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Sample records for anaplasma

  1. Transformation of Anaplasma phagocytophilum

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    Barbet Anthony F

    2006-10-01

    Full Text Available Abstract Background Tick-borne pathogens cause emerging zoonoses, and include fastidious organisms such as Anaplasma phagocytophilum. Because of their obligate intracellular nature, methods for mutagenesis and transformation have not been available. Results To facilitate genetic manipulation, we transformed A. phagocytophilum (Ap to express a green fluorescent protein (GFP with the Himar1 transposase system and selection with the clinically irrelevant antibiotic spectinomycin. Conclusion These transformed bacteria (GFP/Ap grow at normal rates and are brightly fluorescent in human, monkey, and tick cell culture. Molecular characterization of the GFP/Ap genomic DNA confirmed transposition and the flanking genomic insertion locations were sequenced. Three mice inoculated with GFP/Ap by intraperitoneal injection became infected as demonstrated by the appearance of morulae in a peripheral blood neutrophil and re-isolation of the bacteria in culture.

  2. The Life and Death of Anaplasma

    Centers for Disease Control (CDC) Podcasts

    2012-03-30

    Dr. Setu Vora, medical director of critical care and physician director of performance improvement at Backus Hospital in Norwich, Connecticut, reads his poem The Life and Death of Anaplasma and discusses the poem’s origins.  Created: 3/30/2012 by National Center for Emerging and Zoonotic Infectious Diseases (NCEZID).   Date Released: 4/2/2012.

  3. Anaplasma phagocytophilum and Anaplasma marginale Elicit Different Gene Expression Responses in Cultured Tick Cells

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    Zorica Zivkovic

    2009-01-01

    Full Text Available The genus Anaplasma (Rickettsiales: Anaplasmataceae includes obligate tick-transmitted intracellular organisms, Anaplasma phagocytophilum and Anaplasma marginale that multiply in both vertebrate and tick host cells. Recently, we showed that A. marginale affects the expression of tick genes that are involved in tick survival and pathogen infection and multiplication. However, the gene expression profile in A. phagocytophilum-infected tick cells is currently poorly characterized. The objectives of this study were to characterize tick gene expression profile in Ixodes scapularis ticks and cultured ISE6 cells in response to infection with A. phagocypthilum and to compare tick gene expression responses in A. phagocytophilum- and A. marginale-infected tick cells by microarray and real-time RT-PCR analyses. The results of these studies demonstrated modulation of tick gene expression by A. phagocytophilum and provided evidence of different gene expression responses in tick cells infected with A. phagocytophilum and A. marginale. These differences in Anaplasma-tick interactions may reflect differences in pathogen life cycle in the tick cells.

  4. Detection of Rickettsia and Anaplasma from hard ticks in Thailand.

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    Malaisri, Premnika; Hirunkanokpun, Supanee; Baimai, Visut; Trinachartvanit, Wachareeporn; Ahantarig, Arunee

    2015-12-01

    We collected a total of 169 adult hard ticks and 120 nymphs from under the leaves of plants located along tourist nature trails in ten localities. The results present data examining the vector competence of ticks of different genera and the presence of Rickettsia and Anaplasma species. The ticks belonged to three genera, Amblyomma, Dermacentor, and Haemaphysalis, comprising 11 species. Rickettsia bacteria were detected at three collection sites, while Anaplasma bacteria were detected at only one site. Phylogenetic analysis revealed new rickettsia genotypes from Thailand that were closely related to Rickettsia tamurae, Rickettsia monacensis, and Rickettsia montana. This study was also the first to show that Anaplasma bacteria are found in Haemaphysalis shimoga ticks and are closely related evolutionarily to Anaplasma bovis. These results provide additional information for the geographical distribution of tick species and tick-borne bacteria in Thailand and can therefore be applied for ecotourism management.

  5. Molecular survey and sequence analysis of Anaplasma spp. in cattle and ticks in a Malaysian farm.

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    Tay, S T; Koh, F X; Kho, K L; Ong, B L

    2014-12-01

    This study was conducted to determine the occurrence of Anaplasma spp. in the blood samples of cattle, goats, deer and ticks in a Malaysian farm. Using polymerase chain reaction (PCR) and sequencing approach, Anaplasma spp. was detected from 81(84.4%) of 96 cattle blood samples. All blood samples from 23 goats and 22 deer tested were negative. Based on the analysis of the Anaplasma partial 16S ribosomal RNA gene, four sequence types (genotypes 1 to 4) were identified in this study. Genotypes 1-3 showed high sequence similarity to those of Anaplasma platys/ Anaplasma phagocytophilum, whilst genotype 4 was identical to those of Anaplasma marginale/ Anaplasma centrale/ Anaplasma ovis. Anaplasma DNA was detected from six (5.5%) of 109 ticks which were identified as Rhipicephalus (formely known as Boophilus) microplus ticks collected from the cattle. This study reported for the first time the detection of four Anaplasma sequence types circulating in the cattle population in a farm in Malaysia. The detection of Anaplasma DNA in R. microplus ticks in this study provides evidence that the ticks are one of the potential vectors for transmission of anaplasmosis in the cattle.

  6. Anaplasma phagocytophilum in ticks in Slovenia

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    Knap Nataša

    2010-11-01

    Full Text Available Abstract Ticks act as vectors of many pathogens of domestic animals and humans. Anaplasma phagocytophilum in Europe is transmitted by the ixodid tick vector Ixodes ricinus. A. phagocytophilum causes a disease with diverse clinical signs in various hosts. A great genetic diversity of the groESL operon of A. phagocytophilum has been found in ticks elsewhere. In Slovenia, the variety of the groESL operon was conducted only on deer samples. In this study, the prevalence of infected ticks was estimated and the diversity of A. phagocytophilum was evaluated. On 8 locations in Slovenia, 1924 and 5049 (6973 I. ricinus ticks were collected from vegetation in the years 2005 and 2006, respectively. All three feeding stages of the tick's life cycle were examined. The prevalence of ticks infected with A. phagocytophilum in the year 2005 and in the year 2006 was 0.31% and 0.63%, respectively, and it did not differ considerably between locations. The similarity among the sequences of groESL ranged from 95.6% to 99.8%. They clustered in two genetic lineages along with A. phagocytophilum from Slovenian deer. One sequence formed a separate cluster. According to our study, the prevalence of A. phagocytophilum in ticks is comparable to the findings in other studies in Europe, and it does not vary considerably between locations and tick stages. According to groESL operon analysis, two genetic lineages have been confirmed and one proposed. Further studies on other genes would be useful to obtain more information on genetic diversity of A. phagocytophilum in ticks in Slovenia.

  7. Anaplasma phagocytophilum in questing Ixodes ricinus ticks in southwestern Finland.

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    Sormunen, Jani J; Penttinen, Ritva; Klemola, Tero; Vesterinen, Eero J; Hänninen, Jari

    2016-12-01

    Anaplasma phagocytophilum is the causative agent of an emerging tick-borne disease, human granulocytic anaplasmosis. While the bacterium has been reported from questing ticks in neighboring Sweden, Norway and Russia, the few surveys regarding questing ticks in Finland have thus far been negative. In the current study, the prevalence of A. phagocytophilum in Ixodes ricinus populations was evaluated in several study localities around southwestern Finland during 2013-2014. Some of these populations were previously screened and found negative for A. phagocytophilum in 2000. A total of 3158 I. ricinus collected by blanket dragging were screened for Anaplasma spp. using qPCR. Anaplasma were detected in 9.2% of adult ticks (n = 87) and 3.1% of nymphs (n = 979). All larval samples were negative for infection. All Anaplasma-positive samples were identified as A. phagocytophilum by sequencing. This is, to the best of our knowledge, the first report of the pathogen from questing ticks in Finland. Furthermore, the pathogen was detected from several localities found negative during the previous screening 13 years earlier.

  8. Circulation of four Anaplasma phagocytophilum ecotypes in Europe

    NARCIS (Netherlands)

    Jahfari, Setareh; Coipan, E Claudia; Fonville, Manoj; van Leeuwen, Arieke Docters; Hengeveld, Paul; Heylen, Dieter; Heyman, Paul; van Maanen, Cees; Butler, Catherine M; Földvári, Gábor; Szekeres, Sándor; van Duijvendijk, Gilian; Tack, Wesley; Rijks, Jolianne M; van der Giessen, Joke; Takken, Willem; van Wieren, Sipke E; Takumi, Katsuhisa; Sprong, Hein

    2014-01-01

    BACKGROUND: Anaplasma phagocytophilum is the etiological agent of granulocytic anaplasmosis in humans and animals. Wild animals and ticks play key roles in the enzootic cycles of the pathogen. Potential ecotypes of A. phagocytophilum have been characterized genetically, but their host range, zoonoti

  9. Circulation of four Anaplasma phagocytophilum ecotypes in Europe

    NARCIS (Netherlands)

    Jahfari, S.; Coipan, E.C.; Fonville, M.; Leeuwen, van A.D.; Hengeveld, P.; Heylen, D.; Heyman, P.; Maanen, van C.; Butler, C.M.; Foldvari, G.; Szekeres, S.; Duijvendijk, van L.A.G.; Tack, W.; Rijks, J.M.; Giessen, van der J.; Takken, W.; Wieren, van S.E.; Takumi, K.; Sprong, H.

    2014-01-01

    Background: Anaplasma phagocytophilum is the etiological agent of granulocytic anaplasmosis in humans and animals. Wild animals and ticks play key roles in the enzootic cycles of the pathogen. Potential ecotypes of A. phagocytophilum have been characterized genetically, but their host range, zoonoti

  10. Natural Anaplasma phagocytophilum infection in ticks from a forest area of Selenge province, Mongolia

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    G Javkhlan

    2014-03-01

    Full Text Available Anaplasma phagocytophilum is a zoonotic agent of public health importance, infecting both humans and animals. An investigation of the presence of Anaplasma phagocytophilum as well as Anaplasma platys was conducted in a forest area of Selenge province, Mongolia, where ticks are widely distributed and tick-borne diseases are highly endemic. Ticks were collected and tested using polymerase chain reaction based on groEL methodology. Anaplasma phagocytophilum was detected in 14 (6% of Ixodes persulcatus ticks and four (1% Dermacentor nuttalli ticks; infection of Anaplasma platys was detected in 1% of Ixodes persulcatus ticks and 10% of Dermacentor nuttalli ticks. The phylogenetic tree showed that the Anaplasma phagocytophilum clustered with the Russian group, most likely due to similar geographical locations. This finding is significant for both veterinary and public health officials given that these agents can cause both animal and human illness.

  11. Detection of Anaplasma sp. in Korean Native Goats (Capra aegagrus hircus) on Jeju Island, Korea.

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    Seong, Giyong; Han, Yu-Jung; Chae, Jeong-Byoung; Chae, Joon-Seok; Yu, Do-Hyeon; Lee, Young-Sung; Park, Jinho; Park, Bae-Keun; Yoo, Jae-Gyu; Choi, Kyoung-Seong

    2015-12-01

    Anaplasma species are obligate intracellular pathogens that can cause tick-borne diseases in mammalian hosts. To date, very few studies of their occurrence in Korean native goats (Capra aegagrus hircus) have been reported. In the present study, we investigated Anaplasma infection of Korean native goats on Jeju Island, Republic of Korea, and performed phylogenetic analysis based on the 16S rRNA gene sequences. Our results showed that Anaplasma infection was found mostly in adult female goats. The phylogenetic tree revealed that the 7 sequences identified in Korean native goats could belong to Anaplasma sp. and were distinct from A. marginale, A. centrale, and A. ovis. The results indicated that the sequences identified to belong to Anaplasma were closely related to sequences isolated from goats in China and were clustered within the same group. To our knowledge, this is the first study to detect Anaplasma sp. infection in Korean native goats.

  12. Knockdown of the rhipicephalus microplus cytochrome c oxidase subunit III gene is associated with a failure of anaplasma marginale transmission

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    Rhipicephalus microplus is an obligate hematophagous ectoparasite of cattle and an important biological vector of Anaplasma marginale in tropical and subtropical regions. The primary determinants for Anaplasma transmission are infection of tick gut epithelial cells followed by infection of salivary ...

  13. Serologic and molecular characterization of Anaplasma species infection in farm animals and ticks from Sicily.

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    de la Fuente, José; Torina, Alessandra; Caracappa, Santo; Tumino, Giovanni; Furlá, Roberto; Almazán, Consuelo; Kocan, Katherine M

    2005-11-05

    Although Anaplasma marginale was known to be endemic in Italy, the diversity of Anaplasma spp. from this area have not been characterized. In this study, the prevalence of Anaplasma spp. antibodies in randomly selected farm animals collected on the island of Sicily was determined by use of a MSP5 cELISA for Anaplasma spp. and an immunofluorescence test specific for Anaplasma phagocytophilum. Genetic variation among strains of Anaplasma spp. from animals and ticks was characterized using the A. marginale msp1alpha and the Anaplasma spp. msp4 genes. Eight species of ticks were collected and tested by PCR. Seropositivity for Anaplasma spp. and A. phagocytophilum was detected in bovine and ovine samples. All the donkeys were seropositive for A. phagocytophilum but not for Anaplasma spp. Four A. marginale genotypes were identified by msp4 sequences from bovine and tick samples. Two new genotypes of Anaplasma ovis were characterized in sheep. The sequences of A. phagocytophilum from three donkeys proved to be identical to the sequence of the MRK equine isolate from California. Six A. marginale genotypes were found in cattle and one tick using the A. marginale msp1alpha sequences. All genotypes had four repeated sequences in the N-terminal portion of the MSP1a, except for one that had five repeats. The Italian strains of A. marginale contained three repeat sequences that were not reported previously. Definition of the diversity of Anaplasma spp. in Sicily reported, herein is fundamental to development of control strategies for A. marginale, A. ovis and A. phagocytophilum in Sicily.

  14. Anaplasma marginale and Theileria annulata in questing ticks from Portugal.

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    Antunes, S; Ferrolho, J; Domingues, N; Santos, A S; Santos-Silva, M M; Domingos, A

    2016-09-01

    Ticks are ubiquitous arthropods and vectors of several pathogenic agents in animals and humans. Monitoring questing ticks is of great importance to ascertain the occurrence of pathogens and the potential vector species, offering an insight into the risk of disease transmission in a given area. In this study 428 host-seeking ticks, belonging to nine species of Ixodidae and collected from 17 of the 23 Portuguese mainland subregions, were screened for several tick-borne agents with veterinary relevance: Anaplasma marginale, Anaplasma ovis, Anaplasma centrale, Babesia spp., Coxiella burnetii and Theileria spp. Prevalence was assessed by PCR and amplified amplicons sequenced for validation of results. Twenty ticks, in a total of 428, were found positive: one Ixodes ventalloi for Theileria annulata and four Dermacentor marginatus, one Haemaphysalis punctata, five Ixodes ricinus, five I. ventalloi, and four Rhipicephalus sanguineus sensu lato for A. marginale. According to the reviewed literature, this is the first report of A. marginale and T. annulata detection in I. ventalloi. Furthermore, the amplification of A. marginale DNA in several tick species suggests a broad range for this agent in Portugal that might include other uncommon species as R. sanguineus s.l. This work provides new data towards a better understanding of tick-pathogen associations and also contributes to the surveillance of tick-borne agents in geographic areas with limited information.

  15. Comparison of three nucleic acid-based tests for detecting Anaplasma marginale and Anaplasma centrale in cattle

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    Mamohale E. Chaisi

    2017-01-01

    Full Text Available Several nucleic acid-based assays have been developed for detecting Anaplasma marginale and Anaplasma centrale in vectors and hosts, making the choice of method to use in endemic areas difficult. We evaluated the ability of the reverse line blot (RLB hybridisation assay, two nested polymerase chain reaction (nPCR assays and a duplex real-time quantitative polymerase chain reaction (qPCR assay to detect A. marginale and A. centrale infections in cattle (n = 66 in South Africa. The lowest detection limits for A. marginale plasmid DNA were 2500 copies by the RLB assay, 250 copies by the nPCR and qPCR assays and 2500, 250 and 25 copies of A. centrale plasmid DNA by the RLB, nPCR and qPCR assays respectively. The qPCR assay detected more A. marginale- and A. centrale-positive samples than the other assays, either as single or mixed infections. Although the results of the qPCR and nPCR tests were in agreement for the majority (38 of A. marginale-positive samples, 13 samples tested negative for A. marginale using nPCR but positive using qPCR. To explain this discrepancy, the target sequence region of the nPCR assay was evaluated by cloning and sequencing the msp1β gene from selected field samples. The results indicated sequence variation in the internal forward primer (AM100 area amongst the South African A. marginale msp1β sequences, resulting in false negatives. We propose the use of the duplex qPCR assay in future studies as it is more sensitive and offers the benefits of quantification and multiplex detection of both Anaplasma spp.

  16. Molecular Survey of Anaplasma and Ehrlichia of Red Deer and Sika Deer in Gansu, China in 2013.

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    Li, Y; Chen, Z; Liu, Z; Liu, J; Yang, J; Li, Q; Li, Y; Luo, J; Yin, H

    2016-12-01

    Anaplasma and Ehrlichia are important emerging tick-borne pathogens in both humans and animals. Here, we conducted a molecular surveillance study in Gansu, China to assess the prevalence of Anaplasma and Ehrlichia spp. in red deer and sika deer based on polymerase chain reaction (PCR) analysis and sequencing of 16S rRNA or msp genes. PCR revealed that the prevalence of Anaplasma ovis, Anaplasma bovis and Anaplasma platys of the Qilian Mountain samples was 32%, 9% and 9%, respectively; the prevalence of Anaplasma ovis, Anaplasma bovis, Anaplasma platys was 20%, 15% and 15% among the Long Mountain samples, respectively. Of the Long Mountain samples, two (5%) of the 40 samples were positive for Ehrlichia canis, but all 44 of the Qilian Mountain samples were negative for E. canis, and no other Anaplasma or Ehrlichia spp. were found in the samples. The phylogenetic tree showed that the newly isolated Anaplasma and Ehrlichia spp. could be classified as belonging to four clades, including an A. bovis cluster, A. ovis cluster, A. platys cluster and E. canis cluster. In addition, Bartonella schoenbuchensis was firstly identified in blood samples from red deer in Gansu, China. Our results provide important data to increase the understanding of the epidemiology of anaplasmosis and ehrlichiosis of red deer and sika deer and will assist with the implementation of measures to control anaplasmosis and ehrlichiosis transmission to red deer, sika deer and other animals in Gansu, China.

  17. Acute phase response in cattle infected with Anaplasma marginale.

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    Nazifi, S; Razavi, S M; Kaviani, F; Rakhshandehroo, E

    2012-03-23

    This study was undertaken to evaluate the acute phase responses via the assessment of the concentration of serum sialic acids (total, lipid bound and protein bound), inflammatory mediators (IFN-γ and TNF-α) and acute phase proteins (Hp and SAA) in 20 adult crossbred cattle naturally infected by Anaplasma marginale. The infected animals were divided into 2 subgroups on the basis of parasitemia rate (20%). Also, as a control group, 10 clinically healthy cattle from the same farms were sampled. Our data revealed significant decreases in red blood cell count (RBC), hematocrite (PCV) and hemoglobine (Hb) in infected cattle compared to healthy ones. Conversely, the concentrations of Hp, SAA, ceruloplasmin, fibrinogen, serum sialic acids and the circulatory IFN-γ and TNF-α were increased in the diseased cattle (P<0.05). In addition, it was evident that the progression of parasitemia in infected cattle did not induce any significant alterations in the hematological indices (RBCs, PCV and Hb) and the concentrations of Hp, SAA, ceruloplasmin and fibrinogen. SAA was the most sensitive factor to change in the diseased cattle. Therefore, increase in SAA concentration may be a good indicator of inflammatory process in cattle naturally infected with Anaplasma marginale.

  18. Molecular detection of Ehrlichia canis, Anaplasma bovis, Anaplasma platys, Candidatus Midichloria mitochondrii and Babesia canis vogeli in ticks from Israel.

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    Harrus, S; Perlman-Avrahami, A; Mumcuoglu, K Y; Morick, D; Eyal, O; Baneth, G

    2011-03-01

    : Ticks are vectors of important pathogens of human and animals. Therefore, their microbial carriage capacity is constantly being investigated. The aim of this study was to characterize the diversity of domestic animal pathogens in ticks collected from vegetation and the ground, from different parts of Israel. Non-engorged questing adult ticks were collected from 13 localities. A total of 1196 ticks in 131 pools-83 pools of Rhipicephalus turanicus and 48 of Rhipicephalus sanguineus (with two to ten ticks per pool)-were included in this study. In addition, 13 single free-roaming Hyalomma spp. ticks were collected. Screening by molecular techniques revealed the presence of Ehrlichia canis, Anaplasma platys, Anaplasma bovis and Babesia canis vogeli DNA in R. turanicus ticks. E. canis, A. bovis, B. canis vogeli and Candidatus Midichloria mitochondrii DNA sequences were detected in R. sanguineus ticks. Candidatus Midichloria mitochondrii DNA was also detected in Hyalomma spp. ticks. Neither Hepatozoon spp. nor Bartonella spp. DNA was detected in any of the ticks examined. This study describes the first detection of E. canis in the tick R. turanicus, which may serve as a vector of this canine pathogen; E. canis was the most common pathogen detected in the collected questing ticks. It also describes the first detection of A. bovis and Candidatus Midichloria mitochondrii in Israel. To the best of the author's knowledge, this is the first report describing the detection of DNA of the latter two pathogens in R. sanguineus, and of A. bovis in R. turanicus.

  19. Improved molecular detection of Ehrlichia and Anaplasma species applied to Amblyomma ticks collected from cattle and sheep in Ethiopia.

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    Teshale, S; Geysen, D; Ameni, G; Asfaw, Y; Berkvens, D

    2015-02-01

    Detection of Ehrlichia and Anaplasma species in animals and tick vectors is crucial for an understanding of the epidemiology of diseases caused by these pathogens. In this study, a pair of primers designated EBR2 and EBR3 was designed from the Anaplasma 16S rDNA sequence and was used along with a previously described primer EHR 16SD for the simultaneous detection of Ehrlichia and Anaplasma species by nested PCR. The primers were used to amplify 925bp of DNA from known species of Ehrlichia and Anaplasma. Restriction with MboII and MspI enzymes allowed Ehrlichia and Anaplasma speciation. Restriction with MboII differentiated between An. marginale, Anaplasma (formerly Ehrlichia) sp. Omatjenne, and An. centrale with An. marginale and Anaplasma (formerly Ehrlichia) sp. Omatjenne yielding 2 distinct fragments each while An. centrale produced 3 distinct bands. Ehrlichia ruminantium and An. phagocytophylum remained undigested. Subsequent restriction with MspI differentiated E. ruminantium from An. phagocytophylum with 2 and 4 fragments, respectively. When used on tick samples from the field, 63 ticks (16.4%) out of 384 collected from cattle and sheep were positive for one or more species of Ehrlichia and Anaplasma. The positivity ranged from 6.3% at Andasa to 36.7% at Habernosa. Higher overall infection rates were found in Amblyomma lepidum than in Amblyomma variegatum ticks (p=0.009). Amblyomma lepidum from Habernosa were more often infected with all detected species of Anaplasma and Ehrlichia than Am. variegatum. At Bako, however, Anaplasma (formerly Ehrlichia) sp. Omatjenne was detected only in Am. variegatum. A significantly higher proportion of ticks collected from cattle (20.6%) was found positive than in those collected from sheep (3.3%) (p=0.003). Simultaneous detection of Ehrlichia and Anaplasma species and correct identification of mixed infections was possible. Since the ticks were collected from animals, the occurrence of the major species of Ehrlichia and

  20. Molecular Detection of Anaplasma spp. and Ehrlichia spp. in Ruminants from Twelve Provinces of China

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    Qiu, Haixiang; Kelly, Patrick John; Zhang, Jilei; Luo, Qinghua; Yang, Yi; Mao, Yongjiang; Yang, Zhangping; Li, Jing; Wu, Hongzhuan

    2016-01-01

    Anaplasma spp. and Ehrlichia spp. are tick-transmitted bacteria that are of significant economic importance as they can infect large and small ruminants and also people. There is little information on anaplasmosis and ehrlichiosis in ruminants in China. 16S rRNA FRET-qPCRs were used to screen convenience whole blood samples from 2,240 domestic ruminants in 12 provinces of China for Anaplasma spp. and Ehrlichia spp. Positive samples were further analyzed with a standard PCR for the gltA. Anaplasma spp. DNA was detected in the sheep (11.7%; 13/111), goats (81.8%; 219/270), cattle (13.2%; 241/1,830), and water buffaloes (6.9%; 2/29). Ehrlichia spp. DNA was detected in sheep (1.8%; 2/111), goats (1.1%; 3/270), and cattle (3.6%; 65/1830) but not in water buffaloes (0/29). Sequencing of gltA PCR products showed that A. marginale, A. ovis, Ehrlichia canis, and Ehrlichia sp. (JX629807) were present in ruminants from China, while the 16S rRNA FRET-qPCR sequence data indicated that there might also be A. platys, A. phagocytophilum, Anaplasma sp. BL126-13 (KJ410243), and Anaplasma sp. JC3-6 (KM227012). Our study shows that domestic ruminants from China are not uncommonly infected with a variety of Anaplasma spp. and Ehrlichia spp. PMID:28096822

  1. Comparative study of Anaplasma parasites in tick carrying buffaloes and cattle

    Institute of Scientific and Technical Information of China (English)

    RAJPUT Z.I.; HU Song-hua; ARIJO A.G.; HABIB M.; KHALID M.

    2005-01-01

    A comparative study on the prevalence of Anaplasma parasite was conducted on ticks carrying buffaloes and cattle.Five hundred blood samples of both animals (250 of each) were collected during February, March and April. Thin blood smears on glass slides were made, fixed in 100% methyl alcohol and examined. Microscopic examination revealed that 205 (41%) animals had Anaplasma parasites, out of which 89, 44 and 72 animals had Anaplasma marginale, Anaplasma centrale and mixed infection respectively. Infected buffaloes and cattle were 75 and 130 respectively. The infection in female was 53 and 92 in buffaloes and cattle respectively. Twenty-two and 92 blood samples of male were found positive in buffaloes and cattle respectively. Comparative study revealed that the cattle were 26.82% more susceptible than buffaloes. The parasite prevailing percentage in female of both animals was slightly higher than that of the male. This investigation was aimed at studying the comparative prevalence of Anaplasma parasite in tick carrying buffaloes and cattle.

  2. [Microbiological diagnosis of emerging bacterial pathogens: Anaplasma, Bartonella, Rickettsia, and Tropheryma whipplei].

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    Blanco, José Ramón; Jado, Isabel; Marín, Mercedes; Sanfeliu, Isabel; Portillo, Aránzazu; Anda, Pedro; Pons, Immaculada; Oteo, José Antonio

    2008-11-01

    Ehrlichia/Anaplasma, Bartonella, Rickettsia and Tropheryma whipplei (formerly called whippelii) are fastidious bacterial organisms, considered the causative agents of potentially severe emerging and re-emerging diseases with repercussions on public health. The recent availability of advanced molecular biology and cell culture techniques has led to the implication of many of these species in human pathologies. These issues are extensively covered in number 27 of the SEIMC microbiological procedure: Diagnóstico microbiológico de las infecciones por patógenos bacterianos emergentes: Anaplasma, Bartonella, Rickettsia y Tropheryma whippelii (Microbiological diagnosis of Anaplasma, Bartonella, Rickettsia and Tropheryma whippelii infections) (2nd ed., 2007) (www.seimc.org/documentos/protocolos/microbiologia/).

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

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    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.

  4. Anaplasma, Ehrlichia and rickettsial pathogens in ixodid ticks infesting cattle and sheep in western Oromia, Ethiopia.

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    Teshale, Sori; Kumsa, Bersissa; Menandro, Maria Luisa; Cassini, Rudi; Martini, Marco

    2016-10-01

    Although ticks are widely distributed in all agro-ecological zones of Ethiopia, information on tick-borne pathogens is scarce. This study was conducted to determine the presence of Anaplasma spp., Ehrlichia spp., and Rickettsia spp. in Rhipicephalus evertsi and Rhipicephalus (Boophilus) decoloratus collected from cattle and sheep at Bako, western Oromia, Ethiopia, using polymerase chain reaction and sequencing. Anaplasma ovis and Anaplasma spp., Ehrlichia ruminantium and Ehrlichia spp. were detected in Rh. decoloratus, whereas only A. ovis was detected in Rh. evertsi. Both tick species were found to harbor DNA belonging to Rickettsia spp., and Rickettsia africae. Our findings highlight the risk of infection of animals and humans with these zoonotic tick-borne bacteria in Ethiopia.

