant-specific surface protein (VSP) on the cell membrane surface at a given time, but it changes spontaneously every 6 to 13 generations of growth, presumably for evading the host immunity. Only 6 VSPs have been empirically shown to localize to the cell membrane surface thus far. Here, we used mutations of VSPs and methods of identifying their locations in Giardia cells and found that a "motif 1" of 45 residues

  18. 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.

  19. Three-dimensional reconstruction of rabbit-derived Pneumocystis carinii from serial-thin sections. I: Trophozoite.

    Science.gov (United States)

    Palluault, F; Pietrzyk, B; Dei-Cas, E; Slomianny, C; Soulez, B; Camus, D

    1991-01-01

    The highly complex ultrastructural morphology of the endomembrane system in Pneumocystis carinii led us to perform three-dimensional reconstruction from serial-thin sections using the CATIA (Conception Assistée Tridimensionnelle Inter Active) Dassault system program. The three-dimensional reconstruction of a small trophozoite made it possible to better understand the morphological relationship among organelles and to suggest cytophysiological hypotheses. By reconstructing other parasite stages, we gathered information about the evolution of organelles during the life cycle and about their physiology.

  20. Characterization of the products of the heme detoxification pathway in malarial late trophozoites by X-ray diffraction.

    Science.gov (United States)

    Bohle, D S; Dinnebier, R E; Madsen, S K; Stephens, P W

    1997-01-10

    In a process inhibited by the quinoline antimalarial drugs, Plasmodia detoxify heme released during the degradation of hemoglobin by aggregating it into malarial pigment, an insoluble crystalline heme coordination polymer. Synchrotron x-ray powder diffraction patterns for intact desiccated malarial trophozoites and synthetic beta-hematin have been measured; both materials correspond to a single crystalline triclinic lattice with unit cell parameters a = 12.2176(4), b = 14.7184(5), c = 8.0456(3) A; alpha = 90.200(2), beta = 96.806(3), gamma = 97.818(3) degrees and Z = 2. These results unambiguously demonstrate that hemozoin crystallites are identical to synthetic beta-hematin.

  1. An outbreak of Streptococcus equi subspecies zooepidemicus associated with consumption of fresh goat cheese

    Directory of Open Access Journals (Sweden)

    Hakkinen Marjaana

    2006-02-01

    Full Text Available Abstract Background Streptococcus equi subspecies zooepidemicus is a rare infection in humans associated with contact with horses or consumption of unpasteurized milk products. On October 23, 2003, the National Public Health Institute was alerted that within one week three persons had been admitted to Tampere University Central Hospital (TaYS because of S. equi subsp. zooepidemicus septicaemia. All had consumed fresh goat cheese produced in a small-scale dairy located on a farm. We conducted an investigation to determine the source and the extent of the outbreak. Methods Cases were identified from the National Infectious Disease Register. Cases were persons with S. equi subsp. zooepidemicus isolated from a normally sterile site who had illness onset 15.9-31.10.2003. All cases were telephone interviewed by using a standard questionnaire and clinical information was extracted from patient charts. Environmental and food specimens included throat swabs from two persons working in the dairy, milk from goats and raw milk tank, cheeses made of unpasteurized milk, vaginal samples of goats, and borehole well water. The isolates were characterized by ribotyping and pulsed-field gel electrophoresis (PFGE. Results Seven persons met the case definition; six had septicaemia and one had purulent arthritis. Five were women; the median age was 70 years (range 54–93. None of the cases were immunocompromized and none died. Six cases were identified in TaYS, and one in another university hospital in southern Finland. All had eaten goat cheese produced on the implicated farm. S. equi subsp. zooepidemicus was isolated from throat swabs, fresh goat cheese, milk tank, and vaginal samples of one goat. All human and environmental strains were indistinguishable by ribotyping and PFGE. Conclusion The outbreak was caused by goat cheese produced from unpasteurized milk. Outbreaks caused by S. equi subsp. zooepidemicus may not be detected if streptococcal strains are only

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

    Science.gov (United States)

    2014-01-01

    Background 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. Methods 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. Results 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. Conclusions 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

  3. 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)

    Gundelly, Praveen; 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 equi-infected HIV patients was lower in the HAART era (8%) than in pre-HAART era (56%) (P equi infection should include a combination of two 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.

  4. Prevalence and genetic diversity of Rhodococcus equi in wild boars (Sus scrofa), roe deer (Capreolus capreolus) and red deer (Cervus elaphus) in Poland.

    Science.gov (United States)

    Witkowski, Lucjan; Rzewuska, Magdalena; Cisek, Agata Anna; Chrobak-Chmiel, Dorota; Kizerwetter-Świda, Magdalena; Czopowicz, Michał; Welz, Mirosław; Kita, Jerzy

    2015-05-22

    Rhodococcus equi is now considered an emerging zoonotic pathogen. Sources and routes of human infection remain unclear but foodborne transmission seems to be the most probable way. Strains of pig or bovine type are most often isolated from human cases and moreover R. equi is present in submaxillary lymph nodes of apparently healthy pigs and wild boars intended for human consumption. The aim of this study was to estimate the prevalence of R. equi in submaxillary lymph nodes in wild boars, roe deer and red deer. Samples were collected from 936 animals and 27 R. equi strains were isolated, from 5.1 % of wild boars (23/452), 0.7 % of red deer (2/272) and 0.9 % of roe deer (2/212). Genetic diversity of all 27 isolates was studied using VspI-PFGE method, resulting in the detection of 25 PFGE patterns and four PFGE clusters. PFGE patterns of the isolates were compared with virulence plasmid types and no concordance was observed. R. equi was present in wild animal tissues and consumption of the game may be a potential source of R. equi infection for humans. To the authors' best knowledge, this is the first epidemiological report of R. equi prevalence in tissues of roe deer and red deer. However, risk associated with wild ruminant consumption seems marginal. Investigation of R. equi transmission between animals and humans based exclusively on types of virulence plasmids seems to be insufficient to identify sources of R. equi infection for people.

  5. Antibiotic failure in a renal transplant patient with Rhodococcus equi infection: an indication for surgical lobectomy.

    Science.gov (United States)

    Ursales, A; Klein, J A; Beal, S G; Koch, M; Clement-Kruzel, S; Melton, L B; Spak, C W

    2014-12-01

    Rhodococcus equi is an animal pathogen that causes infrequent but challenging infections in immunocompromised individuals, few of which have been described in solid organ transplant recipients. Common clinical presentations include indolent cough, fever, and dyspnea, with necrotizing pneumonia and cavitation. We report a case of a dense right upper lung pneumonia with resultant R. equi bacteremia in a renal transplant recipient. Our patient initially responded to antibiotic treatment with resolution of bacteremia and clinical recovery, followed by interval progression in her right upper lobe consolidation on follow-up computed tomography scans. She underwent lobectomy for definitive therapy with resolution of symptoms. Lobectomy can be utilized in isolated infection after antibiotic failure with excellent clinical outcomes. © 2014 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  6. Induction of morphological and electrophysiological changes in hamster cornea after in vitro interaction with trophozoites of Acanthamoeba spp.

    Science.gov (United States)

    Omaña-Molina, Maritza; Navarro-García, Fernando; González-Robles, Arturo; Serrano-Luna, José de Jesús; Campos-Rodríguez, Rafael; Martínez-Palomo, Adolfo; Tsutsumi, Víctor; Shibayama, Mineko

    2004-06-01

    Acanthamoeba castellani and Acanthamoeba polyphaga are free-living amebae that cause keratitis and granulomatous encephalitis in humans. We have analyzed the early morphological and electrophysiological changes occurring during the in vitro interaction of cultured amebae with intact or physically damaged corneas obtained from hamsters. Both species of Acanthamoeba produced similar cytopathic changes, as seen by light microscopy and scanning electron microscopy. After adhesion to the epithelial surface, trophozoites formed clumps and migrated toward the cell borders, causing the separation of adjacent cells at 1 h of coculture. At later stages (2 to 4 h), some amebae were found under desquamating epithelial cells whereas others were seen associated with damaged cells or forming amebostome-like structures to ingest detached epithelial cells. Control corneas incubated in culture medium conditioned with amebae showed a cytoplasmic vacuolization and blurring of the epithelial-stromal junction. The early stages of corneal epithelial damage caused by amebae were also analyzed by measuring the transepithelial resistance changes in corneas mounted in Ussing chambers. Both species of Acanthamoeba caused a rapid decrease in electrical resistance. The present observations demonstrate that under in vitro conditions, Acanthamoeba trophozoites rapidly cause significant damage to the corneal epithelium. Furthermore, in our experimental model, previous physical damage to the corneas was not a prerequisite for the development of amebic corneal ulcerations.

  7. Entamoeba histolytica P-glycoprotein (EhPgp) inhibition, induce trophozoite acidification and enhance programmed cell death.

    Science.gov (United States)

    Medel Flores, Olivia; Gómez García, Consuelo; Sánchez Monroy, Virgina; Villalba Magadaleno, José D'Artagnan; Nader García, Elvira; Pérez Ishiwara, D Guillermo

    2013-11-01

    Programmed cell death (PCD) is induced in Entamoeba histolytica by a variety of stimuli in vitro and in vivo. In mammals, intracellular acidification serves as a global switch for inactivating cellular processes and initiates molecular mechanisms implicated in the destruction of the genome. In contrast, intracellular alkalinization produced by P-glycoprotein overexpression in multidrug-resistant cells has been related to apoptosis resistance. Our previous studies showed that overexpression of E. histolytica P-glycoprotein (PGP) altered chloride-dependent currents and triggered trophozoite swelling, the reverse process of cell shrinkage produced during PCD. Here we showed that antisense inhibition of PGP expression produced a synchronous death of trophozoites and the enhancement of biochemical and morphological characteristics of PCD induced by G418. The nucleus was contracted, and the nuclear membrane was disrupted. Moreover, chromatin was extensively fragmented. Ca(2+) concentration was increased, while the intracellular pH (ipH) was acidified. In contrast, PGP overexpression prevented intracellular acidification and circumvented the apoptotic effect of G418.

  8. 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.

  9. 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.

  10. 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.

  11. MICs of 32 antimicrobial agents for Rhodococcus equi isolates of animal origin.

    Science.gov (United States)

    Riesenberg, Anne; Feßler, Andrea T; Erol, Erdal; Prenger-Berninghoff, Ellen; Stamm, Ivonne; Böse, Reinhard; Heusinger, Anton; Klarmann, Dieter; Werckenthin, Christiane; Schwarz, Stefan

    2014-04-01

    The aim of this study was to determine the MICs of 32 antimicrobial agents for 200 isolates of Rhodococcus equi of animal origin by applying a recently described broth microdilution protocol, and to investigate isolates with distinctly elevated rifampicin MICs for the genetic basis of rifampicin resistance. The study included 200 R. equi isolates, including 160 isolates from horses and 40 isolates from other animal sources, from the USA and Europe. MIC testing of 32 antimicrobial agents or combinations thereof followed a recently published protocol. A novel PCR protocol for the joint amplification of the three rpoB regions in which rifampicin resistance-mediating mutations have been reported was applied to isolates with elevated rifampicin MICs. The amplicons were sequenced and screened for mutations. Susceptibility testing revealed a rather uniform distribution of MICs for most of the antimicrobial agents tested. The lowest MICs were seen for clarithromycin, rifampicin and imipenem. Six isolates (3%) exhibited distinctly higher MICs of rifampicin than the remaining 194 isolates. In five of these six isolates, single bp exchanges, which resulted in the amino acid exchanges Gln513Leu, Asp516Val, His526Asp or Ser531Leu, were detected in the rifampicin resistance-determining region 1 of the rpoB gene, with Gln513Leu representing a novel substitution for R. equi. This study shows the MIC distribution of 32 antimicrobial agents for a large collection of R. equi isolates of animal origin from two continents. Isolates that exhibited distinctly elevated MICs of rifampicin were only rarely detected.

  12. Systemic and respiratory oxidative stress in the pathogenesis and diagnosis of Rhodococcus equi pneumonia.

    Science.gov (United States)

    Crowley, J; Po, E; Celi, P; Muscatello, G

    2013-12-01

    Oxidative stress (OS) is most simply defined as an imbalance between oxidants and antioxidants. Oxidative stress has been suggested to play roles in various equine respiratory diseases and the significance of OS in the pathogenesis of Rhodococcus equi pneumonia is unknown. To measure and relate biomarkers of OS to lesions consistent with R. equi pneumonia. Case-control study. Various OS biomarkers were measured from blood and exhaled breath condensate (EBC) samples collected from 26 foals between 1 and 2 months of age (n = 12 cases and n = 14 controls) on 2 Thoroughbred farms endemically affected by R. equi pneumonia. Foals were defined as cases (positive) or controls (negative) based on ultrasonographic evidence of pulmonary abscessation (>15 mm in diameter). Haematology and biochemistry testing was also performed on blood samples collected from the foals. Comparison of biomarkers and key haematological and biochemical markers of inflammation between the groups was performed using 2 sample t tests. Derivatives of reactive oxygen metabolites (d-ROMs) were significantly greater in case foals than in control foals (P = 0.027) and the oxidative stress index (OSI) was higher in case foals (P = 0.014). Hydrogen peroxide (H2 O2 ) concentrations in EBC were significantly greater in case foals than in control foals (P = 0.002). Meanwhile, there were no significant differences in traditional measures of inflammation between the 2 groups. Measuring OS in both blood and EBC provided useful information in the early diagnosis of R. equi pneumonia. © 2013 EVJ Ltd.

  13. IcgA is a virulence factor of Rhodococcus equi that modulates intracellular growth.

    Science.gov (United States)

    Wang, Xiaoguang; Coulson, Garry B; Miranda-Casoluengo, Aleksandra A; Miranda-Casoluengo, Raúl; Hondalus, Mary K; Meijer, Wim G

    2014-05-01

    Virulence of the intracellular pathogen Rhodococcus equi depends on a 21.3-kb pathogenicity island located on a conjugative plasmid. To date, the only nonregulatory pathogenicity island-encoded virulence factor identified is the cell envelope-associated VapA protein. Although the pathogenicity islands from porcine and equine R. equi isolates have undergone major rearrangements, the virR operon (virR-icgA-vapH-orf7-virS) is highly conserved in both, suggesting these genes play an important role in pathogenicity. VirR and VirS are transcriptional regulators controlling expression of pathogenicity island genes, including vapA. Here, we show that while vapH and orf7 are dispensable for intracellular growth of R. equi, deletion of icgA, formerly known as orf5, encoding a major facilitator superfamily transport protein, elicited an enhanced growth phenotype in macrophages and a significant reduction in macrophage viability, while extracellular growth in broth remained unaffected. Transcription of virS, located downstream of icgA, and vapA was not affected by the icgA deletion during growth in broth or in macrophages, showing that the enhanced growth phenotype caused by deletion of icgA was not mediated through abnormal transcription of these genes. Transcription of icgA increased 6-fold within 2 h following infection of macrophages and remained significantly higher 48 h postinfection compared to levels at the start of the infection. The major facilitator superfamily transport protein IcgA is the first factor identified in R. equi that negatively affects intracellular replication. Aside from VapA, it is only the second pathogenicity island-encoded structural protein shown to play a direct role in intracellular growth of this pathogenic actinomycete.

  14. Prevalence and Antibiogram study of Rhodococcus equi in equines of Jammu and Kashmir, India.

    Science.gov (United States)

    Mir, Irfan Ahmad; Kumar, Bablu; Taku, Anil; Bhardwaj, Rajinder Kumar; Bhat, Mohd Altaf; Badroo, Gulzar Ahmad

    2015-01-01

    The present study was conducted to determine the prevalence of Rhodococcus equi infection in equines of Jammu and Kashmir, India, and evaluate the zoonotic threat posed by this organism to equine owners and tourists. One hundred and forty-one samples (98 samples from adult animals ≥5 years old and 43 samples from foals less than 6 months old) were collected in duplicate from nasopharyngeal tract of equines for isolation and direct PCR. A total of 12 isolates of R. equi were recovered, of which 9 were from foals and 3 from adult animals. Therefore, the present study recorded prevalence rates of 20.93% and 3.06% among foals and adult equines respectively. The prevalence rates were found to be 25.58% and 4.08% by 16S rRNA species-specific PCR among foals and adult animals respectively. Thus, the PCR-based assay was found to be more sensitive and helped in quick detection of R. equi than the culture based method which is time consuming and laborious. However, the culture-based method is still preferred due to some limitations of PCR. The antibiogram of the isolates revealed that erythromycin and rifampicin were the most effective antimicrobials with 100% sensitivity, followed by amoxicillin (66.67%), lincomycin (58.3%) and kanamycin (58.3%). The results also revealed that resistance was highest for penicillin G (50%), followed by kanamycin (25%) and streptomycin (25%).

  15. Disseminated rhodococcus equi infection in HIV infection despite highly active antiretroviral therapy

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    Ferretti Francesca

    2011-12-01

    Full Text Available Abstract Background Rhodococcus equi (R.equi is an acid fast, GRAM + coccobacillus, which is widespread in the soil and causes pulmonary and extrapulmonary infections in immunocompromised people. In the context of HIV infection, R.equi infection (rhodococcosis is regarded as an opportunistic disease, and its outcome is influenced by highly active antiretroviral therapy (HAART. Case presentation We report two cases of HIV-related rhodococcosis that disseminated despite suppressive HAART and anti-rhodococcal treatment; in both cases there was no immunological recovery, with CD4+ cells count below 200/μL. In the first case, pulmonary rhodococcosis presented 6 months after initiation of HAART, and was followed by an extracerebral intracranial and a cerebral rhodococcal abscess 1 and 8 months, respectively, after onset of pulmonary infection. The second case was characterized by a protracted course with spread of infection to various organs, including subcutaneous tissue, skin, colon and other intra-abdominal tissues, and central nervous system; the spread started 4 years after clinical resolution of a first pulmonary manifestation and progressed over a period of 2 years. Conclusions Our report highlights the importance of an effective immune recovery, despite fully suppressive HAART, along with anti-rhodococcal therapy, in order to clear rhodococcal infection.

  16. The Rhodococcus equi virulence protein VapA disrupts endolysosome function and stimulates lysosome biogenesis.

    Science.gov (United States)

    Rofe, Adam P; Davis, Luther J; Whittingham, Jean L; Latimer-Bowman, Elizabeth C; Wilkinson, Anthony J; Pryor, Paul R

    2017-04-01

    Rhodococcus equi (R. equi) is an important pulmonary pathogen in foals that often leads to the death of the horse. The bacterium harbors a virulence plasmid that encodes numerous virulence-associated proteins (Vaps) including VapA that is essential for intracellular survival inside macrophages. However, little is known about the precise function of VapA. Here, we demonstrate that VapA causes perturbation to late endocytic organelles with swollen endolysosome organelles having reduced Cathepsin B activity and an accumulation of LBPA, LC3 and Rab7. The data are indicative of a loss of endolysosomal function, which leads cells to upregulate lysosome biogenesis to compensate for the loss of functional endolysosomes. Although there is a high degree of homology of the core region of VapA to other Vap proteins, only the highly conserved core region of VapA, and not VapD of VapG, gives the observed effects on endolysosomes. This is the first demonstration of how VapA works and implies that VapA aids R. equi survival by reducing the impact of lysosomes on phagocytosed bacteria. © 2016 The Authors. MicrobiologyOpen published by John Wiley & Sons Ltd.

  17. A novel ROM compression architecture for DDFS utilizing the parabolic approximation of equi-section division.

    Science.gov (United States)

    Jeng, Shiann-Shiun; Lin, Hsing-Chen; Lin, Chi-Huei

    2012-12-01

    In this paper, we propose the parabolic approximation of equi-section division (PAESD) utilizing the symmetry property and amplitude approximation of a sinusoidal waveform to design a direct digital frequency synthesizer (DDFS). The sinusoidal phase of a one-quarter period is divided into equi-sections. The proposed method utilizes the curvature equivalence to derive each parabolic curve function, and then the value of the error function between each parabolic curve function and the sinusoidal function is stored in an error-compensation ROM to reconstruct the real sinusoidal waveform. The upper/lower bound of the maximum error value stored in the error-compensation ROM is derived to determine the minimum required memory word length relative to the number of bits of the equi-sections. Thus, the minimum size of the total ROMs of the DDFS using the PAESD without error-compensation ROM is compressed to 544 bits; the total compression ratio, compared with the minimum size of the total ROMs of the DDFS using the basic look-up table (LUT), is approximately 843:1, achieved by consuming additional circuits [71 adaptive look-up tables (ALUTs), 3 digital signal processor (DSP) block 9-bit elements]. Consequently, the results show that the proposed ROM compression method can effectively achieve a better compression ratio than the state-of-the-art solutions without affecting the spectrum performance of an average spurious-free dynamic range (SFDR) of -85 dBc.

  18. A surface membrane protein of Entamoeba histolytica functions as a receptor for human chemokine IL-8: its role in the attraction of trophozoites to inflammation sites.

    Science.gov (United States)

    Diaz-Valencia, J Daniel; Pérez-Yépez, Eloy Andrés; Ayala-Sumuano, Jorge Tonatiuh; Franco, Elizabeth; Meza, Isaura

    2015-12-01

    Entamoeba histolytica trophozoites respond to the presence of IL-8, moving by chemotaxis towards the source of the chemokine. IL-8 binds to the trophozoite membrane and triggers a response that activates signaling pathways that in turn regulate actin/myosin cytoskeleton organisation to initiate migration towards the chemokine, suggesting the presence of a receptor for IL-8 in the parasite. Antibodies directed to the human IL-8 receptor (CXCR1) specifically recognised a 29 kDa protein in trophozoite membrane fractions. The same protein was immunoprecipitated by this antibody from total amebic extracts. Peptide analysis of the immunoprecipitated protein revealed a sequence with high homology to a previously identified amebic outer membrane peroxiredoxin and a motif within the third loop of human CXCR1, which is an important site for IL-8 binding and activation of signaling processes. Immunodetection assays demonstrated that the anti-human CXCR1 antibody binds to the 29 kDa protein in a different but close site to where IL-8 binds to the trophozoite surface membrane, suggesting that human and amebic receptors for this chemokine share common epitopes. In the context of the human intestinal environment, a receptor for IL-8 could be a great advantage for E. histolytica trophozoite survival, as they could reach an inflammatory milieu containing abundant nutrients. In addition, it has been suggested that the high content of accessible thiol groups of the protein and its peroxidase activity could provide protection in the oxygen rich milieu of colonic lesions, allowing trophozoite invasion of other tissues and escape from the host immune response.

  19. Application and Development of PCR Assay to Detect Babesia caballi%驽巴贝斯虫病PCR检测方法的建立和应用

    Institute of Scientific and Technical Information of China (English)

    罗金; 刘光远; 谢俊仁; 田占成; 沈辉; 王芳芳; 孙晓林; 陈荣贵; 王海军

    2011-01-01

    The 18S rRNA gene recently discovered was shown to be species-specific. A pair of primers was designed to specifically amplified a 452 bp fragment. The PCR result of specificity assay showed that one references B. Caballi could be detected by the PCR test,but no amplification was observed when other five bacterial species isolated from T. Equi,B. Bigimina,B. Motasi, B. Ovata, B. Major tissue were detected. And the sensitivity result showed that the minimum dose of B. Caballi that could be detected by PCR assay was 0. 01 fg/μL,45 clinical sample, from horses farm in China, doubtedly infected with B. Caballi were tested by PCR. The results showed that 12 positive samples could be detected by the PCR assay. At the same time,microscope were also used for the clinical samples. The test revealed that the sensitivity of the PCR assay was higher than that the microscope test.%根据驽巴贝斯虫(Babesia caballi)18S rRNA基因序列设计1对特异性引物,扩增出452 bp核苷酸片段,建立了检测驽巴贝斯虫病的PCR方法.敏感性试验结果表明,该方法最低能检出0.01 fg/μL驽巴贝斯虫DNA模板.特异性试验结果显示,在被检测的6个巴贝斯虫株中,仅驽巴贝斯虫株能扩增出特异性片段,马泰勒虫、双芽巴贝斯虫、莫氏巴贝斯虫、卵形巴贝斯虫、大巴贝斯虫的扩增结果均为阴性.对45份马属动物血样进行检测,本研究建立的PCR方法测得驽巴贝斯虫病的阳性率为26.67%(12/45),与显微镜检测方法进行了比较,结果显示PCR检测方法可显著提高驽巴贝斯虫的检出率.

  20. Increasing awareness of Rhodococcus equi pulmonary infection in the immunocompetent adult: a rare infection with poor prognosis.

    Science.gov (United States)

    Herath, Samantha; Lewis, Christopher; Nisbet, Mitzi

    2013-12-13

    The aim of this case report and review is to increase awareness of this uncommon infection with Rhodococcus equi (R. equi), in immunocompetent adults. R. equi is a soil-dwelling Gram-positive bacillus that frequently causes infection in grazing livestock. Human infection is rare and mostly limited to the immunocompromised hosts. We present a case of pneumonia caused by R. equi infection in a 55-year-old male builder who presented with cough, dyspnoea and night sweats, initially suspected to have pulmonary tuberculosis. Following biopsy of the mediastinal lymph nodes, R. equi was cultured, which is usually not a contaminant. Despite extensive investigations a host immune defect was not identified. The patient recovered after three months of combination antibiotic treatment, initially with intravenous vancomycin and meropenem followed by oral clarithromycin and rifampicin. To further clarify this rare disease we did a literature review that identified 26 adult patients with R. equi infection, without an identified host immunosuppressive condition. In this cohort, the median age at presentation was 53 years and infection holds a strong male predominance 19 (73%). An environmental exposure (e.g. farming, horse breeder) was found in 13 (50%). Ten (38%) of these patients had pulmonary infection. All deaths 3 (12%) occurred in the patients had pulmonary infection. R.equi is an infection that is difficult to diagnose and carries a high mortality if prompt treatment is not established. It is important to realise the potential for this disease to be misdiagnosed as pulmonary tuberculosis or community acquired pneumonia. Clinical suspicion is important especially if an environmental exposure is suspected.

  1. Factors associated to Theileria equi in equids of two microregions from Rio de Janeiro, Brazil Fatores associados à Theileria equi em equídeos de duas microrregiões do Rio de Janeiro, Brasil

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    Tiago Marques dos Santos

    2011-09-01

    Full Text Available Serum samples from 714 equids of Itaguaí and Serrana microregions, Rio de Janeiro, southeastern Brazil, were examined by indirect fluorescent antibody test (titer 1:80 for Theileria equi. The prevalence in the microregions and factors associated with seropositivity were evaluated and the prevalence ratio (PR calculated. The overall prevalence of T. equi infection was 81.09% (n = 579, with higher prevalence (p Amostras de soro de 714 equídeos das microrregiões de Itaguaí e Serrana, Rio de Janeiro, Brasil, foram submetidas ao teste de imunofluorescência indireta (título 1:80 para Theileria equi. A prevalência entre as microrregiões e os fatores associados à soropositividade foram avaliados e a razão de prevalência (RP calculada. A prevalência geral para T. equi foi de 81,09% (n = 579, com maior prevalência (p < 0,05 para microrregião de Itaguaí (85,43%, quando comparado a Serrana (76,92%. A região, altitude, nível da propriedade e origem dos equídeos foram associados (p < 0,05 com a soropositividade para T. equi. Equídeos criados na microrregião de Itaguaí (RP = 1,11; p = 0,003 e em altitudes abaixo de 500 m (RP = 1.10; p = 0,014 apresentaram maior chance de se tornarem soropositivos para T. equi. Além disso, quando são criados em propriedades de nível ruim (RP = 1,13; p = 0,018 e nascidos na propriedade (RP = 1,10; p = 0,008 apresentaram mais chance de terem contato com T. equi. As principais espécies de carrapatos encontradas parasitando os equídeos foram Amblyomma cajennense e Dermacentor (Anocentor nitens. As microrregiões estudadas são endêmicas para theileriose equina e de estabilidade enzoótica para T. equi. Apenas os fatores relacionados à área de coleta das amostras de soro influenciaram a soropositividade dos equídeos para T. equi naquela região.

  2. 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 (Pestimated 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.

  3. 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

    2017-08-10

    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.

  4. The First case of Locally Acquired Tick-Borne Babesia Microti Infection in Canada

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    Jared MP Bullard

    2014-01-01

    Full Text Available A child with a complicated medical history that included asplenia acquired an infection with Babesia microti in the summer of 2013 and had not travelled outside of Manitoba. Although the clinical findings were subtle, astute laboratory work helped to reach a preliminary identification of Babesia species, while reference laboratory testing confirmed the diagnosis. Blacklegged ticks (Ixodes scapularis are known to transmit Borrelia burgdorferi and Anaplasma phagocytophilum in the province; however, the present case represents the first known instance of tick-borne B microti, both in Manitoba and in Canada. The expanding territory of the blacklegged tick increases the relevance of this emerging infection. Clinicians, laboratory medical practitioners and public health officials should be aware of B microti as a potential locally acquired infection in Canada.

  5. 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.

  6. 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 β-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 β-tubulin gene sequence shared conserved regions with previously described Babesia and Theileria species. The intron of the β-tubulin gene was 45 bp. This is the first molecular characterization of an avian piroplasm.

  7. RPS8--a new informative DNA marker for phylogeny of Babesia and Theileria parasites in China.

    Directory of Open Access Journals (Sweden)

    Zhan-Cheng Tian

    Full Text Available Piroplasmosis is a serious debilitating and sometimes fatal disease. Phylogenetic relationships within piroplasmida are complex and remain unclear. We compared the intron-exon structure and DNA sequences of the RPS8 gene from Babesia and Theileria spp. isolates in China. Similar to 18S rDNA, the 40S ribosomal protein S8 gene, RPS8, including both coding and non-coding regions is a useful and novel genetic marker for defining species boundaries and for inferring phylogenies because it tends to have little intra-specific variation but considerable inter-specific difference. However, more samples are needed to verify the usefulness of the RPS8 (coding and non-coding regions gene as a marker for the phylogenetic position and detection of most Babesia and Theileria species, particularly for some closely related species.

  8. Survey of babesiosis in symptomatic dogs from Romania: occurrence of Babesia gibsoni associated with breed.

    Science.gov (United States)

    Imre, Mirela; Farkas, Róbert; Ilie, Marius Stelian; Imre, Kálmán; Dărăbuş, Gheorghe

    2013-12-01

    Blood samples from 49 symptomatic dogs from 5 western and north-western counties of Romania were screened using microscopic examination, polymerase-chain-reaction-restriction-fragment-length-polymorphism procedure (PCR-RFLP), and sequence analysis. Results of the microscopic evaluation of stained blood smears revealed 45 positive samples with the presence of large and small intraerythrocytic piroplasms in 35 and 10 samples, respectively. Babesia canis (35/49, 71.4%) and Babesia gibsoni (14/49, 28.6%) were identified and differentiated by PCR-RFLP targeting the 18S rRNA gene. Results of the sequence analysis of all B. gibsoni and 17 randomly selected B. canis PCR products confirmed the PCR-RFLP-diagnosed species. The distribution of B. gibsoni infection was positively associated (pRomania confirmed by sequencing.

  9. Molecular survey of Babesia infections in cattle from different areas of Myanmar.

    Science.gov (United States)

    Bawm, Saw; Htun, Lat Lat; Maw, Ni Ni; Ngwe, Tin; Tosa, Yusuke; Kon, Tomoyuki; Kaneko, Chiho; Nakao, Ryo; Sakurai, Tatsuya; Kato, Hirotomo; Katakura, Ken

    2016-02-01

    Cattle babesiosis is one of the most important tick-borne diseases worldwide. The present study reports a molecular survey of Babesia infections in cattle in Myanmar. Nested PCR assays based on the Babesia bigemina apical membrane antigen-1 gene (AMA-1) and B. bovis rhoptry associated protein-1 gene (RAP-1) revealed that the overall percentage of B. bigemina and B. bovis infection were 9.8% (70/713) and 17.1% (122/713), respectively. A mixed infection was detected in 4.6% (33/713) of animals. Animals Myanmar, providing useful information for the management and control of the disease. Copyright © 2015 Elsevier GmbH. All rights reserved.