  5. Molecular survey of Rickettsia, Ehrlichia, and Anaplasma infection of domestic cats in Japan.

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    Sasaki, Hiromi; Ichikawa, Yasuaki; Sakata, Yoshimi; Endo, Yasuyuki; Nishigaki, Kazuo; Matsumoto, Kotaro; Inokuma, Hisashi

    2012-12-01

    The prevalence of Rickettsia, Ehrlichia, and Anaplasma in 1764 DNA samples extracted from feline peripheral blood from all 47 prefectures in Japan was evaluated by screening real-time PCR, genus-specific PCR, and DNA nucleotide sequencing. The survey revealed that all cats were negative for Rickettsia infection. Two cats were positive for Ehrlichia or Anaplasma based on the screening PCR assay. Nucleotide sequence analysis of the partial 16S rRNA including the divergent region near the 3'-end revealed that the 2 positives were most similar to Anaplasma bovis with percent identities of 99.8% and 99.2%. This was the first detection of A. bovis DNA fragments in cats. Although these 2 cats showed stomatitis, both were also infected with feline immunodeficiency virus. The relationship between A. bovis carriage and clinical disease is not yet understood.

  6. Anaplasma phagocytophilum in Danish sheep: confirmation by DNA sequencing

    Directory of Open Access Journals (Sweden)

    Thamsborg Stig M

    2009-12-01

    Full Text Available Abstract Background The presence of Anaplasma phagocytophilum, an Ixodes ricinus transmitted bacterium, was investigated in two flocks of Danish grazing lambs. Direct PCR detection was performed on DNA extracted from blood and serum with subsequent confirmation by DNA sequencing. Methods 31 samples obtained from clinically normal lambs in 2000 from Fussingø, Jutland and 12 samples from ten lambs and two ewes from a clinical outbreak at Feddet, Zealand in 2006 were included in the study. Some of the animals from Feddet had shown clinical signs of polyarthritis and general unthriftiness prior to sampling. DNA extraction was optimized from blood and serum and detection achieved by a 16S rRNA targeted PCR with verification of the product by DNA sequencing. Results Five DNA extracts were found positive by PCR, including two samples from 2000 and three from 2006. For both series of samples the product was verified as A. phagocytophilum by DNA sequencing. Conclusions A. phagocytophilum was detected by molecular methods for the first time in Danish grazing lambs during the two seasons investigated (2000 and 2006.

  7. Prevalence of Anaplasma phagocytophilum infection and effect on lamb growth

    Directory of Open Access Journals (Sweden)

    Steinshamn Håvard

    2011-05-01

    Full Text Available Abstract Background A major challenge in sheep farming during the grazing season along the coast of south-western Norway is tick-borne fever (TBF caused by the bacteria Anaplasma phagocytophilum that is transmitted by the tick Ixodes ricinus. Methods A study was carried out in 2007 and 2008 to examine the prevalence of A. phagocytophilum infection and effect on weaning weight in lambs. The study included 1208 lambs from farms in Sunndal Ram Circle in Møre and Romsdal County in Mid-Norway, where ticks are frequently observed. All lambs were blood sampled and serum was analyzed by an indirect fluorescent antibody assay (IFA to determine an antibody status (positive or negative to A. phagocytophilum infection. Weight and weight gain and possible effect of infection were analyzed using ANOVA and the MIXED procedure in SAS. Results The overall prevalence of infection with A. phagocytophilum was 55%. A lower weaning weight of 3% (1.34 kg, p A. phagocytophilum infection compared to seronegative lambs at an average age of 137 days. Conclusions The results show that A. phagocytophilum infection has an effect on lamb weight gain. The study also support previous findings that A. phagocytophilum infection is widespread in areas where ticks are prevalent, even in flocks treated prophylactic with acaricides.

  8. Anaplasma phagocytophilum infection in moose (Alces alces) in Norway.

    Science.gov (United States)

    Pūraitė, Irma; Rosef, Olav; Paulauskas, Algimantas; Radzijevskaja, Jana

    2015-01-01

    Anaplasma phagocytophilum is a tick-borne bacterium that infects a wide range of animal species. The aim of our study was to investigate the prevalence of A. phagocytophilum in Norwegian moose Alces alces and to characterize the bacteria by sequencing of partial msp4 and 16S rRNA genes. Hunters collected spleen samples from 99 moose of different ages during 2013 and 2014 in two areas: Aust-Agder County (n = 70) where Ixodes ricinus ticks are abundant and Oppland County (n = 29) where ticks were either absent, or abundance very low. A. phagocytophilum was detected only in moose from the I. ricinus - abundant area. The overall prevalence of infection according to 16S rRNA and msp4 gene-based PCR was 41.4% and 31.4% respectively. Sequence analysis of the partial 16S rRNA and msp4 gene revealed two and eight different sequence types respectively. Four of eight msp4 sequence types determined in this study were unique, while others were identical to sequences derived from other ruminants and ticks. The present study indicates that moose could be a potential wildlife reservoir of A. phagocytophilum in Norway.

  9. Experimental transmission of Anaplasma marginale by male Dermacentor reticulatus

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    Kocan Katherine M

    2007-11-01

    Full Text Available Abstract Background Bovine anaplasmosis has been reported in several European countries, but the vector competency of tick species for Anaplasma marginale from these localities has not been determined. Because of the wide distributional range of Dermacentor reticulatus within Europe and the major role of Dermacentor spp. as a vector of A. marginale in the United States, we tested the vector competency of D. reticulatus for A. marginale. Results Male D. reticulatus were allowed to feed for 7 days on a calf persistently infected with a Zaria isolate of A. marginale, after which they were removed and held off-host for 7 days. The ticks were then allowed to feed a second time for 7 days on a susceptible tick-naïve calf. Infection of calf No. 4291 was detected 20 days post exposure (p.i. and confirmed by msp4 PCR. Thirty percent of the dissected acquisition fed ticks was infected. In addition, A. marginale colonies were detected by light microscopy in the salivary glands of the acquisition fed ticks. Transmission of A. marginale to calf No. 9191 was confirmed by examination of Giemsa-stained blood smears and msp4 PCR. Ticks were dissected after transmission feeding and presence of A. marginale was confirmed in 18.5% of the dissected ticks. Conclusion This study demonstrates that D. reticulatus males are competent vectors of A. marginale. Further studies are needed to confirm the vector competency of D. reticulatus for other A. marginale strains from geographic areas in Europe.

  10. First molecular evidence for the presence of Anaplasma DNA in milk from sheep and goats in China.

    Science.gov (United States)

    Zhang, Yan; Lv, Yali; Cui, Yanyan; Wang, Jinhong; Cao, Shuxuan; Jian, Fuchun; Wang, Rongjun; Zhang, Longxian; Ning, Changshen

    2016-07-01

    Anaplasmosis, a disease caused by bacteria in the genus of Anaplasma, imposes economic constraints on animal breeders and also threatens human health. In the present study, we investigated the presence of Anaplasma spp. DNA in milk collected from sheep and goats in China. A total of 120 milk samples and 414 field-sampled blood specimens from sheep and goats were analyzed by PCR and DNA sequencing. Anaplasma ovis was detected in 12 milk samples (three from sheep and nine from goats). The blood specimens corresponding to the A. ovis DNA-positive milk were analyzed for Anaplasma DNA presence, and A. ovis DNA was identified in 10 out of the 12 blood specimens. One goat's milk sample was Anaplasma bovis DNA-positive, as was the corresponding blood sample. Anaplasma phagocytophilum was found in a milk sample and blood sample from one goat. One milk sample from Xinmi in Henan province was simultaneously positive for A. bovis and A. phagocytophilum; however, the corresponding blood was negative for both species. DNA sequencing of the PCR products and phylogenetic analysis confirmed that the sequences from the milk samples matched those of the corresponding blood samples. This is the first report to detect the Anaplasma DNA in milk samples under natural condition, and represents the first evidence of the presence of A. ovis, A. bovis and A. phagocytophilum DNA in milk from sheep and goats.

  11. Identification of Anaplasma marginale type IV secretion system effector proteins.

    Directory of Open Access Journals (Sweden)

    Svetlana Lockwood

    Full Text Available BACKGROUND: Anaplasma marginale, an obligate intracellular alphaproteobacterium in the order Rickettsiales, is a tick-borne pathogen and the leading cause of anaplasmosis in cattle worldwide. Complete genome sequencing of A. marginale revealed that it has a type IV secretion system (T4SS. The T4SS is one of seven known types of secretion systems utilized by bacteria, with the type III and IV secretion systems particularly prevalent among pathogenic Gram-negative bacteria. The T4SS is predicted to play an important role in the invasion and pathogenesis of A. marginale by translocating effector proteins across its membrane into eukaryotic target cells. However, T4SS effector proteins have not been identified and tested in the laboratory until now. RESULTS: By combining computational methods with phylogenetic analysis and sequence identity searches, we identified a subset of potential T4SS effectors in A. marginale strain St. Maries and chose six for laboratory testing. Four (AM185, AM470, AM705 [AnkA], and AM1141 of these six proteins were translocated in a T4SS-dependent manner using Legionella pneumophila as a reporter system. CONCLUSIONS: The algorithm employed to find T4SS effector proteins in A. marginale identified four such proteins that were verified by laboratory testing. L. pneumophila was shown to work as a model system for A. marginale and thus can be used as a screening tool for A. marginale effector proteins. The first T4SS effector proteins for A. marginale have been identified in this work.

  12. Molecular detection of Ehrlichia canis and Anaplasma platys in dogs in Southern Brazil.

    Science.gov (United States)

    Lasta, Camila Serina; dos Santos, Andrea Pires; Messick, Joanne Belle; Oliveira, Simone Tostes; Biondo, Alexander Welker; Vieira, Rafael Felipe da Costa; Dalmolin, Magnus Larruscaim; González, Félix Hilario Diaz

    2013-01-01

    The aims of this study were to determine the occurrence of Anaplasma platys and Ehrlichia canis infection in dogs in Porto Alegre, Southern Brazil; and to investigate their association with hematological abnormalities. Serum samples from 196 dogs were first tested using dot-ELISA for antibodies against Anaplasma spp. and Ehrlichia canis. Peripheral blood samples from 199 dogs were subjected to 16S rRNA nested PCR (nPCR) for A. platys and E. canis, followed by DNA sequencing to ensure pathogen identity. A total of 19/196 samples (9.69%) were positive for Anaplasma spp. using ELISA and 28/199 (14.07%) samples were positive for A. platys by nested PCR. All the dog samples were negative for E. canis, both in anti-E. canis antibody tests and in nested PCR. There were no significant differences in hematological parameters between A. platys-PCR positive and negative dogs and Anaplasma spp. serologically positive dogs, except for basophil counts, which were higher in nPCR-positive dogs. This is the first report showing A. platys presence in dogs in Southern Brazil. In conclusion, hematological parameters may not be sufficient to diagnose A. platys infection in dogs in Southern Brazil, probably due either to low pathogenicity or to chronic infection. On the other hand, E. canis may either have very low occurrence or be absent in dogs in Porto Alegre.

  13. Evaluation of Gulf Coast Ticks (Acari: Ixodidae) for Ehrlichia and Anaplasma Species.

    Science.gov (United States)

    Allerdice, Michelle E J; Hecht, Joy A; Karpathy, Sandor E; Paddock, Christopher D

    2016-12-28

    Amblyomma maculatum Koch (the Gulf Coast tick) is an aggressive, human-biting ixodid tick distributed throughout much of the southeastern United States and is the primary vector for Rickettsia parkeri, an emerging human pathogen. Amblyomma maculatum has diverse host preferences that include white-tailed deer, a known reservoir for Ehrlichia and Anaplasma species, including the human pathogens E. ewingii and E. chaffeensis To examine more closely the potential role of A. maculatum in the maintenance of various pathogenic Ehrlichia and Anaplasma species, we screened DNA samples from 493 questing adult A. maculatum collected from six U.S. states using broad-range Anaplasmataceae and Ehrlichia genus-specific PCR assays. Of the samples tested, four (0.8%) were positive for DNA of Ehrlichia ewingii, one (0.2%) was positive for Anaplasma platys, and one (0.2%) was positive for a previously unreported Ehrlichia species closely related to Ehrlichia muris and an uncultivated Ehrlichia species from Haemaphysalis longicornis ticks in Japan. No ticks contained DNA of Ehrlichia chaffeensis, Ehrlichia canis, the Panola Mountain Ehrlichia, or Anaplasma phagocytophilum This is the first identification of E. ewingii, A. platys, and the novel Ehrlichia in questing Gulf Coast ticks; nonetheless the low prevalence of these agents suggests that A. maculatum is not likely an important vector of these zoonotic pathogens.

  14. Investigation of Anaplasma marginale Seroprevalence in a Traditionally Managed Large California Beef Herd

    Science.gov (United States)

    Foley, Janet E.

    2016-01-01

    Recent observations by stakeholders suggested that ecosystem changes may be driving an increased incidence of bovine erythrocytic anaplasmosis, resulting in a reemerging cattle disease in California. The objective of this prospective cohort study was to estimate the incidence of Anaplasma marginale infection using seroconversion in a northern California beef cattle herd. A total of 143 Black Angus cattle (106 prebreeding heifers and 37 cows) were enrolled in the study. Serum samples were collected to determine Anaplasma marginale seroprevalence using a commercially available competitive enzyme-linked immunosorbent assay test kit. Repeat sampling was performed in seronegative animals to determine the incidence density rate from March through September (2013). Seroprevalence of heifers was significantly lower than that of cows at the beginning of the study (P < 0.001) but not at study completion (P = 0.075). Incidence density rate of Anaplasma marginale infection was 8.17 (95% confidence interval: 6.04, 10.81) cases per 1000 cow-days during the study period. Study cattle became Anaplasma marginale seropositive and likely carriers protected from severe clinical disease that might have occurred had they been first infected as mature adults. No evidence was found within this herd to suggest increased risk for clinical bovine erythrocytic anaplasmosis. PMID:27656312

  15. Knockout of an outer membrane protein operon of anaplasma marginale by transposon mutagenesis

    Science.gov (United States)

    Large amounts of data generated by genomics, transcriptomics and proteomics technologies have increased our understanding of the biology of Anaplasma marginale. However, these data have also led to new assumptions that require testing, ideally through classic genetic mutation. One example is the def...

  16. Molecular detection of novel Anaplasmataceae closely related to Anaplasma platys and Ehrlichia canis in the dromedary camel (Camelus dromedarius).

    Science.gov (United States)

    Bastos, Armanda D S; Mohammed, Osama B; Bennett, Nigel C; Petevinos, Charalambos; Alagaili, Abdulaziz N

    2015-09-30

    Serological surveys have confirmed Anaplasma marginale and Anaplasma phagocytophilum infections in dromedary camels, but molecular surveys and genetic characterisation of camel-associated Anaplasma species are lacking. In this study, we detected tick-borne Anaplasmataceae in 30 of 100 (30%) healthy dromedary camels screened using a combined 16S rRNA-groEL PCR-sequencing approach. Nucleotide sequencing confirmed Anaplasmataceae genome presence in 28 of the 33 16S rRNA PCR-positive samples, with two additional positive samples, for which 16S rRNA sequence data were ambiguous, being identified by groEL gene characterisation. Phylogenetic analyses of a 1289 nt segment of the 16S rRNA gene confirmed the presence of a unique Ehrlichia lineage and a discrete Anaplasma lineage, comprising three variants, occurring at an overall prevalence of 4% and 26%, respectively. Genetic characterisation of an aligned 559 nt groEL gene region revealed the camel-associated Anaplasma and Ehrlichia lineages to be novel and most closely related to Anaplasma platys and Ehrlichia canis. Based on the confirmed monophyly, minimum pairwise genetic distances between each novel lineage and its closest sister taxon, and the inability to isolate the bacteria, we propose that Candidatus status be assigned to each. This first genetic characterisation of Anaplasmataceae from naturally infected, asymptomatic dromedary camels in Saudi Arabia confirms the presence of two novel lineages that are phylogenetically linked to two pathogenic canid species of increasing zoonotic concern.

  17. Detection and identification of Anaplasma phagocytophilum, Borrelia burgdorferi, and Rickettsia helvetica in Danish Ixodes ricinus ticks

    DEFF Research Database (Denmark)

    Skarphédinsson, Sigurdur; Lyholm, Birgitte Fjendbo; Ljungberg, Marianne

    2007-01-01

    Jutland and Funen, while 11% were positive for Borrelia burgdorferi. The Borrelia genotype B. afzelii was most prevalent, followed by B. valaisiana, B. burgdorferi s.s. and B. garinii.A. phagocytophilum was found in 14.5% of nymphs and 40.5% of adult ticks, while Borrelia was found in 13% of nymphs and 8......Borreliosis is an endemic infection in Denmark. Recent serosurveys have indicated that human anaplasmosis may be equally common. The aim of this study was to look for Anaplasma phagocytophilum and related pathogens in Ixodes ricinus ticks and estimate their prevalence, compared to Borrelia, using......% of adult ticks. The difference in prevalence between Anaplasma and Borrelia in adult ticks supports the idea that their maintenance cycles in nature may be different. Ticks were also infected with Rickettsia helvetica. Our study indicates that A. phagocytophilum prevalence in ticks in Denmark is as high...

  18. Infection with Anaplasma phagocytophilum in a seronegative patient in Sicily, Italy: Case Report

    OpenAIRE

    2005-01-01

    Abstract Background Anaplasma phagocytophilum causes human granulocytic anaplasmosis (HGA) in humans, which has been recognized as an emerging tick-borne disease in the United States and Europe. Although about 65 cases of HGA have been reported in Europe, some of them do not fulfill the criteria for confirmed HGA. Confirmation of HGA requires A. phagocytophilum isolation from blood, and/or identification of morulae in granulocytes and/or positive PCR results with subsequent sequencing of the ...

  19. 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...

  20. Molecular detection of Ehrlichia canisand Anaplasma platys in dogs in Southern Brazil

    Directory of Open Access Journals (Sweden)

    Camila Serina Lasta

    Full Text Available The aims of this study were to determine the occurrence ofAnaplasma platys and Ehrlichia canisinfection in dogs in Porto Alegre, Southern Brazil; and to investigate their association with hematological abnormalities. Serum samples from 196 dogs were first tested using dot-ELISA for antibodies against Anaplasmaspp. and Ehrlichia canis. Peripheral blood samples from 199 dogs were subjected to 16S rRNA nested PCR (nPCR for A. platysand E. canis, followed by DNA sequencing to ensure pathogen identity. A total of 19/196 samples (9.69% were positive forAnaplasma spp. using ELISA and 28/199 (14.07% samples were positive for A. platys by nested PCR. All the dog samples were negative for E. canis, both in anti-E. canisantibody tests and in nested PCR. There were no significant differences in hematological parameters between A. platys-PCR positive and negative dogs and Anaplasma spp. serologically positive dogs, except for basophil counts, which were higher in nPCR-positive dogs. This is the first report showing A. platys presence in dogs in Southern Brazil. In conclusion, hematological parameters may not be sufficient to diagnose A. platys infection in dogs in Southern Brazil, probably due either to low pathogenicity or to chronic infection. On the other hand, E. canis may either have very low occurrence or be absent in dogs in Porto Alegre.

  1. Molecular diagnosis and species identification of Ehrlichia and Anaplasma infections in dogs from Panama, Central America.

    Science.gov (United States)

    Santamaria, Annamaria; Calzada, Jose E; Saldaña, Azael; Yabsley, Michael J; Gottdenker, Nicole L

    2014-05-01

    The prevalence and distribution of Anaplasma and Ehrlichia were estimated in 201 symptomatic dogs from Panama by nested PCR and DNA sequencing of the 16S rRNA gene. In total, 163 dogs (70.6%; 163/201) were infected with either Anaplasma or Ehrlichia. On the basis of PCR results, the majority of cases were infected with E. canis (64.2%; 129/201) followed by 21.4% (43/201) with A. platys, whereas 7.5% (15/201) had Anaplasma/Ehrlichia co-infections. Further analyses of 16S rDNA partial sequences show sequence homology with E. canis and A. platys from other countries. Hematology findings from 79 E. canis PCR-positive dogs included anemia (74.7%), thrombocytopenia (81.9%), macroplatelets (29.1%), and leukopenia (6.3%). Among 16 A. platys PCR-positive dogs with available hematology, 62.5% were anemic, 75% had thrombocytopenia, and 100% had macroplatelets. On the basis of E.canis serology (enzyme-linked immunosorbent assay [ELISA]) (n=92 dogs), 30 dogs that were seropositive for E. canis were also PCR-positive, whereas among seronegatives (n=62), 10 were PCR-positive for E. canis. This study provides the first characterization of canine anaplasmosis and ehrlichiosis infections in Panama and is important to veterinary public health and comparative studies of these pathogens in the Americas.

  2. First case of Anaplasma platys infection in a dog from Croatia

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    Dyachenko Viktor

    2012-03-01

    Full Text Available Abstract Background It is known that Anaplasma (A. platys, the causative agent of infectious canine cyclic thrombocytopenia, is endemic in countries of the Mediterranean basin. However, few reports are available from the Balkans. This case report describes a dog, which was imported from Croatia to Germany in May 2010. One month later the dog was presented to a local veterinarian in Germany due to intermittent/recurrent diarrhoea. Diagnostic tests were performed to identify infections caused by Anaplasma spp., Ehrlichia spp., Hepatozoon canis, Babesia spp., Leishmania spp., Borrelia burgdorferi and/or Dirofilaria immitis. Findings Haematological examination of a blood smear revealed basophilic inclusions in thrombocytes, which were confirmed as A. platys with a species-specific real-time PCR. Additionally, an infection with Babesia (B. vogeli was also detected (PCR and serology. No specific antibodies against Anaplasma antigen were detectable. Although the dog showed no specific clinical signs, thrombocytopenia, anaemia and elevated C-reactive protein (CRP were observed. Sequencing of a 1,348-bp partial ribosomal RNA gene revealed highest homology to A. platys sequences from Thailand, Japan and France. Conclusions A. platys was detected for first time in a dog imported from Croatia. As the dog was also co-infected by B. vogeli, unique serological and haematological findings were recorded. Thrombocytopenia, anaemia and elevated values of C-reactive protein were the laboratory test abnormalities observed in this case. A. platys infections should be considered in dogs coming from Croatia and adjacent regions.

  3. Prevalence of Anaplasma and Bartonella spp. in Ticks Collected from Korean Water Deer (Hydropotes inermis argyropus).

    Science.gov (United States)

    Kang, Jun-Gu; Ko, Sungjin; Kim, Heung-Chul; Chong, Sung-Tae; Klein, Terry A; Chae, Jeong-Byoung; Jo, Yong-Sun; Choi, Kyoung-Seong; Yu, Do-Hyeon; Park, Bae-Keun; Park, Jinho; Chae, Joon-Seok

    2016-02-01

    Deer serve as reservoirs of tick-borne pathogens that impact on medical and veterinary health worldwide. In the Republic of Korea, the population of Korean water deer (KWD, Hydropotes inermis argyropus) has greatly increased from 1982 to 2011, in part, as a result of reforestation programs established following the Korean War when much of the land was barren of trees. Eighty seven Haemaphysalis flava, 228 Haemaphysalis longicornis, 8 Ixodes nipponensis, and 40 Ixodes persulcatus (21 larvae, 114 nymphs, and 228 adults) were collected from 27 out of 70 KWD. A total of 89/363 ticks (266 pools, 24.5% minimum infection rate) and 5 (1.4%) fed ticks were positive for Anaplasma phagocytophilum using nested PCR targeting the 16S rRNA and groEL genes, respectively. The 16S rRNA gene fragment sequences of 88/89 (98.9%) of positive samples for A. phagocytophilum corresponded to previously described gene sequences from KWD spleen tissues. The 16S rRNA gene fragment sequences of 20/363 (5.5%) of the ticks were positive for A. bovis and were identical to previously reported sequences. Using the ITS specific nested PCR, 11/363 (3.0%) of the ticks were positive for Bartonella spp. This is the first report of Anaplasma and Bartonella spp. detected in ticks collected from KWD, suggesting that ticks are vectors of Anaplasma and Bartonella spp. between reservoir hosts in natural surroundings.

  4. Development of duplex PCR for simultaneous detection of Theileria spp. and Anaplasma spp. in sheep and goats.

    Science.gov (United States)

    Cui, Yanyan; Zhang, Yan; Jian, Fuchun; Zhang, Longxian; Wang, Rongjun; Cao, Shuxuan; Wang, Xiaoxing; Yan, Yaqun; Ning, Changshen

    2017-01-31

    Theileria spp. and Anaplasma spp., which are important tick-borne pathogens (TBPs), impact the health of humans and animals in tropical and subtropical areas. Theileria and Anaplasma co-infections are common in sheep and goats. Following alignment of the relevant DNA sequences, two primer sets were designed to specifically target the Theileria spp. 18S rRNA and Anaplasma spp. 16S rRNA gene sequences. Genomic DNA from the two genera was serially diluted tenfold for testing the sensitivities of detection of the primer sets. The specificities of the primer sets were confirmed when DNA from Anaplasma and Theileria (positive controls), other related hematoparasites (negative controls) and ddH2O were used as templates. Fifty field samples were also used to evaluate the utility of single PCR and duplex PCR assays, and the detection results were compared with those of the PCR methods previously published. An optimized duplex PCR assay was established from the two primer sets based on the relevant genes from the two TBPs, and this assay generated products of 298-bp (Theileria spp.) and 139-bp (Anaplasma spp.). The detection limit of the assay was 29.4 × 10(-3) ng per μl, and there was no cross-reaction with the DNA from other hematoparasites. The results showed that the newly developed duplex PCR assay had an efficiency of detection (P > 0.05) similar to other published PCR methods. In this study, a duplex PCR assay was developed that can simultaneously identify Theileria spp. and Anaplasma spp. in sheep and goats. This duplex PCR is a potentially valuable assay for epidemiological studies of TBPs in that it can detect cases of mixed infections of the pathogens.

  5. Molecular identification and characterization of Anaplasma platys and Ehrlichia canis in dogs in Mexico.

    Science.gov (United States)

    Almazán, Consuelo; González-Álvarez, Vicente H; Fernández de Mera, Isabel G; Cabezas-Cruz, Alejandro; Rodríguez-Martínez, Rafael; de la Fuente, José

    2016-03-01

    The tick-borne pathogens Ehrlichia canis and Anaplasma platys are the causative agents of canine monocytic ehrlichiosis (CME) and canine cyclic thrombocytopenia (CCT). Although molecular evidence of E. canis has been shown, phylogenetic analysis of this pathogen has not been performed and A. platys has not been identified in Mexico, where the tick vector Rhipicephalus sanguineus sensu lato (s.l.) is common. The aim of this research was to screen, identify and characterize E. canis and A. platys by PCR and phylogenetic analysis in dogs from La Comarca Lagunera, a region formed by three municipalities, Torreon, Gomez-Palacio and Lerdo, in the Northern states of Coahuila and Durango, Mexico. Blood samples and five engorged R. sanguineus s.l. ticks per animal were collected from 43 females and 57 male dogs presented to veterinary clinics or lived in the dog shelter from La Comarca Lagunera. All the sampled dogs were apparently healthy and PCR for Anaplasma 16S rRNA, Ehrlichia 16S rRNA, and E. canis trp36 were performed. PCR products were sequenced and used for phylogenetic analysis. PCR products were successfully amplified in 31% of the samples using primers for Anaplasma 16S rRNA, while 10% and 4% amplified products using primers for Ehrlichia 16S rRNA and E. canis trp36 respectively. Subsequent sequencing and phylogenetic analyses of these products showed that three samples corresponded to A. platys and four to E. canis. Based on the analysis of trp36 we confirmed that the E. canis strains isolated from Mexico belong to a conservative clade of E. canis and are closely related to strains from USA. In conclusion, this is the first molecular identification of A. platys and the first molecular characterization and phylogenetic study of both A. platys and E. canis in dogs in Mexico.

  6. 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.

  7. Survey of Ticks Collected from Tennessee Cattle and Their Pastures for Anaplasma and Ehrlichia Species.

    Science.gov (United States)

    Pompo, K; Mays, S; Wesselman, C; Paulsen, D J; Fryxell, R T Trout

    2016-02-01

    Anaplasma marginale is the causative agent for bovine anaplasmosis (BA) and Ehrlichia ruminantium is the causative agent for heartwater, 2 devastating diseases of cattle. BA is common in the United States and frequently reported in western Tennessee cattle; however, cases of heartwater are not yet established in the continental United States. Because both pathogens are transmitted via the bites of infected ticks, the objective of this study was to survey cattle and pastures for ticks and for each pathogen. University of Tennessee AgResearch has 7 research and education centers (REC) located throughout the state at which they manage cattle. Ticks were collected from selected cattle (every fourth to sixth animal) and pastures (via dragging) associated with the herd from each REC during the summer of 2013. A total of 512 ticks were collected from cattle (n = 386) and pastures (n = 126) and were PCR-screened for Anaplasma and Ehrlichia using genus-specific primers. Collections consisted of 398 (77.7%) Amblyomma americanum, 84 (16.4%) Amblyomma maculatum, and 30 (5.9%) Dermacentor variabilis. Ticks were not recovered from pastures or cattle east of the Tennessee Plateau. The North American vectors for An. marginale and E. ruminantium were identified (D. variabilis and A. maculatum, respectively), but neither pathogen was recovered. A large proportion of ticks were collected from cattle and, of these, a majority were attached to their host (compared to questing on their host or engorged on the host). Four A. americanum were positive for Ehrlichia spp. (Ehrlichia ewingii, Ehrlichia chaffeensis, and Panola Mountain Ehrlichia), all in western Tennessee. With the identification of a few Ehrlichia infections in cattle-associated ticks and current A. marginale rates in Tennessee beef cattle nearing 11%, additional research is needed to establish baseline tick, Anaplasma, and Ehrlichia data for future management studies.