  10. Detection of Babesia bigemina in cattle of different genetic groups and in Rhipicephalus (Boophilus) microplus tick.

    Science.gov (United States)

    Oliveira, M C S; Oliveira-Sequeira, T C G; Regitano, L C A; Alencar, M M; Néo, T A; Silva, A M; Oliveira, H N

    2008-08-17

    Babesia bigemina infections were investigated in four genetic groups of beef cattle and in Rhipicephalus (Boophilus) microplus engorged female ticks. Blood samples and engorged female ticks were collected from 15 cows and 15 calves from each of the following genetic groups: Nelore, Angus x Nelore, Canchim x Nelore, and Simmental x Nelore. Microscopic examination of blood smears and tick hemolymph revealed that merozoites of B. bigemina (6/60) as well as kinetes of Babesia spp. (9/549) were only detected in samples (blood and ticks, respectively) originated from calves. PCR-based methods using primers for specific detection of B. bigemina revealed 100% infection in both calves and cows, regardless the genetic group. Tick infection was detected by nested-PCR amplifications showing that the frequency of B. bigemina was higher (P0.05).

  11. Characterization of the Products of the Heme Detoxification Pathway in Malarial Late Trophozoites by X-ray Diffraction

    Energy Technology Data Exchange (ETDEWEB)

    Bohle,D.; Dinnebier, R.; Madsen, S.; Stephens, P.

    2007-01-01

    Heme is a potent multifunction regulator whose biochemical levels and distribution are precisely controlled on both intra- and extracellular levels. Efficient regulation of heme is particularly critical for intra-erythrocytic parasites such as plasmodia which process large quantities of heme in the post-invasion digestion of the erythrocyte's hemoglobin. Plasmodia, which lack heme oxygenases, detoxify heme by sequestering it into an insoluble heme aggregate termed malarial pigment or hemozoin. The quinoline-based family of antimalarials interfere with this process by an as yet unknown mechanism that has recently come under intense scrutiny as part of the effort to combat the spread of chloroquine-resistant strains of Plasmodium. A variety of spectroscopic and bioanalytical techniques indicate that hemozoin is similar to the synthetic aggregated heme phase {beta}-hematin, which is thought to form strands of hemes linked by propionate oxygen-iron bonds as well as interstrand propionate hydrogen bonds. Characterization of the carboxylate stretching bands for the propionic acid side chains by IR and Raman spectroscopy provides the best evidence for the presence of iron-oxygen bonds to the propionate side chains. Unfortunately, crystallographic characterization of these heme aggregates has been hampered by the phase heterogeneity of many synthetic preparations as well as by the small size of the synthetic and natural crystallites isolated from either trophozoites and infected hosts. High resolution powder diffraction has been used extensively for the solution of many structural problems, and it can solve problems posed by diffraction from microcrystalline phases. In this communication we describe the characterization of {beta}-hematin derived from both synthetic and natural sources and provide new unambiguous evidence that the heme aggregate present in late stage trophozoites is {beta}-hematin. In a process inhibited by the quinoline antimalarial drugs, Plasmodia

  12. 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.

  13. 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.

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

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    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.

  15. Human Coinfection with Borrelia burgdorferi and Babesia microti in the United States

    OpenAIRE

    2015-01-01

    Borrelia burgdorferi, the causative agent of Lyme disease, and Babesia microti, a causative agent of babesiosis, are increasingly implicated in the growing tick-borne disease burden in the northeastern United States. These pathogens are transmitted via the bite of an infected tick vector, Ixodes scapularis, which is capable of harboring and inoculating a host with multiple pathogens simultaneously. Clinical presentation of the diseases is heterogeneous and ranges from mild flu-like symptoms t...

  16. Prevalence of Babesia and Anaplasma in ticks infesting dogs in Great Britain.

    Science.gov (United States)

    Smith, Faith D; Wall, Lauren Ellse Richard

    2013-11-15

    Ticks are important vectors of disease in companion animals and transmit an extensive range of viral, bacterial and protozoan pathogens to dogs and cats. They may also be vectors of zoonotic pathogens which affect the health of in-contact owners. In recent years, babesiosis, and anaplasmosis have all shown signs of increased prevalence and distribution in various parts of Europe. Here, the prevalence of Anaplasma spp. and Babesia spp. pathogens in Ixodes ticks, collected from dogs in the UK in 2009, were evaluated using PCR and sequence analysis of the 16S rDNA or 18S rDNA regions respectively. Species identification was performed by alignment with existing sequences in GenBank. After sequencing, 5 out of 677 tick samples (0.74%) contained rDNA which shared 97-100%% sequence homology with Anaplasma phagocytophilum. Of these, three samples came from Ixodes ricinus and two from Ixodes hexagonus. Sixteen out of 742 ticks (2.4%) were positive for Babesia and of these 11 showed 97-100% homology with B. gibsoni. All of these 11 samples were derived from I. ricinus. One sample, again from I. ricinus, showed 99% homology for B. divergens. Four of the Babesia spp sequences were of the "venatorum" or EU1 type, three of which came from I. ricinus and one from an Ixodes canisuga. This strain has been associated with severe human cases of babeisiosis. A further 246 positive results, which appeared to show the presence of Anaplasma following PCR, were shown by sequence analysis to be derived from the bacterium Candidatus "Midichloria mitochondrii", which to date has been assumed to be non-pathogenic. The results are of interest because the presence of B. gibsoni in the UK further confirms the worldwide distribution of this piroplasm and supports the inference that I. ricinus may act as a vector for Babesia of the gibsoni-complex.

  17. Serum protein response and renal failure in canine Babesia annae infection

    OpenAIRE

    Camacho, Angel; Guitian, Francisco; Pallas, Estrella; Gestal, Juan; Olmeda, Sonia; Goethert, Heidi; III, Sam; Spielman, Andrew

    2005-01-01

    International audience; Babesia annae piroplasms have recently been recognised as a cause of infection and disease among dogs in Europe. The pathogenesis and clinical implications of this emerging disease remain poorly understood. We conducted this study to describe the electrophoretic profiles associated with the infection and to determine if B. annae associated azotaemia is caused by renal failure. We examined by microscopy 2 979 canine blood samples submitted to a diagnostic laboratory in ...

  18. Vertical Transmission of Babesia microti in BALB/c Mice: Preliminary Report.

    Science.gov (United States)

    Bednarska, Malgorzata; Bajer, Anna; Drozdowska, Anna; Mierzejewska, Ewa J; Tolkacz, Katarzyna; Welc-Falęciak, Renata

    2015-01-01

    Babesia spp. (Apicomplexa, Piroplasmida) are obligate parasites of many species of mammals, causing a malaria-like infection- babesiosis. Three routes of Babesia infection have been recognized to date. The main route is by a tick bite, the second is via blood transfusion. The third, vertical route of infection is poorly recognized and understood. Our study focused on vertical transmission of B. microti in a well-established mouse model. We assessed the success of this route of infection in BALB/c mice with acute and chronic infections of B. microti. In experimental groups, females were mated on the 1st day of Babesia infection (Group G0); on the 28th day post infection (dpi) in the post- acute phase of the parasite infection (G28); and on the 90th and 150th dpi (G90 and G150 group, respectively), in the chronic phase of the parasite infection. Pups were obtained from 58% of females mated in the post-acute phase (G28) and from 33% of females in groups G90 and G150. Mice mated in the pre-acute phase of infection (G0) did not deliver pups. Congenital B. microti infections were detected by PCR amplification of Babesia 18S rDNA in almost all pups (96%) from the experimental groups G28, G90 and G150. Parasitaemia in the F1 generation was low and varied between 0.01-0.001%. Vertical transmission of B. microti was demonstrated for the first time in BALB/c mice.

  19. Vertical Transmission of Babesia microti in BALB/c Mice: Preliminary Report.

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    Malgorzata Bednarska

    Full Text Available Babesia spp. (Apicomplexa, Piroplasmida are obligate parasites of many species of mammals, causing a malaria-like infection- babesiosis. Three routes of Babesia infection have been recognized to date. The main route is by a tick bite, the second is via blood transfusion. The third, vertical route of infection is poorly recognized and understood. Our study focused on vertical transmission of B. microti in a well-established mouse model. We assessed the success of this route of infection in BALB/c mice with acute and chronic infections of B. microti. In experimental groups, females were mated on the 1st day of Babesia infection (Group G0; on the 28th day post infection (dpi in the post- acute phase of the parasite infection (G28; and on the 90th and 150th dpi (G90 and G150 group, respectively, in the chronic phase of the parasite infection. Pups were obtained from 58% of females mated in the post-acute phase (G28 and from 33% of females in groups G90 and G150. Mice mated in the pre-acute phase of infection (G0 did not deliver pups. Congenital B. microti infections were detected by PCR amplification of Babesia 18S rDNA in almost all pups (96% from the experimental groups G28, G90 and G150. Parasitaemia in the F1 generation was low and varied between 0.01-0.001%. Vertical transmission of B. microti was demonstrated for the first time in BALB/c mice.

  20. 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)

    Awinda, Peter O; Mealey, Robert H; Williams, Laura B A; Conrad, Patricia A; Packham, Andrea E; Reif, Kathryn E; Grause, Juanita F; Pelzel-McCluskey, Angela M; Chung, Chungwon; Bastos, Reginaldo G; Kappmeyer, Lowell S; Howe, Daniel K; Ness, SallyAnne L; Knowles, Donald P; Ueti, Massaro W

    2013-11-01

    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 reemergence of Theileria equi in the United States prompted a widespread national survey resulting in identification of limited distribution of equine piroplasmosis (EP) in the U.S. horse population. This program identified Babesia caballi-seropositive horses using rhoptry-associated protein 1 (RAP-1)-competitive enzyme-linked immunosorbent assay (cELISA), despite B. caballi being considered nonendemic on the U.S. mainland. The purpose of the present study was to evaluate the suitability of RAP-1-cELISA as a single serological test to determine the infection status of B. caballi in U.S. horses. Immunoblotting indicated that sera from U.S. horses reacted with B. caballi lysate and purified B. caballi RAP-1 protein. Antibody reactivity to B. caballi lysate was exclusively directed against a single ∼50-kDa band corresponding to a native B. caballi RAP-1 protein. In contrast, sera from experimentally and naturally infected horses from regions where B. caballi is endemic bound multiple proteins ranging from 30 to 50 kDa. Dilutions of sera from U.S. horses positive by cELISA revealed low levels of antibodies, while sera from horses experimentally infected with B. caballi and from areas where B. caballi is endemic had comparatively high antibody levels. Finally, blood transfer from seropositive U.S. horses into naive horses demonstrated no evidence of B. caballi transmission, confirming that antibody reactivity in cELISA-positive U.S. horses was not consistent with infection. Therefore, we conclude that a combination of cELISA and immunoblotting is required for the accurate serodiagnosis of B. caballi.

  1. Identification of pathogens and virulence profile of Rhodococcus equi and Escherichia coli strains obtained from sand of parks

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    M.C. Fernandes

    2013-01-01

    Full Text Available The identification of pathogens of viral (Rotavirus, Coronavirus, parasitic (Toxocara spp. and bacterial (Escherichia coli, Salmonella spp., Rhodococcus equi origin shed in feces, and the virulence profile of R. equi and E. coli isolates were investigated in 200 samples of sand obtained from 40 parks, located in central region of state of Sao Paulo, Brazil, using different diagnostic methods. From 200 samples analyzed, 23 (11.5% strains of R. equi were isolated. None of the R. equi isolates showed a virulent (vapA gene or intermediately virulent (vapB gene profiles. Sixty-three (31.5% strains of E. coli were identified. The following genes encoding virulence factors were identified in E. coli: eae, bfp, saa, iucD, papGI, sfa and hly. Phylogenetic classification showed that 63 E. coli isolates belonged to groups B1 (52.4%, A (25.4% and B2 (22.2%. No E. coli serotype O157:H7 was identified. Eggs of Toxocara sp. were found in three parks and genetic material of bovine Coronavirus was identified in one sample of one park. No Salmonella spp. and Rotavirus isolates were identified in the samples of sand. The presence of R. equi, Toxocara sp, bovine Coronavirus and virulent E. coli isolates in the environment of parks indicates that the sanitary conditions of the sand should be improved in order to reduce the risks of fecal transmission of pathogens of zoonotic potential to humans in these places.

  2. Identification of pathogens and virulence profile of Rhodococcus equi and Escherichia coli strains obtained from sand of parks.

    Science.gov (United States)

    Fernandes, M C; Takai, S; Leite, D S; Pinto, J P A N; Brandão, P E; Santarém, V A; Listoni, F J P; Da Silva, A V; Ribeiro, M G

    2013-01-01

    The identification of pathogens of viral (Rotavirus, Coronavirus), parasitic (Toxocara spp.) and bacterial (Escherichia coli, Salmonella spp., Rhodococcus equi) origin shed in feces, and the virulence profile of R. equi and E. coli isolates were investigated in 200 samples of sand obtained from 40 parks, located in central region of state of Sao Paulo, Brazil, using different diagnostic methods. From 200 samples analyzed, 23 (11.5%) strains of R. equi were isolated. None of the R. equi isolates showed a virulent (vapA gene) or intermediately virulent (vapB gene) profiles. Sixty-three (31.5%) strains of E. coli were identified. The following genes encoding virulence factors were identified in E. coli: eae, bfp, saa, iucD, papGI, sfa and hly. Phylogenetic classification showed that 63 E. coli isolates belonged to groups B1 (52.4%), A (25.4%) and B2 (22.2%). No E. coli serotype O157:H7 was identified. Eggs of Toxocara sp. were found in three parks and genetic material of bovine Coronavirus was identified in one sample of one park. No Salmonella spp. and Rotavirus isolates were identified in the samples of sand. The presence of R. equi, Toxocara sp, bovine Coronavirus and virulent E. coli isolates in the environment of parks indicates that the sanitary conditions of the sand should be improved in order to reduce the risks of fecal transmission of pathogens of zoonotic potential to humans in these places.

  3. 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.

  4. Identification and characterization of the RouenBd1987 Babesia divergens Rhopty-Associated Protein 1.

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    Marilis Rodriguez

    Full Text Available Human babesiosis is caused by one of several babesial species transmitted by ixodid ticks that have distinct geographical distributions based on the presence of competent animal hosts. The pathology of babesiosis, like malaria, is a consequence of the parasitaemia which develops through the cyclical replication of Babesia parasites in a patient's red blood cells, though symptoms typically are nonspecific. We have identified the gene encoding Rhoptry-Associated Protein -1 (RAP-1 from a human isolate of B. divergens, Rouen1987 and characterized its protein product at the molecular and cellular level. Consistent with other Babesia RAP-1 homologues, BdRAP-1 is expressed as a 46 kDa protein in the parasite rhoptries, suggesting a possible role in red cell invasion. Native BdRAP-1 binds to an unidentified red cell receptor(s that appears to be non-sialylated and non-proteinacious in nature, but we do not find significant reduction in growth with anti-rRAP1 antibodies in vitro, highlighting the possibility the B. divergens is able to use alternative pathways for invasion, or there is an alternative, complementary, role for BdRAP-1 during the invasion process. As it is the parasite's ability to recognize and then invade host cells which is central to clinical disease, characterising and understanding the role of Babesia-derived proteins involved in these steps are of great interest for the development of an effective prophylaxis.

  5. Molecular and serological detection of Babesia spp. in neotropical and exotic carnivores in Brazilian zoos.

    Science.gov (United States)

    André, Marcos Rogério; Adania, Cristina Harumi; Teixeira, Rodrigo Hidalgo Friciello; Allegretti, Silmara Marques; Machado, Rosangela Zacarias

    2011-03-01

    Large and small piroplasms have been observed in the blood smears of various wild carnivores, but few studies utilizing molecular characterization have been done. The goal of this present study was to investigate the presence of Babesia sp. by molecular and serologic techniques in exotic and neotropical carnivores maintained in captivity at Brazilian zoos. Blood and sera samples were collected from 146 Brazilian wild felids, 21 exotic felids, 1 genet (Genetta tigrina), 3 European wolves (Canis lupus), and 94 Brazilian wild canids in Brazilian zoos in the São Paulo and Mato Grosso states and in the Federal District. A total of 53 wild felids (31.74%) and 10 wild canids (10.31%) were seropositive for Babesia canis by Indirect Fluorescent Antibody Test (IFAT). Antibodies were detected in ocelots, little-spotted cats, margays, pampas cats, jaguars, pumas, jaguarundis, crab-eating foxes, and bush dogs. Babesia sp. DNA, with high similarity to B. leo, was detected in one pampas cat and one genet.