  8. Serological evidence of exposure to Ehrlichia canis and Anaplasma phagocytophilum in Central Italian healthy domestic cats.

    Science.gov (United States)

    Ebani, Valentina V; Bertelloni, Fabrizio

    2014-10-01

    The aim of the present survey was to estimate the seroprevalences of Ehrlichia canis and Anaplasma phagocytophilum in the Central Italian feline population. Serum samples of 560 healthy domestic cats were examined by indirect immunofluorescence assay (IFAT), considering an antibody titre of 1:40 as cut-off. Seroprevalences of 6.4% and 4.5% were found for E. canis and A. phagocytophilum, respectively. Adult, mixed breed cats showed seroprevalences higher than younger and purebred subjects, whereas no differences were observed in relation to gender and living conditions.

  9. [First serological study of the prevalence of Anaplasma phagocytophilum in dromedary (Camelus dromedarius) in Tunisia].

    Science.gov (United States)

    Ben Said, M; Belkahia, H; Sayahi, L; Aloui, M; Jemli, M H; Hadj Mohamed, B; Sassi, L; Darghouth, M A; Djaïem, A A; Bayoudh, M; Messadi, L

    2014-02-01

    The aim of the study was to evaluate the seroprevalence of Anaplasma phagocytophilum in dromedary (Camelus dromedarius). Sera of 226 healthy dromedaries from three regions of Tunisia (Sidi Bouzid, Bouficha and Douz) were tested by indirect immunofluorescence (IFA). The overall infection rate was estimated at 29.2%. The study of risk factors showed that region, age, gender, presence of ticks and types of breeding had no influence on the seroprevalence of A. phagocytophilum. This study indicates for the first time in Tunisia that dromedary may be involved in the natural cycle of A. phagocytophilum.

  10. 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...

  11. First description of natural Ehrlichia canis and Anaplasma platys infections in dogs from Argentina.

    Science.gov (United States)

    Eiras, Diego Fernando; Craviotto, María Belén; Vezzani, Darío; Eyal, Osnat; Baneth, Gad

    2013-03-01

    Bacteria belonging to the Anaplasmataceae family are vector transmitted agents that affect a variety of vertebrate hosts including the tick-borne pathogens Ehrlichia canis and Anaplasma platys, which cause canine monocytic ehrlichiosis and cyclic thrombocytopenia, respectively. These two infections, typically reported from tropical and sub-tropical regions, have not been previously reported in dogs from Argentina. A total of 86 blood samples from dogs with suspected rickettsial disease and 28 non-suspected dogs were studied. Analysis included evaluation of hematological findings, PCR for Ehrlichia and Anaplasma species and sequencing of the positive PCR products. E. canis was detected in the blood of six dogs and A. platys in eighteen. All the dogs categorized as non-suspected were negative by PCR. Co-infection with Hepatozoon canis and Babesia vogeli was documented. This first report of E. canis and A. platys infections in dogs from Argentina indicates that these tick-borne infections have a considerably broader range than previously recognized in South America.

  12. In vitro cultivation of Anaplasma marginale and A. phagocytophilum in tick cell lines: a review.

    Science.gov (United States)

    Passos, Lygia Maria Friche

    2012-01-01

    Continuous cell lines have been established from several ixodid and argasid tick species, representing an excellent tool suitable for the isolation of pathogens and their subsequent propagation, which in turn allows the production of antigenic material for diagnostic tests, antibody and vaccine production, and also for studies on host-vector-pathogen relationships. This paper reviews the use of tick cells for culture initiation and maintenance of two obligate intracellular bacterial pathogens, Anaplasma marginale and Anaplasma phagocytophilum. These in vitro cultivation systems have been used in a wide range of studies, covering morphological ultrastructural analysis, genetics, proteomics and biological differences between strains, including genome transcriptional and protein expression approaches, enabling comparisons between host and vector cells. Thus, such systems open a new window for a better understanding of interactions between pathogens and tick cells. Last but not least, such systems contribute to the reduction in usage of animals for experimental research, as antigenic material can be produced in reasonably large quantities without the use of in vivo species-specific systems.

  13. 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.

  14. A Molecular survey of Anaplasma spp., Rickettsia spp., Ehrlichia canis and Babesia microti in foxes and fleas from Sicily.

    Science.gov (United States)

    Torina, A; Blanda, V; Antoci, F; Scimeca, S; D'Agostino, R; Scariano, E; Piazza, A; Galluzzo, P; Giudice, E; Caracappa, S

    2013-11-01

    Fleas (Insecta: Siphonaptera) are obligate bloodsucking insects, which parasitize birds and mammals, and are distributed throughout the world. Several species have been implicated in pathogen transmission. This study aimed to monitor red foxes and the fleas isolated from them in the Palermo and Ragusa provinces of Sicily, Italy, as these organisms are potential reservoirs and vectors of pathogens. Thirteen foxes (Vulpes vulpes) and 110 fleas were analysed by polymerase chain reaction (PCR) to detect DNA of the pathogens Ehrlichia canis, Babesia microti, Rickettsia spp., Anaplasma phagocytophilum, Anaplasma platys, Anaplasma marginale and Anaplasma ovis. In the foxes, A. ovis was detected in only one animal, whereas the prevalence of the E. canis pathogen was 31%. B. microti and Rickettsia spp. were not detected. Of all of the collected fleas, 75 belonged to the species Xenopsylla cheopis, 32 belonged to Ctenocephalides canis, two belonged to Ctenocephalides felis and one belonged to Cediopsylla inaequalis. In the fleas, the following pathogens were found: A. ovis (prevalence 25%), A. marginale (1%), A. phagocytophilum (1%), Rickettsia felis (2%) and E. canis (3%). X. cheopis was the flea species most frequently infected with Anaplasma, in particular A. ovis (33%), A. marginale (1%) and A. phagocytophilum (1%). Both C. felis exemplars were positive for R. felis. E. canis was found in the lone C. inaequalis and also in 3% of the X. cheopis specimens. No fleas were positive for B. microti or A. platys. As foxes often live in proximity to domestic areas, they may constitute potential reservoirs for human and animal parasites. Further studies should be performed on fleas to determine their vectorial capacity.

  15. Expression of Heat Shock and Other Stress Response Proteins in Ticks and Cultured Tick Cells in Response to Anaplasma spp. Infection and Heat Shock

    Directory of Open Access Journals (Sweden)

    Margarita Villar

    2010-01-01

    Full Text Available Ticks are ectoparasites of animals and humans that serve as vectors of Anaplasma and other pathogens that affect humans and animals worldwide. Ticks and the pathogens that they transmit have coevolved molecular interactions involving genetic traits of both the tick and the pathogen that mediate their development and survival. In this paper, the expression of heat shock proteins (HSPs and other stress response proteins (SRPs was characterized in ticks and cultured tick cells by proteomics and transcriptomics analyses in response to Anaplasma spp. infection and heat shock. The results of these studies demonstrated that the stress response was activated in ticks and cultured tick cells after Anaplasma spp. infection and heat shock. However, in the natural vector-pathogen relationship, HSPs and other SRPs were not strongly activated, which likely resulted from tick-pathogen coevolution. These results also demonstrated pathogen- and tick-specific differences in the expression of HSPs and other SRPs in ticks and cultured tick cells infected with Anaplasma spp. and suggested the existence of post-transcriptional mechanisms induced by Anaplasma spp. to control tick response to infection. These results illustrated the complexity of the stress response in ticks and suggested a function for the HSPs and other SRPs during Anaplasma spp. infection.

  16. Babesia canis vogeli, Ehrlichia canis, and Anaplasma platys infection in a dog.

    Science.gov (United States)

    Al Izzi, Salah; Martin, Donald S; Chan, Roxanne Y Y; Leutenegger, Christian M

    2013-12-01

    A 12-month-old male neutered mixed breed dog was presented with a history of diarrhea, lethargy, emaciation, polydypsia, and sniffling. Physical examination findings included pale mucous membranes, increased heart and respiratory rates, and normal rectal temperature (38°C). Hematologic abnormalities included anemia and thrombocytopenia. Biochemical abnormalities included hypoalbuminemia, hyperbilirubinemia, and elevated ALP and ALT activities. A SNAP 4Dx test result was positive for Ehrlichia canis. Babesia canis vogeli organisms were found in the peripheral blood films, while morulae of E canis were not seen. Real-time polymerase chain reaction testing confirmed the presence of both B c vogeli and E canis organisms, and also was positive for Anaplasma platys infection. The dog recovered following treatment with doxycycline and imidocarb dipropionate, with normal hematology and biochemical profiles.

  17. First molecular detection of Ehrlichia canis and Anaplasma platys in ticks from dogs in Cebu, Philippines.

    Science.gov (United States)

    Ybañez, Adrian P; Perez, Zandro O; Gabotero, Shirleny R; Yandug, Ryan T; Kotaro, Matsumoto; Inokuma, Hisashi

    2012-12-01

    Ehrlichia canis infection of dogs in the Philippines has been detected by serological and peripheral blood smear examination methods, but not by molecular means. Anaplasma platys infection in dogs has not yet been officially reported, although it is suspected to occur in the country. Thus, sensitive and specific molecular techniques were used in this study to demonstrate the presence of both E. canis and A. platys in the Philippines. A total of 164 Rhipicephalus sanguineus ticks was collected from 36 dogs. Seven tick samples were found positive with E. canis and one sample with A. platys. To further characterize these pathogens, molecular analyses based on citrate synthase and heat-shock operon genes were also performed. Philippine strains were found to be not divergent from strains from other countries. The present results are the first molecular detection and analyses of E. canis and A. platys in ticks from dogs in the Philippines.

  18. First identification of Anaplasma platys in the blood of dogs from French Guiana.

    Science.gov (United States)

    Dahmani, Mustapha; Marié, Jean-Lou; Mediannikov, Oleg; Raoult, Didier; Davoust, Bernard

    2015-02-01

    Anaplasma platys is the causative agent of infectious cyclic thrombocytopenia in dogs. This infection is worldwide and reported with a higher incidence in tropical and subtropical areas such as South America. Until now, there has been no report of this bacterium in French Guiana. The aim of this study was molecular investigation of A. platys occurrence in the blood of autochthonous dogs in this region. A total 65 blood samples were taken from the shelter dogs in the cities of Cayenne and Kourou, and from dogs of private owners in the city of Cayenne. The results show that at least 15.38% (10/65) were positive to this pathogen. The strain identified in this study has been reported worldwide. These findings should be considered in the way that local veterinarians handle suspected cases of canine anaplasmosis and ehrlichiosis.

  19. Anaplasma phagocytophilum AptA modulates Erk1/2 signalling.

    Science.gov (United States)

    Sukumaran, Bindu; Mastronunzio, Juliana E; Narasimhan, Sukanya; Fankhauser, Sarah; Uchil, Pradeep D; Levy, Roie; Graham, Morven; Colpitts, Tonya Michelle; Lesser, Cammie F; Fikrig, Erol

    2011-01-01

    Anaplasma phagocytophilum causes human granulocytic anaplasmosis, one of the most common tick-borne diseases in North America. This unusual obligate intracellular pathogen selectively persists within polymorphonuclear leucocytes. In this study, using the yeast surrogate model we identified an A. phagocytophilum virulence protein, AptA (A. phagocytophilum toxin A), that activates mammalian Erk1/2 mitogen-activated protein kinase. This activation is important for A. phagocytophilum survival within human neutrophils. AptA interacts with the intermediate filament protein vimentin, which is essential for A. phagocytophilum-induced Erk1/2 activation and infection. A. phagocytophilum infection reorganizes vimentin around the bacterial inclusion, thereby contributing to intracellular survival. These observations reveal a major role for the bacterial protein, AptA, and the host protein, vimentin, in the activation of Erk1/2 during A. phagocytophilum infection.

  20. 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.

  1. Integrated Metabolomics, Transcriptomics and Proteomics Identifies Metabolic Pathways Affected by Anaplasma phagocytophilum Infection in Tick Cells.

    Science.gov (United States)

    Villar, Margarita; Ayllón, Nieves; Alberdi, Pilar; Moreno, Andrés; Moreno, María; Tobes, Raquel; Mateos-Hernández, Lourdes; Weisheit, Sabine; Bell-Sakyi, Lesley; de la Fuente, José

    2015-12-01

    Anaplasma phagocytophilum is an emerging zoonotic pathogen that causes human granulocytic anaplasmosis. These intracellular bacteria establish infection by affecting cell function in both the vertebrate host and the tick vector, Ixodes scapularis. Previous studies have characterized the tick transcriptome and proteome in response to A. phagocytophilum infection. However, in the postgenomic era, the integration of omics datasets through a systems biology approach allows network-based analyses to describe the complexity and functionality of biological systems such as host-pathogen interactions and the discovery of new targets for prevention and control of infectious diseases. This study reports the first systems biology integration of metabolomics, transcriptomics, and proteomics data to characterize essential metabolic pathways involved in the tick response to A. phagocytophilum infection. The ISE6 tick cells used in this study constitute a model for hemocytes involved in pathogen infection and immune response. The results showed that infection affected protein processing in endoplasmic reticulum and glucose metabolic pathways in tick cells. These results supported tick-Anaplasma co-evolution by providing new evidence of how tick cells limit pathogen infection, while the pathogen benefits from the tick cell response to establish infection. Additionally, ticks benefit from A. phagocytophilum infection by increasing survival while pathogens guarantee transmission. The results suggested that A. phagocytophilum induces protein misfolding to limit the tick cell response and facilitate infection but requires protein degradation to prevent ER stress and cell apoptosis to survive in infected cells. Additionally, A. phagocytophilum may benefit from the tick cell's ability to limit bacterial infection through PEPCK inhibition leading to decreased glucose metabolism, which also results in the inhibition of cell apoptosis that increases infection of tick cells. These results

  2. Comparison of Microscopy and PCR-RFLP for detection of Anaplasma marginale in carrier cattle

    Directory of Open Access Journals (Sweden)

    P Shayan

    2010-09-01

    Full Text Available Background and Objectives: In Iran, anaplasmosis is normally diagnosed with traditional Giemsa staining method. This is not applicable for identification of the carrier animals. The aim of this study was to compare the detection of Anaplasma marginale in two different numbers of microscopic fields (50 and 100 using conventional Giemsa staining method compared with the PCR-RFLP technique."nMaterials and Methods: In this study, examinations were performed on 150 blood samples from cattle without clinical signs. Sensitivity and specificity of two microscopic fields (50 and 100 fields were compared with A. marginale specific PCR-RFLP. The degree of agreement between PCR-RFLP and the two microscopic tests was determined by Kappa (κ values with 95% confidence intervals."nResults: PCR-RFLP showed that 58 samples were A. marginale, while routine microscopy showed erythrocytes harboring Anaplasma like structures in 16 and 75 blood samples determined in 50 and 100 microscopic fields respectively. Examination of 50 and 100 microscopic fields showed 25.8% and 91.4% sensitivity and 99% and 76.1% specificity compared to 100% sensitivity and specificity by PCR-RFLP. The Kappa coefficient between PCR-RFLP and Microscopy (50 fields indicated a fair level of agreement (0.29. The Kappa coefficient between PCR-RFLP and Microscopy (100 fields indicated a good level of agreement (0.64"nConclusion: Our results showed that the microscopic examination remains the convenient technique for day-to-day diagnosis of clinical cases in the laboratory but for the detection of carrier animal with low bacteremia, microscopy with 100 fields is preferable to Microscopy with 50 fields and molecular methods such as PCR-RFLP can be used as a safe method for identifying cattle persistently infected with A. marginale.

  3. Bacterial Profiling Reveals Novel "Ca. Neoehrlichia", Ehrlichia, and Anaplasma Species in Australian Human-Biting Ticks.

    Directory of Open Access Journals (Sweden)

    Alexander W Gofton

    Full Text Available In Australia, a conclusive aetiology of Lyme disease-like illness in human patients remains elusive, despite growing numbers of people presenting with symptoms attributed to tick bites. In the present study, we surveyed the microbial communities harboured by human-biting ticks from across Australia to identify bacteria that may contribute to this syndrome. Universal PCR primers were used to amplify the V1-2 hyper-variable region of bacterial 16S rRNA genes in DNA samples from individual Ixodes holocyclus (n = 279, Amblyomma triguttatum (n = 167, Haemaphysalis bancrofti (n = 7, and H. longicornis (n = 7 ticks. The 16S amplicons were sequenced on the Illumina MiSeq platform and analysed in USEARCH, QIIME, and BLAST to assign genus and species-level taxonomies. Nested PCR and Sanger sequencing were used to confirm the NGS data and further analyse novel findings. All 460 ticks were negative for Borrelia spp. by both NGS and nested PCR analysis. Two novel "Candidatus Neoehrlichia" spp. were identified in 12.9% of I. holocyclus ticks. A novel Anaplasma sp. was identified in 1.8% of A. triguttatum ticks, and a novel Ehrlichia sp. was identified in both A. triguttatum (1.2% ticks and a single I. holocyclus (0.6% tick. Further phylogenetic analysis of novel "Ca. Neoehrlichia", Anaplasma and Ehrlichia based on 1,265 bp 16S rRNA gene sequences suggests that these are new species. Determining whether these newly discovered organisms cause disease in humans and animals, like closely related bacteria do abroad, is of public health importance and requires further investigation.

  4. Bacterial Profiling Reveals Novel "Ca. Neoehrlichia", Ehrlichia, and Anaplasma Species in Australian Human-Biting Ticks.

    Science.gov (United States)

    Gofton, Alexander W; Doggett, Stephen; Ratchford, Andrew; Oskam, Charlotte L; Paparini, Andrea; Ryan, Una; Irwin, Peter

    2015-01-01

    In Australia, a conclusive aetiology of Lyme disease-like illness in human patients remains elusive, despite growing numbers of people presenting with symptoms attributed to tick bites. In the present study, we surveyed the microbial communities harboured by human-biting ticks from across Australia to identify bacteria that may contribute to this syndrome. Universal PCR primers were used to amplify the V1-2 hyper-variable region of bacterial 16S rRNA genes in DNA samples from individual Ixodes holocyclus (n = 279), Amblyomma triguttatum (n = 167), Haemaphysalis bancrofti (n = 7), and H. longicornis (n = 7) ticks. The 16S amplicons were sequenced on the Illumina MiSeq platform and analysed in USEARCH, QIIME, and BLAST to assign genus and species-level taxonomies. Nested PCR and Sanger sequencing were used to confirm the NGS data and further analyse novel findings. All 460 ticks were negative for Borrelia spp. by both NGS and nested PCR analysis. Two novel "Candidatus Neoehrlichia" spp. were identified in 12.9% of I. holocyclus ticks. A novel Anaplasma sp. was identified in 1.8% of A. triguttatum ticks, and a novel Ehrlichia sp. was identified in both A. triguttatum (1.2%) ticks and a single I. holocyclus (0.6%) tick. Further phylogenetic analysis of novel "Ca. Neoehrlichia", Anaplasma and Ehrlichia based on 1,265 bp 16S rRNA gene sequences suggests that these are new species. Determining whether these newly discovered organisms cause disease in humans and animals, like closely related bacteria do abroad, is of public health importance and requires further investigation.

  5. Mechanisms of infection by pathogens transmitted by ticks on the example of bacteria: Anaplasma phagocytophilum and Borrelia burgdorferi

    Directory of Open Access Journals (Sweden)

    Paula Wróblewska

    2016-06-01

    Full Text Available Tick-borne diseases are transmission diseases belonging to the group of zoonoses but carried by ticks. These diseases are a major public health problem but also a problem for groups occupationally exposed to tick bites. Ixodes ricinus is a species of ticks which is the most common reservoir and the vector of a large number of microorganisms pathogenic to humans. It transfers, among others, bacteria of the species: Anaplasma phagocytophilum and Borrelia burgdorferi. The article discusses the mechanisms of infection with Borrelia burgdorferi and Anaplasma phagocytophilum for both ticks as well as for animals and humans. The two microorganisms discussed have developed many characteristics and mechanisms of adaptation to the environment, as well as defense mechanisms against the body's immune response. Understanding the biology of ticks and the function of proteins produced by ticks and pathogenic microorganisms is the key in the development of effective treatments and prevention of Lyme disease and anaplasmosis.

  6. Mechanisms of infection by pathogens transmitted by ticks on the example of bacteria: Anaplasma phagocytophilum and Borrelia burgdorferi

    OpenAIRE

    2016-01-01

    Tick-borne diseases are transmission diseases belonging to the group of zoonoses but carried by ticks. These diseases are a major public health problem but also a problem for groups occupationally exposed to tick bites. Ixodes ricinus is a species of ticks which is the most common reservoir and the vector of a large number of microorganisms pathogenic to humans. It transfers, among others, bacteria of the species: Anaplasma phagocytophilum and Borrelia burgdorferi. The...

  7. Anaplasma marginale infection in cattle from south-western Amazonia Infecção por Anaplasma marginale em bovinos na Amazônia Sul Ocidental, Brasil

    Directory of Open Access Journals (Sweden)

    Luciana Gatto Brito

    2010-03-01

    Full Text Available The present study provides the first epidemiological data regarding infection by Anaplasma marginale in cattle reared in south-western Brazilian Amazonia. One simple procedure was adapted for the extraction of DNA from blood clots collected in seven microregions of Rondônia State and two mesoregions of Acre State. PCR method was used to asses the frequency of A. marginale infections in 4 to12-month-old cattle. The cattle infection was investigated by polymerase chain reaction (PCR using the specific primer "msp5" for A. marginale. The DNA amplifications revealed that the mean frequency of A. marginale infection was 98.6% (1,627/1,650 in samples from Rondonia, and 92.87% (208/225 in samples from Acre. The high frequency of A. marginale infections in 4 to 12-month-old cattle indicate a situation of enzootic stability in the studied areas and are comparable to those detected by immunodiagnosis in different endemic regions in Brazil. The DNA extraction of clotted blood method described here can be used for epidemiological studies on anaplasmosis and other bovine hemoparasites.O presente estudo fornece os primeiros dados epidemiológicos relativos a infecção por Anaplasma marginale em bovinos criados na Amazônia Sul Ocidental brasileira. Foi adaptado um procedimento simples para a extração de DNA a partir de coágulos sanguíneos coletados em sete microrregiões do estado de Rondônia e duas mesoregiões do estado do Acre. A técnica da reação em cadeia da polimerase (PCR foi aplicada para avaliar a freqüência da infecção por A. marginale em bovinos com idade entre 4 e 12 meses. Após a extração do DNA de cada amostra, a infecção nos bovinos foi investigada pela amplificação do gene "msp5" de A. marginale. As técnicas de amplificação do DNA revelaram que a freqüência de infecção por A. marginale foi de 98,6% (1.627/1.650 nas amostras provenientes de Rondônia e de 92,87% (208/225 nas amostras do Acre. A alta freqüência da

  8. Molecular detection of Anaplasma platys and Ehrlichia canis in dogs from Kabylie, Algeria.

    Science.gov (United States)

    Dahmani, Mustapha; Loudahi, Abdelghani; Mediannikov, Oleg; Fenollar, Florence; Raoult, Didier; Davoust, Bernard

    2015-03-01

    Ehrlichia canis and Anaplasma platys are bacteria belonging to the Anaplasmataceae family that cause acute, self-limiting and sometimes fatal vector-borne infections in dogs. These bacteria have been reported worldwide and are transmitted mainly by Rhipicephalus sanguineus. Aside from a report on E. canis once in 1935, no other Anaplasmataceae bacteria have been reported in Algeria to date. The aim of this study was to identify the microbial species implicated in ehrlichiosis and anaplasmosis by a molecular epidemiological survey in dogs. The study was carried out in Kabylie, in northeast Algeria. Sampling was performed in 11 municipalities in the province of Tizi Ouzou and 2 municipalities in the province of Béjaïa. Peripheral blood samples from 110 dogs were screened by qPCR, which is capable of identifying most Anaplasmataceae bacteria. Out of 110, a total of 13 samples screened positive (7/110 E. canis and 6/110 A. platys), and two genetic variants of A. platys and one of E. canis were identified. This is the first study to report the presence of A. platys in dogs from Algeria using a molecular investigative method. This survey was conducted in early spring. As tick activity can affect the prevalence of these pathogens in dogs, further investigations are needed to establish the year-round prevalence of these infections.

  9. The occurrence of Dirofilaria immitis, Borrelia burgdorferi, Ehrlichia canis and Anaplasma phagocytophium in dogs in China.

    Science.gov (United States)

    Xia, Z; Yu, D; Mao, J; Zhang, Z; Yu, J

    2012-06-01

    A survey of the occurrence of Dirofilaria immitis, Borrelia burgdorferi, Ehrlichia canis and Anaplasma phagocytophium in dogs was undertaken in the People's Republic of China between October 2008 and October 2009. A total of 600 blood samples were taken from dogs in four cities in China: 300 in Beijing, 150 in Shenzhen, 30 in Shanghai and 120 in Zhengzhou. All samples were tested for the heartworm antigen and antibodies of canine B. burgdorferi, E. canis and A. phagocytophium by using the canine SNAP® 4Dx® test kit. The occurrence of D. immitis, B. burgdorferi, E. canis and A. phagocytophium was 1.17% (7/600), 0.17% (1/600), 2.17% (13/600) and 0.5% (3/600), respectively. In Shenzhen city 2% (3/150), 8.67% (13/150) and 2% (3/150) of samples were positive for D. immitis, E. canis and A. phagocytophium, respectively. The occurrence of heartworm antigen was 0.33% (1/300) in Beijing, 2.00% (3/150) in Shenzhen, 3.33% (1/30) in Shanghai and 1.67% (2/120) in Zhengzhou. We found E. canis and A. phagocytophium only at one site, Shenzhen, while the only occurrence of B. burgdorferi was at Beijing. In conclusion, the dog population in China is at potential risk for D. immitis, B. burgdorferi, E. canis and A. phagocytophium infection, the risk being especially high in southern China.

  10. Occurrence of Ehrlichia canis and Anaplasma platys in household dogs from northern Parana.

    Science.gov (United States)

    da Silva, Gislaine Cristina Ferreira; Benitez, Aline do Nascimento; Girotto, Aline; Taroda, Alessandra; Vidotto, Marilda Carlos; Garcia, João Luis; de Freitas, Julio Cesar; Arlington, Selwyn Headley; Vidotto, Odilon

    2012-01-01

    Canine monocytic ehrlichiosis caused primarily by Ehrlichia canis and canine thrombocytic anaplasmosis induced by Anaplasma platys are important emerging zoonotic tick-borne diseases of dogs. There is evidence that these pathogens can also affect humans. This study evaluated the presence of E. canis and A. platys in blood samples collected from 256 domiciled dogs in the municipality of Jataizinho, located in north region of the State of Parana, Brazil, by PCR assay. The occurrence of E. canis and A. platys was 16.4% (42/256) and 19.4% (49/256), respectively; while 5.47% (14/256) of the dogs evaluated were co-infected by these two organisms. The presence of E. canis and A. platys was not significantly associated with the variables evaluated (sex, age, outdoor access, and presence of ticks during blood collection). Infection of dogs by E. canis was associated with anemia and thrombocytopenia, while infection induced by A. platys was related only to thrombocytopenia. Canine monocytic ehrlichiosis and canine thrombocytic anaplasmosis should be included in the differential diagnoses when these hematological alterations are observed during routine laboratory evaluation of dogs.

  11. Detection of Ehrlichia canis and Anaplasma platys DNA using multiplex PCR.

    Science.gov (United States)

    Rufino, Claudia Pinheiro; Moraes, Pablo Henrique Gonçalves; Reis, Thais; Campos, Ruan; Aguiar, Délia Cristina Figueira; McCulloch, John Anthony; Meneses, Andre Marcelo Conceição; Gonçalves, Evonnildo Costa

    2013-12-01

    We hereby propose a novel sensitive, specific, and cost-efficient method to detect Ehrlichia canis and Anaplasma platys DNA from canine whole blood samples by multiplex PCR. Blood samples from hemoparasited dogs attending the Veterinary Hospital at the Universidade Federal Rural da Amazônia-UFRA, Belém, Brazil, were collected in tubes containing EDTA. Amplification of E. canis and A. platys 16S rDNA by nested (n) PCR was successfully achieved by using primers specific to the Anaplasmataceae in the first round of PCR, followed by a second round of PCR using E. canis-specific primers in conjunction with A. platys-specific primers. The amplicons obtained were cloned and sequenced, yielding sequences of 478 and 473 bp (including primers) pertaining to regions of the 16S rDNA of E. canis and A. platys, respectively. The protocol we here propose may help to measure the prevalence of canine monocytic ehrlichiosis (CME) and canine cyclic thrompocytopenia, not only in northern Brazil, where there is no data available, but also elsewhere.

  12. First report of Anaplasma phagocytophilum and Babesia microti in rodents in Finland.

    Science.gov (United States)

    Kallio, Eva R; Begon, Michael; Birtles, Richard J; Bown, Kevin J; Koskela, Esa; Mappes, Tapio; Watts, Phillip C

    2014-06-01

    Tick-borne diseases pose an increasingly important public health problem in Europe. Rodents are the reservoir host for many tick-transmitted pathogens, including Anaplasma phagocytophilum and Babesia microti, which can cause human granulocytic anaplasmosis and babesiosis, respectively. To estimate the presence of these pathogens in rodents in Finland, we examined blood samples from 151 bank voles (Myodes glareolus) and demonstrate, for the first time, that A. phagocytophilum and B. microti commonly infect bank voles (in 22% and 40% of animals, respectively) in Finland. Sequence analysis of a fragment of 18S rRNA showed that the B. microti strain isolated was identical to the Munich strain, which is considered to be nonzoonotic. The A. phagocytophilum strain (based on a fragment of the msp4 gene) was identical to one found earlier in rodents in the United Kingdom that is transmitted by the tick Ixodes trianguliceps, all the life stages of which feed on small mammals. The infection probability of B. microti in the bank voles was the greater the older the individual was, and males were more often infected than females. A. phagocytophilum infection probability first increased and then decreased with the age of individual without any difference between sexes. While these pathogens presumably pose a limited zoonotic risk to humans in Finland, they might have important interactions with other rodent pathogens and therefore affect infection dynamics of, for example, zoonotic pathogens.

  13. Superinfection occurs in Anaplasma phagocytophilum infected sheep irrespective of infection phase and protection status

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    Bergström Karin

    2009-10-01

    Full Text Available Abstract Background Anaplasma phagocytophilum infection in domestic ruminants is widespread in the coastal areas of southern Norway. The bacteria may persist in mammalian hosts. Several genetic variants of A. phagocytophilum exist. In the present study, we investigate whether superinfection occurs in the acute and persistent phase of the infection. Methods Five-month-old lambs of the Norwegian Dala breed were experimentally infected with two 16S rRNA gene variants of A. phagocytophilum, i.e. A. phagocytophilum variant 1 (GenBank accession number M73220 and variant 2 (GenBank acc. no. AF336220. Eighteen lambs were used, two lambs in each group. Eight groups were experimentally inoculated with either variant 1 or 2 on day 0. Six of these groups were then challenged with the other variant on either days 7, 42 or 84, respectively. One group was left uninfected. The occurrence of A. phagocytophilum in blood samples was determined using semi-nested PCR analysis and gene sequencing. Specific antibodies were measured by an indirect immunofluorescence antibody assay (IFA. Results A. phagocytophilum variant 1 and 2 differed significantly with regards to clinical reaction and cross-immunity in infected lambs. Both variants were found in the blood after challenge. However, variant 1 was detected most frequently. Conclusion The present experiment indicates that superinfection of different genotypes occurs during the acute as well as the persistent phase of an A. phagocytophilum infection, even in lambs protected against the challenged infection.

  14. Identification of Anaplasma phagocytophilum in tick populations in Estonia, the European part of Russia and Belarus.

    Science.gov (United States)

    Katargina, O; Geller, J; Alekseev, A; Dubinina, H; Efremova, G; Mishaeva, N; Vasilenko, V; Kuznetsova, T; Järvekülg, L; Vene, S; Lundkvist, A; Golovljova, I

    2012-01-01

    Anaplasma phagocytophilum is associated with diseases of goats, sheep, cattle, dogs and horses. In the beginning of the 1990s it was identified as a human pathogen, causing human granulocytic anaplasmosis (HGA) in the USA, Europe and the far east of Russia. A. phagocytophilum is maintained in nature in an enzootic cycle including ticks as the main vector and a wide range of mammalian species as reservoirs. Ixodes ricinus and I. persulcatus ticks were collected in Estonia, Belarus and the European part of Russia and screened for the presence of A. phagocytophilum by real-time PCR. Positive samples were found only among I. ricinus, in 13.4% in the European part of Russia, 4.2% in Belarus, 1.7% in mainland Estonia and 2.6% on Saaremaa Island. Positive samples were sequenced for partial 16S rRNA, groESL and ankA genes and phylogenetic analyses were performed. The results showed that A. phagocytophilum circulating in Eastern Europe belongs to different groESL lineages and 16S rRNA gene variants and also consists of variable numbers of repetitive elements within the ankA gene.

  15. Association of Anaplasma marginale strain superinfection with infection prevalence within tropical regions.

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    Elizabeth J Castañeda-Ortiz

    Full Text Available Strain superinfection occurs when a second strain infects a host already infected with and having mounted an immune response to a primary strain. The incidence of superinfection with Anaplasma marginale, a tick-borne rickettsial pathogen of domestic and wild ruminants, has been shown to be higher in tropical versus temperate regions. This has been attributed to the higher prevalence of infection, with consequent immunity against primary strains and thus greater selective pressure for superinfection with antigenically distinct strains. However an alternative explanation would be the differences in the transmitting vector, Dermacentor andersoni in the studied temperate regions and Rhipicephalus microplus in the studied tropical regions. To address this question, we examined two tropical populations sharing the same vector, R. microplus, but with significantly different infection prevalence. Using two separate markers, msp1α (one allele per genome and msp2 (multiple alleles per genome, there were higher levels of multiple strain infections in the high infection prevalence as compared to the low prevalence population. The association of higher strain diversity with infection prevalence supports the hypothesis that high levels of infection prevalence and consequent population immunity is the predominant driver of strain superinfection.

  16. Detection and phylogenetic characterization of Theileria spp. and Anaplasma marginale in Rhipicephalus bursa in Portugal.

    Science.gov (United States)

    Ferrolho, Joana; Antunes, Sandra; Santos, Ana S; Velez, Rita; Padre, Ludovina; Cabezas-Cruz, Alejandro; Santos-Silva, Maria Margarida; Domingos, Ana

    2016-04-01

    Ticks are obligatory blood-sucking arthropod (Acari:Ixodida) ectoparasites of domestic and wild animals as well as humans. The incidence of tick-borne diseases is rising worldwide, challenging our approach toward diagnosis, treatment and control options. Rhipicephalus bursa Canestrini and Fanzago, 1877, a two-host tick widely distributed in the Palearctic Mediterranean region, is considered a multi-host tick that can be commonly found on sheep, goats and cattle, and occasionally on horses, dogs, deer and humans. R. bursa is a species involved in the transmission of several tick-borne pathogens with a known impact on animal health and production. The aim of this study was to estimate R. bursa prevalence in Portugal Mainland and circulating pathogens in order to contribute to a better knowledge of the impact of this tick species. Anaplasma marginale and Theileria spp. were detected and classified using phylogenetic analysis. This is the first report of Theileria annulata and Theileria equi detection in R. bursa ticks feeding on cattle and horses, respectively, in Portugal. This study contributes toward the identification of currently circulating pathogens in this tick species as a prerequisite for developing future effective anti-tick control measures.

  17. 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.

  18. Theileria, Babesia, and Anaplasma detected by PCR in ruminant herds at Bié Province, Angola.

    Science.gov (United States)

    Kubelová, M; Mazancová, J; Siroký, P

    2012-11-01

    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 one specimen at Andulo, whereas B. bovis was not detected in Bié. We did not detected T. parva, the causative agent of serious diseases in cattle; nevertheless, infection by T. velifera was quite frequent (14% of examined animals, and five out of seven farms). Causative agent of heartwater disease - E. ruminantium, was not detected. Taking into account short-term perspective of PCR methods in monitoring of epidemiological status in herds, the number of infected animals and distribution of detected pathogens should not be ignored.

  19. Theileria, Babesia, and Anaplasma detected by PCR in ruminant herds at Bié Province, Angola

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    Kubelová M.

    2012-11-01

    Full Text Available 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 one specimen at Andulo, whereas B. bovis was not detected in Bié. We did not detected T. parva, the causative agent of serious diseases in cattle; nevertheless, infection by T. velifera was quite frequent (14% of examined animals, and five out of seven farms. Causative agent of heartwater disease – E. ruminantium, was not detected. Taking into account short-term perspective of PCR methods in monitoring of epidemiological status in herds, the number of infected animals and distribution of detected pathogens should not be ignored.

  20. Prevalence and Diversity among Anaplasma phagocytophilum Strains Originating from Ixodes ricinus Ticks from Northwest Norway

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    Ann-Kristin Tveten

    2014-01-01

    Full Text Available The tick-borne pathogen Anaplasma phagocytophilum causes great concern for livestock farmers. Tick-borne fever is a widespread disease in Norway, and antibodies have been produced amongst sheep, roe deer, red deer, and moose. The main vector Ixodes ricinus is found along the Norwegian coastline as far north as the Arctic Circle. A total number of 1804 I. ricinus ticks were collected and the prevalence of the pathogen was determined by species-specific qPCR. The overall infection rate varied from 2.83% to 3.32%, but there were no significant differences (p=0.01 in the overall infection rate in 2010, 2011, or 2012. A multilocus sequencing analysis was performed to further characterise the isolates. The genotyping of 27 strains resulted in classification into 19 different sequences types (ST, none of which was found in the MLST database. The nucleotide diversity was for every locus <0.01, and the number of SNPs was between 1 and 2.8 per 100 bp. The majority of SNPs were synonymous. A goeBURST analysis demonstrated that the strains from northwest Norway cluster together with other Norwegian strains in the MLST database and the strains that are included in this study constitute clonal complexes (CC 9, 10, and 11 in addition to the singleton.

  1. Global proteomic analysis of two tick-borne emerging zoonotic agents: Anaplasma phagocytophilum and Ehrlichia chaffeensis

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    Lin, Mingqun ..; Kikuchi, Takane; Brewer, Heather M.; Norbeck, Angela D.; Rikihisa, Yasuko

    2011-02-17

    Anaplasma phagocytophilum and Ehrlichia chaffeensis are obligatory intracellular {alpha}-proteobacteria that infect human leukocytes and cause potentially fatal emerging zoonoses. In the present study, we determined global protein expression profiles of these bacteria cultured in the human promyelocytic leukemia cell line, HL-60. Mass spectrometric (MS) analyses identified a total of 1,212 A. phagocytophilum and 1,021 E. chaffeensis proteins, representing 89.3 and 92.3% of the predicted bacterial proteomes, respectively. Nearly all bacterial proteins ({approx}99%) with known functions were expressed, whereas only approximately 80% of hypothetical proteins were detected in infected human cells. Quantitative MS/MS analyses indicated that highly expressed proteins in both bacteria included chaperones, enzymes involved in biosynthesis and metabolism, and outer membrane proteins, such as A. phagocytophilum P44 and E. chaffeensis P28/OMP-1. Among 113 A. phagocytophilum p44 paralogous genes, 110 of them were expressed and 88 of them were encoded by pseudogenes. In addition, bacterial infection of HL-60 cells up-regulated the expression of human proteins involved mostly in cytoskeleton components, vesicular trafficking, cell signaling, and energy metabolism, but down regulated some pattern recognition receptors involved in innate immunity. Our proteomics data represent a comprehensive analysis of A. phagocytophilum and E. chaffeensis proteomes, and provide a quantitative view of human host protein expression profiles regulated by bacterial infection. The availability of these proteomic data will provide new insights into biology and pathogenesis of these obligatory intracellular pathogens.

  2. Anaplasma phagocytophilum Manipulates Host Cell Apoptosis by Different Mechanisms to Establish Infection

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    Pilar Alberdi

    2016-07-01

    Full Text Available Anaplasma phagocytophilum is an emerging zoonotic pathogen that causes human and animal granulocytic anaplasmosis and tick-borne fever of ruminants. This obligate intracellular bacterium evolved to use common strategies to establish infection in both vertebrate hosts and tick vectors. Herein, we discuss the different strategies used by the pathogen to modulate cell apoptosis and establish infection in host cells. In vertebrate neutrophils and human promyelocytic cells HL-60, both pro-apoptotic and anti-apoptotic factors have been reported. Tissue-specific differences in tick response to infection and differential regulation of apoptosis pathways have been observed in adult female midguts and salivary glands in response to infection with A. phagocytophilum. In tick midguts, pathogen inhibits apoptosis through the Janus kinase/signal transducers and activators of transcription (JAK/STAT pathway, while in salivary glands, the intrinsic apoptosis pathways is inhibited but tick cells respond with the activation of the extrinsic apoptosis pathway. In Ixodes scapularis ISE6 cells, bacterial infection down-regulates mitochondrial porin and manipulates protein processing in the endoplasmic reticulum and cell glucose metabolism to inhibit apoptosis and facilitate infection, whereas in IRE/CTVM20 tick cells, inhibition of apoptosis appears to be regulated by lower caspase levels. These results suggest that A. phagocytophilum uses different mechanisms to inhibit apoptosis for infection of both vertebrate and invertebrate hosts.

  3. Ixodes scapularis saliva mitigates inflammatory cytokine secretion during Anaplasma phagocytophilum stimulation of immune cells

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    Chen Gang

    2012-10-01

    Full Text Available Abstract Background Ixodes scapularis saliva enables the transmission of infectious agents to the mammalian host due to its immunomodulatory, anesthetic and anti-coagulant properties. However, how I. scapularis saliva influences host cytokine secretion in the presence of the obligate intracellular rickettsial pathogen Anaplasma phagocytophilum remains elusive. Methods Bone marrow derived macrophages (BMDMs were stimulated with pathogen associated molecular patterns (PAMPs and A. phagocytophilum. Cytokine secretion was measured in the presence and absence of I. scapularis saliva. Human peripheral blood mononuclear cells (PBMCs were also stimulated with Tumor Necrosis Factor (TNF-α in the presence and absence of I. scapularis saliva and interleukin (IL-8 was measured. Results I. scapularis saliva inhibits inflammatory cytokine secretion by macrophages during stimulation of Toll-like (TLR and Nod-like receptor (NLR signaling pathways. The effect of I. scapularis saliva on immune cells is not restricted to murine macrophages because decreasing levels of interleukin (IL-8 were observed after TNF-α stimulation of human peripheral blood mononuclear cells. I. scapularis saliva also mitigates pro-inflammatory cytokine response by murine macrophages during challenge with A. phagocytophilum. Conclusions These findings suggest that I. scapularis may inhibit inflammatory cytokine secretion during rickettsial transmission at the vector-host interface.

  4. Transplacental transmission of Anaplasma marginale in beef cattle chronically infected in southern Brazil

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    Hermann Eduardo Gonzalez Grau

    Full Text Available In this study, we have investigated the incidence of transplacental transmission of Anaplasma marginale in chronically infected cows with no history of acute anaplasmosis during gestation. In addition, we evaluated various techniques for detection of transplacental transmission ofA. marginale. Blood samples were collected from 30 cows at three different periods: at the time of artificial insemination, at gestational diagnosis, and after calving. Also, blood was collected from the newborn calves, including one sample before colostrum intake, and another three days after birth. A. marginale-specific antibodies were detected in 100% of the cows with an indirect fluorescent antibody test (IFAT, and in 97% of them, using an indirect enzyme-linked immunosorbent assay (ELISA. Also, we observed that all of the three-day-old newborn calves were seropositive by IFAT. According to polymerase chain reaction, 63.3% of the cows were carriers of A. marginale, as well as 6.7% of the newborn calves. This represented a transplacental transmission rate of 10.5%. Furthermore, a correlation of 93.3% was observed between the two serodiagnostic techniques, demonstrating that both ELISA and IFAT can be used in epidemiological surveys of A. marginale. These results confirm the occurrence of transplacental transmission of A. marginale in chronically infected cows and suggest the importance of this transmission route in areas of enzootic instability.

  5. Flåtten Ixodes ricinus som sykdomsvektor i Sør-Norge. Etablering og utvikling av PCR-baserte påvisningsmetoder og påvisning av Babesia, Borrelia og Anaplasma

    OpenAIRE

    2004-01-01

    NORSK SAMMENDRAG: Hensikten med denne undersøkelsen var å undersøke flåtten Ixodes ricinus som smittebærer av Ehrlichia/ Anaplasma, Borrelia burgdorferi sensu lata og Babesia i Sør-Norge, og å etablere DNA-basert metode for påvisning av Babesia og forbedre metode for påvisning av Ehrlichia/ Anaplasma. For å kontrollere variasjon i følsomheten av Ehrlichia/ Anaplasma PCR med 16s rDNA primere Ehr 521/747, ble en internkontroll konstruert. Internkontrollen er et DNA-fragment so...

  6. Wild boar (Sus scrofa) - reservoir host of Toxoplasma gondii, Neospora caninum and Anaplasma phagocytophilum in Slovakia.

    Science.gov (United States)

    Reiterová, Katarína; Špilovská, Silvia; Blaňarová, Lucia; Derdáková, Markéta; Čobádiová, Andrea; Hisira, Vladimír

    2016-03-01

    In Central Europe the wild boar population is permanently growing and consequently Cf foodborne infections. In this study serological and molecular detection of Toxoplasma gondii and Neospora caninum in wild boars was evaluated. Moreover, same samples were screened for the presence and genetic variability of tick-borne bacterium Anaplasma phagocytophilum. Blood samples collected from 113 wild boars from Southern Slovakia were examined for antibodies to T. gondii by indirect and to N. caninum by competitive ELISA. The presence of parasitic DNA in blood samples was determined by standard or real time PCR techniques. Antibodies against T. gondii and N. caninum were detected in 45 (39.8%) and 38 (33.6%) animals, respectively. Females were more frequently infected for both pathogens than males. The high seropositivity against both coccidia indicates a permanent occurrence of these pathogens in the studied locality. T. gondii DNA was confirmed in five seropositive boars (4.4%) and N. caninum in 23 blood samples (20.4%). Three out of 23 N. caninum PCR positive animals did not show seropositivity. Three out of 113 blood samples of wild boars were positive for A. phagocytophilum (2.7%). The obtained A. phagocytophilum sequences were 100% identical with GenBankTM isolates from Slovak dog (KC985242); German horse (JF893938) or wild boar (EF143810) and red deer (EF143808) from Poland. Coinfections of T. gondii with N. caninum and N. caninum with A. phagocytophilum were detected in single cases. Results suggest a potential zoonotic risk of toxoplasmosis transmission to humans and the spread of neosporosis to farm animals.

  7. Analysis of the population structure of Anaplasma phagocytophilum using multilocus sequence typing.

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    Christian Huhn

    Full Text Available Anaplasma phagocytophilum is a Gram-negative obligate intracellular bacterium that replicates in neutrophils. It is transmitted via tick-bite and causes febrile disease in humans and animals. Human granulocytic anaplasmosis is regarded as an emerging infectious disease in North America, Europe and Asia. However, although increasingly detected, it is still rare in Europe. Clinically apparent A. phagocytophilum infections in animals are mainly found in horses, dogs, cats, sheep and cattle. Evidence from cross-infection experiments that A. phagocytophilum isolates of distinct host origin are not uniformly infectious for heterologous hosts has led to several approaches of molecular strain characterization. Unfortunately, the results of these studies are not always easily comparable, because different gene regions and fragment lengths were investigated. Multilocus sequence typing is a widely accepted method for molecular characterization of bacteria. We here provide for the first time a universal typing method that is easily transferable between different laboratories. We validated our approach on an unprecedented large data set of almost 400 A. phagocytophilum strains from humans and animals mostly from Europe. The typability was 74% (284/383. One major clonal complex containing 177 strains was detected. However, 54% (49/90 of the sequence types were not part of a clonal complex indicating that the population structure of A. phagocytophilum is probably semiclonal. All strains from humans, dogs and horses from Europe belonged to the same clonal complex. As canine and equine granulocytic anaplasmosis occurs frequently in Europe, human granulocytic anaplasmosis is likely to be underdiagnosed in Europe. Further, wild boars and hedgehogs may serve as reservoir hosts of the disease in humans and domestic animals in Europe, because their strains belonged to the same clonal complex. In contrast, as they were only distantly related, roe deer, voles and

  8. Sp110 transcription is induced and required by Anaplasma phagocytophilum for infection of human promyelocytic cells

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    Naranjo Victoria

    2007-09-01

    Full Text Available Abstract Background The tick-borne intracellular pathogen, Anaplasma phagocytophilum (Rickettsiales: Anaplasmataceae causes human granulocytic anaplasmosis after infection of polymorphonuclear leucocytes. The human Sp110 gene is a member of the nuclear body (NB components that functions as a nuclear hormone receptor transcriptional coactivator and plays an important role in immunoprotective mechanisms against pathogens in humans. In this research, we hypothesized that Sp110 may be involved in the infection of human promyelocytic HL-60 cells with A. phagocytophilum. Methods The human Sp110 and A. phagocytophilum msp4 mRNA levels were evaluated by real-time RT-PCR in infected human HL-60 cells sampled at 0, 12, 24, 48, 72 and 96 hours post-infection. The effect of Sp110 expression on A. phagocytophilum infection was determined by RNA interference (RNAi. The expression of Sp110 was silenced in HL-60 cells by RNAi using pre-designed siRNAs using the Nucleofector 96-well shuttle system (Amaxa Biosystems, Gaithersburg, MD, USA. The A. phagocytophilum infection levels were evaluated in HL-60 cells after RNAi by real-time PCR of msp4 and normalizing against human Alu sequences. Results While Sp110 mRNA levels increased concurrently with A. phagocytophilum infections in HL-60 cells, the silencing of Sp110 expression by RNA interference resulted in decreased infection levels. Conclusion These results demonstrated that Sp110 expression is required for A. phagocytophilum infection and multiplication in HL-60 cells, and suggest a previously undescribed mechanism by which A. phagocytophilum modulates Sp110 mRNA levels to facilitate establishment of infection of human HL-60 cells.

  9. Hematologic and Clinical Aspects of Experimental Ovine Anaplasmosis Caused by Anaplasma Ovis in Iran

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    A Gharabaghi

    2012-12-01

    Full Text Available Background: Anaplasma ovis infections can cause clinical symptoms in acute phase and lead to huge economic losses in flocks. The aim of the present study was to investigate the hematological and parasito­logical changes in experimental anaplasmosis in sheep with Iranian strain of A. ovis.Method: Five male sheep without any blood parasite infection were selected. One hundred ml hepari­nized blood was collected from splenectomised sheep that showed 6% A. ovis parasitemia. Inocu­lums of 20 ml blood were administered intravenously to each test animal. Hematological, parasito­logical and clinical changes of experimental anaplasmosis were studied in 0-38 days post infec­tion.Result: Parasitemia was detected 3 days post infection and reached its maximum level on the day 12 of experiment in test animals. Then the parasitemia was declined, but the organism could be found persistently until the last day of study. The red cell counts, packed cell volume and hemoglobin concentra­tion were decreased and mean corpuscular volume was increased significantly during the infection period. Reticulocytosis and basophilic stippling were also detected. No significant changes were observed in total and differential leukocyte count and animal body temperature.Conclusion: Experimental A. ovis infection in sheep resulted in marked normocytic normochromic anemia at the beginning of the infection which became macrocytic normochromic by the develop­ment of the disease. There were negative correlations between parasitemia and RBC, PCV and Hb values, therefore hematological assessment can be considered as a practical diagnostic tool in ovine anaplasmosis.

  10. Anaplasma phagocytophilum Infection Subverts Carbohydrate Metabolic Pathways in the Tick Vector, Ixodes scapularis

    Science.gov (United States)

    Cabezas-Cruz, Alejandro; Alberdi, Pilar; Valdés, James J.; Villar, Margarita; de la Fuente, José

    2017-01-01

    The obligate intracellular pathogen, Anaplasma phagocytophilum, is the causative agent of human, equine, and canine granulocytic anaplasmosis and tick-borne fever (TBF) in ruminants. A. phagocytophilum has become an emerging tick-borne pathogen in the United States, Europe, Africa, and Asia, with increasing numbers of infected people and animals every year. It has been recognized that intracellular pathogens manipulate host cell metabolic pathways to increase infection and transmission in both vertebrate and invertebrate hosts. However, our current knowledge on how A. phagocytophilum affect these processes in the tick vector, Ixodes scapularis is limited. In this study, a genome-wide search for components of major carbohydrate metabolic pathways was performed in I. scapularis ticks for which the genome was recently published. The enzymes involved in the seven major carbohydrate metabolic pathways glycolysis, gluconeogenesis, pentose phosphate, tricarboxylic acid cycle (TCA), glyceroneogenesis, and mitochondrial oxidative phosphorylation and β-oxidation were identified. Then, the available transcriptomics and proteomics data was used to characterize the mRNA and protein levels of I. scapularis major carbohydrate metabolic pathway components in response to A. phagocytophilum infection of tick tissues and cultured cells. The results showed that major carbohydrate metabolic pathways are conserved in ticks. A. phagocytophilum infection inhibits gluconeogenesis and mitochondrial metabolism, but increases the expression of glycolytic genes. A model was proposed to explain how A. phagocytophilum could simultaneously control tick cell glucose metabolism and cytoskeleton organization, which may be achieved in part by up-regulating and stabilizing hypoxia inducible factor 1 alpha in a hypoxia-independent manner. The present work provides a more comprehensive view of the major carbohydrate metabolic pathways involved in the response to A. phagocytophilum infection in ticks

  11. Genetic variants of Anaplasma phagocytophilum from 14 equine granulocytic anaplasmosis cases

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    Pfister Kurt

    2011-08-01

    Full Text Available Abstract Background Equine Granulocytic Anaplasmosis (EGA is caused by Anaplasma phagocytophilum, a tick-transmitted, obligate intracellular bacterium. In Europe, it is transmitted by Ixodes ricinus. A large number of genetic variants of A. phagocytophilum circulate in nature and have been found in ticks and different animals. Attempts have been made to assign certain genetic variants to certain host species or pathologies, but have not been successful so far. The purpose of this study was to investigate the causing agent A. phagocytophilum of 14 cases of EGA in naturally infected horses with molecular methods on the basis of 4 partial genes (16S rRNA, groEL, msp2, and msp4. Results All DNA extracts of EDTA-blood samples of the horses gave bands of the correct nucleotide size in all four genotyping PCRs. Sequence analysis revealed 4 different variants in the partial 16S rRNA, groEL gene and msp2 genes, and 3 in the msp4 gene. One 16S rRNA gene variant involved in 11 of the 14 cases was identical to the "prototype" variant causing disease in humans in the amplified part [GenBank: U02521]. Phylogenetic analysis revealed as expected for the groEL gene that sequences from horses clustered separately from roe deer. Sequences of the partial msp2 gene from this study formed a separate cluster from ruminant variants in Europe and from all US variants. Conclusions The results show that more than one variant of A. phagocytophilum seems to be involved in EGA in Germany. The comparative genetic analysis of the variants involved points towards different natural cycles in the epidemiology of A. phagocytophilum, possibly involving different reservoir hosts or host adaptation, rather than a strict species separation.

  12. Demonstration of transplacental transmission of a human isolate of Anaplasma phagocytophilum in an experimentally infected sheep.

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    Reppert, E; Galindo, R C; Breshears, M A; Kocan, K M; Blouin, E F; de la Fuente, J

    2013-11-01

    Anaplasma phagocytophilum, first identified as a pathogen of sheep in Europe, more recently has been recognized as an emerging tick-borne pathogen of humans in the U.S. and Europe. Transmission of A. phagocytophilum is reported to be by ticks, primarily of the genus Ixodes. While mechanical and transplacental transmission of the type genus organism, A. marginale, occur in addition to tick transmission, these modes of transmission have not been considered for A. phagocytophilum. Recently, we developed a sheep model for studying host-tick-pathogen interactions of the human NY-18 A. phagocytophilum isolate. Sheep were susceptible to infection with this human isolate and served as a source of infection for I. scapularis ticks, but they did not display clinical signs of disease, and the pathogen was not apparent in stained blood smears. In the course of these experiments, one sheep unexpectedly gave birth to a lamb 5 weeks after being experimentally infected by inoculation with the pathogen propagated in HL-60 cells. The lamb was depressed and not feeding and was subsequently euthanized 18 h after birth. Tissues were collected at necropsy for microscopic examination and PCR to confirm A. phagocytophilum infection. At necropsy, the stomach contained colostrum, the spleen was moderately enlarged and thickened with conspicuous lymphoid follicles, and mesenteric lymph nodes were mildly enlarged and contained moderate infiltrates of eosinophils and neutrophils. Blood, spleen, heart, skin and cervical and mesenteric lymph nodes tested positive for A. phagocytophilum by PCR, and sequence analysis confirmed that the lamb was infected with the NY-18 isolate. Transplacental transmission should therefore be considered as a means of A. phagocytophilum transmission and may likely contribute to the epidemiology of tick-borne fever in sheep and other mammals, including humans.