  6. Zoonotic Babesia: possibly emerging pathogens to be considered for tick-infested humans in Central Europe.

    Science.gov (United States)

    Hunfeld, K P; Brade, V

    2004-04-01

    The three host-tick Ixodes (I.) ricinus is regarded as an important vector of tick-borne microorganisms pathogenic for humans in central Europe and is primarily known as the main vector of Borrelia (B.) burgdorferi and the virus causing tick-borne encephalitis (TBE), the most clinically relevant tick transmitted pathogens for humans in European countries. Furthermore, it is now well established that I. ricinus also transmits Ehrlichia (E.) phagocytophila, Babesia (Ba.) divergens, and Ba. microti, all agents of zoonotic infections in dear, sheep, cattle, dogs, and horses. In addition to their known zoonotic potential, recent molecular-epidemiological and seroepidemiological surveys as well as increasingly reported clinical cases of infections caused by these tick-borne organisms other than B. burgdorferi (TOBB) also strongly suggest a possible relevance of Babesia, Ehrlichia and Rickettsia for humans at risk in Europe. However, there are few medical microbiological investigations and epidemiological data on the distribution and relevance of Babesia for humans in our part of the northern hemisphere. There is also very little diagnostic and clinical knowledge on human babesiosis in many regions of Europe. Furthermore, sophisticated diagnostic tools designed for the reliable detection of the underlying pathogens, are not yet generally available to the microbiological laboratory. This review aims to provide basic information on human babesiosis and the most relevant causative pathogens of the disease in Europe and to draw attention to this parasitic infection as a possibly emerging and probably under-diagnosed disease in this part of the northern hemisphere.

  7. A Historical Overview of Research on Babesia orientalis, a Protozoan Parasite Infecting Water Buffalo

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    Lan He

    2017-07-01

    Full Text Available Babesiosis is a globally important zoonotic disease caused by tick-borne intraerythrocytic protozoan of the genus Babesia (phylum apicomplexa. In China, there are five species that infect cattle buffalo and cause great economic loss, which include Babesia bigemina, B. bovis, B. major, B. ovata, and B. orientalis. Among them, B. orientalis is the most recently identified new Babesia species epidemic in China. This review summarized the work done in the past 33 years to give an overview of what learned about this parasite. This parasitic protozoan was found in 1984 in Central and South China and then named as B. orientalis in 1997 based on its differences in transmitting host, morphology, pathogenicity and characteristics of in vitro cultivation when compared with B. bigemina and B. bovis. It was found that Rhipicephalus haemaphysaloides is the transmitting vector and water buffalo is the only reported host. Phylogenetic analysis based on the 18S rRNA gene also confirmed that B. orientalis is a new species. After species verification, four diagnostic methods including semi-nest PCR, loop-mediated isothermal amplification assay, reverse line blot hybridization assay, and real-time PCR were established for lab and field use purposes. Genomic sequencing was conducted and the complete genomes of mitochondria and apicoplast were annotated. Future work will be focused on developing effective vaccines, identifying drug targets and screening useful drugs for controlling B. orientalis in water buffalo.

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

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    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.

  9. 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.

  10. Association of airborne concentration of virulent Rhodococcus equi with location (stall versus paddock) and month (January through June) on 30 horse breeding farms in central Kentucky.

    Science.gov (United States)

    Cohen, Noah D; Kuskie, Kyle R; Smith, Jacqueline L; Slovis, Nathan M; Brown, Stuart E; Stepusin, Randolph S; Chaffin, M Keith; Takai, Shinji; Carter, Craig N

    2012-10-01

    To determine whether the concentration of airborne virulent Rhodococcus equi varied by location (stall vs paddock) and month on horse farms. Air samples from stalls and paddocks used to house mares and foals on 30 horse breeding farms in central Kentucky. Air samples from 1 stall and 1 paddock were obtained monthly from each farm from January through June 2009. Concentrations of airborne virulent R equi were determined via a modified colony immunoblot assay. Random-effects logistic regression was used to determine the association of the presence of airborne virulent R equi with location from which air samples were obtained and month during which samples were collected. Of 180 air samples, virulent R equi was identified in 49 (27%) and 13 (7%) obtained from stalls and paddocks, respectively. The OR of detecting virulent R equi in air samples from stalls versus paddocks was 5.2 (95% confidence interval, 2.1 to 13.1). Of 60 air samples, virulent R equi was identified in 25 (42%), 18 (30%), and 6 (10%) obtained from stalls during January and February, March and April, and May and June, respectively. The OR of detecting virulent R equi from stall air samples collected during May and June versus January and February was 0.22 (95% confidence interval, 0.08 to 0.63). Foals were more likely to be exposed to airborne virulent R equi when housed in stalls versus paddocks and earlier (January and February) versus later (May and June) during the foaling season.

  11. Characterization of Rhodococcus equi isolates from submaxillary lymph nodes of wild boars (Sus scrofa), red deer (Cervus elaphus) and roe deer (Capreolus capreolus).

    Science.gov (United States)

    Rzewuska, Magdalena; Witkowski, Lucjan; Cisek, Agata A; Stefańska, Ilona; Chrobak, Dorota; Stefaniuk, Elżbieta; Kizerwetter-Świda, Magdalena; Takai, Shinji

    2014-08-06

    Rhodococcus equi is a soil saprophyte and an opportunistic pathogen causing infections in animals, and rarely in humans. The presence of R. equi in tissues and faeces of some wild animal species was demonstrated previously. In this study we characterized R. equi isolates from submaxillary lymph nodes of free-living wild boars (n=23), red deer (n=2) and roe deer (n=2). This is the first description of R. equi strains isolated from tissues of the Cervidae. All isolates were initially recognized as R. equi based on the phenotypic properties. Their identification was confirmed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, detection of the choE gene and by sequence analysis of the 16S rRNA and rpoB genes. The presence of three plasmidic genes (traA, vapA and vapB) associated with R. equi virulence was investigated by PCR. In 16 wild boar isolates the traA and vapB genes were detected and they were located on virulence plasmids type 5, 7 or 11. The isolates from cervids and the remaining wild boar isolates were classified as avirulent based on a genotype traA(-)/vapA(-)B(-). In summary, these results confirm that wild boars can be a source of intermediately virulent R. equi strains, and indicate that red deer and roe deer can be a reservoir of avirulent R. equi strains. Copyright © 2014 Elsevier B.V. All rights reserved.

  12. Minimal inhibitory concentration of azithromycin in Rhodococcus equi strains isolated from foals Concentração inibitória mínima da azitromicina em linhagens de Rhodococcus equi isoladas de potros

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    M.G. Ribeiro

    2006-12-01

    Full Text Available O perfil de sensibilidade microbiana e a concentração inibitória mínima (MIC da azitromicina para 42 cepas de Rhodococcus equi isoladas de potros, no Brasil, e em uma cepa-controle, foi avaliado, respectivamente, pelos métodos de difusão com discos e E-test. A azitromicina apresentou 100% de efetividade in vitro para todas as cepas em ambos os testes. As cepas de R. equi apresentaram MIC90 para azitromicina em valores <1.5µg/ml. Este estudo mostra a alta efetividade da azitromicina em linhagens de R. equi isoladas no Brasil, sugerindo o uso dessa droga como alternativa na terapia da rodococose em potros.

  13. 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

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    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",

  14. Coexistence of sense and anti-sense mRNAs of variant surface protein in Giardia lamblia trophozoites.

    Science.gov (United States)

    Guo, Junli; Zheng, Wenyu; Wang, Yuehua; Li, Yao; Lu, Siqi; Feng, Xianmin

    2014-02-14

    A strategy of the parasitic protozoan Giardia lamblia to evade attack from the host immune system is periodic changes of its surface antigen, a member of the variant surface protein (VSP) family. A post-transcriptional gene silencing mechanism has been proposed to explain the presence of only one among many possible VSPs at any time. To investigate this phenomenon further, we extracted total RNA from cultured trophozoites of the G. lamblia C2 isolate, and cDNA was reverse-transcribed from the RNA. Sense and anti-sense VSPs were amplified from the total cDNA using nested PCR with primers designed from the 3'-conserved region and the known 5' or 3' end of the cDNA library. Sequence analyses of the amplified products revealed more than 34 full-length antisense VSPs and a smear of sense VSPs. Sequence alignments and comparisons revealed that these VSPs contained variable N-termini and conserved C-termini, and could be classified into 5 clades based on the sizes and variations of the N-terminal sequence. All antisense VSPs existed in the sense forms, but no corresponding antisense VSP existed for sense RNA (snsRNA) 16. The coexistence of sense and antisense VSP mRNAs in cultured G. lamblia supports the post-transcriptional regulation of VSP expression. We propose that VSPs transcribed simultaneously in the sense and antisense forms form double-stranded RNAs (dsRNAs) which are degraded by the Dicer endonuclease, while a VSP without an antisense transcription (e.g., snsRNA16) will be expressed on the surface of Giardia. In addition, in the course of this investigation VSPs were identified that were previously not known. PCR-based amplification of specific sense and antisense VSP cDNAs can be used to identify the specific VSP on G. lamblia trophozoites, which is easier than using specific monoclonal antibody approaches.

  15. Babesia bovis expresses Bbo-6cys-E, a member of a novel gene family that is homologous to the 6-cys family of Plasmodium

    Science.gov (United States)

    A novel Babesia bovis gene family encoding proteins with similarities to the Plasmodium 6cys protein family was identified by TBLASTN searches of the Babesia bovis genome using the sequence of the P. falciparum PFS230 protein as query, and was termed Bbo-6cys gene family. The Bbo-cys6 gene family co...

  16. Transmission of Babesia caballi by Dermacentor nitens (Acari: Ixodidae) Is Restricted to One Generation in the Absense of Alimentary Reinfection on a Susceptible Equine Host

    Science.gov (United States)

    The tropical horse tick, Dermacentor nitens, is the natural vector of Babesia caballi in the Americas; the distribution of this tick in the United States is limited to the southernmost parts of Florida and Texas. Babesia caballi, one of the etiologic agents of equine babesiosis, occurs widely throug...

  17. Immunolocalization of Delta-Giardin within the Ventral Disc in of Trophozoites and in cysts of Giardia duodenalis using the Multiplex Laser Scanning Confocal Microscopy

    Science.gov (United States)

    Immunolocalization of alpha2-, beta- and delta-giardin in Giardia showed that in the trophozoites and cysts delta-giardin it strictly associated with the ventral disc. Optical sectioning of the ventral discs, together with quantitative colocalization of the immunoreactivity for delta- and beta-giard...

  18. 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.

  19. Study on the production of IL-12, IL-10 in early stage after infection with Babesia microti and Babesia rodhaini in mice

    Institute of Scientific and Technical Information of China (English)

    LIU Yun; Hachikuchi Rie; Kenichiro Ono

    2007-01-01

    The serum levels of IL-12 and IL-10 in mice after infected with Babesia microti (B. microti) and Babesia rodhaini(B. rodhaini) were examined. Collected the mice serum and examined the concentration of IL-12 and IL-10 by using ELISA after infection with B. microti and B. rodhaini at 0, 3, 6, 9, 12, 18, 24, 36, 72, 96 h in mice. The results showed that B. microti infection resulted in IL-12 increasing, which peaked at 3 and 24 h after the infection, while same infection did not induce a significant change in IL-10 compared to uninfected mice. When mice were infected with B. rodhaini, any significant changes were not decteted both in IL-12and IL-10 in comparison with uninfected animals during the period of 3-72 h after infection. Instead, a significant decline in IL-12 and IL-10 was found compared to uninfected mice 96 h after infection with B. rodhaini. It indicates that the mutagenetic cytokine is IL-12in the serum of mice after infection with B. microti, and no any significant changes were detected in both IL-12 and IL-10 from 0 to 72 h after infected with B. rodhaini.

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

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    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.

  1. Sympatric occurrence of Ixodes ricinus, Dermacentor reticulatus and Haemaphysalis concinna ticks and Rickettsia and Babesia species in Slovakia.

    Science.gov (United States)

    Svehlová, Andrea; Berthová, Lenka; Sallay, Balázs; Boldiš, Vojtech; Sparagano, Olivier A E; Spitalská, Eva

    2014-09-01

    Vojka nad Dunajom in the south-west of the Slovak Republic is a locality with sympatric occurrence of 3 species of ticks. This study investigated the spatial distribution of Dermacentor reticulatus, Ixodes ricinus, and Haemaphysalis concinna ticks in this area and determined the prevalence of Babesia and Rickettsia species in questing adults of these tick species considered as potential risk for humans and animals. Ticks were collected by blanket dragging over the vegetation from September 2011 to October 2012. All ticks were subjected to DNA extraction and individually assayed with PCR-based methods targeting the gltA, sca4, 23S rRNA genes of Rickettsia spp. and the 18S rRNA gene of Babesia spp. D. reticulatus was the dominant species occurring in this area (67.7%, n=600), followed by I. ricinus (31.8%, n=282) and H. concinna (0.5%, n=4) ticks. Rickettsial infection was determined in 10.8% (n=65) and 11.7% (n=33) of D. reticulatus and I. ricinus ticks, respectively. Babesia spp. infection was confirmed in 1.8% (n=11) of D. reticulatus and 0.4% (n=1) of I. ricinus ticks. DNA of 6 different pathogenic tick-borne species, Rickettsia helvetica, Rickettsia monacensis, Rickettsia slovaca, Rickettsia raoultii, Babesia canis, and Babesia venatorum were identified in this locality with sympatric occurrence of I. ricinus, D. reticulatus, and H. concinna ticks.

  2. 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

    OpenAIRE

    Jaime Galvão Dias Júnior; Fernando José Paganini Listoni; Jane Megid; Aldo Santana de Carvalho Filho; Márcio Garcia Ribeiro

    2000-01-01

    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...

  3. Prevalence of Streptococcus equi subsp. equi in horses and associated risk factors in the State of Rio Grande do Sul, Brazil.

    Science.gov (United States)

    Libardoni, Felipe; Machado, Gustavo; Gressler, Letícia Trevisan; Kowalski, Ananda Paula; Diehl, Gustavo Nogueira; dos Santos, Lucila Carboneiro; Corbellini, Luis Gustavo; de Vargas, Agueda Castagna

    2016-02-01

    The aim of this study was to estimate the prevalence of equine strangles and to identify associated risk factors for this disease through a cross-sectional study of nasal swabs. Nasal swabs (n=1010) from healthy equines (absence of nasal discharge, lymphadenopathy and cough) from 341 farms were plated on 5% blood agar; of these horses, 24 were identified as positive for Streptococcus equi through isolation, PCR and DNA sequencing. The estimated prevalence for individual animals was 2.3%, and for herds, it was 5.86%. Statistical analysis identified the following as associated risk factors: the number of group events that were attended by the equines (PR: 1.06); the sharing of food containers (PR: 3.74); and at least one previous positive diagnosis of strangles on the farm (PR: 3.20). These results constitute an epidemiological contribution to the horse industry and may support measures for the future control of the disease.