  13. Gene expression profile suggests that pigs (Sus scrofa are susceptible to Anaplasma phagocytophilum but control infection

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    Galindo Ruth C

    2012-08-01

    Full Text Available Abstract Background Anaplasma phagocytophilum infects a wide variety of hosts and causes granulocytic anaplasmosis in humans, horses and dogs and tick-borne fever in ruminants. Infection with A. phagocytophilum results in the modification of host gene expression and immune response. The objective of this research was to characterize gene expression in pigs (Sus scrofa naturally and experimentally infected with A. phagocytophilum trying to identify mechanisms that help to explain low infection prevalence in this species. Results For gene expression analysis in naturally infected pigs, microarray hybridization was used. The expression of differentially expressed immune response genes was analyzed by real-time RT-PCR in naturally and experimentally infected pigs. Results suggested that A. phagocytophilum infection affected cytoskeleton rearrangement and increased both innate and adaptive immune responses by up regulation of interleukin 1 receptor accessory protein-like 1 (IL1RAPL1, T-cell receptor alpha chain (TCR-alpha, thrombospondin 4 (TSP-4 and Gap junction protein alpha 1 (GJA1 genes. Higher serum levels of IL-1 beta, IL-8 and TNF-alpha in infected pigs when compared to controls supported data obtained at the mRNA level. Conclusions These results suggested that pigs are susceptible to A. phagocytophilum but control infection, particularly through activation of innate immune responses, phagocytosis and autophagy. This fact may account for the low infection prevalence detected in pigs in some regions and thus their low or no impact as a reservoir host for this pathogen. These results advanced our understanding of the molecular mechanisms at the host-pathogen interface and suggested a role for newly reported genes in the protection of pigs against A. phagocytophilum.

  14. The efficacy of three chlortetracycline regimens in the treatment of persistent Anaplasma marginale infection.

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    Reinbold, James B; Coetzee, Johann F; Hollis, Larry C; Nickell, Jason S; Riegel, Casey; Olson, K C; Ganta, Roman R

    2010-09-28

    Chemosterilization is reported in cattle fed chlortetracycline hydrochloride (CTC) at dosages ranging from 1.1mg/kg for 120 days to 11 mg/kg for 30-60 days. The relationship between plasma CTC drug concentration and carrier clearance has not been described. Chronic carrier status was established in 21 steers with a Virginia isolate of Anaplasma marginale and confirmed by cELISA and an A. marginale-specific RT-PCR. Four negative, splenectomized steers served as active disease transmission sentinels. Steers were randomized to receive 4.4 mg/kg/day (LD); 11 mg/kg/day (MD); or 22 mg/kg/day (HD) of oral chlortetracycline; or placebo (CONTROL) for 80 days. The LD, MD and HD treatment groups consisted of 5 infected steers and 1 splenectomized steer; CONTROL group had six infected steers and 1 splenectomized steer. The daily treatments and ration were divided equally and fed twice daily. Blood samples were collected semi-weekly for determining plasma drug concentration by ultrahigh performance liquid chromatography-mass spectrometry/mass spectrometry method and assessment of disease status by both cELISA and RT-PCR. Mean (CV%) chlortetracycline plasma drug concentrations in the LD, MD, and HD groups were 85.3 (28%), 214.5 (32%) and 518.9 (40%)ng/mL during days 4 through 53 of treatment. A negative RT-PCR assay result was confirmed in all CTC-treated groups within 49 days of treatment; however, cELISA required an additional 49 to 88 days before similar results. Subinoculation of splenectomized steers confirmed chemosterilization. These results are important for influencing future chemosterilization strategies and impacting free trade policy among countries and regions of contrasting endemicity.

  15. Longitudinal analysis of tick densities and Borrelia, Anaplasma, and Ehrlichia infections of Ixodes ricinus ticks in different habitat areas in The Netherlands.

    NARCIS (Netherlands)

    Wielinga, Peter R; Gaasenbeek, Cor; Fonville, Manoj; Boer, Albert de; Vries, Ankje de; Dimmers, Wim; Akkerhuis Op Jagers, Gerard; Schouls, Leo M; Borgsteede, Fred; Giessen, Joke W B van der

    2006-01-01

    From 2000 to 2004, ticks were collected by dragging a blanket in four habitat areas in The Netherlands: dunes, heather, forest, and a city park. Tick densities were calculated, and infection with Borrelia burgdorferi and Anaplasma and Ehrlichia species was investigated by reverse line blot analysis.

  16. Longitudinal analysis of tick densities and Borrelia, Anaplasma, and Ehrlichia infections of Ixodes ricinus ticks in different habitat areas in the Netherlands

    NARCIS (Netherlands)

    Wielinga, P.R.; Gaasenbeek, C.P.H.; Fonville, M.; Boer, de A.G.; Vries, de A.; Dimmers, W.J.; Jagers op Akkerhuis, G.A.J.M.; Schouls, L.M.; Borgsteede, F.H.M.; Giessen, van der J.W.B.

    2006-01-01

    From 2000 to 2004, ticks were collected by dragging a blanket in four habitat areas in The Netherlands: dunes, heather, forest, and a city park. Tick densities were calculated, and infection with Borrelia burgdorferi and Anaplasma and Ehrlichia species was investigated by reverse line blot analysis.

  17. Detection of Anaplasma phagocytophilum in red foxes (Vulpes vulpes) and raccoon dogs (Nyctereutes procyonoides) from Brandenburg, Germany.

    Science.gov (United States)

    Härtwig, Vera; von Loewenich, Friederike D; Schulze, Christoph; Straubinger, Reinhard K; Daugschies, Arwid; Dyachenko, Viktor

    2014-04-01

    Anaplasma phagocytophilum is an obligate intracellular and tick-transmitted bacterium, which causes granulocytic anaplasmosis in animals and humans. Although infection with A. phagocytophilum in domestic animals and vector ticks is documented, there is sparse information on the occurrence of A. phagocytophilum in wild animals. Red foxes (Vulpes vulpes) as well as raccoon dogs (Nyctereutes procyonoides) are wildlife species highly abundant in certain areas of Germany and represent a potential wildlife reservoir for zoonotic diseases. To obtain data about the occurrence of A. phagocytophilum in these animals, red fox and raccoon dog carcasses (hunted or found dead) were collected from January to September 2009 in the Federal State of Brandenburg, Germany. Lung tissue samples were subjected to DNA extraction and were examined for the presence of A. phagocytophilum DNA by means of real-time PCR. Anaplasma phagocytophilum was detected in 10 out of 122 (8.2%) lungs of red foxes and in 3 out of 13 (23%) lungs of raccoon dogs. To the best of our knowledge, A. phagocytophilum was detected for the first time in red foxes and raccoon dogs in Germany.

  18. Molecular characterization of Rickettsia massiliae and Anaplasma platys infecting Rhipicephalus sanguineus ticks and domestic dogs, Buenos Aires (Argentina).

    Science.gov (United States)

    Cicuttin, Gabriel L; Brambati, Diego F; Rodríguez Eugui, Juan I; Lebrero, Cecilia González; De Salvo, María N; Beltrán, Fernando J; Gury Dohmen, Federico E; Jado, Isabel; Anda, Pedro

    2014-09-01

    Rickettsioses, ehrlichioses and anaplasmoses are emerging diseases that are mainly transmitted by arthropods and that affect humans and animals. The aim of the present study was to use molecular techniques to detect and characterize those pathogens in dogs and ticks from Buenos Aires city. We studied 207 Rhipicephalus sanguineus ticks and 52 canine blood samples from poor neighborhoods of Buenos Aires city. The samples were molecularly screened for the genera Rickettsia, Ehrlichia, and Anaplasma by PCR and sequencing. DNA of Rickettsia massiliae (3.4%) and Anaplasma platys (13.5%) was detected in ticks and blood samples, respectively. For characterization, the positive samples were subjected to amplification of a fragment of the 190-kDa outer membrane protein gene (spotted fever group rickettsiae) and a fragment of the groESL gene (specific for A. platys). A phylogenetic tree was constructed using the neighbor-joining method, revealing that the sequences were closely related to those of strains from other geographic regions. The results indicate that human and animal pathogens are abundant in dogs and their ticks in Buenos Aires city and portray the potentially high risk of human exposure to infection with these agents, especially in poor neighborhoods, where there is close contact with animals in an environment of poor health conditions.

  19. Surveillance of Egyptian fleas for agents of public health significance: Anaplasma, Bartonella, Coxiella, Ehrlichia, Rickettsia, and Yersinia pestis.

    Science.gov (United States)

    Loftis, Amanda D; Reeves, Will K; Szumlas, Daniel E; Abbassy, Magda M; Helmy, Ibrahim M; Moriarity, John R; Dasch, Gregory A

    2006-07-01

    Serologic surveys in Egypt have documented human and animal exposure to vector-borne bacterial pathogens, but the presence and distribution of these agents in arthropods has not been determined. Between July 2002 and July 2003, fleas were collected from 221 mammals trapped in 17 cities throughout Egypt. A total of 987 fleas were collected, representing four species (Ctenocephalides felis, Echidnophaga gallinacea, Leptopsylla segnis, and Xenopsylla cheopis); 899 of these fleas were X. cheopis from rats (Rattus spp.). Fleas were tested for DNA from Anaplasma spp., Bartonella spp., Coxiella burnetii, Ehrlichia spp., Rickettsia spp., and Yersinia pestis. Rickettsia typhi, the agent of murine typhus, was detected in X. cheopis and L. segnis from rats from nine cities. A spotted-fever group Rickettsia sp. similar to "RF2125" was detected in E. gallinacea, and two unidentified spotted fever group Rickettsia were detected in two X. cheopis. Novel Bartonella genotypes were detected in X. cheopis and L. segnis from three cities. Coxiella burnetii was detected in two fleas. Anaplasma, Ehrlichia, and Y. pestis were not detected.

  20. DETECTION OF ANTIBODIES TO ANAPLASMA, BARTONELLA AND COXIELLA IN RURAL INHABITANTS OF THE CARIBBEAN AREA OF COLOMBIA

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    Salim Máttar

    2006-12-01

    Full Text Available Objetivo. Establecer la seroprevalencia de Bartonella spp, Anaplasma phagocytophilum (antesErlichia y Coexiella burnetii. Materiales y métodos. Se analizaron sueros representativos de unsector de la población en el año 2003, recolectados de personas que trabajan en actividades delcampo en los departamentos de Córdoba y Sucre que sirvieron como población base de las muestrasque se obtuvieron. Los trabajadores rurales elegidos a participar tenían entra 16 – 65 años deedad. Los sueros fueron examinados por IFA para detección de anticuerpos contra IgG para Bartonellaspp, Erlichia Anaplasma phagocytophilum y Coexiella burnetii. Resultados. La seroprevalencia deanticuerpos de todos los microorganismos estudiados fue de 56.8%. De 81 muestras de sueroanalizadas el 26.6% fueron seropositivas contra C. burnetii, el 37.7% tuvieron anticuerpos contraBartonella y el 20% de los individuos evaluados fueron seropositivos para Anaplasmaphagocytophilum. Conclusiones. Nuestros datos indican que la prevalencia de anticuerpos contraBartonella, A. phagocytophilum y C. burnetii son altos en nuestra región. Los resultados indicanque estas enfermedades zoonoticas son muy comunes en las personas que residen en el área delcaribe colombiano. Este estudio demuestra por primera vez la presencia de estos microorganismosen Colombia.

  1. Simultaneous detection of Anaplasma and Ehrlichia species in ruminants and detection of Ehrlichia ruminantium in Amblyomma variegatum ticks by reverse line blot hybridization.

    Science.gov (United States)

    Bekker, Cornelis P J; de Vos, Sander; Taoufik, Amar; Sparagano, Olivier A E; Jongejan, Frans

    2002-10-22

    The detection of Anaplasma and Ehrlichia species is usually based on species-specific PCR assays, since no assay is yet available which can detect and identify these species simultaneously. To this end, we developed a reverse line blot (RLB) assay for simultaneous detection and identification of Anaplasma and Ehrlichia species in domestic ruminants and ticks. In a PCR the hypervariable V1 region of the 16S ribosomal RNA (rRNA) gene was amplified with a set of primers unique for members of the genera Anaplasma and Ehrlichia [Int. J. Syst. Evol. Microbiol. 51 (2001) 2145]. Amplified PCR products from blood of domestic ruminants or Amblyomma variegatum tick samples were hybridized onto a membrane to which eight species-specific oligonucleotide probes and one Ehrlichia and Anaplasma catch-all oligonucleotide probe were covalently linked. No DNA was amplified from uninfected blood, nor from other hemoparasites such as Theileria annulata, or Babesia bigemina. The species-specific probes did not cross-react with DNA amplified from other species. E. ruminantium, A. ovis and another Ehrlichia were identified by RLB in blood samples collected from small ruminants in Mozambique. Finally, A. variegatum ticks were tested after feeding on E. ruminantium infected sheep. E. ruminantium could be detected in adult ticks even if feeding of nymphs was carried out 3.5 years post-infection. In conclusion, the developed species-specific oligonucleotide probes used in an RLB assay can simultaneously detect and identify several Ehrlichia and Anaplasma species. However, as no quantitative data for the detection limit are available yet, only positive results are interpretable at this stage.

  2. Subdominant Outer Membrane Antigens in Anaplasma marginale: Conservation, Antigenicity, and Protective Capacity Using Recombinant Protein.

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    Deirdre R Ducken

    Full Text Available Anaplasma marginale is a tick-borne rickettsial pathogen of cattle with a worldwide distribution. Currently a safe and efficacious vaccine is unavailable. Outer membrane protein (OMP extracts or a defined surface protein complex reproducibly induce protective immunity. However, there are several knowledge gaps limiting progress in vaccine development. First, are these OMPs conserved among the diversity of A. marginale strains circulating in endemic regions? Second, are the most highly conserved outer membrane proteins in the immunogens recognized by immunized and protected animals? Lastly, can this subset of OMPs recognized by antibody from protected vaccinates and conserved among strains recapitulate the protection of outer membrane vaccines? To address the first goal, genes encoding OMPs AM202, AM368, AM854, AM936, AM1041, and AM1096, major subdominant components of the outer membrane, were cloned and sequenced from geographically diverse strains and isolates. AM202, AM936, AM854, and AM1096 share 99.9 to 100% amino acid identity. AM1041 has 97.1 to 100% and AM368 has 98.3 to 99.9% amino acid identity. While all four of the most highly conserved OMPs were recognized by IgG from animals immunized with outer membranes, linked surface protein complexes, or unlinked surface protein complexes and shown to be protected from challenge, the highest titers and consistent recognition among vaccinates were to AM854 and AM936. Consequently, animals were immunized with recombinant AM854 and AM936 and challenged. Recombinant vaccinates and purified outer membrane vaccinates had similar IgG and IgG2 responses to both proteins. However, the recombinant vaccinates developed higher bacteremia after challenge as compared to adjuvant-only controls and outer membrane vaccinates. These results provide the first evidence that vaccination with specific antigens may exacerbate disease. Progressing from the protective capacity of outer membrane formulations to

  3. Essential domains of Anaplasma phagocytophilum invasins utilized to infect mammalian host cells.

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    David Seidman

    2015-02-01

    Full Text Available Anaplasma phagocytophilum causes granulocytic anaplasmosis, an emerging disease of humans and domestic animals. The obligate intracellular bacterium uses its invasins OmpA, Asp14, and AipA to infect myeloid and non-phagocytic cells. Identifying the domains of these proteins that mediate binding and entry, and determining the molecular basis of their interactions with host cell receptors would significantly advance understanding of A. phagocytophilum infection. Here, we identified the OmpA binding domain as residues 59 to 74. Polyclonal antibody generated against a peptide spanning OmpA residues 59 to 74 inhibited A. phagocytophilum infection of host cells and binding to its receptor, sialyl Lewis x (sLe(x-capped P-selectin glycoprotein ligand 1. Molecular docking analyses predicted that OmpA residues G61 and K64 interact with the two sLe(x sugars that are important for infection, α2,3-sialic acid and α1,3-fucose. Amino acid substitution analyses demonstrated that K64 was necessary, and G61 was contributory, for recombinant OmpA to bind to host cells and competitively inhibit A. phagocytophilum infection. Adherence of OmpA to RF/6A endothelial cells, which express little to no sLe(x but express the structurally similar glycan, 6-sulfo-sLe(x, required α2,3-sialic acid and α1,3-fucose and was antagonized by 6-sulfo-sLe(x antibody. Binding and uptake of OmpA-coated latex beads by myeloid cells was sensitive to sialidase, fucosidase, and sLe(x antibody. The Asp14 binding domain was also defined, as antibody specific for residues 113 to 124 inhibited infection. Because OmpA, Asp14, and AipA each contribute to the infection process, it was rationalized that the most effective blocking approach would target all three. An antibody cocktail targeting the OmpA, Asp14, and AipA binding domains neutralized A. phagocytophilum binding and infection of host cells. This study dissects OmpA-receptor interactions and demonstrates the effectiveness of binding

  4. Subdominant Outer Membrane Antigens in Anaplasma marginale: Conservation, Antigenicity, and Protective Capacity Using Recombinant Protein.

    Science.gov (United States)

    Ducken, Deirdre R; Brown, Wendy C; Alperin, Debra C; Brayton, Kelly A; Reif, Kathryn E; Turse, Joshua E; Palmer, Guy H; Noh, Susan M

    2015-01-01

    Anaplasma marginale is a tick-borne rickettsial pathogen of cattle with a worldwide distribution. Currently a safe and efficacious vaccine is unavailable. Outer membrane protein (OMP) extracts or a defined surface protein complex reproducibly induce protective immunity. However, there are several knowledge gaps limiting progress in vaccine development. First, are these OMPs conserved among the diversity of A. marginale strains circulating in endemic regions? Second, are the most highly conserved outer membrane proteins in the immunogens recognized by immunized and protected animals? Lastly, can this subset of OMPs recognized by antibody from protected vaccinates and conserved among strains recapitulate the protection of outer membrane vaccines? To address the first goal, genes encoding OMPs AM202, AM368, AM854, AM936, AM1041, and AM1096, major subdominant components of the outer membrane, were cloned and sequenced from geographically diverse strains and isolates. AM202, AM936, AM854, and AM1096 share 99.9 to 100% amino acid identity. AM1041 has 97.1 to 100% and AM368 has 98.3 to 99.9% amino acid identity. While all four of the most highly conserved OMPs were recognized by IgG from animals immunized with outer membranes, linked surface protein complexes, or unlinked surface protein complexes and shown to be protected from challenge, the highest titers and consistent recognition among vaccinates were to AM854 and AM936. Consequently, animals were immunized with recombinant AM854 and AM936 and challenged. Recombinant vaccinates and purified outer membrane vaccinates had similar IgG and IgG2 responses to both proteins. However, the recombinant vaccinates developed higher bacteremia after challenge as compared to adjuvant-only controls and outer membrane vaccinates. These results provide the first evidence that vaccination with specific antigens may exacerbate disease. Progressing from the protective capacity of outer membrane formulations to recombinant vaccines

  5. An O-Methyltransferase Is Required for Infection of Tick Cells by Anaplasma phagocytophilum.

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    Adela S Oliva Chávez

    Full Text Available Anaplasma phagocytophilum, the causative agent of Human Granulocytic Anaplasmosis (HGA, is an obligately intracellular α-proteobacterium that is transmitted by Ixodes spp ticks. However, the pathogen is not transovarially transmitted between tick generations and therefore needs to survive in both a mammalian host and the arthropod vector to complete its life cycle. To adapt to different environments, pathogens rely on differential gene expression as well as the modification of proteins and other molecules. Random transposon mutagenesis of A. phagocytophilum resulted in an insertion within the coding region of an o-methyltransferase (omt family 3 gene. In wild-type bacteria, expression of omt was up-regulated during binding to tick cells (ISE6 at 2 hr post-inoculation, but nearly absent by 4 hr p.i. Gene disruption reduced bacterial binding to ISE6 cells, and the mutant bacteria that were able to enter the cells were arrested in their replication and development. Analyses of the proteomes of wild-type versus mutant bacteria during binding to ISE6 cells identified Major Surface Protein 4 (Msp4, but also hypothetical protein APH_0406, as the most differentially methylated. Importantly, two glutamic acid residues (the targets of the OMT were methyl-modified in wild-type Msp4, whereas a single asparagine (not a target of the OMT was methylated in APH_0406. In vitro methylation assays demonstrated that recombinant OMT specifically methylated Msp4. Towards a greater understanding of the overall structure and catalytic activity of the OMT, we solved the apo (PDB_ID:4OA8, the S-adenosine homocystein-bound (PDB_ID:4OA5, the SAH-Mn2+ bound (PDB_ID:4PCA, and SAM- Mn2+ bound (PDB_ID:4PCL X-ray crystal structures of the enzyme. Here, we characterized a mutation in A. phagocytophilum that affected the ability of the bacteria to productively infect cells from its natural vector. Nevertheless, due to the lack of complementation, we cannot rule out secondary

  6. In vitro cultivation of Anaplasma marginale and A. phagocytophilum in tick cell lines: a review Cultivo in vitro de Anaplasma marginale e A. phagocytophilum em células de carrapatos: uma revisão

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    Lygia Maria Friche Passos

    2012-06-01

    Full Text Available Continuous cell lines have been established from several ixodid and argasid tick species, representing an excellent tool suitable for the isolation of pathogens and their subsequent propagation, which in turn allows the production of antigenic material for diagnostic tests, antibody and vaccine production, and also for studies on host-vector-pathogen relationships. This paper reviews the use of tick cells for culture initiation and maintenance of two obligate intracellular bacterial pathogens, Anaplasma marginale and Anaplasma phagocytophilum. These in vitro cultivation systems have been used in a wide range of studies, covering morphological ultrastructural analysis, genetics, proteomics and biological differences between strains, including genome transcriptional and protein expression approaches, enabling comparisons between host and vector cells. Thus, such systems open a new window for a better understanding of interactions between pathogens and tick cells. Last but not least, such systems contribute to the reduction in usage of animals for experimental research, as antigenic material can be produced in reasonably large quantities without the use of in vivo species-specific systems.Linhagens contínuas de células já foram estabelecidas a partir de várias espécies de carrapatos ixodídeos e argasídeos e representam uma ferramenta excelente para o isolamento e propagação de patógenos, permitindo a produção de material antigênico para testes diagnósticos, produção de anticorpos e vacinas, e também para estudos das relações entre hospedeiro-vetor-patógenos. Este artigo revisa o uso de células de carrapatos para estabelecimento e manutenção in vitro de dois patógenos intracelulares, Anaplasma marginale e Anaplasma phagocytophilum. Estes sistemas de cultivo in vitro, têm sido utilizados em vários estudos, tais como análises morfológicas, genéticas, proteômicas e estudos diferenciais entre isolados, incluindo gen

  7. Clinical and molecular features of one case of human infection with Anaplasma phagocytophilum from Podlaskie Province in eastern Poland

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    Renata Welc-Falęciak

    2015-09-01

    Full Text Available The article focuses on the clinical and laboratory diagnosis of human granulocytic anaplasmosis (HGA caused by [i]Anaplasma phagocytophilum[/i] infection in one of 28 patients (3.6%; n=1/28 tested samples with early Lyme borreliosis. The clinical and laboratory results of a 42-year-old patient fulfilled criteria of confirm anaplasmosis and suggest an acute stage of illness. The described case provides strong presumptive evidence that infection in this patient was acquired with a pathogenic strain of [i]A. phagocytophilum[/i] through a tick bite. A positive DNA with PCR for A. phagocytophilum infection was sequenced and analyzed phylogenetically. Physicians should consider the possibility of anaplasmosis in patients with early Lyme borreliosis, and [i]A. phagocytophilum[/i] should be considered as a differential diagnosis in all patients from an endemic region of potential high risk factors for tick-borne diseases.

  8. The potential role of migratory birds in transmission cycles of Babesia spp., Anaplasma phagocytophilum, and Rickettsia spp.

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    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.

  9. Development of enzyme-linked immunosorbent assays based on recombinant MSP1a and MSP2 of Anaplasma marginale

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    Flábio R Araújo

    2005-11-01

    Full Text Available Indirect enzyme-linked immunosorbent assays (ELISAs based on recombinant MSP1a and MSP2 from a Brazilian isolate of Anaplasma marginale were developed to detect antibodies against this rickettsia in cattle. The high sensitivities (99% for both tests and specificities (100% for both tests were confirmed with sera from cattle positive or negative for A. marginale antibodies, respectively, by immunofluorescent antibody test. By the analysis of 583 sera from cattle of three regions of the state of Pernambuco, Brazil, the agreement between both tests was high, with a kappa index of 0.89. The similar performances of the ELISAs suggest that both tests can be used in epidemiological surveys for detection of antibodies to A. marginale in cattle.

  10. Seroprevalence of antibodies to tick-borne encephalitis virus and Anaplasma phagocytophilum in healthy adults from western Norway.

    Science.gov (United States)

    Hjetland, Reidar; Henningsson, Anna J; Vainio, Kirsti; Dudman, Susanne G; Grude, Nils; Ulvestad, Elling

    2015-01-01

    The aim of this study was to assess the seroprevalence of antibodies to tick-borne encephalitis virus (TBEV) and Anaplasma phagocytophilum in a healthy adult population from Sogn and Fjordane county in western Norway. Sera from 1, 213 blood donors were analysed for IgG-antibodies to TBEV, and a random subgroup of 301 donors for IgG to A. phagocytophilum. In the TBEV ELISA, five (0.4%) sera were positive. These were all interpreted as "false" positives, as four had received vaccines against flaviviruses, and the remaining was negative for neutralizing antibodies to TBEV. Antibodies to A. phagocytophilum were detected by indirect immunofluorescence in 49 (16.2%) subjects (titer range 80-1280). The results indicate that TBE currently is not endemic in this part of western Norway. However, there is serological evidence of the existence of human granulocytic anaplasmosis in the population.

  11. Association of Ehrlichia canis, Hemotropic Mycoplasma spp. and Anaplasma platys and severe anemia in dogs in Thailand.

    Science.gov (United States)

    Kaewmongkol, Gunn; Lukkana, Nicha; Yangtara, Sarawut; Kaewmongkol, Sarawan; Thengchaisri, Naris; Sirinarumitr, Theerapol; Jittapalapong, Sathaporn; Fenwick, Stanley G

    2017-03-01

    Canine tick-borne bacteria; Ehrlichia canis, hemotropic Mycoplasma spp. and Anaplasma spp., are organisms transmitted by Rhipicephalus sanguineus ticks. However, only a few clinical studies evaluating dogs infected with these organisms and anemia condition have been published. In this study, the potential tick-borne bacteria linked to anemia were investigated in eighty-one blood samples selected from anemic dogs using a broad range nested-PCR of the 16S rRNA gene. Positive results were shown in 12/81 blood specimens (14.81%). Nucleotide sequences from the PCR products were analyzed using BLAST and resulted in identification of Ehrlichia canis (8), Candidatus Mycoplasma haematoparvum (1) and Anaplasma platys (3). Two other PCR assays were used to detect and identify the positive results of these pathogens including a specific PCR for Ehrlichia canis (gp36) and a specific nested-PCR for hemoplasma species (16S rRNA) and the phylogenetic analyses of E. canis and canine hemoplasmas were performed using these two loci. These specific PCRs revealed co-infection of E. canis and Mycoplasma haemocanis in two cases. These two male dogs had presented with jaundice, severe hemolytic anemia, severe thrombocytopenia, leukocytosis, mild azotemia and hepatitis. Ehrlichia canis was detected in a significantly greater number of severe anemia cases (PCVcanis infections (odds ratio: 7.11, p=0.020). However, no statistical differences were detected between E. canis detection and degrees of thrombocytopenia or leukopenia. From the results of this study, we conclude that the severity of anemia is associated with E. canis infections rather than the severity of thrombocytopenia.

  12. Sheep experimentally infected with a human isolate of Anaplasma phagocytophilum serve as a host for infection of Ixodes scapularis ticks.

    Science.gov (United States)

    Kocan, Katherine M; Busby, Ann T; Allison, Robin W; Breshears, Melanie A; Coburn, Lisa; Galindo, Ruth C; Ayllón, Nieves; Blouin, Edmour F; de la Fuente, José

    2012-06-01

    Anaplasma phagocytophilum, first identified as a pathogen of ruminants in Europe, has more recently been recognized as an emerging tick-borne pathogen of humans in the U.S. and Europe. A. phagocytophilum is transmitted by Ixodes spp., but the tick developmental cycle and pathogen/vector interactions have not been fully described. In this research, we report on the experimental infection of sheep with the human NY-18 isolate of A. phagocytophilum which then served as a host for infection of I. scapularis nymphs and adults. A. phagocytophilum was propagated in the human promyelocytic cell line, HL-60, and the infected cell cultures were then used to infect sheep by intravenous inoculation. Infections in sheep were confirmed by PCR and an Anaplasma-competitive ELISA. Clinical signs were not apparent in any of the infected sheep, and only limited hematologic and mild serum biochemical abnormalities were identified. While A. phagocytophilum morulae were rarely seen in neutrophils, blood film evaluation revealed prominent large granular lymphocytes, occasional plasma cells, and rare macrophages. Upon necropsy, gross lesions were restricted to the lymphoid system. Mild splenomegaly and lymphadenomegaly with microscopic evidence of lymphoid hyperplasia was observed in all infected sheep. Female I. scapularis that were allowed to feed and acquire infection on each of the 3 experimentally infected sheep became infected with A. phagocytophilum as determined by PCR of guts (80-87%) and salivary glands (67-100%). Female I. scapularis that acquired infection as nymphs on an experimentally infected sheep transmitted A. phagocytophilum to a susceptible sheep, thus confirming transstadial transmission. Sheep proved to be a good host for the production of I. scapularis infected with this human isolate of A. phagocytophilum, which can be used as a model for future studies of the tick/pathogen interface.

  13. Detecção de anticorpos para Anaplasma sp. em pequenos ruminantes no semi-árido do estado de Pernambuco, Brasil Detection of antibodies against Anaplasma sp. in small ruminants from the semi-arid region against Pernambuco State, Brazil

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    Rafael A. N. Ramos

    2008-06-01

    Full Text Available Neste trabalho é descrita a detecção de anticorpos para Anaplasma sp. em caprinos e ovinos da região do semi-árido do Estado de Pernambuco, Brasil, utilizando-se um ensaio de imunoadsorção enzimática baseado em MSP5 recombinante de Anaplasma marginale. Foram analisados soros de 243 caprinos e 68 ovinos provenientes do município de Ibimirim, e observadas freqüências de anticorpos de 11,93% (29/243 e 16,17% (11/68 para caprinos e ovinos, respectivamente. A importância epidemiológica dos achados foi discutida.This paper reports the detection of antibodies against Anaplasma sp. in goats and sheep from the semi-arid region from Pernambuco State, Brazil, using an enzyme-linked immunosorbent assay with recombinant MSP5 of Anaplasma marginale. Sera from 243 goats and 68 sheep from Ibimirim municipality were analyzed and frequencies of antibodies of 11.93% (29/243 and 16.17% (11/68 were found for goats and sheep, respectively. The epidemiological relevance of the findings was discussed.

  14. Infections and risk factors for livestock with species of Anaplasma, Babesia and Brucella under semi-nomadic rearing in Karamoja Region, Uganda.

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    Lolli, Chiara; Marenzoni, Maria Luisa; Strona, Paolo; Lappo, Pier Giorgio; Etiang, Patrick; Diverio, Silvana

    2016-03-01

    A survey was conducted to estimate the prevalence of Anaplasma, Babesia and Brucella spp. infections in cattle, goats and sheep in the Karamoja Region of Uganda and to identify possible risk factors existing in this semi-nomadic and pastoral area. Low cost laboratory tests were used to diagnose infections (Rose Bengal test for Brucella spp. antibodies and direct microscopic examination for Anaplasma and Babesia spp.). Multivariable logistic regression models were applied to identify possible risk factors linked to gender, animal species, age (only for cattle) and districts. A total of 3935 cattle, 729 goats and 306 sheep of five districts of the Karamoja Region were tested. Seroprevalence for Brucella was 9.2 % (CI, 95 %: 8.4-10), for Anaplasma 19.5 % (CI 95 %: 18.4-20.6) and for Babesia 16 % (CI 95 %: 15-17.1). Significant differences in infections prevalence were observed against risk factors associated with districts and species. Cattle were the species with higher risk of the infections. Female gender was identified as at risk only for Brucella spp. infection. Cattle more than one year old had greater likelihood to be Brucella seropositive. Co-infections of Anaplasma and Babesia spp. were statistically associated, especially in goats and sheep. Further studies to identify risk factors related to host species and geographical districts are needed. The influence on the semi-nomadic agro-pastoral system in Karamoja of animal raids and animal mixing should be further investigated. Findings were important to sensitize Karamojong undertaking measures on infection control, especially on cattle, which are their main source of food.

  15. Prevalence and first molecular characterization of Anaplasma phagocytophilum, the agent of human granulocytic anaplasmosis, in Rhipicephalus sanguineus ticks attached to dogs from Egypt

    OpenAIRE

    Mohamed W. Ghafar; Sayed A. Amer

    2012-01-01

    PCR targeting 16S rRNA gene integrated with sequence analysis were performed to investigate the prevalence and the molecular identity of Anaplasma phagocytophilum in Egyptian Rhipicephalus sanguineus ticks attached to dogs. A total of 413 adult and nymphal R. sanguineus ticks were collected while attached to 72 free-roaming dogs from four locations (Imbaba, Boulaq, Haram, Monib) in Giza Governorate, Egypt. DNA was successfully extracted from 401 specimens (133 nymphs and 268 adults). The over...

  16. Vector-Borne Diseases in Stray Dogs in Peninsular Malaysia and Molecular Detection of Anaplasma and Ehrlichia spp. from Rhipicephalus sanguineus (Acari: Ixodidae) Ticks.

    Science.gov (United States)

    Koh, Fui Xian; Panchadcharam, Chandrawathani; Tay, Sun Tee

    2016-01-01

    Little data are available on the prevalence and transmission of vector-borne diseases in stray dogs in Peninsular Malaysia. This study was designed to determine the occurrence of vector-borne pathogens in Malaysian stray dogs using serological and molecular approaches. In total, 48 dog blood samples were subjected to serological analysis using SNAP 4Dx kit (IDEXX Laboratories, Westbrook, ME). The presence of Ehrlichia and Anaplasma DNA in the dog blood samples and Rhipicephalus sanguineus (Latreille) ticks was detected using nested polymerase chain reaction assays. Positive serological findings against Ehrlichia canis and Anaplasma phagocytophilum were obtained in 17 (39.5%) and four (9.3%) of 43 dog samples, respectively. None of the dog blood samples were positive for Borrelia burgdorferi and Dirofilaria immitis. DNA of E. canis and A. phagocytophilum was detected in 12 (25.5%) and two (4.3%) of 47 dog blood samples, and 17 (51.5%) and one (3.0%) of 33 R. sanguineus ticks, respectively. Additionally, DNA of Ehrlichia spp. closely related to Ehrlichia chaffeensis was detected in two (6.1%) R. sanguineus ticks. This study highlights the prevalence of anaplasmosis and ehrlichiosis in dogs in Malaysia. Due to the zoonotic potential of Ehrlichia and Anaplasma spp., appropriate measures should be instituted for prevention and control of vector-borne diseases in dogs.

  17. A loop-mediated isothermal amplification (LAMP) assay targeting 16S rRNA gene for rapid detection of Anaplasma phagocytophilum infection in sheep and goats.

    Science.gov (United States)

    Ning, Changshen; Wang, Jinhong; Zhang, Yan; Wang, Xiaoxing; Cui, Yanyan; Yan, Yaqun; Wang, Rongjun; Jian, Fuchun; Zhang, Longxian

    2017-01-24

    Anaplasma phagocytophilum is a zoonotic pathogen and the causative agent of human granulocytic anaplasmosis (HGA) in humans and tick-borne fever in various kinds of animals. In the present study, a loop-mediated isothermal amplification (LAMP) assay for rapid detection of A. phagocytophilum was developed using primers specific to 16S rRNA gene of this organism. The LAMP assay was performed at 65 C for 60 min and terminated at 80 C for 10 min. The optimal reaction conditions, under which no cross-reaction was observed with other closely related tick borne parasites (Anaplasma bovis, Anaplasma ovis, Theileria luwenshuni, Babesia motasi and Schistosoma japonicum) was established. The assay exhibited much higher sensitivity when compared with conventional PCR (1 copy vs 1000 copies). To evaluate the applicability of the LAMP assay, 94 sheep field blood samples were analyzed for A. phagocytophilum infection using LAMP, nested PCR and conventional PCR assay at the same time. A positive LAMP result was obtained from 53 of the 94 samples (56.4%), while only 12 (12.8%) and 3 (3.2%) were tested positive by nested PCR and conventional PCR, respectively. In conclusion, this LAMP assay is a specific, sensitive, and rapid method for the detection of A. phagocytophilum in sheep.

  18. Ehrlichia canis and Anaplasma platys in ticks of dogs in Cuiaba, Mato GrossoEhrlichia canis e Anaplasma platys em carrapatos de cães de Cuiabá, Mato Grosso

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    Valéria Dutra

    2012-06-01

    Full Text Available Diseases transmitted by arthropods such as Rhipicephalus sanguineus, are caused by a spectrum of pathogens. Among these are the canine monocytic ehrlichiosis and cyclical thrombocytopenia with a cosmopolitan distribution. Aiming to verify the presence of DNA of Anaplasma platys and Ehrlichia canis in ticks R. sanguineus collected in the period 2008 to 2009 of 380 infected dogs. Ticks, after maceration, were subjected to DNA extraction and then nested PCR was performed for amplification of A. platys and E. canis. Of these, 81 (29.7% amplified DNA from ehrlichiais agents, where 38 (17.9% amplified in E. canis and 32 (15.7% for A. platys. The observation of two pathogens, combined with worldwide distribution of the tick R. sanguineus, demonstrates the high risk of infection with these pathogens in dogs in the city of Cuiaba. Doenças transmitidas por artrópodes, como o Rhipicephalus sanguineus, são causadas por um espectro de patógenos. Dentre estas, estão a erliquiose monocítica canina e trombocitopenia cíclica com distribuição cosmopolita. Com o objetivo de verificar a presença de DNA de Anaplasma platys e Ehrlichia canis em carrapatos R. sanguineus coletados no período de 2008 a 2009 de 380 cães infestados. Os carrapatos, após a maceração, foram submetidos a extração de DNA e, em seguida, foi realizada a Nested PCR para a amplificação da espécie A. platys e E. canis. Destes, 81 (29.7% amplificaram o DNA dos agentes ehrlichiais, onde 38 (17.9% amplificaram para E. canis e 32 (15.7% para A. platys. A observação dos dois patógenos, combinado com distribuição mundial do carrapato R. sanguineus, demonstra o elevado risco de infecção por esses patógenos de cães na cidade de Cuiabá.

  19. Prevalence and molecular analysis of Anaplasma platys in dogs in Lara, Venezuela Prevalência e análise molecular de Anaplasma platys em cães da Venezuela

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    Haibin Huang

    2005-09-01

    Full Text Available Blood specimens from clinically normal military dogs and their trainers, in Lara, Venezuela were screened for Anaplasma platys, A. phagocytophilum, or Ehrlichia ewingii using 16S rRNA PCR tests. Sixteen percent (7/43 of dog specimens were positive by A. platys PCR test followed by sequencing of the PCR products, and all human blood specimens [25] were negative. All specimens from these dogs and humans were PCR negative for E. ewingii or A. phagocytophilum. Twelve Rhipicephalus sanguineus ticks removed from these dogs were negative for A. platys by reverse transcription PCR test. Almost the entire 16S rRNA gene (1,364 bp and groESL operon (1,646 bp sequences of A. platys isolated from a dog were determined, revealing that both sequences were closely related to the sequences of an A. platys strain detected in R. sanguineus ticks from the Democratic Republic of Congo.Amostras de sangue coletadas de cães clinicamente sadios pertencentes ao exército da Venezuela e de seus treinadores foram analisadas pela técnica de PCR 16S rRNA específica para Anaplasma platys, A. phagocytophilum ou Ehrlichia ewingii. Dezesseis por cento (7/43 dos cães foram positivos, enquanto que todas as amostras de origem humana [25] foram negativas para A. platys. Todas as amostras, tanto de humanos quanto de caninos, foram negativas para E. ewingii ou A. phagocytophilum. Doze carrapatos da espécie Rhipicephalus sanguineus, coletados dos cães, foram negativos para A. platys pelo teste de PCR de transcrição reversa. As seqüênciasquase inteiras do gene 16S rRNA (1.364 pb e do operon groESL (1.646 pb de A. platys isolado de um cão foram determinadas, revelando que ambas as seqüências estão estreitamente relacionadas às seqüências de A. platys detectadas em carrapatos R. sanguineus na República Democrática do Congo.

  20. Molecular detection and genetic identification of Babesia bigemina, Theileria annulata, Theileria orientalis and Anaplasma marginale in Turkey.

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    Zhou, Mo; Cao, Shinuo; Sevinc, Ferda; Sevinc, Mutlu; Ceylan, Onur; Moumouni, Paul Franck Adjou; Jirapattharasate, Charoonluk; Liu, Mingming; Wang, Guanbo; Iguchi, Aiko; Vudriko, Patrick; Suzuki, Hiroshi; Xuan, Xuenan

    2016-02-01

    Babesia spp., Theileria spp. and Anaplasma spp. are significant tick-borne pathogens of livestock globally. In this study, we investigated the presence and distribution of Babesia bigemina, Theileria annulata, Theileria orientalis and Anaplasma marginale in cattle from 6 provinces of Turkey using species-specific PCR assays. The PCR were conducted using the primers based on the B. bigemina rhoptry-associated protein 1a (BbiRAP-1a), T. annulata merozoite surface antigen-1 (Tams-1), T. orientalis major piroplasm surface protein (ToMPSP) and A. marginale major surface protein 4 (AmMSP4) genes, respectively. Fragments of B. bigemina internal transcribed spacer (BbiITS), T. annulata internal transcribed spacer (TaITS), ToMPSP and AmMSP4 genes were sequenced for phylogenetic analysis. PCR results revealed that the overall infections of A. marginale, T. annulata, B. bigemina and T. orientalis were 29.1%, 18.9%, 11.2% and 5.6%, respectively. The co-infection of two or three pathogens was detected in 29/196 (15.1%) of the cattle samples. The results of sequence analysis indicated that BbiRAP-1a, BbiITS, Tams-1, ToMPSP and AmMSP4 were conserved among the Turkish samples, with 99.76%, 99-99.8%, 99.34-99.78%, 96.9-99.61% and 99.42-99.71% sequence identity values, respectively. In contrast, the Turkish TaITS gene sequences were relatively diverse with 92.3-96.63% identity values. B. bigemina isolates from Turkey were found in the same clade as the isolates from other countries in phylogenetic analysis. On the other hand, phylogenetic analysis based on T. annulata ITS sequences revealed significant differences in the genotypes of T. annulata isolates from Turkey. Additionally, the T. orientalis isolates from Turkish samples were classified as MPSP type 3 genotype. This is the first report of type 3 MPSP in Turkey. Moreover, AmMSP4 isolates from Turkey were found in the same clade as the isolates from other countries. This study provides important data for understanding the

  1. The first clinical and laboratory evidence of co-infection by Anaplasma phagocytophilum and Ehrlichia canis in a Brazilian dog.

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    Silveira, Júlia A G; Valente, Pâmela C L G; Paes, Paulo R O; Vasconcelos, Artur V; Silvestre, Bruna T; Ribeiro, Múcio F B

    2015-04-01

    Information on Anaplasma phagocytophilum in Brazil is very restricted. The aim of this study was to report clinical, parasitological, hematological and molecular evidence of a natural A. phagocytophilum infection of an urban Brazilian dog. The dog was an eight-month-old male French bulldog. Veterinary clinical examinations were performed three times: in April, June and December 2013. Biochemical and hematological analyses were performed during all examinations, and blood samples were collected for parasitological surveys in June and December. Morulae were present within neutrophils in blood smears from June. Both samples were PCR positive for A. phagocytophilum and Ehrlichia spp. Phylogenetic analysis revealed that the phylogenetic topology placed samples from this study in close proximity to other A. phagocytophilum isolates. Ehrlichia isolates from this dog were 100% identical to E. canis isolates, thus E. canis and A. phagocytophilum co-infection was diagnosed in this dog. Lethargy and skin lesions were the clinical signs observed in this dog. Abnormal hematological parameters, among those, severe thrombocytopenia, were observed in all three occasions. This finding highlights the growing importance of A. phagocytophilum in South America.

  2. Infection of water buffalo in Rio de Janeiro Brazil with Anaplasma marginale strains also reported in cattle.

    Science.gov (United States)

    Silva, Jenevaldo B; Cabezas-Cruz, Alejandro; Fonseca, Adivaldo H; Barbosa, José D; de la Fuente, José

    2014-10-15

    Anaplasma marginale is the most prevalent pathogen of cattle in tropical and subtropical regions of the world and causes the disease bovine anaplasmosis. The importance of water buffalo in the world economy is increasing. In addition, while water buffalo may serve as a reservoir host for A. marginale, the susceptibility of this host for A. marginale cattle strains in Brazil has not been reported. The major surface protein 1 alpha (msp1α) gene has been shown to be a stable genetic marker for identification of A. marginale strains. Herein, we analyzed blood samples from 200 water buffalo and identified the A. marginale strains in an endemic area of Rio de Janeiro, Brazil, where ticks were present and water buffalo and cattle co-mingled. Ticks that were feeding on the study buffalo were collected and identified. The prevalence of A. marginale in water buffalo in this study was low (10%). Sequence analysis of the msp1α gene demonstrated the presence of 8 different A. marginale strains. Two A. marginale strains in the water buffalo, (α-β-β-β-Γ) and (α-β-β-Γ), were similar to those reported in cattle from nearby regions. The results of this study suggested that water buffalo in this region are naturally infected with the same strains of A. marginale found in cattle.

  3. Knockdown of the Rhipicephalus microplus cytochrome c oxidase subunit III gene is associated with a failure of Anaplasma marginale transmission.

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    Thais D Bifano

    Full Text Available Rhipicephalus microplus is an obligate hematophagous ectoparasite of cattle and an important biological vector of Anaplasma marginale in tropical and subtropical regions. The primary determinants for A. marginale transmission are infection of the tick gut, followed by infection of salivary glands. Transmission of A. marginale to cattle occurs via infected saliva delivered during tick feeding. Interference in colonization of either the tick gut or salivary glands can affect transmission of A. marginale to naïve animals. In this study, we used the tick embryonic cell line BME26 to identify genes that are modulated in response to A. marginale infection. Suppression-subtractive hybridization libraries (SSH were constructed, and five up-regulated genes {glutathione S-transferase (GST, cytochrome c oxidase sub III (COXIII, dynein (DYN, synaptobrevin (SYN and phosphatidylinositol-3,4,5-triphosphate 3-phosphatase (PHOS} were selected as targets for functional in vivo genomic analysis. RNA interference (RNAi was used to determine the effect of tick gene knockdown on A. marginale acquisition and transmission. Although RNAi consistently knocked down all individually examined tick genes in infected tick guts and salivary glands, only the group of ticks injected with dsCOXIII failed to transmit A. marginale to naïve calves. To our knowledge, this is the first report demonstrating that RNAi of a tick gene is associated with a failure of A. marginale transmission.

  4. Detection and prevalence of Anaplasma phagocytophilum and Rickettsia helvetica in Ixodes ricinus ticks in seven study areas in Sweden

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    Falk Kerstin

    2010-08-01

    Full Text Available Abstract Background Tick-borne Anaplasma phagocytophilum and Rickettsia spp. are considered to be emerging human pathogens, but only limited data are available on their occurrence in Sweden. Two real-time PCR assays followed by nested PCR and sequence analysis were carried out to investigate the prevalence of A. phagocytophilum and spotted fever rickettsiae in ticks from seven areas in Sweden. Results In 139 pooled samples, representing a total of 1245 Ixodes ricinus ticks (204 larvae, 963 nymphs, 38 males, 40 females, the overall positive mean infection prevalence was 1.3-15.0% for A. phagocytophilum and 1.5-17.3% for R. helvetica. A. phagocytophilum was only detected in nymphs (1.7-19.4%, whereas R. helvetica was detected in all three tick stages. Support for vertical and transstadial transmission was only obtained for R. helvetica. Both agents showed similar infection rates across study areas, although infection rates were greater in coastal areas. Conclusions The results show that both pathogens occurred in all seven locations, indicating that they are prevalent in Sweden and should be considered etiological agents in patients recently bitten by ticks.

  5. Evaluation of an indirect enzyme-linked immunosorbent assay (ELISA) for the detection of antibodies against Anaplasma phagocytophilum in sheep.

    Science.gov (United States)

    Woldehiwet, Z; Yavari, C

    2012-01-01

    An indirect enzyme-linked immunosorbent assay (ELISA) for detection of antibodies against Anaplasma phagocytophilum in ovine serum samples was evaluated. The assay used purified A. phagocytophilum grown in tick cell cultures as antigen. Serum samples were diluted 1 in 200 and binding was detected with anti-sheep IgG conjugated to horseradish peroxidase. All tests were carried out in the presence of positive and negative control samples. Optical density (OD) values obtained for each test sample at 490 nm were used to calculate percentage positivity (PP) of each sample based on the ratio of the OD of the test sample that of the positive reference sample. Known negative samples (n=69) obtained from uninfected sheep bred and maintained in a tick-free environment and subsequently shown to be susceptible to A. phagocytophilum were used to establish the cut-off point between negative and positive samples and to establish the specificity of the test. Serum samples obtained from 92 animals 14-21 days after infection were used to establish the sensitivity of the test. Using a cut-off point of 20PP (mean+2 standard deviations of the PP of 69 control samples) the test was shown to have a sensitivity of 84.8% and a specificity of 95.7%. Lowering the cut-off point to 15PP increased the sensitivity to 94.6%, but reduced the specificity to 92.8%.

  6. Molecular characterization and specific detection of Anaplasma species (AP-sd) in sika deer and its first detection in wild brown bears and rodents in Hokkaido, Japan.

    Science.gov (United States)

    Moustafa, Mohamed Abdallah Mohamed; Lee, Kyunglee; Taylor, Kyle; Nakao, Ryo; Sashika, Mariko; Shimozuru, Michito; Tsubota, Toshio

    2015-12-01

    A previously undescribed Anaplasma species (herein referred to as AP-sd) has been detected in sika deer, cattle and ticks in Japan. Despite being highly similar to some strains of A. phagocytophilum, AP-sd has never been detected in humans. Its ambiguous epidemiology and the lack of tools for its specific detection make it difficult to understand and interpret the prevalence of this Anaplasma species. We developed a method for specific detection, and examined AP-sd prevalence in Hokkaido wildlife. Our study included 250 sika deer (Cervus nippon yesoensis), 13 brown bears (Ursus arctos yesoensis) and 252 rodents including 138 (Apodemus speciosus), 45 (Apodemus argenteus), 42 (Myodes rufocanus) and 27 (Myodes rutilus) were collected from Hokkaido island, northern Japan, collected during 2010 to 2015. A 770 bp and 382 bp segment of the 16S rRNA and gltA genes, respectively, were amplified by nested PCR. Results were confirmed by cloning and sequencing of the positive PCR products. A reverse line blot hybridization (RLB) based on the 16S rRNA gene was then developed for the specific detection of AP-sd. The prevalence of AP-sd by nested PCR in sika deer was 51% (128/250). We detected this Anaplasma sp. for the first time in wild brown bears and rodents with a prevalence of 15% (2/13) and 2.4% (6/252), respectively. The sequencing results of the 16S rRNA and gltA gene amplicons were divergent from the selected A. phagocytophilum sequences in GenBank. Using a newly designed AP-sd specific probe for RLB has enabled us to specifically detect this Anaplasma species. Besides sika deer and cattle, wild brown bears and rodents were identified as potential reservoir hosts for AP-sd. This study provided a high throughput molecular method that specifically detects AP-sd, and which can be used to investigate its ecology and its potential as a threat to humans in Japan.

  7. Occurrence of Ehrlichia canis and Anaplasma platys in household dogs from northern Parana Ocorrência de Ehrlichia canis e Anaplasma platys em cães domiciliados da região norte do Paraná

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    Gislaine Cristina Ferreira da Silva

    2012-12-01

    Full Text Available Canine monocytic ehrlichiosis caused primarily by Ehrlichia canis and canine thrombocytic anaplasmosis induced by Anaplasma platys are important emerging zoonotic tick-borne diseases of dogs. There is evidence that these pathogens can also affect humans. This study evaluated the presence of E. canis and A. platys in blood samples collected from 256 domiciled dogs in the municipality of Jataizinho, located in north region of the State of Parana, Brazil, by PCR assay. The occurrence of E. canis and A. platys was 16.4% (42/256 and 19.4% (49/256, respectively; while 5.47% (14/256 of the dogs evaluated were co-infected by these two organisms. The presence of E. canis and A. platys was not significantly associated with the variables evaluated (sex, age, outdoor access, and presence of ticks during blood collection. Infection of dogs by E. canis was associated with anemia and thrombocytopenia, while infection induced by A. platys was related only to thrombocytopenia. Canine monocytic ehrlichiosis and canine thrombocytic anaplasmosis should be included in the differential diagnoses when these hematological alterations are observed during routine laboratory evaluation of dogs.Erliquiose monocítica canina, causada principalmente por Ehrlichia canis, e anaplasmose trombocítica canina, devida a infecção com Anaplasma platys, são importantes doenças transmitidas por carrapatos que acometem os cães, com evidências que podem também acometer o homem. O presente estudo avaliou a ocorrência desses agentes em amostras de sangue de 256 cães domiciliados na cidade de Jataizinho, na região Norte do Paraná, Brasil, utilizando a técnica da Reação em Cadeia da Polimerase (PCR. A ocorrência de E. canis e A. platys foi de 16,4% (42/256 e 19,4% (49/256, respectivamente, com 5,47% (14/256 dos animais apresentando coinfecção. Não foi observada associação significativa com as variáveis sexo, idade, acesso à rua e presença de carrapatos no momento da

  8. Comparação entre diversos antígenos para o diagnóstico de Anaplasma marginale por ELISA Comparison between several antigens for diagnosis of Anaplasma marginale by ELISA

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    Carlos A.N. Ramos

    2010-01-01

    Full Text Available Anaplasmose bovina é uma doença com grande importância nas regiões tropicais e subtropicais do mundo por determinar perdas econômicas devido à mortalidade e redução da produtividade. É causada por Anaplasma marginale, uma riquétsia intraeritrocítica obrigatória cujo controle requer, além de uma vacina eficiente, uma acurada identificação de bovinos cronicamente infectados. Apesar de existirem atualmente diversos métodos de diagnóstico dessa riquétsia, os métodos sorológicos, em particular o ensaio de imunoadsorção enzimática-ELISAs, são os mais utilizados devido à sua versatilidade e praticidade. No entanto, devido ao grande número de antígenos disponíveis, atualmente torna-se necessária uma avaliação para definir quais antígenos apresentam um melhor desempenho no diagnóstico da anaplasmose. Soros de bovinos positivos e negativos para A. marginale por PCR, e soros de animais provenientes do Brasil e Costa Rica, foram testados em ELISAs baseados em MSP1a, MSP2 e MSP5 recombinantes, um pool das três proteínas recombinantes, e antígeno de lisado de corpúsculos iniciais da riquétsia (CI. Utilizando soro de bovinos positivos para A. marginale por PCR, uma maior sensibilidade foi observada no ELISA CI. No entanto, uma maior especificidade, com soro de bovinos negativos a PCR, foi observada com os ELISAs recombinantes. O porcentual de bovinos positivos do Brasil e Costa Rica foi maior com ELISA CI. Razões para essas diferenças são discutidas.Bovine anaplasmosis is a major disease in tropical and subtropical regions of the world by determine economical loss due mortality and productive reduction. The disease is caused by Anaplasma marginale, an intraerythrocytic rickettsia whose control requires, besides an efficient vaccine, the accurate identification of chronically infected cattle. Although the existence of diverse methods of diagnosis of this rickettsia, the serological methods, in particular the enzyme

  9. Genetic variants of Anaplasma phagocytophilum in wild caprine and cervid ungulates from the Alps in Tyrol, Austria.

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    Silaghi, Cornelia; Hamel, Dietmar; Thiel, Claudia; Pfister, Kurt; Passos, Lygia Maria Friche; Rehbein, Steffen

    2011-04-01

    The occurrence of genetic variants of Anaplasma phagocytophilum was studied in wild ungulates from the northern and central eastern Alps in Tyrol, Austria. For this purpose, spleen samples collected from 53 game animals during the hunting season 2008/2009 (16 roe deer [Capreolus capreolus], 10 red deer [Cervus elaphus], 16 Alpine chamois [Rupicapra r. rupicapra], 7 Alpine ibex [Capra i. ibex], and 4 European mouflons [Ovis orientalis musimon]) were analyzed. Thirty-five animals originated from the Karwendel mountains, 12 from the Kaunertal area (Ötztal Alps), and the remaining from other mountainous areas in Tyrol. DNA extracts were screened with a real-time polymerase chain reaction targeting the msp2 gene of A. phagocytophilum. A total of 23 (43.4%) samples, from all ungulate species studied, were A. phagocytophilum positive. As of the date of this article, A. phagocytophilum has not been reported in the Alpine ibex. The positive samples were investigated further with polymerase chain reactions for amplification of the partial 16S rRNA, groEL, and msp4 genes. Sequence analysis using forward and reverse primers revealed seven different 16S rRNA gene variants. No variant could be attributed to any particular ungulate species. The groEL gene revealed 11 different variants, which grouped in the phylogenetic analysis into two distinct clusters: one cluster contained the sequences from roe deer, whereas the sequences of the other species formed the second cluster. The msp4 gene showed a high degree of variability in the amplified part with a total of 10 different sequence types. The results show that the wild mountain ungulates were infected to a considerable extent with various variants of A. phagocytophilum. The pathogenicity of the variants and the reservoir competence of the species investigated in this study deserve further attention in future studies.

  10. Development of a multilocus sequence typing scheme for the study of Anaplasma marginale population structure over space and time.