  4. Affects of N-terminal variation in the SeM protein of Streptococcus equi on antibody and fibrinogen binding.

    Science.gov (United States)

    Timoney, John F; DeNegri, Rafaela; Sheoran, Abhineet; Forster, Nathalie

    2010-02-10

    The clonal Streptococcus equi causes equine strangles, a highly contagious suppurative lymphadenopathy and rhinopharyngitis. An important virulence factor and vaccine component, the antiphagocytic fibrinogen binding SeM of S. equi is a surface anchored fibrillar protein. Two recent studies of N. American, Japanese and European isolates have revealed a high frequency of N-terminal amino acid variation in SeM of S. equi CF32 that suggests this region of the protein is subject to immunologic selection pressure. The aims of the present study were firstly to map regions of SeM reactive with convalescent equine IgG and IgA and stimulatory for lymph node cells and secondly to determine effects of N-terminal variation on the functionality of SeM. Variation did not significantly affect fibrinogen binding or susceptibility of S. equi to an opsonic equine serum. Linear epitopes reactive with convalescent IgG and mucosal IgA were concentrated toward the conserved center of SeM. However, IgA but not IgG from every horse reacted with at least one peptide that contained variable sequence. Lymph node cells (CD4+) from horses immunized with SeM were strongly responsive to a peptide (alphaalpha36-138) encoding the entire variable region. SeM (CF32) specific mouse Mab 04D11 which reacted strongly with this larger peptide but not with shorter peptides within that sequence reacted strongly with whole cells of S. equi CF32 but only weakly with cells of any of 14 isolates of S. equi expressing different variants of SeM. These results in combination suggest that N-terminal variation alters a conformational epitope of significance in mucosal IgA and systemic T cell responses but does not affect antibody mediated phagocytosis and killing.

  5. Policies and Processes for Social Inclusion: Using EquiFrame and EquIPP for Policy Dialogue

    Science.gov (United States)

    MacLachlan, Malcolm; Mannan, Hasheem; Huss, Tessy; Munthali, Alister; Amin, Mutamad

    2016-01-01

    The application of EquiFrame in the analysis of sexual and reproductive health policies by Ivanova et al to a new thematic area, their selection of only some of the Core Concepts of human rights in health service provision and the addition of new vulnerable groups relevant to the purpose of their analysis, are all very welcome developments. We also applaud their application of EquiFrame to policies in countries where it has not previously been used, along with their use of interviews with policy-makers to produce a deeper understanding of policy processes. We argue that clear justification for the inclusion of additional, or replacement of some exiting vulnerable groups within EquiFrame should be accompanied by clear definitions of such groups, along with the evidence-base that justifies their classification as a vulnerable or marginalised group. To illustrate the versatility of EquiFrame, we summarise a range of ways in which it has been used across a number of regions; including a brief Case Study of its use to develop the National Health Policy of Malawi. While EquiFrame focuses on policy content, we preview a new policy analysis tool – Equity and Inclusion in Policy Processes (EquIPP) – which assesses the extent of equity and inclusion in broader policy processes. Together, EquiFrame and EquIPP can be used to help governments and civil society ensure that policies are addressing the much stronger emphasis on social inclusion, now apparent in the Sustainable Development Goals (SDGs). PMID:26927591

  6. Rhodococcus equi's extreme resistance to hydrogen peroxide is mainly conferred by one of its four catalase genes.

    Directory of Open Access Journals (Sweden)

    Pauline Bidaud

    Full Text Available Rhodococcus equi is one of the most widespread causes of disease in foals aged from 1 to 6 months. R. equi possesses antioxidant defense mechanisms to protect it from reactive oxygen metabolites such as hydrogen peroxide (H(2O(2 generated during the respiratory burst of phagocytic cells. These defense mechanisms include enzymes such as catalase, which detoxify hydrogen peroxide. Recently, an analysis of the R. equi 103 genome sequence revealed the presence of four potential catalase genes. We first constructed ΔkatA-, ΔkatB-, ΔkatC-and ΔkatD-deficient mutants to study the ability of R. equi to survive exposure to H(2O(2in vitro and within mouse peritoneal macrophages. Results showed that ΔkatA and, to a lesser extent ΔkatC, were affected by 80 mM H(2O(2. Moreover, katA deletion seems to significantly affect the ability of R. equi to survive within murine macrophages. We finally investigated the expression of the four catalases in response to H(2O(2 assays with a real time PCR technique. Results showed that katA is overexpressed 367.9 times (± 122.6 in response to exposure to 50 mM of H(2O(2 added in the stationary phase, and 3.11 times (± 0.59 when treatment was administered in the exponential phase. In untreated bacteria, katB, katC and katD were overexpressed from 4.3 to 17.5 times in the stationary compared to the exponential phase. Taken together, our results show that KatA is the major catalase involved in the extreme H(2O(2 resistance capability of R. equi.

  7. Age-related changes following in vitro stimulation with Rhodococcus equi of peripheral blood leukocytes from neonatal foals.

    Science.gov (United States)

    Kachroo, Priyanka; Ivanov, Ivan; Seabury, Ashley G; Liu, Mei; Chowdhary, Bhanu P; Cohen, Noah D

    2013-01-01

    Rhodococcus equi is an intracellular bacterium primarily known as an equine pathogen that infects young foals causing a pyogranulomatuous pneumonia. The molecular mechanisms mediating the immune response of foals to R. equi are not fully elucidated. Hence, global genomic high-throughput tools like gene expression microarrays might identify age-related gene expression signatures and molecular pathways that contribute to the immune mechanisms underlying the inherent susceptibility of foals to disease caused by R. equi. The objectives of this study were 2-fold: 1) to compare the expression profiles at specific ages of blood leukocytes from foals stimulated with virulent R. equi with those of unstimulated leukocytes; and, 2) to characterize the age-related changes in the gene expression profile associated with blood leukocytes in response to stimulation with virulent R. equi. Peripheral blood leukocytes were obtained from 6 foals within 24 hours (h) of birth (day 1) and 2, 4, and 8 weeks after birth. The samples were split, such that half were stimulated with live virulent R. equi, and the other half served as unstimulated control. RNA was extracted and the generated cDNA was labeled with fluorescent dyes for microarray hybridizations using an equine microarray. Our findings suggest that there is age-related differential expression of genes involved in host immune response and immunity. We found induction of genes critical for host immunity against pathogens (MHC class II) only at the later time-points (compared to birth). While it appears that foals up to 8-weeks of age are able to initiate a protective inflammatory response against the bacteria, relatively decreased expression of various other immune-related genes points toward inherent diminished immune responses closer to birth. These genes and pathways may contribute to disease susceptibility in foals if infected early in life, and might thus be targeted for developing preventative or therapeutic strategies.

  8. Effects of age and macrophage lineage on intracellular survival and cytokine induction after infection with Rhodococcus equi.

    Science.gov (United States)

    Berghaus, Londa J; Giguère, Steeve; Sturgill, Tracy L

    2014-07-15

    Rhodococcus equi, a facultative intracellular pathogen of macrophages, causes life-threatening pneumonia in foals and in people with underlying immune deficiencies. As a basis for this study, we hypothesized that macrophage lineage and age would affect intracellular survival of R. equi and cytokine induction after infection. Monocyte-derived and bronchoalveolar macrophages from 10 adult horses and from 10 foals (sampled at 1-3 days, 2 weeks, 1 month, 3 months, and 5 months of age) were infected ex vivo with virulent R. equi. Intracellular R. equi were quantified and mRNA expression of IL-1β, IL-4, IL-6, IL-8, IL-10, IL-12 p40, IL-18, IFN-γ, and TNF-α was measured. Intracellular replication of R. equi was significantly (Pequi was significantly (P=0.002) higher in 3-month-old foals than in 3-day old foals, 2-week-old foals, 1-month-old foals, and adult horses. Expression of IL-4 mRNA was significantly higher in monocyte-derived macrophages whereas expression of IL-6, IL-18, and TNF-α was significantly higher in bronchoalveolar macrophages. Induction of IL-1β, IL-10, IL-12 p40, and IL-8 mRNA in bronchoalveolar macrophages of 1-3-day old foals was significantly higher than in older foals or adult horses. Preferential intracellular survival of R. equi in bronchoalveolar macrophages of juvenile horses may play a role in the pulmonary tropism of the pathogen and in the window of age susceptibility to infection. Copyright © 2014 Elsevier B.V. All rights reserved.

  9. Age-related changes following in vitro stimulation with Rhodococcus equi of peripheral blood leukocytes from neonatal foals.

    Directory of Open Access Journals (Sweden)

    Priyanka Kachroo

    Full Text Available Rhodococcus equi is an intracellular bacterium primarily known as an equine pathogen that infects young foals causing a pyogranulomatuous pneumonia. The molecular mechanisms mediating the immune response of foals to R. equi are not fully elucidated. Hence, global genomic high-throughput tools like gene expression microarrays might identify age-related gene expression signatures and molecular pathways that contribute to the immune mechanisms underlying the inherent susceptibility of foals to disease caused by R. equi. The objectives of this study were 2-fold: 1 to compare the expression profiles at specific ages of blood leukocytes from foals stimulated with virulent R. equi with those of unstimulated leukocytes; and, 2 to characterize the age-related changes in the gene expression profile associated with blood leukocytes in response to stimulation with virulent R. equi. Peripheral blood leukocytes were obtained from 6 foals within 24 hours (h of birth (day 1 and 2, 4, and 8 weeks after birth. The samples were split, such that half were stimulated with live virulent R. equi, and the other half served as unstimulated control. RNA was extracted and the generated cDNA was labeled with fluorescent dyes for microarray hybridizations using an equine microarray. Our findings suggest that there is age-related differential expression of genes involved in host immune response and immunity. We found induction of genes critical for host immunity against pathogens (MHC class II only at the later time-points (compared to birth. While it appears that foals up to 8-weeks of age are able to initiate a protective inflammatory response against the bacteria, relatively decreased expression of various other immune-related genes points toward inherent diminished immune responses closer to birth. These genes and pathways may contribute to disease susceptibility in foals if infected early in life, and might thus be targeted for developing preventative or therapeutic

  10. 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.

  11. Efeito in vitro da josamicina em cepas de rhodocaccus equi isoladas de afecções pulmonares em potros

    OpenAIRE

    Ribeiro, Márcio Garcia; Universidade Estadual Paulista; Filho, Aldo Santana de Carvalho; Universidade Estadual Paulista; Megid, Jane; Universidade Estadual Paulista; Listoni, Fernando José Paganini; Universidade Estadual Paulista; Dias Júnior, Jaime Galvão

    2000-01-01

    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...

  12. 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;

    for economic evaluation in a small array deployment. This progression covers development Stages 3 to 4 in the 5 Stage development programme on which the EquiMar technical protocols are based. This report aims at providing a methodology for the analysis and presentation of data obtained from sea trials...... 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....

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

    Science.gov (United States)

    van der Geize, R; Grommen, A W F; Hessels, G I; Jacobs, A A C; Dijkhuizen, L

    2011-08-01

    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.

  15. Complex unconstrained three-dimensional hand movement and constant equi-affine speed.

    Science.gov (United States)

    Maoz, Uri; Berthoz, Alain; Flash, Tamar

    2009-02-01

    One long-established simplifying principle behind the large repertoire and high versatility of human hand movements is the two-thirds power law-an empirical law stating a relationship between local geometry and kinematics of human hand trajectories during planar curved movements. It was further generalized not only to various types of human movements, but also to motion perception and prediction, although it was unsuccessful in explaining unconstrained three-dimensional (3D) movements. Recently, movement obeying the power law was proved to be equivalent to moving with constant planar equi-affine speed. Generalizing such motion to 3D space-i.e., to movement at constant spatial equi-affine speed-predicts the emergence of a new power law, whose utility for describing spatial scribbling movements we have previously demonstrated. In this empirical investigation of the new power law, subjects repetitively traced six different 3D geometrical shapes with their hand. We show that the 3D power law explains the data consistently better than both the two-thirds power law and an additional power law that was previously suggested for spatial hand movements. We also found small yet systematic modifications of the power-law's exponents across the various shapes, which further scrutiny suggested to be correlated with global geometric factors of the traced shape. Nevertheless, averaging over all subjects and shapes, the power-law exponents are generally in accordance with constant spatial equi-affine speed. Taken together, our findings provide evidence for the potential role of non-Euclidean geometry in motion planning and control. Moreover, these results seem to imply a relationship between geometry and kinematics that is more complex than the simple local one stipulated by the two-thirds power law and similar models.

  16. Gallium maltolate as an alternative to macrolides for treatment of presumed Rhodococcus equi pneumonia in foals.

    Science.gov (United States)

    Cohen, Noah D; Slovis, Nathan M; Giguère, Steeve; Baker, Samantha; Chaffin, M Keith; Bernstein, Lawrence R

    2015-01-01

    Macrolide-resistant isolates of Rhodococcus equi are emerging, prompting the search for clinically effective alternative antimicrobials. The proportion of foals with ultrasonographic evidence of pneumonia presumed to be caused by R. equi that had a successful outcome when administered gallium maltolate (GaM) PO would not be more than 10% inferior (ie, lower) than that of foals receiving standard treatment. Fifty-four foals with subclinical pulmonary abscesses among 509 foals at 6 breeding farms in Kentucky. Controlled, randomized, prospective noninferiority study. Foals with ultrasonographic lesions >1 cm in diameter (n = 54) were randomly allocated to receive per os either clarithromycin combined with rifampin (CLR+R) or GaM, and followed up for 28 days by daily physical inspections and weekly (n = 1 farm) or biweekly (n = 4 farms) thoracic ultrasound examinations by individuals unaware of treatment-group assignments. Treatment success was defined as resolution of ultrasonographically identified pulmonary abscesses within 28 days of initiating treatment. Noninferiority was defined as a 90% confidence interval for the observed difference in CLR+R minus GaM that was ≤10%. The proportion of GaM-treated foals that resolved (70%; 14/20) was similar to that of foals treated with CLR+R (74%; 25/34), but we failed to demonstrate noninferiority for GaM relative to CLR+R; however, GaM was noninferior to CLR+R treatment when results from a noncompliant farm were excluded. Gallium maltolate is not inferior to macrolides for treating foals with subclinical pneumonia. Use of GaM might reduce pressure for macrolide-resistance in R. equi. Copyright © 2015 The Authors. Journal of Veterinary Internal Medicine published by Wiley Periodicals, Inc. on behalf of the American College of Veterinary Internal Medicine.

  17. The potential role of migratory birds in transmission cycles of Babesia spp., Anaplasma phagocytophilum, and Rickettsia spp.

    Science.gov (United States)

    Hildebrandt, Anke; Franke, Jan; Meier, Frank; Sachse, Svea; Dorn, Wolfram; Straube, Eberhard

    2010-06-01

    Babesia spp., Anaplasma phagocytophilum, and Rickettsia spp. are potentially emerging tick-borne pathogens, whereas many issues about their ecology, e.g. reservoir host specificity, are still unclear. In spring 2007, we collected 191 feeding Ixodes ricinus ticks from 99 birds of 11 different species on a German bird conservation island in the Baltic Sea. Babesia spp. were detected in 4.7% (9/191), A. phagocytophilum was present in 2.6% (5/191), and Rickettsia spp. were identified in 7.3% (14/191) of the investigated ticks. Further characterization of Babesia spp. infections resulted in B. divergens and B. microti. Among the Rickettsia spp. infections, we identified at least 2 different species: R. monacensis and R. helvetica. Furthermore, 2 ticks harboured mixed infections. Our study provides first interesting insights into the role of migratory birds in the distribution of several emerging tick-borne pathogens.

  18. 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.

  19. 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).

  20. Classification of Babesia canis strains in Europe based on polymorphism of the Bc28.1-gene from the Babesia canis Bc28 multigene family.