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    Guillemi, Eliana C; Ruybal, Paula; Lia, Verónica; Gonzalez, Sergio; Lew, Sergio; Zimmer, Patricia; Lopez Arias, Ludmila; Rodriguez, Jose L; Rodriguez, Sonia Y; Frutos, Roger; Wilkowsky, Silvina E; Farber, Marisa D

    2015-03-01

    Bovine Anaplasmosis caused by Anaplasma marginale is a worldwide disease prevalent in tropical and subtropical regions where Rhipicephalus microplus is considered the most significant biological vector. Molecular markers previously applied for A. marginale typing are efficient for isolate discrimination but they are not a suitable tool for studying population structure and dynamics. Here we report the development of an MLST scheme based on the study of seven genes: dnaA, ftsZ, groEl, lipA, recA, secY and sucB. Five annotated genomes (Saint Maries, Florida, Mississippi, Puerto Rico and Virginia) and 53 bovine blood samples from different world regions were analyzed. High nucleotide diversity and a large proportion of synonymous substitutions, indicative of negative selection resulted from DnaSP 5.00.02 package application. Recombination events were detected in almost all genes, this evidence together with the coexistence of more than one A. marginale strain in the same sample might suggest the superinfection phenomena as a potential source of variation. The allelic profile analysis performed through GoeBURST shown two main CC that did not support geography. In addition, the AMOVA test confirmed the occurrence of at least two main genetically divergent groups. The composition of the emergent groups reflected the impact of both historical and environmental traits on A. marginale population structure. Finally, a web-based platform "Galaxy MLST-Pipeline" was developed to automate DNA sequence editing and data analysis that together with the Data Base are freely available to users. The A. marginale MLST scheme developed here is a valuable tool with a high discrimination power, besides PCR based strategies are still the better choice for epidemiological intracellular pathogens studies. Finally, the allelic profile describe herein would contribute to uncover the mechanisms in how intracellular pathogens challenge virulence paradigm.

  11. Infection of Ixodes spp. tick cells with different Anaplasma phagocytophilum isolates induces the inhibition of apoptotic cell death.

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    Alberdi, Pilar; Ayllón, Nieves; Cabezas-Cruz, Alejandro; Bell-Sakyi, Lesley; Zweygarth, Erich; Stuen, Snorre; de la Fuente, José

    2015-09-01

    Anaplasma phagocytophilum is an intracellular rickettsial pathogen transmitted by Ixodes spp. ticks, which causes granulocytic anaplasmosis in humans, horses and dogs and tick-borne fever (TBF) in ruminants. In the United States, human granulocytic anaplasmosis (HGA) is highly prevalent while TBF has not been reported. However, in Europe the situation is the opposite, with high prevalence for TBF in sheep and low prevalence of HGA. The origin of these differences has not been identified and our hypothesis is that different A. phagocytophilum isolates impact differently on tick vector capacity through inhibition of apoptosis to establish infection of the tick vector. In this study we used three different isolates of A. phagocytophilum of human, canine and ovine origin to infect the Ixodes ricinus-derived cell line IRE/CTVM20 and the Ixodes scapularis-derived cell line ISE6 in order to characterize the effect of infection on the level of tick cell apoptosis. Inhibition of apoptosis was observed by flow cytometry as early as 24h post-infection for both tick cell lines and all three isolates of A. phagocytophilum, suggesting that pathogen infection inhibits apoptotic pathways to facilitate infection independently of the origin of the A. phagocytophilum isolate and tick vector species. However, infection with A. phagocytophilum isolates inhibited the intrinsic apoptosis pathway at different levels in I. scapularis and I. ricinus cells. These results suggested an impact of vector-pathogen co-evolution on the adaptation of A. phagocytophilum isolates to grow in tick cells as each isolate grew better in the tick cell line derived from its natural vector species. These results increase our understanding of the mechanisms of A. phagocytophilum infection and multiplication and suggest that multiple mechanisms may affect disease prevalence in different geographical regions.

  12. Systems biology of tissue-specific response to Anaplasma phagocytophilum reveals differentiated apoptosis in the tick vector Ixodes scapularis.

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    Nieves Ayllón

    2015-03-01

    Full Text Available Anaplasma phagocytophilum is an emerging pathogen that causes human granulocytic anaplasmosis. Infection with this zoonotic pathogen affects cell function in both vertebrate host and the tick vector, Ixodes scapularis. Global tissue-specific response and apoptosis signaling pathways were characterized in I. scapularis nymphs and adult female midguts and salivary glands infected with A. phagocytophilum using a systems biology approach combining transcriptomics and proteomics. Apoptosis was selected for pathway-focused analysis due to its role in bacterial infection of tick cells. The results showed tissue-specific differences in tick response to infection and revealed differentiated regulation of apoptosis pathways. The impact of bacterial infection was more pronounced in tick nymphs and midguts than in salivary glands, probably reflecting bacterial developmental cycle. All apoptosis pathways described in other organisms were identified in I. scapularis, except for the absence of the Perforin ortholog. Functional characterization using RNA interference showed that Porin knockdown significantly increases tick colonization by A. phagocytophilum. Infection with A. phagocytophilum produced complex tissue-specific alterations in transcript and protein levels. In tick nymphs, the results suggested a possible effect of bacterial infection on the inhibition of tick immune response. In tick midguts, the results suggested that A. phagocytophilum infection inhibited cell apoptosis to facilitate and establish infection through up-regulation of the JAK/STAT pathway. Bacterial infection inhibited the intrinsic apoptosis pathway in tick salivary glands by down-regulating Porin expression that resulted in the inhibition of Cytochrome c release as the anti-apoptotic mechanism to facilitate bacterial infection. However, tick salivary glands may promote apoptosis to limit bacterial infection through induction of the extrinsic apoptosis pathway. These dynamic

  13. Cattle experimentally infected by Anaplasma marginale: Influence of splenectomy on disease pathogenesis, oxidative profile, and antioxidant status.

    Science.gov (United States)

    Doyle, Rovaina L; França, Raqueli T; Oliveira, Camila B; Rezer, João F P; Klafke, Guilherme M; Martins, João R; Santos, Andrea P; do Nascimento, Naíla C; Mesick, Joanne B; Lopes, Sonia T A; Leal, Daniela B R; Da Silva, Aleksandro S; Andrade, Cinthia M

    2016-06-01

    Bovine anaplasmosis is caused by the obligate intraerythrocytic bacteria Anaplasma marginale. These bacteria are transmitted by tick species such as Rhipicephalus (Boophilus) microplus, blood-sucking insects, and fomites (needles, clippers, and other blood contaminated equipment). During the acute phase of infection, animals may develop fever, anemia, jaundice, and hepatosplenomegaly. The aims of this study are to quantify the bacteremia by quantitative PCR in eight naïve calves experimentally infected by A. marginale [splenectomized (n = 4), and intact/non-splenectomized (n = 4)], and to correlate these findings with markers of oxidative stress on days 0, 8, 15, 21 and 23 post-infection. Complete blood counts (CBC) were performed in both groups. Lipid peroxidation was estimated by quantifying thiobarbituric acid reactive substances (TBARS); and non-enzymatic antioxidants were assessed by erythrocyte content of non-protein thiols (NPSH). There were no significant differences in complete blood counts (CBC) between the two groups. However, both groups had a slight decrease on packet cell volume (PCV), erythrocytes and hemoglobin concentration, as well as an increase in total leukocyte counts due to elevated lymphocytes when comparing pre and post-infection with A. marginale. Progressive increase on TBARS levels and concomitant decrease on NPSH content were observed in all animals, without significant differences between splenectomized and intact animals. A positive correlation between bacteremia and TBARS, and a negative correlation between bacteremia and NPSH were observed in both groups with higher correlation for NPSH in splenectomized animals. A negative correlation between TBARS and NPSH levels was observed in both groups indicating lipid peroxidation without a non-enzymatic antioxidant response. The results of experimental infection by A. marginale in cattle showed that bacteremia has an impact on lipid peroxidation regardless of the splenectomy.

  14. Monoinfections caused by Borrelia burgdorferi and Borrelia burgdorferi / Anaplasma phagocytophilum co-infections in forestry workers and farmers

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    Małgorzata Tokarska-Rodak

    2015-10-01

    Full Text Available Background: The presence of co-infections induced by tick-borne pathogens in humans is an important epidemiological phenomenon. This issue has attracted growing attention of doctors and people working under conditions of an increased risk of being exposed to tick bites. Material and Methods: The research group consisted of 93 individuals with current anti-immunoglobulin M/G (IgM/ IgG Borrelia burgdorferi or IgG anti-Anaplasma phagocytophilum. The respondents were identified during the screening survey in a group of farmers and foresters occupationally exposed to tick bites. The aim of the work was to analyse the frequency of antibodies to specific antigens of B. burgdorferi and the levels of cytokines in forestry workers and farmers with B. burgdorferi monoinfections and B. burgdorferi / A. phagocytophilum co-infections. Statistical analysis was performed using the Chi2, Mann-Whitney U and Kruskal-Wallis tests. Results: There is a stronger generation of IgG antibodies to B. burgdorferi antigens in patients with B. burgdorferi / A. phagocytophilum co-infections, such as variable major protein-like sequence expressed (VlsE (p < 0.05, p19 (p < 0.02, p17 (p < 0.05 and complement regulator-acquiring surface protein 3 (CRASP3 (p < 0.02 compared to persons with B. burgdorferi monoinfections. The discrepancies in the synthesis of cytokines interleukin 6 (IL-6, IL-10, and tumor necrosis factor α (TNF-α have not been found in persons with B. burgdorferi monoinfections and B. burgdorferi / A. phagocytophilum co-infection. Conclusions: The immune response directed against B. burgdorferi is stronger in patients co-infected with B. burgdorferi and A. phagocytophilum than in those with monoinfection. Med Pr 2015;66(5:645–651

  15. The incidence of Borrelia burgdorferi, Anaplasma phagocytophilum and Babesia microti coinfections among foresters and farmers in eastern Poland

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    Anna Pañczuk

    2016-01-01

    Full Text Available Background & objectives: Lyme borreliosis is the most common tick-borne disease in Europe and the USA. However, a great variety of pathogens are transmitted by ticks, which results in mixed infections, with Lyme borreliosis. The aim of the present study was to show the incidence of Borrelia burgdorferi, Anaplasma phagocytophilum, and Babesia microti coinfections among the population of foresters and farmers, as these people, due to their profession, are particularly exposed to tick contact. Methods: The study was carried out in eastern Poland (the northern part of the Lublin Province in 2013. The study was performed in a group of 93 individuals occupationally exposed to tick bites (foresters and farmers, whose blood serum showed the presence of IgG anti-B. burgdorferi antibodies. Blood serum in this group were evaluated for IgG anti-A. phagocytophilum and IgG anti-B. microti antibodies by means of IFA IgG indirect immunofluorescence tests. Information related to age, sex, number of tick bite episodes, presence of various symptoms related to the tick bites, and antibiotic therapy applied as treatment for diagnosed Lyme borreliosis were obtained from the subjects through a structured questionnaire. The results were analyzed in Statistica v. 7.1 statistical analysis software. Results: The presence of IgG antibodies against the analyzed pathogens revealed B. burgdorferi and A. phagocytophilum coinfection in 26 (28% of the examined subjects and B. burgdorferi and B. microti coinfection in one person (1.1%. No coinfection with all the three pathogens was observed in any individual. The co-occurrence of headache plus bone, joint and muscle pain was noted significantly more often among individuals diagnosed with B. burgdorferi and A. phagocytophilum coinfection. Interpretation & conclusion: Foresters and farmers are exposed to B. burgdorferi and A. phagocytophilum coinfection in the study area. Therefore, it is probable that these pathogens may severely

  16. Anaplasma phagocytophilum infection in American robins and gray catbirds: an assessment of reservoir competence and disease in captive wildlife.

    Science.gov (United States)

    Johnston, Emily; Tsao, Jean I; Muñoz, Juan David; Owen, Jen

    2013-01-01

    Anaplasma phagocytophilum (Dumler et al.) is the bacterial agent of human granulocytic anaplasmosis, an emerging infectious disease. The main vector of A. phagocytophilum in the United States is the blacklegged tick (Ixodes scapularis (Say)) and various small and medium-sized mammals are reservoirs. Previous studies indicate that birds are exposed to A. phagocytophilum; however, because no studies have directly investigated avian susceptibility, reservoir competence, and morbidity for A. phagocytophilum, uncertainty remains as to what role birds could play in its transmission ecology. In a controlled laboratory study, we tested whether two species, the American robin (Turdus migratorius (L.)) and the gray catbird (Dumetella carolinensis (L.)), can become infected with and then transmit A. phagocytophilum to feeding ticks, and whether exposed birds develop disease. Wild caught, seronegative birds (n = 10 per species) were exposed to A. phagocytophilum-infected I. scapularis nymphs (day 0). Transmission was assessed by xenodiagnosis on days 7, 14, 42, and 77; blood was assayed for bacteremia and serology. A. phagocytophilum was detected using quantitative polymerase chain reaction targeting the 16s rRNA gene. One robin infected 2 of 13 larval ticks (15%) on day 7; no other birds were found to infect feeding ticks at any time. Birds did not develop bacteremia, specific antibodies or significant illness because of exposure. Mouse controls became bacteremic, infected feeding ticks, and seroconverted. Our results suggest that these two avian species are unlikely to play a significant role in the maintenance of the agent of human granulocytic anaplasmosis and that avian serosurveys may not be a reliable indicator of A. phagocytophilum exposure.

  17. Whole genome transcription profiling of Anaplasma phagocytophilum in human and tick host cells by tiling array analysis

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    Chavez Adela

    2008-07-01

    Full Text Available Abstract Background Anaplasma phagocytophilum (Ap is an obligate intracellular bacterium and the agent of human granulocytic anaplasmosis, an emerging tick-borne disease. Ap alternately infects ticks and mammals and a variety of cell types within each. Understanding the biology behind such versatile cellular parasitism may be derived through the use of tiling microarrays to establish high resolution, genome-wide transcription profiles of the organism as it infects cell lines representative of its life cycle (tick; ISE6 and pathogenesis (human; HL-60 and HMEC-1. Results Detailed, host cell specific transcriptional behavior was revealed. There was extensive differential Ap gene transcription between the tick (ISE6 and the human (HL-60 and HMEC-1 cell lines, with far fewer differentially transcribed genes between the human cell lines, and all disproportionately represented by membrane or surface proteins. There were Ap genes exclusively transcribed in each cell line, apparent human- and tick-specific operons and paralogs, and anti-sense transcripts that suggest novel expression regulation processes. Seven virB2 paralogs (of the bacterial type IV secretion system showed human or tick cell dependent transcription. Previously unrecognized genes and coding sequences were identified, as were the expressed p44/msp2 (major surface proteins paralogs (of 114 total, through elevated signal produced to the unique hypervariable region of each – 2/114 in HL-60, 3/114 in HMEC-1, and none in ISE6. Conclusion Using these methods, whole genome transcription profiles can likely be generated for Ap, as well as other obligate intracellular organisms, in any host cells and for all stages of the cell infection process. Visual representation of comprehensive transcription data alongside an annotated map of the genome renders complex transcription into discernable patterns.

  18. Toxoplasma gondii, Neospora caninum and tick-transmitted bacterium Anaplasma phagocytophilum infections in one selected goat farm in Slovakia.

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    Čobádiová, Andrea; Reiterová, Katarina; Derdáková, Markéta; Špilovská, Silvia; Turčeková, Ludmila; Hviščová, Ivana; Hisira, Vladimir

    2013-12-01

    Parasitic diseases of livestock together with poor welfare conditions can negatively affect the health status and production of small ruminants. Protozoan parasites and tick-borne infectious agents are common threat of livestock including small ruminants mostly during the pasture season. Therefore the priority of the study was to analyse the circulation and presence of two protozoan parasites Toxoplasma gondii and Neospora caninum as well as tick-transmitted bacterium Anaplasma phagocytophilum in one selected goat farm in Eastern Slovakia. Throughout a three-year study period we have repeatedly screened the sera and blood of goats and dogs from monitored farm. In total, 343 blood serum samples from 116 goats were examined by ELISA. The mean seropositivity for T. gondii was 56.9% (66/116, CI (95%) = 48-66.0) and 15.5% (18/116, CI (95%) = 9.3-22.7) for N. caninum. The permanent occurrence of anti-Toxoplasma and anti-Neospora antibodies was detected in repeatedly examined goats during the whole monitored period. The presence of both parasites in the flock was analysed by PCR. DNA of T. gondii was confirmed in 12 out of 25 Toxoplasma-seropositive goats and N. caninum in 14 samples out of 18 Neospora-seropositive animals; four goats were co-infected with both pathogens. The risk of endogenous transmission of both parasites was pursued by examination of 41 kid's sera, where seropositivity for toxoplasmosis was 31.7% and for neosporosis 14.6%. In dogs 61.1% seropositivity for T. gondii and 38.9% for N. caninum was found, however, their faeces were negative for coccidian oocysts. Eight out of 108 tested animals were infected with A. phagocytophilum, the causative agent of tick-borne fever. Seven of them were simultaneously infected with T. gondii and A. phagocytophilum, out of which four goats were concurrently infected with all three pathogens.

  19. Prevalence and haemato-biochemical profile ofAnaplasma marginale infection in dairy animals of Punjab (India)

    Institute of Scientific and Technical Information of China (English)

    Ashuma; Amrita Sharma; Lachhman Das Singla; Paramjit Kaur; Mandeep Singh Bal; Balwinder Kaur Batth; Prayag Dutt Juyal

    2013-01-01

    Objective:To do the systematic comparison of prevalence of anaplasmosis byPCR andGiemsa stained thin blood smear (GSTBS) based parasitological assays in dairy cattle ofPunjab, which has not been reported yet.To analyse the haematobiochemical alterations in infected animals to arrive at the conclusion regarding the pathogenicity induced byAnaplasma marginale (A. marginale) in latent and patent infection.Methods:Study was conducted on320 animals (236 cows,62 calves and22 buffaloes) ofPunjab,India.PCR on genome ofA. marginale was performed by targeting msp1β gene using specific primersBAP-2/AL34S, amplifies products of size407 bp.Questionnaires based data on the characteristics of the infected animals and management strategies of the farm were collected and correlated.Results:Higher prevalence and more significant association was observed in thePCR based molecular diagnosis (P=0.00012) as compared to that inGSTBS (P=0.0288) based diagnosis with various regions under study.With respect to the regions, highest prevalence was recorded inFerozepur byPCR based diagnosis, while that inJalandhar byGSTBS examination.Similar marked significant association of thePCR based diagnosis with the age of the animals under study (P=0.00013) was observed elucidating no inverse age resistance toA. marginale in cow calves.Haematobiochemical profile of infected animals revealed marked anemia, liver dysfunction and increase globulin concentrate indicating rise in immunoglobulin level to counteract infection.Conclusions:PCR is far more sensitive in detecting the disease even in latent infection which may act as nidus for spread of anaplasmosis to susceptible animals in endemic areas.Severity of anaemia and liver dysfunction were comparable both in patent as well as latent infection indicating pathogenicity of both.

  20. Molecular biological identification of Babesia, Theileria, and Anaplasma species in cattle in Egypt using PCR assays, gene sequence analysis and a novel DNA microarray.

    Science.gov (United States)

    El-Ashker, Maged; Hotzel, Helmut; Gwida, Mayada; El-Beskawy, Mohamed; Silaghi, Cornelia; Tomaso, Herbert

    2015-01-30

    In this preliminary study, a novel DNA microarray system was tested for the diagnosis of bovine piroplasmosis and anaplasmosis in comparison with microscopy and PCR assay results. In the Dakahlia Governorate, Egypt, 164 cattle were investigated for the presence of piroplasms and Anaplasma species. All investigated cattle were clinically examined. Blood samples were screened for the presence of blood parasites using microscopy and PCR assays. Seventy-one animals were acutely ill, whereas 93 were apparently healthy. In acutely ill cattle, Babesia/Theileria species (n=11) and Anaplasma marginale (n=10) were detected. Mixed infections with Babesia/Theileria spp. and A. marginale were present in two further cases. A. marginale infections were also detected in apparently healthy subjects (n=23). The results of PCR assays were confirmed by DNA sequencing. All samples that were positive by PCR for Babesia/Theileria spp. gave also positive results in the microarray analysis. The microarray chips identified Babesia bovis (n=12) and Babesia bigemina (n=2). Cattle with babesiosis were likely to have hemoglobinuria and nervous signs when compared to those with anaplasmosis that frequently had bloody feces. We conclude that clinical examination in combination with microscopy are still very useful in diagnosing acute cases of babesiosis and anaplasmosis, but a combination of molecular biological diagnostic assays will detect even asymptomatic carriers. In perspective, parallel detection of Babesia/Theileria spp. and A. marginale infections using a single microarray system will be a valuable improvement.

  1. Experimental infection and co-infection of dogs with Anaplasma platys and Ehrlichia canis: hematologic, serologic and molecular findings

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    Diniz PPVP

    2010-04-01

    Full Text Available Abstract Background Rhipicephalus sanguineus is a ubiquitous tick responsible for transmitting Ehrlichia canis and most likely Anaplasma platys to dogs, as either single or co-infections. The objective of this study was to assess the effects of either simultaneous or sequential experimental infections with E. canis and A. platys on hematological and serological parameters, duration of infection, and efficacy of doxycycline therapy in dogs infected with one or both organisms. Six dogs per group were either uninfected, A. platys infected, E. canis infected, A. platys and E. canis co-infected, A. platys infected and E. canis challenged or E. canis infected and A. platys challenged at day 112 post-infection (PI. Doxycycline treatment was initiated at 211 days PI, followed by dexamethasone immunosuppression beginning 410 days PI. Results Initially, transient decreases in hematocrit occurred in all groups infected with E. canis, but the mean hematocrit was significantly lower in the A. platys and E. canis co-infected group. All dogs except the controls developed marked thrombocytopenia after initial infection followed by gradually increased platelet counts by 112 days PI in groups with the single infections, while platelet counts remained significantly lower in the A. platys and E. canis co-infected group. Both sequential and simultaneous infections of A. platys and E. canis produced an enhanced humoral immune response to A. platys when compared to infection with A. platys alone. Likewise, co-infection with E. canis and A. platys resulted in a more persistent A. platys infection compared to dogs infected with A. platys only, but nearly all A. platys infected dogs became A. platys PCR negative prior to doxycycline treatment. E. canis infected dogs, whether single or co-infected, remained thrombocytopenic and E. canis PCR positive in blood for 420 days. When treated with doxycycline, all E. canis infected dogs became E. canis PCR negative and the

  2. Prevalence of Rickettsiales (Anaplasma phagocytophilum and Rickettsia spp.) in hard ticks (Ixodes ricinus) in the city of Hamburg, Germany.

    Science.gov (United States)

    May, Kathrin; Strube, Christina

    2014-06-01

    To narrow the gap of missing knowledge on Rickettsia spp. and Anaplasma phagocytophilum infections in ticks in northwestern Germany and, at the same time, to provide first prevalence data on these pathogens in the city of Hamburg, a total of 1,400 questing Ixodes ricinus ticks were collected at ten different public green areas from April until October 2011. Ticks were examined using probe-based quantitative real-time PCR. A percentage of 3.6% (51/1,400) ticks were tested positive for A. phagocytophilum infections divided into 2.1% (3/141) adults [1.7% (1/60) females and 2.5% (2/81) males] and 3.8% (48/1,259) nymphs. The percentage of infected ticks per sampling site varied statistically significantly from 0.7% (1/140) to 12.1% (17/140), whereas between sampling months, no statistically significant differences were observed (2.0-6.5%, 4-13/140). The overall Rickettsia spp. infection rate was 52.5% (735/1,400). In adult ticks, Rickettsia spp. infection rate was 56% (79/141) divided into 61.7% (37/60) infected females and 51.9% (42/81) infected males. Nymphs showed an infection rate of 52.1% (656/1,259). In contrast to A. phagocytophilum infections, no statistically significant differences in Rickettsia spp. infection rates among sampling sites (44.3-63.6%, 62-89/140) were observed, whereas seasonal variations were obvious: the percentage of Rickettsia-positive ticks was significantly lower in April (36.5%, 73/200) and May (29.5%, 59/200) compared to the summer and fall months (55.0-64.5%, 110-129/200). Rickettsia species differentiation via real-time pyrosequencing revealed Rickettsia helvetica as the only occurring species. Co-infections with both Rickettsia spp. and A. phagocytophilum were detected in 2.0% (28/1,400) of the ticks. The present study revealed that in the city of Hamburg, the tick infection rate with A. phagocytophilum is comparable with other German data, whereas the Rickettsia spp. infection rate of 52.5% is by far the highest prevalence detected in

  3. Molecular detection of Anaplasma platys, Ehrlichia canis, Hepatozoon canis and Rickettsia monacensis in dogs from Maio Island of Cape Verde archipelago.

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    Lauzi, Stefania; Maia, João P; Epis, Sara; Marcos, Ricardo; Pereira, Cristina; Luzzago, Camilla; Santos, Marta; Puente-Payo, Pablo; Giordano, Alessia; Pajoro, Massimo; Sironi, Giuseppe; Faustino, Augusto

    2016-07-01

    Tick-borne diseases are emerging worldwide and have an important zoonotic relevance. Dogs play an important role in the epidemiology of several zoonotic tick-borne pathogens acting as sentinels and/or reservoirs. This study focused on the molecular identification of tick-borne pathogens in blood samples of 153 autochthonous asymptomatic dogs in Maio Island, Cape Verde archipelago. Eighty-four (54.9%) dogs were positive for one or more pathogens. Fifty-five (35.9%) dogs were infected with Hepatozoon canis, 53 (34.6%) with Anaplasma platys, five (3.3%) with Ehrlichia canis and Rickettsia monacensis, an emerging human pathogen, was also identified in a single dog (0.7%). The former three pathogens cause important canine tick-borne diseases that are transmitted or potentially transmitted by Rhipicephalus sanguineus s.l., the only hard tick identified in Cape Verde. Furthermore, Wolbachia spp. was amplified from the blood of one dog. None of the dogs were positive for Anaplasma phagocytophilum, Borrelia burgdorferi sensu lato, Midichloria mitochondrii, Bartonella spp., Babesia spp. or Theileria spp. Fifty-four (35.3%) animals showed single infections and 30 (19.6%) co-infections, with A. platys and H. canis co-infection being the most frequent (28 dogs, 18.3%). The frequency of E. canis infection was statistically different among age groups (P=0.017), being higher among dogs older than 4 years compared to younger dogs. Infection by A. platys was also statistically different among age groups (P=0.031), being higher in dogs younger than 2 years compared to older dogs. The statistical analyses showed no significant association of PCR positivity with gender or location. The frequency of tick-borne pathogens detected in dogs in Maio Island, including R. monacensis, highlights the need to improve diagnosis and control in order to prevent the risk of transmission of these pathogens among dogs and humans living in or travelling to this touristic island.

  4. Detection of Anaplasma antibodies in wildlife and domestic species in wildlife-livestock interface areas of Kenya by major surface protein 5 competitive inhibition enzyme-linked immunosorbent assay

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    J.J.N. Ngeranwa

    2008-09-01

    Full Text Available The seroprevalence of Anaplasma antibodies in wildlife (eland, blue wildebeest, kongoni, impala, Thomson's gazelle, Grant's gazelle, giraffe and plains zebra and domestic animal (cattle, sheep and goat populations was studied in wildlife / livestock interface areas of Kenya. Serum samples were analyzed by competitive inhibition enzyme-linked immunosorbent assay (CI-ELISA, using a recombinant antigen (MSP-5 from Anaplasma marginale surface membrane. A monoclonal antibody, FC-16, was used as the primary antibody, while anti-mouse conjugated to horseradish peroxidase was used as the secondary antibody. The results indicate a high seroprevalence in both wildlife and livestock populations, in contrast to earlier reports from Kenya, which indicated a low seroprevalence. The differences are attributed to the accurate analytical method used (CI-ELISA, as compared with agglutination techniques, clinical signs and microscopy employed by the earlier workers.

  5. Eficácia do dipropionato de imidocarb, da enrofloxacina e do cloridrato de oxitetraciclina no tratamento de bovinos naturalmente infectados por Anaplasma marginale

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    L.R. Alberton

    2015-08-01

    Full Text Available O agente de maior importância, em relação à anaplasmose bovina, é o Anaplasma marginale. Os principais sinais clínicos dessa enfermidade são anemia hemolítica, icterícia, dispneia, taquicardia, febre, fadiga, lacrimejamento, sialorreia, micção frequente, anorexia, perda de peso, aborto e morte. A terapia antimicrobiana é o principal protocolo terapêutico. O objetivo do presente trabalho foi avaliar a eficácia do dipropionato de imidocarb, da enrofloxacina e do cloridrato de oxitetraciclina no tratamento de bovinos leiteiros naturalmente infectados por Anaplasma marginale. Para isso, foram avaliados 48 zebuínos mestiços que apresentavam os sinais clínicos sugestivos da doença. Os animais foram submetidos à coleta de sangue para a realização de hemograma e à extração de DNA para a confirmação da presença de A. marginale, por meio da reação em cadeia pela polimerase (PCR. Os animais foram divididos em três grupos experimentais, para realização dos protocolos terapêuticos, utilizando-se dipropionato de imidocarb, oxitetraciclina e enrofloxacina. Trinta e seis animais (75% apresentaram reação positiva ao PCR. Os animais positivos não apresentaram diferenças significativas quanto ao hemograma e ao leucograma quando comparados com os negativos, no entanto os níveis de proteínas séricas foram inferiores nos animais positivos (P<0,05. Os três protocolos terapêuticos foram capazes de reduzir a infecção ao longo do tratamento (P<0,01, porém, após cinco dias de tratamento, a enrofloxacina apresentou maior efetividade em relação aos demais (P<0,01. Após o final do tratamento, nenhum protocolo foi capaz de eliminar totalmente a infecção pelo A. marginale em bovinos naturalmente infectados e manejados a campo.