    Science.gov (United States)

    Carcy, B; Randazzo, S; Depoix, D; Adaszek, L; Cardoso, L; Baneth, G; Gorenflot, A; Schetters, T P

    2015-07-30

    The vast majority of clinical babesiosis cases in dogs in Europe is caused by Babesia canis. Although dogs can be vaccinated, the level of protection is highly variable, which might be due to genetic diversity of B. canis strains. One of the major merozoite surface antigens of B. canis is a protein with a Mr of 28 kDa that belongs to the Bc28 multigene family, that comprises at least two genes, Bc28.1 and a homologous Bc28.2 gene. The two genes are relatively conserved but they are very distinct in their 3' ends, enabling the design of specific primers. Sequencing of the Bc28.1 genes from 4 genetically distinct B. canis laboratory strains (A8, B, 34.01 and G) revealed 20 mutations at conserved positions of which three allowed the classification of B. canis strains into three main groups (A, B and 34.01/G) by RFLP. This assay was subsequently used to analyze blood samples of 394 dogs suspected of clinical babesiosis from nine countries in Europe. All blood samples were first analyzed with a previously described assay that allowed detection of the different Babesia species that infect dogs. Sixty one percent of the samples contained detectable levels of Babesia DNA. Of these, 98.3% were positive for B. canis, the remaining cases were positive for B. vogeli. Analysis of the Bc28.1 gene, performed on 178 of the B. canis samples, revealed an overall dominance of genotype B (62.4%), followed by genotypes A (37.1%) and 34 (11.8%). Interestingly, a great variation in the geographical distribution and prevalence of the three B. canis genotypes was observed; in the North-East genotype A predominated (72.1% A against 27.9% B), in contrast to the South-West where genotype B predominated (10.3% A against 89.7% B). In the central part of Europe intermediate levels were found (26.0-42.9% A against 74.0-57.1% B, from West to East). Genotype 34 was only identified in France (26.9% among 78 samples) and mostly as co-infection with genotypes A or B (61.9%). A comparative analysis of

  1. EhNPC1 and EhNPC2 Proteins Participate in Trafficking of Exogenous Cholesterol in Entamoeba histolytica Trophozoites: Relevance for Phagocytosis

    Science.gov (United States)

    Bolaños, Jeni; Betanzos, Abigail; Javier-Reyna, Rosario; García- Rivera, Guillermina; Huerta, Miriam; Pais-Morales, Jonnatan; González-Robles, Arturo; Rodríguez, Mario A.

    2016-01-01

    Entamoeba histolytica, the highly phagocytic protozoan causative of human amoebiasis lacks the machinery to synthesize cholesterol. Here, we investigated the presence of NPC1 and NPC2 proteins in this parasite, which are involved in cholesterol trafficking in mammals. Bioinformatics analysis revealed one Ehnpc1 and two Ehnpc2 genes. EhNPC1 appeared as a transmembrane protein and both EhNPC2 as peripheral membrane proteins. Molecular docking predicted that EhNPC1 and EhNPC2 bind cholesterol and interact with each other. Genes and proteins were identified in trophozoites. Serum pulse-chase and confocal microscopy assays unveiled that after trophozoites sensed the cholesterol source, EhNPC1 and EhNPC2 were organized around the plasma membrane in a punctuated pattern. Vesicles emerged and increased in number and size and some appeared full of cholesterol with EhNPC1 or EhNPC2 facing the extracellular space. Both proteins, but mostly EhNPC2, were found out of the cell associated with cholesterol. EhNPC1 and cholesterol formed networks from the plasma membrane to the nucleus. EhNPC2 appeared in erythrocytes that were being ingested by trophozoites, co-localizing with cholesterol of erythrocytes, whereas EhNPC1 surrounded the phagocytic cup. EhNPC1 and EhNPC2 co-localized with EhSERCA in the endoplasmic reticulum and with lysobisphosphatidic acid and EhADH (an Alix protein) in phagolysosomes. Immunoprecipitation assays confirmed the EhNPC1 and EhNPC2 association with cholesterol, EhRab7A and EhADH. Serum starved and blockage of cholesterol trafficking caused a low rate of phagocytosis and incapability of trophozoites to produce damage in the mouse colon. Ehnpc1 and Ehnpc2 knockdown provoked in trophozoites a lower intracellular cholesterol concentration and a diminished rate of phagocytosis; and Ehnpc1 silencing also produced a decrease of trophozoites movement. Trafficking of EhNPC1 and EhNPC2 during cholesterol uptake and phagocytosis as well as their association with

  2. EhNPC1 and EhNPC2 Proteins Participate in Trafficking of Exogenous Cholesterol in Entamoeba histolytica Trophozoites: Relevance for Phagocytosis.

    Directory of Open Access Journals (Sweden)

    Jeni Bolaños

    2016-12-01

    Full Text Available Entamoeba histolytica, the highly phagocytic protozoan causative of human amoebiasis lacks the machinery to synthesize cholesterol. Here, we investigated the presence of NPC1 and NPC2 proteins in this parasite, which are involved in cholesterol trafficking in mammals. Bioinformatics analysis revealed one Ehnpc1 and two Ehnpc2 genes. EhNPC1 appeared as a transmembrane protein and both EhNPC2 as peripheral membrane proteins. Molecular docking predicted that EhNPC1 and EhNPC2 bind cholesterol and interact with each other. Genes and proteins were identified in trophozoites. Serum pulse-chase and confocal microscopy assays unveiled that after trophozoites sensed the cholesterol source, EhNPC1 and EhNPC2 were organized around the plasma membrane in a punctuated pattern. Vesicles emerged and increased in number and size and some appeared full of cholesterol with EhNPC1 or EhNPC2 facing the extracellular space. Both proteins, but mostly EhNPC2, were found out of the cell associated with cholesterol. EhNPC1 and cholesterol formed networks from the plasma membrane to the nucleus. EhNPC2 appeared in erythrocytes that were being ingested by trophozoites, co-localizing with cholesterol of erythrocytes, whereas EhNPC1 surrounded the phagocytic cup. EhNPC1 and EhNPC2 co-localized with EhSERCA in the endoplasmic reticulum and with lysobisphosphatidic acid and EhADH (an Alix protein in phagolysosomes. Immunoprecipitation assays confirmed the EhNPC1 and EhNPC2 association with cholesterol, EhRab7A and EhADH. Serum starved and blockage of cholesterol trafficking caused a low rate of phagocytosis and incapability of trophozoites to produce damage in the mouse colon. Ehnpc1 and Ehnpc2 knockdown provoked in trophozoites a lower intracellular cholesterol concentration and a diminished rate of phagocytosis; and Ehnpc1 silencing also produced a decrease of trophozoites movement. Trafficking of EhNPC1 and EhNPC2 during cholesterol uptake and phagocytosis as well as their

  3. Occurrence of Theileria equi in horses raised in the Jaboticabal microregion, São Paulo State, Brazil Ocorrência de Theileria equi em equinos criados na microrregião de Jaboticabal, Estado de São Paulo, Brasil

    Directory of Open Access Journals (Sweden)

    Cristiane Divan Baldani

    2010-12-01

    Full Text Available Blood and serum samples from 170 horses raised in the Jaboticabal microregion, São Paulo State, Brazil, were collected and tested by microscopic examination of blood smears, indirect fluorescent antibody test (IFAT and nested polymerase chain reaction (nPCR for Theileria equi infections. The association among the test results was verified by the McNemar test. During the examination of thin blood smears, parasites were detected in six (3.52% horses. Anti-T. equi antibodies were detected in 100% sera samples, with titers ranging between 1:80 and 1:5120. The nPCR based on the T. equi merozoite antigen gene (EMA-1 allowed the visualization of specie-specific amplified product in 108 (63.53% horses. All six samples judged positive microscopically were also positive for nPCR. Statistical analysis indicated general disagreement (p Amostras de sangue e soro de 170 equinos criados na microrregião de Jaboticabal, Estado de São Paulo, Brasil, foram coletadas e avaliadas pelo exame direto em esfregaço sanguíneo, reação de imunofluorescência indireta (RIFI e nested reação em cadeia da polimerase (nPCR para a detecção de infecções por Theileria equi. A concordância dos resultados entre os testes de diagnóstico foi verificada pelo teste de McNemar. Durante o exame dos esfregaços sanguíneos, parasitos foram detectados em seis (3,52% equinos. Anticorpos anti-T. equi foram detectados em 100% das amostras de soros, com títulos variando entre 1:80 e 1:5120. O nPCR, baseado na sequência do gene do antígeno de merozoíto de T. equi (EMA-1, permitiu a visualização de produtos de amplificação espécie-específico em 108 (63,53% equinos. Houve diferença altamente significativa (p < 0,0001 entre RIFI e nPCR; RIFI e esfregaço sanguíneo; e nPCR e esfregaço na detecção do parasito. O resultado do presente estudo indica que a infecção por T. equi está amplamente distribuída entre os equinos na microrregião de Jaboticabal, região Nordeste

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

    Science.gov (United States)

    Ramsay, Joshua D; Ueti, Massaro W; Johnson, Wendell C; Scoles, Glen A; Knowles, Donald P; Mealey, Robert H

    2013-01-01

    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, breed

  5. 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

  6. Estimating the Sensitivity and Specificity of Real-Time Quantitative PCR of Fecal Samples for Diagnosis of Rhodococcus equi Pneumonia in Foals.

    Science.gov (United States)

    Shaw, S D; Cohen, N D; Chaffin, M K; Blodgett, G P; Syndergaard, M; Hurych, D

    2015-01-01

    Real-time, quantitative PCR (qPCR) methods for detecting Rhodococcus equi in feces have been developed as a noninvasive, rapid diagnostic test for R. equi pneumonia, but have not been evaluated in a large population of foals. The objective of this study was to evaluate the clinical utility of fecal PCR as a diagnostic test for R. equi pneumonia in foals using receiver operating characteristic (ROC) methods. 186 foals born in 2011 at an R. equi-endemic ranch in Texas. Fecal samples were collected at the time of onset of clinical signs for pneumonic foals (n = 31). Foals with pneumonia were matched by age and birth date to healthy (n = 31) and subclinical (n = 124) control foals; fecal samples were collected from these controls. DNA was extracted from feces using commercial kits and concentration of virulent R. equi in feces was determined by qPCR. Concentration of R. equi in feces differed significantly (P equi pneumonia was 89% (95% CI, 83-99), with a sensitivity of 94% and specificity of 72%. qPCR of feces can be useful as an alternative to tracheobronchial aspiration for the diagnosis of R. equi in foals with clinical signs of pneumonia. Caution should be used in extrapolating results of this study to other populations because fecal concentration of R. equi might vary by geographic location or management practices. Copyright © 2015 The Authors. Journal of Veterinary Internal Medicine published by Wiley Periodicals, Inc. on behalf of the American College of Veterinary Internal Medicine.

  7. ANTIBODIES DETECTION TO RHODOCOCCUS EQUI IN VACCINATED MARES AND FOALS BY INDIRECT ENZYME IMMUNOASSAY DETECÇÃO DE ANTICORPOS ANTI-Rhodococcus equi EM ÉGUAS VACINADAS E POTROS PELO ENSAIO IMUNOENZIMÁTICO INDIRETO

    Directory of Open Access Journals (Sweden)

    Carla Braga Martins

    2010-04-01

    Full Text Available The humoral immune response in ‘Brasileiro de Hipismo’ (BH breed and Breton mares was compared after using the Rhodococcus equi vaccine, and the effect of maternal immunoprophylaxis on antibody transfer to newborn foals through the colostrum was evaluated. Blood samples were obtained from 16 pregnant mares vaccinated against R. equi, 16 foals (offspring from vaccinated mares, 8 unvaccinated pregnant mares and 8 foals (offspring from control mares. R equi serum antibody titers were determined by enzyme-linked immunosorbant assay (ELISA after the immunization of pregnant mares using two different antigens, APTX and the commercial vaccine. There was no difference in antibody production between the two breeds. Significant increase in R. equi antibody titers was observed in mares after vaccination (p<0.01, reaching a peak at foaling. Afterward, titers tended to decrease for up to 60 days after birth (dab and then remained constant until 150 dab. Significant antibody transfer to the vaccinated mares newborn foal occurred through the colostrum. A slight reduction in antibody titer was observed at 60 dab, after which titers remained constant for up to 150 dab. The commercial antigen detected significantly higher antibody titers than did APTX (p<0.01.

    KEY WORDS: Antibodies, ELISA, Rhodococcus equi, immunization, foals.
    Comparou-se a resposta imune humoral em éguas da raça Brasileiro de Hipismo (BH e Bretão, após a imunização com a vacina anti-Rhodococcus equi, bem como avaliou-se o efeito da imunoprofilaxia ativa materna na transferência de anticorpos pelo colostro em equinos recém-nascidos. Coletaram-se amostras sanguíneas de dezesseis éguas prenhes vacinadas contra R. equi, dezesseis potros filhos das éguas vacinadas, oito éguas prenhes não vacinadas e oito potros filhos das éguas não vacinadas. Determinou-se a titulação de anticorpos anti-R. equi utilizando-se o ensaio imunoenzimático indireto (ELISA com os dois

  8. 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

  9. 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.

  10. 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.

  11. 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.

  12. Genetic characterization of Babesia and Theileria parasites in water buffaloes in Sri Lanka.

    Science.gov (United States)

    Sivakumar, Thillaiampalam; Tattiyapong, Muncharee; Fukushi, Shintaro; Hayashida, Kyoko; Kothalawala, Hemal; Silva, Seekkuge Susil Priyantha; Vimalakumar, Singarayar Caniciyas; Kanagaratnam, Ratnam; Meewewa, Asela Sanjeewa; Suthaharan, Kalpana; Puvirajan, Thamotharampillai; de Silva, Weligodage Kumarawansa; Igarashi, Ikuo; Yokoyama, Naoaki

    2014-02-24

    Water buffaloes are thought to be the reservoir hosts for several hemoprotozoan parasites that infect cattle. In the present study, we surveyed Sri Lankan bred water buffaloes for infections with Babesia bovis, Babesia bigemina, Theileria annulata, and Theileria orientalis using parasite-specific PCR assays. When 320 blood-derived DNA samples from water buffaloes reared in three different districts (Polonnaruwa, Mannar, and Mullaitivu) of Sri Lanka were PCR screened, B. bovis, B. bigemina, and T. orientalis were detected. While T. orientalis was the predominant parasite (82.5%), low PCR-positive rates were observed for B. bovis (1.9%) and B. bigemina (1.6%). Amplicons of the gene sequences of the Rhoptry Associated Protein-1 (RAP-1) of B. bovis, the Apical Membrane Antigen-1 (AMA-1) of B. bigemina, and the Major Piroplasm Surface Protein (MPSP) of T. orientalis were compared with those characterized previously in Sri Lankan cattle. While the B. bigemina AMA-1 sequences from water buffaloes shared high identity values with those from cattle, B. bovis RAP-1 sequences from water buffaloes diverged genetically from those of cattle. For T. orientalis, none of the MPSP sequence types reported previously in Sri Lankan cattle (types 1, 3, 5, and 7) were detected in the water buffaloes, and the MPSP sequences analyzed in the present study belonged to types N1 or N2. In summary, in addition to reporting the first PCR-based survey of Babesia and Theileria parasites in water buffaloes in Sri Lanka, the present study found that the predominant variants of water buffalo-derived B. bovis RAP-1 and T. orientalis MPSP sequences were different from those previously described from cattle in this country.

  13. 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.

  14. Babesia canis and other tick-borne infections in dogs in Central Poland.

    Science.gov (United States)

    Welc-Faleciak, Renata; Rodo, Anna; Siński, Edward; Bajer, Anna

    2009-12-23

    Vector-borne infections constitute increasing health problem in dogs worldwide, including sled dogs, dramatically decreasing the fitness of working dogs and even leading to death. In the period 2006-2008 eighty-two blood samples were collected from eight sled dog kennels in Central Poland. The prevalence of four vector-borne infections (Babesia canis, Bartonella sp., Anaplasma/Ehrlichia and Borrelia burgdorferi) was estimated in 82 sled dogs using PCR and nested PCR for diagnosis and the same methods were used to identify the vector-borne pathogens in 26 dogs presenting at veterinary clinics with symptoms of vector-borne diseases. None of four studied vector-borne pathogens was detected in samples originating from veterinary clinics. Among the remaining 82 dogs B. canis infections were confirmed in three dogs undergoing treatment for babesiosis. The DNA of tick-borne pathogens was also found among 22 (27.8%) of the 79 apparently healthy dogs, including 20 cases of B. canis infection (25.3%), one case of B. burgdorferi s.l. infection and one case of Anaplasma phagocytophilum infection. No evidence of Bartonella spp. and Ehrlichia canis infections were found in this set of samples. Sequencing of a Babesia fragment of 18S rDNA amplified from acute (n=5) and asymptomatic (n=5) cases revealed that all isolates were identical to the Babesia canis canis sequence, originally isolated from Dermacentor reticulatus ticks in Poland. A range of factors was shown to affect the distribution of babesiosis in sled dogs. The data are also discussed in respect to the health risk factors generated by asymptomatic B. canis infections and the efficiency of chemoprophylaxis measures taken by sled dog owners.

  15. Primary Babesia rodhaini infection followed by recovery confers protective immunity against B. rodhaini reinfection and Babesia microti challenge infection in mice.

    Science.gov (United States)

    Wang, Guanbo; Efstratiou, Artemis; Adjou Moumouni, Paul Franck; Liu, Mingming; Jirapattharasate, Charoonluk; Guo, Huanping; Gao, Yang; Cao, Shinuo; Zhou, Mo; Suzuki, Hiroshi; Igarashi, Ikuo; Xuan, Xuenan

    2016-10-01

    In the present study, we investigated the protective immunity against challenge infections with Babesia rodhaini and Babesia microti in the mice recovered from B. rodhaini infection. Six groups with 5 test mice in each group were used in this study, and were intraperitoneally immunized with alive and dead B. rodhaini. The challenge infections with B. rodhaini or B. microti were performed using different time courses. Our results showed that the mice recovered from primary B. rodhaini infection exhibited low parasitemia and no mortalities after the challenge infections, whereas mock mice which had received no primary infection showed a rapid increase of parasitemia and died within 7 days after the challenge with B. rodhaini. Mice immunized with dead B. rodhaini were not protected against either B. rodhaini or B. microti challenge infections, although high titers of antibody response were induced. These results indicate that only mice immunized with alive B. rodhaini could acquire protective immunity against B. rodhaini or B. microti challenge infection. Moreover, the test mice produced high levels of antibody response and low levels of cytokines (INF-γ, IL-4, IL-12, IL-10) against B. rodhaini or B. microti after challenge infection. Mock mice, however, showed rapid increases of these cytokines, which means disordered cytokines secretion occurred during the acute stage of challenge infection. The above results proved that mice immunized with alive B. rodhaini could acquire protective immunity against B. rodhaini and B. microti infections.

  16. 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...

  17. Theileria equi isolates vary in susceptibility to imidocarb dipropionate but demonstrate uniform in vitro susceptibility to a bumped kinase inhibitor

    Science.gov (United States)

    The apicomplexan hemoparasite Theileria equi is a causative agent of equine piroplasmosis, eradicated from the United States in 1988. However, recent outbreaks have sparked renewed interest in treatment options for infected horses. Imidocarb dipropionate is the current drug of choice, however variat...

  18. Genetic characterization of theileria equi infecting horses in North America: evidence for a limited source of U.S. introductions

    Science.gov (United States)

    Theileria equi is a tick-borne Apicomplexan hemoparasite that causes equine piroplasmosis (EP). This parasite has a worldwide distribution, but until recent outbreaks the United States has been considered to be free of EP. Maximum parsimony analysis of 18S rRNA gene sequences of North American T. eq...

  19. 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...

  20. A novel method to generate unmarked gene deletions in the intracellular pathogen Rhodococcus equi using 5-fluorocytosine conditional lethality

    NARCIS (Netherlands)

    van der Geize, R.; de Jong, W.; Hessels, G. I.; Grommen, A. W. F.; Jacobs, A. A. C.; Dijkhuizen, L.

    2008-01-01

    A novel method to efficiently generate unmarked in-frame gene deletions in Rhodococcus equi was developed, exploiting the cytotoxic effect of 5-fluorocytosine (5-FC) by the action of cytosine deaminase (CD) and uracil phosphoribosyltransferase (UPRT) enzymes. The opportunistic, intracellular

  1. Babesia crassa n.sp. (Sporozoa, Babesiidae) of domestic sheep in Iran.

    Science.gov (United States)

    Hashemi-Fesharki, R; Uilenberg, G

    1981-01-01

    A large Babesia sp., isolated from a sheep in Iran, provided to be serologically and morphologically different from B, motasi and B. ovis. The parasite, designated B. crassa n. sp., is characterized by the frequent occurrence of four organisms in one erythrocyte, which is the result of quadruple division and, in other cases, of two successive binary divisions. Parasited resulting from the first of two successive binary divisions are exceptionally broad. B. crassa appears to be of low pathogenicity for sheep and goats, to which it is also infective. The vector is unknown.

  2. Theileria, Babesia, and Anaplasma detected by PCR in ruminant herds at Bié Province, Angola

    OpenAIRE

    2012-01-01

    Distribution of Anaplasma spp., Babesia spp., Theileria spp., and Ehrlichia ruminantium, was for the first time studied in Bié Province, central Angola. We examined 76 blood samples of cattle originated from seven farms, and 13 blood samples of goats from two farms employing molecular genetic tools (PCR). Most prevalent was A. ovis-infection in goats (100%) and A. marginale-infection in cattle (38% of examined animals, and six out of seven farms). B. bigemina-infection was detected in only on...

  3. Characterizations of individual mouse red blood cells parasitized by Babesia microti using 3-D holographic microscopy

    CERN Document Server

    Park, HyunJoo; Kim, Kyoohyun; Cho, Shin-Hyeong; Lee, Won-Ja; Kim, Youngchan; Lee, SangEun; Park, YongKeun

    2015-01-01

    Babesia microti causes emergency human babesiosis. However, little is known about the alterations in B. microti invaded red blood cells (Bm-RBCs) at the individual cell level. Through quantitative phase imaging techniques based on laser interferometry, we present the simultaneous measurements of structural, chemical, and mechanical modifications in individual mouse Bm-RBCs. 3-D refractive index maps of individual RBCs and in situ parasite vacuoles are imaged, from which total contents and concentration of dry mass are also precisely quantified. In addition, we examine the dynamic membrane fluctuation of Bm-RBCs, which provide information on cell membrane deformability.

  4. Efficacy of Malarone(®) in dogs naturally infected with Babesia gibsoni.

    Science.gov (United States)

    Iguchi, Aiko; Shiranaga, Nobuyuki; Matsuu, Aya; Hikasa, Yoshiaki

    2014-09-01

    The efficacy of Malarone(®) alone and in combination with doxycycline (DOXY) against Babesia gibsoni infections was examined in 8 dogs. In all dogs except one treated with Malarone(®), parasitemia decreased, and anemia improved soon after initiation of treatment. However, 3 of 4 dogs treated with Malarone(®) relapsed, and relapse was inhibited in 2 of 4 dogs treated with Malarone(®) and DOXY. All relapsed dogs responded well to the second treatment, but 1 dog relapsed again and did not respond to the third treatment. Malarone(®) may be useful for acute stage of B. gibsoni infections, and at least second repeating treatment might be effective.

  5. 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.

  6. 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.

  7. Production of Nfa1-specific monoclonal antibodies that influences the in vitro cytotoxicity of Naegleria fowleri trophozoites on microglial cells.

    Science.gov (United States)

    Lee, Yang-Jin; Kim, Jong-Hyun; Jeong, Seok-Ryoul; Song, Kyoung-Ju; Kim, Kyongmin; Park, Sun; Park, Moon-Sung; Shin, Ho-Joon

    2007-10-01

    Naegleria fowleri, agent of fatal primary amoebic meningoencephalitis, appears to induce cytotoxicity mechanically through its contact with the cell. The nfa1 gene cloned from a cDNA library of pathogenic N. fowleri by immunoscreening consists of 360 bp and expresses a 13.1-kDa recombinant protein (rNfa1) that demonstrated localization in the pseudopodia when examined using immunocytochemistry. To study the mechanisms involved in N. fowleri cytotoxicity, we developed a large volume of rNfa1-specific monoclonal antibody (McAb) against a 17-kDa His-tag fusion rNfa1 protein using a cell fusion technique. We established eight McAb-producing hybridoma cells. The antibodies were all immunoglobulin G2b and reacted strongly with a 17-kDa band representing the rNfa1 fusion protein in Western blotting, demonstrating immunoreactivity to the Nfa1 protein in pseudopodia (especially in the food cups) of N. fowleri trophozoites. A 51Cr-release assay indicated N. fowleri cytotoxicity by demonstrating that it eliminated 37.8, 60.6, and 98.8% of the target (microglial) cells 6, 12, and 24 h after co-incubation, respectively. When an anti-Nfa1 McAb was added to the coculture system, N. fowleri cytotoxicity decreased to 29.8, 44.1, and 66.3%, respectively.

  8. Synthesis, characterization and amoebicidal potential of locally synthesized TiO2 nanoparticles against pathogenic Acanthamoeba trophozoites in vitro.

    Science.gov (United States)

    Imran, Muhammad; Muazzam, Ambreen Gul; Habib, Amir; Matin, Abdul

    2016-06-01

    Acanthamoeba is an opportunistic protozoan pathogen that plays a pivotal role in the ecosystem. It may cause blinding keratitis and fatal encephalitis involving the central nervous system. Here we synthesized pure and Zn doped TiO2 nanoparticles (~10-30nm) via sol-gel and sol-hydrothermal methods and demonstrated its impact on the biological characteristics of pathogenic Acanthamoeba castellanii. Our results revealed that pure and Zn doped TiO2 nanoparticles synthesized by sol-hydrothermal methods (ranging 5, 10, 25 and 50μg/ml) exhibited amoebicidal effects i.e., >60% of trophozoites executed under normal light at maximum dose (50μg/ml) within 1h incubation. In contrast pure/doped TiO2 obtained via sol gel method showed ~40% amoeba damage. Furthermore, amoebae growth assay demonstrated that Zn doped TiO2 also inhibited Acanthamoeba numbers up to 7days in dose dependent manner. It was interesting to note that all the tested TiO2 nanoparticles have shown maximum amoebicidal effects at pH7 which is quite relevant to amoebic growth favorable conditions. Our results confirmed that TiO2 has inhibitory effects on Acanthamoeba growth and viability. Overall, we reported the amoebicidal and amoebic growth inhibition potential of pure and Zn doped TiO2 nanoparticles against Acanthamoeba due to attached OH(-) groups, reduced size and decreased band gap of sol hydrothermally synthesized TiO2 nanoparticles.

  9. Re-emergence of the apicomplexan Theileria equi in the United States: elimination of persistent infection and transmission risk.

    Directory of Open Access Journals (Sweden)

    Massaro W Ueti

    Full Text Available Arthropod-borne apicomplexan pathogens that cause asymptomatic persistent infections present a significant challenge due to their life-long transmission potential. Although anti-microbials have been used to ameliorate acute disease in animals and humans, chemotherapeutic efficacy for apicomplexan pathogen elimination from a persistently infected host and removal of transmission risk is largely unconfirmed. The recent re-emergence of the apicomplexan Theileria equi in U.S. horses prompted testing whether imidocarb dipropionate was able to eliminate T. equi from naturally infected horses and remove transmission risk. Following imidocarb treatment, levels of T. equi declined from a mean of 10(4.9 organisms/ml of blood to undetectable by nested PCR in 24 of 25 naturally infected horses. Further, blood transfer from treated horses that became nested PCR negative failed to transmit to naïve splenectomized horses. Although these results were consistent with elimination of infection in 24 of 25 horses, T. equi-specific antibodies persisted in the majority of imidocarb treated horses. Imidocarb treatment was unsuccessful in one horse which remained infected as measured by nested PCR and retained the ability to infect a naïve recipient via intravenous blood transfer. However, a second round of treatment eliminated T. equi infection. These results support the utility of imidocarb chemotherapy for assistance in the control and eradication of this tick-borne pathogen. Successful imidocarb dipropionate treatment of persistently infected horses provides a tool to aid the global equine industry by removing transmission risk associated with infection and facilitating international movement of equids between endemic and non-endemic regions.

  10. 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.

  11. An Invertron-Like Linear Plasmid Mediates Intracellular Survival and Virulence in Bovine Isolates of Rhodococcus equi.

    Science.gov (United States)

    Valero-Rello, Ana; Hapeshi, Alexia; Anastasi, Elisa; Alvarez, Sonsiray; Scortti, Mariela; Meijer, Wim G; MacArthur, Iain; Vázquez-Boland, José A

    2015-07-01

    We report a novel host-associated virulence plasmid in Rhodococcus equi, pVAPN, carried by bovine isolates of this facultative intracellular pathogenic actinomycete. Surprisingly, pVAPN is a 120-kb invertron-like linear replicon unrelated to the circular virulence plasmids associated with equine (pVAPA) and porcine (pVAPB variant) R. equi isolates. pVAPN is similar to the linear plasmid pNSL1 from Rhodococcus sp. NS1 and harbors six new vap multigene family members (vapN to vapS) in a vap pathogenicity locus presumably acquired via en bloc mobilization from a direct predecessor of equine pVAPA. Loss of pVAPN rendered R. equi avirulent in macrophages and mice. Mating experiments using an in vivo transconjugant selection strategy demonstrated that pVAPN transfer is sufficient to confer virulence to a plasmid-cured R. equi recipient. Phylogenetic analyses assigned the vap multigene family complement from pVAPN, pVAPA, and pVAPB to seven monophyletic clades, each containing plasmid type-specific allelic variants of a precursor vap gene carried by the nearest vap island ancestor. Deletion of vapN, the predicted "bovine-type" allelic counterpart of vapA, essential for virulence in pVAPA, abrogated pVAPN-mediated intramacrophage proliferation and virulence in mice. Our findings support a model in which R. equi virulence is conferred by host-adapted plasmids. Their central role is mediating intracellular proliferation in macrophages, promoted by a key vap determinant present in the common ancestor of the plasmid-specific vap islands, with host tropism as a secondary trait selected during coevolution with specific animal species. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  12. Cytopathic changes and pro-inflammatory cytokines induced by Naegleria fowleri trophozoites in rat microglial cells and protective effects of an anti-Nfa1 antibody.

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    Oh, Y-H; Jeong, S-R; Kim, J-H; Song, K-J; Kim, K; Park, S; Sohn, S; Shin, H-J

    2005-12-01

    Naegleria fowleri, a free-living amoeba, causes fatal primary amoebic meningoencephalitis in experimental animals and humans. The nfa1 gene (360 bp) was previously cloned from a cDNA library of pathogenic N. fowleri by immunoscreening, and produced a 13.1-kDa recombinant protein that showed pseudopodia-specific localization by immunocytochemistry. On the basis of an idea that the pseudopodia-specific Nfa1 protein seems to be involved in the pathogenicity of N. fowleri, the cytopathic activity of N. fowleri trophozoites co-cultured with rat microglial cells was observed, and the effects of an anti-Nfa1 antibody in a co-culture system were elucidated. Using light, scanning and transmission electron microscopy, it was seen that N. fowleri trophozoites in contact with microglial cells produced vigorous pseudopodia and a food-cup structure. Microglial cells were destroyed by N. fowleri trophozoites as seen from necrotic cell death in a time-dependent manner. In a(51)Cr release assay, N. fowleri showed 17.8%, 24.9%, 54.6% and 98% cytotoxicity against microglial cells at 3, 6, 12 and 24 h post-incubation, respectively. However, when anti-Nfa1 antibody was added in a coculture system, N. fowleri cytotoxicity was reduced to 15.5%, 20.3%, 46.7% and 66.9%, respectively. Moreover, microglial cells co-cultured with N. fowleri trophozoites secreted the pro-inflammatory cytokines, TNF-alpha, IL-1beta and IL-6. In the presence of anti-Nfa1 antibody, the secretion of TNF-alpha was slightly, but not significantly, decreased.

  13. Characterization of recombinase DMC1B and its functional role as Rad51 in DNA damage repair in Giardia duodenalis trophozoites.

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    Torres-Huerta, Ana Laura; Martínez-Miguel, Rosa María; Bazán-Tejeda, María Luisa; Bermúdez-Cruz, Rosa María

    2016-08-01

    Homologous recombination (HR) is a highly conserved pathway for the repair of chromosomes that harbor DNA double-stranded breaks (DSBs). The recombinase RAD51 plays a key role by catalyzing the pairing of homologous DNA molecules and the exchange of information between them. Two putative DMC1 homologs (DMC1A and DMC1B) have been identified in Giardia duodenalis. In terms of sequences, GdDMC1A and GdDMC1B bear all of the characteristic recombinase domains: DNA binding domains (helix-turn-helix motif, loops 1 and 2), an ATPcap and Walker A and B motifs associated with ATP binding and hydrolysis. Because GdDMC1B is expressed at the trophozoite stage and GdDMC1A is expressed in the cyst stage, we cloned the giardial dmc1B gene and expressed and purified its protein to determine its activities, including DNA binding, ATP hydrolysis, and DNA strand exchange. Our results revealed that it possessed these activities, and they were modulated by divalent metal ions in different manners. GdDMC1B expression at the protein and transcript levels, as well as its subcellular localization in trophozoites upon DNA damage, was assessed. We found a significant increase in GdDMC1B transcript and protein levels after ionizing radiation treatment. Additionally, GdDMC1B protein was mostly located in the nucleus of trophozoites after DNA damage. These results indicate that GdDMC1B is the recombinase responsible for DSBs repair in the trophozoite; therefore, a functional Rad51 role is proposed for GdDMC1B.

  14. Identifying differentially expressed genes in trophozoites and cysts of Acanthamoeba T4 genotype: Implications for developing new treatments for Acanthamoeba keratitis.

    Science.gov (United States)

    Abedkhojasteh, Hoda; Niyyati, Maryam; Rezaei, Sasan; Mohebali, Mehdi; Farnia, Shohreh; Kazemi-Rad, Elham; Roozafzoon, Reza; Sianati, Hamed; Rezaeian, Mostafa; Heidari, Mansour

    2015-02-01

    Acanthamoeba T4 genotype is the most prevalent genotype associated with amoebic keratitis. Acanthamoeba keratitis therapy is difficult due to transformation of trophozoite to cyst stage, which hinders the treatment of the disease. Although encystation assists the organism to survive against the chemotherapeutic compounds, the precise mechanism of encystation remains poorly understood. The purpose of this work was to identify differentially expressed genes in Acanthamoeba T4 genotype which might be useful for understanding of the encystment process and may thus help develop more efficient treatment. The mRNA profile of trophozoite and cyst of Acanthamoeba T4 genotype isolated from a soft contact lens wearer were analyzed using a cDNA amplified fragment length polymorphism (cDNA-AFLP) technique. Subsequently, a real time reverse transcriptase-PCR was performed to validate the cDNA-AFLP results. Three genes, heat shock protein70 (hsp70), actin-I and elongation factor-1alpha (EF-1α) were differentially expressed during Acanthamoeba differentiation. An in silico result predicted that transformation of trophozoite to cyst could be mediated through their cooperation with the protein partners interaction. Taken together, our experimental and bioinformatics findings suggested potential functions of hsp70, EF-1α and actin-I in differentiation of Acanthamoeba T4 genotype which may be useful in the design of an efficient therapeutic strategy in AK.

  15. The Effects of Babesiosis on Oxidative Stress and DNA Damage in Anatolian Black Goats Naturally Infected with Babesia ovis.

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    Ismail Kucukkurt

    2014-03-01

    Full Text Available A reactive oxygen and nitrogen intermediate produced during an inflammatory response is the important part of host-defense strategies of organisms to kill the parasite. However, it is not well known whether these intermediates cause DNA damage and oxidative stress in goats infected with Babesia ovis. The purpose of this study was to clarify the effects of babesiosis on basal levels of DNA damage and oxidative status of goats naturally infected with B.ovis.DNA damage and antioxidant parameters were determined in B. ovis infected goats. Ten infected Anatolian Black Goats with B. ovis diagnosed via clinical signs and microscopic findings and ten healthy were used in the study.The Babesia infection increased the levels of DNA damage, malondialdehyde (MDA, protein carbonyl content (PCO and plasma concentration of nitric oxide metabolites (NOx, and decreased total antioxidant activities (AOA and reduced glutathione (GSH. A significant positive correlation between DNA damage, MDA, PCO, and NOx concentrations was found in the infected goats. DNA damage showed a negative association with AOA and GSH concentrations in the infected goats.The Babesia infection increases oxidative stress markers and DNA damage and decreases AOA in goats. These results suggest that the increases in the production of free radicals due to Babesia infection not only contribute to host-defense strategies of organisms to kill the parasite but also induce oxidative damage in other cells.

  16. 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.

  17. Identification, genetic diversity and prevalence of Theileria and Babesia species in a sheep population from Northern Spain.

    Science.gov (United States)

    Nagore, Daniel; García-Sanmartín, Josune; García-Pérez, Ana L; Juste, Ramón A; Hurtado, Ana

    2004-08-01

    The genetic diversity and prevalence of virtually all Theileria and Babesia species in a sheep population were studied using a specifically designed reverse line blot macroarray. The amplified hypervariable V4 region of the 18S rRNA gene was hybridised against generic and species-specific probes. In a first screening (Study I), 320 apparently healthy animals corresponding to 32 flocks located in the Basque Country (Northern Spain) were analysed. The survey demonstrated a high prevalence of subclinical infections (64.7%). Three Theileria genotypes were identified, sharing 96.7-97.0% similarity between their 18S rRNA gene sequences: Theileria ovis, Theileria sp. OT1 (99.6% similarity with the recently described pathogenic piroplasm Theileria sp. China 1), and Theileria sp. OT3. Two Babesia species sharing 91.5% similarity were also detected: Babesia ovis and Babesia motasi. The complete 18S rRNA gene sequences of these and other piroplasm species were phylogenetically analysed. Prevalence of piroplasms was also investigated in a second group of 80 sheep from 16 flocks reared in mountain areas that had been heavily exposed to ticks and had suffered a recent abortion episode (Study II). The screening revealed a significantly higher (P distribution of Theileria spp. in the studied sheep population suggests that the parasites involved are of relatively low pathogenicity, in contrast to what has been reported for Theileria sp. China 1 in other countries.

  18. Detection and molecular characterization of canine babesiosis causative agent Babesia canis in the naturally infected dog in Lithuania.

    Science.gov (United States)

    Paulauskas, Algimantas; Radzijevskaja, Jana; Karvelienė, Birutė; Grigonis, Aidas; Aleksandravičienė, Asta; Zamokas, Gintaras; Babickaitė, Lina; Sabūnas, Vytautas; Petkevičius, Saulius

    2014-10-15

    Canine babesiosis caused by Babesia canis is an emerging infectious disease in Europe. Although previously uncommon, canine babesiosis has become quite frequent in Lithuania during the past decade. In the last few years an increasing number of cases with a wide variety of clinical signs have been recorded throughout the country. In Lithuania the identification of the disease agent in veterinarian clinics is based on a microscopic analysis of size and morphology. To date, no data on the genetic characterization of Babesia species in dogs have been documented for Lithuania. A total of 123 blood samples from dogs showing clinical signs of babesiosis on the basis of veterinary examination were tested for the presence of babesial parasites. Babesia isolated from dogs were detected and characterized by nested-PCR and sequence analysis of a fragment of the 18S rRNA gene. Babesia parasites were detected in blood smears of 94 dogs (76.4%). The molecular analysis revealed the presence of B. canis in 108 dogs (87.8%). Two genotypes of B. canis were distinguished on the basis on two nucleotide (GA → AG) substitutions observed in 18S rRNA gene sequences. The results of the present study provide knowledge of the distribution of B. canis genotypes in dogs in Lithuania, and show the necessity to use a molecular analysis for an accurate diagnosis of canine babesiosis.

  19. Expressed gene sequences from larval genes over-expressed upon Babesia bovis infection of Rhipecephalus (Boophilus) microplus

    Science.gov (United States)

    Boophilus ticks are vectors of Babesia bovis, the protozoan causative agent of cattle fever, a disease which is responsible for significant production losses to cattle producers in much of Africa, Central and South America and Australia. We utilized subtractive cDNA library synthesis techniques to o...

  20. High-Quality Draft Genome Sequence of Babesia divergens, the Etiological Agent of Cattle and Human Babesiosis

    Science.gov (United States)

    Cuesta, Isabel; González, Luis M.; Estrada, Karel; Grande, Ricardo; Zaballos, Ángel; Lobo, Cheryl A.; Barrera, Jorge

    2014-01-01

    Babesia divergens causes significant morbidity and mortality in cattle and splenectomized or immunocompromised individuals. Here, we present a 10.7-Mb high-quality draft genome of this parasite close to chromosome resolution that will enable comparative genome analyses and synteny studies among related parasites. PMID:25395649

  1. Evaluation of the growth-inhibitory effect of trifluralin analogues on in vitro cultured babesia bovis parasites

    Science.gov (United States)

    Babesia bovis caused bovine babesiosis is a world tick borne hemoprotozoan disease leading to fever, anemia, weight losses and ultimately death. Several babesicidal drugs that have been in use in cattle for years have proven to be partially ineffective and the development of alternative highly speci...

  2. The Ovarian transcriptome of the cattle tick, Rhipicephalus (Boophilus) microplus, feeding upon a bovine host infected with Babesia bovis

    Science.gov (United States)

    Background: Cattle babesiosis is a tick-borne disease of cattle with the most severe form of the disease caused by the apicomplexan, Babesia bovis. Babesiosis is transmitted to cattle through the bite of infected cattle ticks of the genus Rhipicephalus. The most prevalent species is Rhipicephalus (B...

  3. 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.

  4. 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

  5. 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.

  6. 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.

  7. Morphological and Molecular Descriptors of the Developmental Cycle of Babesia divergens Parasites in Human Erythrocytes.

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    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.

  8. First Molecular Detection of Babesia gibsoni in Dogs from Wuhan, China

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    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.

  9. [The prevalence of Babesia canis canis in marsh ticks (Dermacentor reticulatus) in the Saarland].

    Science.gov (United States)

    Beelitz, Pamela; Schumacher, Stefan; Marholdt, Fritz; Pfister, Kurt; Silaghi, Cornelia

    2012-01-01

    An accumulation of autochthonous cases of canine babesiosis caused by Babesia canis has been registered in a small animal clinic in the Saarland since the beginning of 2006, some cases with fatal outcome. This study aims to contribute to the explanation of strong focal occurrence of infections with B. canis in this region.Therefore, patient owners who had presented their dogs in the years 2006 and 2007 because of babesiosis and who had claimed not having left the Saarland with their dogs at least six months before the outbreak of Babesiosis, were asked for their dog walking habits. Accordingly, a selection often tick collection sites of various landscape structures was made.Tick sampling by flagging the vegetation took place every month from March to December 2008. The collected ticks were identified morphologically. In eight of ten collecting sites a total of 397 adult Dermacentor reticulatus ticks were collected from March to December with the highest frequencies during the months of May, October and November. All collected specimens were examined by genus-specific conventional PCR for the presence of Babesia-DNA. In positive samples, the PCR-products were differentiated by sequencing. ten D. reticulatus (2.5%) ticks examined were found positive for DNA of B. canis canis originating from three out of eight collection sites. Consequently, an endemic distribution of D. reticulatus was confirmed and a natural

  10. Detection of Leishmania infantum, Babesia canis, and rickettsiae in ticks removed from dogs living in Italy.

    Science.gov (United States)

    Trotta, Michele; Nicetto, Martina; Fogliazza, Alessandro; Montarsi, Fabrizio; Caldin, Marco; Furlanello, Tommaso; Solano-Gallego, Laia

    2012-12-01

    The aims of this study were to determine natural infections by Anaplasma phagocytophilum/Anaplasma platys, Bartonella henselae, Ehrlichia canis, Leishmania infantum, Rickettsia spp., Babesia spp., and Hepatozoon spp. by molecular methods in ticks (n=91) removed from dogs with clinical signs and laboratory abnormalities compatible with tick-borne diseases (n=22) living in Italy and to assess the distribution and species of ticks encountered. Ticks from dogs living in southern Italy were all identified as Rhipicephalus sanguineus (n=25), ticks from central Italy included Rh. sanguineus (n=8) and Ixodes ricinus (n=9), ticks from northern Italy included Rh. sanguineus (n=45), Dermacentor marginatus (n=3), and one I. ricinus. Leishmania infantum, Rickettsia spp., and Babesia canis were the only pathogens detected in 7 (8%), 4 (4%), and 2 (2%) out of 91 ticks, respectively. L. infantum was detected in I. ricinus from central Italy and in Rh. sanguineus from northern and central Italy. Rickettsia conorii and Ri. massiliae were detected in Rh. sanguineus ticks from central and southern Italy (Sicily), respectively. Bab. canis was detected in D. marginatus ticks from northern Italy.

  11. Detection of Babesia vogeli in stray cats of metropolitan Bangkok, Thailand.

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    Simking, Patcharathorn; Wongnakphet, Sirichai; Stich, Roger W; Jittapalapong, Sathaporn

    2010-10-11

    The combination of a rapidly growing stray animal population and the lack of animal control in Bangkok has resulted in a unique opportunity to evaluate the potential role of companion animals as sentinels and reservoirs of infectious diseases, including several of those caused by vector-borne parasites. The purpose of this study was to determine the prevalence and factors associated with the distribution of Babesia species infections among stray cats in Bangkok. Blood samples were collected from 1490 stray cats residing in 140 monasteries of 50 metropolitan districts of Bangkok, and assayed with light microscopy and PCR for evidence of Babesia spp. Pear-shaped merozoites were observed microscopically from two (0.13%) of these cats, while a nested 18S rDNA-based PCR assay detected babesial infections in 21 (1.4%) of the cats tested. The prevalence of infection was significantly different between sexes (ppopulations in Thailand, and suggest that the presence of pet companion animals could be a risk factor for exposure of stray cats to vector-borne parasites.

  12. Defective production of interferon-γ and tumour necrosis factor-α by AIDS mononuclear cells after in vitro exposure to Rhodococcus equi

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    S. Delia

    1995-01-01

    Full Text Available The production of interferon-γ and tumour necrosis factor-α was evaluated in the peripheral blood mononuclear cells (PBMCs from healthy donors and AIDS patients after Rhodococcus equi infection in vitro. PBMCs from healthy donors secreted elevated levels of IFN-γ and TNF-α when challenged in vitro with killed R. equi, whereas the release of both cytokines was impaired in supernatant cultures from AIDS patients. We conclude that the failure of IFN-γ generation in AIDS patients in response to R. equi is not antigen-specific but it may reflect the global impairment of T-cell function. In such patients, however, the infection with R. equi, a facultative intracellular pathogen which survives and replicates within macrophages, may be responsible for the impairment in the TNF-α release, possibly enhancing the HIV-induced macrophage dysftmction.

  13. Phylogenetic characterization of Babesia canis vogeli in dogs in the state of Goiás, Brazil Caracterização filogenética de Babesia canis vogeli em cães do estado de Goiás, Brasil

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    Sabrina Castilho Duarte

    2011-12-01

    Full Text Available The genus Babesia comprises protozoa that cause diseases known as babesiosis. Dogs are commonly affected by Babesia canis or Babesia gibsoni. Babesia canis is divided into the subspecies Babesia canis canis, Babesia canis vogeli and Babesia canis rossi. Among these, Babesia canis vogeli predominates in Brazil. The objective of this study was to conduct a phylogenetic analysis on Babesia isolates from dogs in Goiânia, Goiás. Blood samples were obtained from 890 dogs presenting clinical signs suggestive of canine babesiosis that were attended at a veterinary hospital of Goiás. Only samples presenting typical intraerythrocytic parasites were used in the study. These were subjected to DNA extraction and amplification of a fragment of the 18S rRNA, by means of PCR. The PCR products were purified and sequenced. Sequences were obtained from 35 samples but only 17 of these were kept after quality assessment. Similarity analysis using BLASTn demonstrated that all 17 sequences corresponded to B. canis vogeli. Analysis using the Mega4 software showed that the isolates of B. canis vogeli from dogs in Goiânia present a high degree of molecular similarity (99.2 to 100% in comparison with other reference isolates from other regions of Brazil and worldwide, deposited in GenBank.O gênero Babesia compreende protozoários causadores de enfermidades denominadas babesioses. Cães geralmente são acometidos por Babesia canis ou Babesia gibsoni, sendo a primeira classificada em subespécies Babesia canis canis, Babesia canis vogeli e Babesia canis rossi. Entre essas, Babesia canis vogeli predomina no Brasil. O objetivo desse trabalho foi realizar estudo filogenético de amostras de Babesia em cães, em Goiânia, Goiás. Amostras de sangue foram obtidas de 890 cães ate