  6. Real time polymerase chain reaction to diagnose Anaplasma marginale in cattle and deer (Ozotoceros bezoarticus leucogaster of the Brazilian Pantanal Reação da polimerase em cadeia tempo real para diagnóstico de Anaplasma marginale em bovino e veado campeiro do Pantanal brasileiro Ozotoceros bezoarticus leucogaster

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    Graziela Picoloto

    2010-09-01

    Full Text Available Epizootiological study of Anaplasma marginale in regions that contain various reservoir hosts, co-existence of rickettsia pathogens, and common vectors is a complicated task. To achieve diagnosis of this rickettsia in cattle and campeiro deer of Brazilian Pantanal, a comparison was made between a real time polymerase chain reaction (RT-PCR with intercalating Sybr Green fluorochrome and primers based on msp5 gene of A. marginale; a conventional PCR (C-PCR; and parasitological examination using thin blood smear stained with Giemsa-MayGrunwald. Both PCRs showed good performance in the diagnosis of A. marginale in cattle, and were superior to the parasitological exam. The RT-PCR detected seven positive campeiro deer (16.3%. This rate was significantly higher compared to C-PCR, which identified one animal as positive (2.3%, and also compared to parasitological diagnosis, which did not find any positive animals. The dissociation temperature average of positive reactions in cattle (81.72 ºC ± 0.20 was identical to dissociation temperature found in the cervids (81.72 ºC ± 0.12, suggesting that both animal species were infected with A. marginale. We concluded that RT-PCR can be used for A. marginale diagnosis and in epizootiological studies of cattle and cervids; in spite of the small number of campeiro deer samples, the results indicated that this wildlife species has importance in the Anaplasma epizootiology in the Brazilian Pantanal.O estudo epizootiológico de Anaplasma marginale em regiões que existem vários reservatórios, co-existência de espécies de riquétsias patógenas e vetores comuns é uma tarefa complicada. Com o objetivo de obter o diagnóstico dessa riquétsia em bovinos e veado campeiro do Pantanal brasileiro foi avaliada uma reação da polimerase em cadeia em tempo real (PCR-TR com o fluoróforo intercalante de fita dupla de DNA Sybr Green e iniciadores baseados na seqüência do gene msp5 de A. marginale comparando-a a uma

  7. Prevalence of antibodies against Rickettsia conorii, Babesia canis, Ehrlichia canis, and Anaplasma phagocytophilum antigens in dogs from the Stretto di Messina area (Italy).

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    Pennisi, Maria-Grazia; Caprì, Alessandra; Solano-Gallego, Laia; Lombardo, Gabriella; Torina, Alessandra; Masucci, Marisa

    2012-12-01

    The aims of this study were to determine the seroprevalence for Rickettsia conorii, Ehrlichia canis, Anaplasma phagocytophilum, and Babesia canis in outdoor-kennelled dogs (n=249) from the Stretto di Messina (Italy) and to compare seroprevalence in 2 public shelters and 4 privately-owned kennels where different tick-preventive measures were implemented in order to focus on the specific sanitary risk posed by public shelters in southern Italy for tick-borne pathogens. R. conorii (72%) and B. canis (70%) were the most prevalent infections when compared to E. canis (46%) and A. phagocytophilum (38%). Seroprevalence for R. conorii, E. canis, and A. phagocytophilum was significantly higher in public shelters than in private kennels. However, B. canis seropositivity was similar in both types of kennels. In addition, in private kennels where a regular ectocide treatment was carried out by means of spot-on devices, dogs did not present E. canis and A. phagocytophilum antibodies. One hundred fifty-one dogs out of 249 (61%) were seropositive to more than one pathogen with R. conorii and B. canis the most common ones. Coinfections were more frequently found in public-shelter dogs. This study demonstrated high seroprevalences against R. conorii, B. canis, E. canis, and A. phagocytophilum in kennelled dogs from both coastal sites of the Stretto di Messina and the importance of regular tick-bite prevention by means of individual spot-on devices.

  8. High throughput pyrosequencing technology for molecular differential detection of Babesia vogeli, Hepatozoon canis, Ehrlichia canis and Anaplasma platys in canine blood samples.

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    Kaewkong, Worasak; Intapan, Pewpan M; Sanpool, Oranuch; Janwan, Penchom; Thanchomnang, Tongjit; Kongklieng, Amornmas; Tantrawatpan, Chairat; Boonmars, Thidarut; Lulitanond, Viraphong; Taweethavonsawat, Piyanan; Chungpivat, Sudchit; Maleewong, Wanchai

    2014-06-01

    Canine babesiosis, hepatozoonosis, ehrlichiosis, and anaplasmosis are tick-borne diseases caused by different hemopathogens. These diseases are causes of morbidity and mortality in dogs. The classic method for parasite detection and differentiation is based on microscopic observation of blood smears. The limitations of the microscopic method are that its performance requires a specially qualified person with professional competence, and it is ineffective in differentiating closely related species. This study applied PCR amplification with high throughput pyrosequencing for molecular differential detection of the following 4 hemoparasites common to tropical areas in dog blood samples: Babesia vogeli, Hepatozoon canis, Ehrlichia canis, and Anaplasma platys. PCR was initially used to amplify specific target regions of the ribosomal RNA genes of each parasite using 2 primer pairs that included 18S rRNA for protozoa (B. vogeli and H. canis) and 16S rRNA for rickettsia (E. canis and A. platys). Babesia vogeli and H. canis were discriminated using 9 nucleotide positions out of 30 base pairs, whereas E. canis and A. platys were differentiated using 15 nucleotide positions out of 34 base pairs that were determined from regions adjacent to 3' ends of the sequencing primers. This method provides a challenging alternative for a rapid diagnosis and surveillance of these tick-borne diseases in canines.

  9. Infection with Ehrlichia canis and Anaplasma platys (Rickettsiales: Anaplasmataceae) in two lineages of Rhipicephalus sanguineus sensu lato (Acari: Ixodidae) from Argentina.

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    Cicuttin, Gabriel L; Tarragona, Evelina L; De Salvo, M Nazarena; Mangold, Atilio J; Nava, Santiago

    2015-09-01

    Natural infection with Ehrlichia canis and Anaplasma platys in ticks belonging to the tropical and temperate lineages of Rhipicephalus sanguineus sensu lato from Argentina was evaluated. Samples were tested for Ehrlichia canis infection by PCR assays using 16S rRNA, dsb and p28 gene, while detection of A. platys was performed with 16S rRNA and groESL gene. The assignment of the ticks to each lineage was corroborated with 16S rDNA sequences. All ticks infected with E. canis and A. platys belonged to the tropical lineage. These results constitute the first record of E. canis infection in R. sanguineus s.l ticks from Argentina. No ticks from the temperate lineage were found to be infected with E. canis, coinciding with previous studies performed in Argentina and Uruguay where E. canis infection was not detected in R. sanguineus s.l from the temperate lineage. Because the presence of the tropical lineage of R. sanguineus s.l has been documented in tropical areas of northern Argentina between 22° and 24° of south latitude, the findings of this work indicate that transmission of E. canis and A. platys to dogs by R. sanguineus s.l probably occurs along this region.

  10. Occurrence of ticks and prevalence of Anaplasma phagocytophilum and Borrelia burgdorferi s.l. in three types of urban biotopes: forests, parks and cemeteries.

    Science.gov (United States)

    Hornok, Sándor; Meli, Marina L; Gönczi, Enikő; Halász, Edina; Takács, Nóra; Farkas, Róbert; Hofmann-Lehmann, Regina

    2014-10-01

    The aim of the present study was to compare different urban biotopes for the occurrence of ixodid tick species, for the population density of Ixodes ricinus and for the prevalence rates of two emerging, zoonotic pathogens. Altogether 2455 ticks were collected from the vegetation on 30 places (forests, parks, cemeteries) of Budapest, Hungary. I. ricinus and Haemaphysalis concinna were collected in all three biotope types, but Dermacentor reticulatus only in parks and forests, and D. marginatus only in a forest. Highest population density of I. ricinus was observed in neglected parts of cemeteries. In females of this tick species the prevalence rates of Anaplasma phagocytophilum and Borrelia burgdorferi s.l. were significantly lower in cemeteries, than in parks or forests. In conclusion, risks associated with the presence of ticks and tick-borne pathogens may be high in a city, but this depends on biotope types, due to habitat-related differences in the vegetation, as well as in the availability of tick hosts and pathogen reservoirs.

  11. Prevalence and first molecular characterization of Anaplasma phagocytophilum, the agent of human granulocytic anaplasmosis, in Rhipicephalus sanguineus ticks attached to dogs from Egypt

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    Mohamed W. Ghafar

    2012-04-01

    Full Text Available PCR targeting 16S rRNA gene integrated with sequence analysis were performed to investigate the prevalence and the molecular identity of Anaplasma phagocytophilum in Egyptian Rhipicephalus sanguineus ticks attached to dogs. A total of 413 adult and nymphal R. sanguineus ticks were collected while attached to 72 free-roaming dogs from four locations (Imbaba, Boulaq, Haram, Monib in Giza Governorate, Egypt. DNA was successfully extracted from 401 specimens (133 nymphs and 268 adults. The overall prevalence rate was 13.7% and adult ticks showed a significantly higher infection rate (16.4% compared to nymphs (8.3%. Sequence comparisons of 218-bp showed that detected organism belongs to A. phagocytophilum. The sequence showed 99.1% similarity (2 nucleotide differences with some strains described as human pathogens and with that detected in the established tick vectors. Phylogenetic analysis placed the bacteria on a separate branch with that found in R. annulatus from Egypt (DQ379972 (99.5% similarity. Our variant strain was designated as A. phagocytophilum-Ghafar-EGY (AB608266. This report is the first molecular characterization of A. phagocytophilum in R. sanguineus in Egypt, suggesting that this tick species may act as a competent vector for a variant strain of human granulocytic anaplasmosis agent.

  12. Larvae of chigger mites Neotrombicula spp. (Acari: Trombiculidae) exhibited Borrelia but no Anaplasma infections: a field study including birds from the Czech Carpathians as hosts of chiggers.

    Science.gov (United States)

    Literak, Ivan; Stekolnikov, Alexandr A; Sychra, Oldrich; Dubska, Lenka; Taragelova, Veronika

    2008-04-01

    Chigger mites were collected from 1,080 wild birds of 37 species at Certak (Czech Republic), in the western Carpathian Mountains, from 29 July to 24 September 2005. The prevalence of infestation with chigger larvae was 7%. A total of 325 chigger specimens from 10 bird species was identified and three chigger species were found: Neotrombicula autumnalis, N. carpathica, and N. inopinata, the latter two species being reported on new hosts. Neotrombicula carpathica is reported in the Czech Republic for the first time. A total of 509 chigger larvae found on 79 host specimens were examined by polymerase chain reaction (PCR) for the presence of Borrelia burgdorferi s.l. DNA (fragments of the rrf (5S)--rrl (23S) intergenic spacer), and Anaplasma phagocytophilum DNA (epank1 gene). A fragment of specific Borrelia DNA was amplified through PCR in one sample, and the PCR product was further analyzed by reverse line blotting assay, whereby both genospecies of B. garinii and B. valaisiana were proved. This sample pooled five chigger larvae collected from one Sylvia atricapilla on 11 August 2005. No A. phagocytophilum DNA was amplified. We conclude that larvae of the genus Neotrombicula can be infected with Borrelia genospecies originated from their present or former hosts.

  13. Molecular diagnosis of Anaplasma marginale in cattle: quantitative evaluation of a real-time PCR (Polymerase Chain Reaction based on msp5 gene

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    Gisele M. Bacanelli

    2014-01-01

    Full Text Available The rickettsia Anaplasma marginale is considered the main agent of bovine anaplasmosis. Due the nonspecific clinical signs of the anaplasmosis, the diagnosis of infection depends of laboratory confirmation. In recent years, molecular diagnostic methods have been used to detect A. marginale in cattle. However, the existence of a large number of assays of different sensitivity and cost makes the choice of an appropriate test difficult. In the present study, a real-time Polymerase Chain Reaction (PCR based on the msp5 target gene was quantitatively assessed and compared to an end point PCR. Both reactions were subjected to sensitivity and specificity evaluation using plasmid DNA and samples from cattle experimentally infected with A. marginale. A comparative field trial of the tests was carried out using samples of cattle from a stable enzootic area for A. marginale. The real-time PCR showed a higher sensitivity than the end point PCR. This reaction (i.e. real-time PCR was able to detect one copy of the msp5 gene in 100 ηg of plasmidial DNA, and more than 80% of its results were positive among experimentally infected animals seven days after infection. In addition, based on in silico analysis, the real-time PCR evaluated in the present study appears to be useful for the detection of A. ovis.

  14. Characterization of two strains of Anaplasma marginale isolated from cattle in Rio de Janeiro, Brazil, after propagation in tick cell culture.

    Science.gov (United States)

    Baêta, Bruna A; Ribeiro, Carla C D U; Teixeira, Rafaella C; Cabezas-Cruz, Alejandro; Passos, Lygia M F; Zweygarth, Erich; Fonseca, Adivaldo H

    2015-03-01

    IDE8 tick cell cultures have been used for the isolation and propagation of several isolates of Anaplasma marginale. The genetic heterogeneity of A. marginale strains in cattle is diverse in endemic regions worldwide and the analyses of msp1α (major surface protein 1 alpha) gene sequences have allowed the identification of different strains. This study reports the isolation and propagation of two new isolates of A. marginale in IDE8 cells from blood of two cattle and their morphological and molecular characterization using light microscopy and the msp1α gene, respectively. Small colonies were observed in cytospin smears of each of the isolates 60 days after culture initiation. Based on msp1α sequence variation, the two isolates were found to be separate strains and were named AmRio1 and AmRio2. Analysis of msp1α microsatellite in both strains resulted in a single genotype, genotype E. The amino acid sequence of one MSP1α tandem repeat from the strain AmRio1 resulted in a new sequence (named 162) with one amino acid change. The results of these phylogenetic analyses demonstrated that A. marginale strains from Brazil and Argentina formed two large clusters of which one was less divergent that the other.

  15. Comparative analysis of the infectivity rate of both Borrelia burgdorferi and Anaplasma phagocytophilum in humans and dogs in a New Jersey community

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    Gaito A

    2014-08-01

    Full Text Available Andrea Gaito,1 Vedrana Gjivoje,2 Sebastian Lutz,1 Ben Baxter2 1Private medical practice, Somerset County, NJ, USA; 2Bernardsville Animal Hospital, Somerset County, NJ, USA Abstract: Ticks are important vectors of disease and transmit an extensive array of bacterial, viral and protozoan diseases to both humans and dogs within a community. Borrelia burgdorferi, the causative agent of Lyme disease, has been extensively studied within both the human and veterinary population. Anaplasma phagocytophilum, an intracellular rickettsial pathogen also transmitted by ixodid ticks, has emerged as an important zoonotic infection with significant veterinary and medical implications, and is responsible for both canine granulocytic anaplasmosis and human granulocytic anaplasmosis. Multiple surveys exist in the international literature referencing infectivity rates of both of these diseases separately in both the dog and human populations. This is the first study to simultaneously examine the infectivity rate of both anaplasmosis and Lyme disease in humans and dogs in a community endemic for tick-borne diseases. Keywords: Lyme disease, anaplasmosis, dogs, humans 

  16. Host, habitat and climate preferences of Ixodes angustus (Acari: Ixodidae) and infection with Borrelia burgdorferi and Anaplasma phagocytophilum in California, USA.

    Science.gov (United States)

    Stephenson, Nicole; Wong, Johnny; Foley, Janet

    2016-10-01

    The Holarctic tick Ixodes angustus is a competent vector for Borrelia burgdorferi, the etiologic agent of Lyme disease, and possibly Anaplasma phagocytophilum, the etiologic agent of granulocytic anaplasmosis, as well. From 2005 to 2013, we collected host-feeding I. angustus individuals from live-trapped small mammals and by flagging vegetation from 12 study sites in northern and central California, and tested for B. burgdorferi sensu lato, A. phagocytophilum, and Rickettsia spp. DNA by real-time PCR. Among 261 I. angustus collected (259 from hosts and two by flagging), the most common hosts were tree squirrels (20 % of ticks) and chipmunks (37 %). The PCR-prevalence for A. phagocytophilum and B. burgdorferi in ticks was 2 % and zero, respectively. The minimum infection prevalence on pooled DNA samples was 10 % for Rickettsia spp. DNA sequencing of the ompA gene identified this rickettsia as Candidatus Rickettsia angustus, a putative endosymbiont. A zero-inflated negative binomial mixed effects model was used to evaluate geographical and climatological predictors of I. angustus burden. When host species within study site and season within year were included in the model as nested random effects, all significant variables revealed that I. angustus burden increased as temperature decreased. Together with published data, these findings suggest that I. angustus is a host generalist, has a broad geographic distribution, is more abundant in areas with lower temperature within it's range, and is rarely infected with the pathogens A. phagocytophilum and B. burgdorferi.

  17. Prevalence of Babesia canis, Borrelia burgdorferi sensu lato, and Anaplasma phagocytophilum in hard ticks collected from meadows of Lubelskie Voivodship (eastern Poland

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    Dzięgiel Beata

    2014-03-01

    Full Text Available The aim of the study was to assess the distribution of Anaplasma phagocytophilum, Borrelia burgdorferi sensu lato, and Babesia canis in adult females and males of Ixodes ricinus and Dermacentor reticulatus ticks, inhabiting meadows near large forest complexes throughout the Lubelskie Voivodship (eastern region of Poland. Ticks were collected using the flagging method. Among 720 ticks collected, 506 were identified as D. reticulatus, and 214 as I. ricinus. DNA of B. canis and B. burgdorferi s.l. was detected in 21.3% and 0.6% of D. reticulatus ticks, respectively. In I. ricinus ticks, DNA specific to B. burgdorferi s.l. and A. phagocytophilum was detected in 5.6% and 10.3%, respectively. Co-infections of B. burgdorferi s.l. and A. phagocytophilum were found in two I. ricinus ticks. These results indicate that the Lublin region is an area at risk of tick-borne diseases of humans and animals, which must be considered in clinical practice.

  18. Molecular evidence of Anaplasma phagocytophilum and Babesia microti co-infections in Ixodes ricinus ticks in central-eastern region of Poland.

    Science.gov (United States)

    Sytykiewicz, Hubert; Karbowiak, Grzegorz; Hapunik, Joanna; Szpechciński, Adam; Supergan-Marwicz, Marta; Goławska, Sylwia; Sprawka, Iwona; Czerniewicz, Paweł

    2012-01-01

    The aim of the study was to elucidate the distribution of Anaplasma phagocytophilum and Babesia microti co-infection in Ixodes ricinus populations within the central-eastern region of Poland. The prevalence of analysed tick-borne human pathogens in single and polymicrobial infections in I. ricinus ticks were analysed using the conventional and nested PCR techniques. A total number of 1,123 questing tick individuals (291 females, 267 males and 565 nymphs) were collected at different ecosystems (municipal parks, suburban forests, and woodlands). In the presented study, 95 samples of ticks (8.5%) were infected with A.phagocytophilum, 3.1% (n=35) with B. microti, whereas the co-existence status of these human pathogens was detected in 1.8% (n=20) of all tested samples. It has been demonstrated that the prevalence of co-infection status was the highest among females of I. ricinus (11 samples, 3.8%), whereas the lowest within tested nymphs (5 samples, 0.9%). Ticks collected at city parks in Warsaw and suburban areas of this town characterized the highest prevalence of co-infections (3.3 and 4.8%, respectively). Furthermore, it was established that co-infection rates of ticks inhabiting woodlands within Kampinos National Park and Nadbużański Landscape Park were similar and reached the levels of 1.4% (n=5) and 1.1% (n=4), respectively.

  19. Prevalence Rates of Borrelia burgdorferi (Spirochaetales: Spirochaetaceae), Anaplasma phagocytophilum (Rickettsiales: Anaplasmataceae), and Babesia microti (Piroplasmida: Babesiidae) in Host-Seeking Ixodes scapularis (Acari: Ixodidae) from Pennsylvania.

    Science.gov (United States)

    Hutchinson, M L; Strohecker, M D; Simmons, T W; Kyle, A D; Helwig, M W

    2015-07-01

    The etiological agents responsible for Lyme disease (Borrelia burgdorferi), human granulocytic anaplasmosis (Anaplasma phagocytophilum), and babesiosis (Babesia microti) are primarily transmitted by the blacklegged tick, Ixodes scapularis Say. Despite Pennsylvania having in recent years reported the highest number of Lyme disease cases in the United States, relatively little is known regarding the geographic distribution of the vector and its pathogens in the state. Previous attempts at climate-based predictive modeling of I. scapularis occurrence have not coincided with the high human incidence rates in parts of the state. To elucidate the distribution and pathogen infection rates of I. scapularis, we collected and tested 1,855 adult ticks statewide from 2012 to 2014. The presence of I. scapularis and B. burgdorferi was confirmed from all 67 Pennsylvania counties. Analyses were performed on 1,363 ticks collected in the fall of 2013 to avoid temporal bias across years. Infection rates were highest for B. burgdorferi (47.4%), followed by Ba. microti (3.5%) and A. phagocytophilum (3.3%). Coinfections included B. burgdorferi+Ba. microti (2.0%), B. burgdorferi+A. phagocytophilum (1.5%) and one tick positive for A. phagocytophilum+Ba. microti. Infection rates for B. burgdorferi were lower in the western region of the state. Our findings substantiate that Lyme disease risk is high throughout Pennsylvania.

  20. Comparison between indirect enzyme-linked immunosorbent assays for Anaplasma marginale antibodies with recombinant major surface protein 5 and initial body antigens

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    Virgínia MG Silva

    2006-08-01

    Full Text Available Indirect enzyme-linked immunosorbent assays (ELISAs based on recombinant major surface protein 5 (rMSP5 and initial body (IB antigens from a Brazilian isolate of Anaplasma marginale were developed to detect antibodies against this rickettsia in cattle. Both tests showed the same sensitivity (98.2% and specificities (100% for rMSP5 and 93.8% for IB ELISA which did not differ statistically. No cross-reactions were detected with Babesia bigemina antibodies, but 5 (rMSP5 ELISA to 15% (IB ELISA of cross-reactions were detected with B. bovis antibodies. However, such difference was not statistically significant. Prevalences of seropositive crossbred beef cattle raised extensively in Miranda county, state of Mato Grosso do Sul, Brazil, were 78.1% by rMSP5 ELISA and 79.7% by IB ELISA. In the analysis of sera from dairy calves naturally-infected with A. marginale, the dynamics of antibody production was very similar between both tests, with maternal antibodies reaching the lowest levels at 15-30 days, followed by an increase in the mean optical densities in both ELISAs, suggesting the development of active immunity against A. marginale. Results showed that all calves were seropositive by one-year old, characterizing a situation of enzootic stability. The similar performances of the ELISAs suggest that both tests can be used in epidemiological surveys for detection of antibodies to A. marginale in cattle.

  1. Immunogenicity of Outer Membrane Proteins VirB9-1 and VirB9-2, a Novel Nanovaccine against Anaplasma marginale.

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    Liang Zhao

    Full Text Available Anaplasma marginale is the most prevalent tick-borne livestock pathogen and poses a significant threat to cattle industry. In contrast to currently available live blood-derived vaccines against A. marginale, alternative safer and better-defined subunit vaccines will be of great significance. Two proteins (VirB9-1 and VirB9-2 from the Type IV secretion system of A. marginale have been shown to induce humoral and cellular immunity. In this study, Escherichia coli were used to express VirB9-1 and VirB9-2 proteins. Silica vesicles having a thin wall of 6 nm and pore size of 5.8 nm were used as the carrier and adjuvant to deliver these two antigens both as individual or mixed nano-formulations. High loading capacity was achieved for both proteins, and the mouse immunisation trial with individual as well as mixed nano-formulations showed high levels of antibody titres over 107 and strong T-cell responses. The mixed nano-formulation also stimulated high-level recall responses in bovine T-cell proliferation assays. These results open a promising path towards the development of efficient A. marginale vaccines and provide better understanding on the role of silica vesicles to deliver multivalent vaccines as mixed nano-formulations able to activate both B-cell and T-cell immunity, for improved animal health.

  2. Identificación Hematológica y Molecular de Anaplasma platys en Caninos Domésticos de Lima Metropolitana con Signos Clínicos Compatibles con Anaplasmosis

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    Tateishi T., Viviana; Laboratorio de Patología Clínica y Biología Molecular, Facultad de Medicina Veterinaria, Universidad Nacional Mayor de San Marcos, Lima; Lí E., Olga; Laboratorio de Patología Clínica y Biología Molecular, Facultad de Medicina Veterinaria, Universidad Nacional Mayor de San Marcos, Lima; Hoyos S., Luis; Laboratorio de Patología Clínica y Biología Molecular, Universidad Nacional Mayor de San Marcos, Lima; Rivera G., Hermelinda; Laboratorio de Microbiología y Parasitología Veterinaria, Facultad de Medicina Veterinaria, Universidad Nacional Mayor de San Marcos, Lima Perú.; Manchego S., Alberto; Laboratorio de Microbiología y Parasitología Veterinaria, Facultad de Medicina Veterinaria, Universidad Nacional Mayor de San Marcos, Lima; Barrios A., Luis; Laboratorio de Patología Clínica y Biología Molecular, Facultad de Medicina Veterinaria, Universidad Nacional Mayor de San Marcos, Lima; More B., Juan; Laboratorio de Microbiología y Parasitología Veterinaria, Facultad de Medicina Veterinaria, Universidad Nacional Mayor de San Marcos, Lima

    2015-01-01

    El presente estudio tuvo como objetivo determinar la presencia de Anaplasma platys en caninos domésticos de Lima Metropolitana con signos clínicos compatibles con anaplasmosis, mediante la identificación de corpúsculos de inclusión en plaquetas y a través de la técnica Hemi-Nested PCR en muestras de sangre periférica. Se recolectaron 144 muestras de sangre entre enero y diciembre de 2012. De estas, el 29.2% (42/144) fue positiva (individuos trombocitopénicos con presencia de corpúsculos de in...

  3. Induced immune response of Escherichia coli BL21 expressing recombinant MSP1a and MSP1b proteins of Anaplasma marginale

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    Katia Tamekuni

    2009-11-01

    Full Text Available This work aims to evaluate the potential of immunization with E. coli BL21 expressing the recombinant rMSP1a and rMSP1b proteins of Anaplasma marginale. E. coli BL21 was transformed with recombinant plasmids pET102/msp1α and pET101/msp1β, and rMSP1a and rMSP1b were expressed after induction by IPTG. BALB/c mice were vaccinated with formolized BL21/rMSP1a and BL21/rMSP1b, and the production in mice sera of whole IgG was determined by ELISA. The mice immunized with BL21/rMSP1a showed a better humoral response for whole IgG when compared to the mice immunized with BL21/rMSP1b; these mice exhibited a small response after the second vaccination. Sera of mice immunized with BL21/rMSP1a reacted via western blot with BL21 and rMSP1a, with molecular masses varying from 70 to 105 kDa. Sera of mice immunized with BL21/rMSP1b reacted with BL21 and rMSP1b with a molecular mass of 100 kDa. These results demonstrate that BL21 containing rMSP1a and rMSP1b in the outer membrane were able to produce an immune response in mice, reinforcing its use in vaccine models against bovine anaplasmosis.Esse trabalho avaliou o potencial de imunização de Escherichia coli BL21 expressando as proteínas recombinantes rMSP1a e rMSP1b de Anaplasma marginale. A E. coli BL21 foi transformada com os plasmídios recombinantes pET102/msp1α e pET101/msp1β e as proteínas rMSP1a e rMSP1b foram expressas após indução com IPTG. Camundongos BALB/c foram vacinados com BL21/rMSP1a e BL21/rMSP1b formolisadas, e a produção de IgG total foi determinada pelo teste de ELISA nos soros dos camundongos imunizados. Os camundongos imunizados com a BL21/rMSP1a mostraram uma melhor resposta humoral para IgG total, comparada à resposta apresentada pelos camundongos imunizados com BL21/rMSP1b; estes camundongos exibiram uma menor resposta após a segunda vacinação. Soros de camundongos imunizados BL21/rMSP1a reagiram pelo western blot com BL21 e rMSP1a, com massa molecular variando de 70 a

  4. Retrospective analysis of clinical and laboratory findings in hunting dogs with serologic reactions to tick-borne pathogens (Anaplasma phagocytophilum, Borrelia burgdorferi, Babesia canis, Ehrlichia canis, Ricketsia conorii

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    Spasojević-Kosić Ljubica

    2015-01-01

    Full Text Available Seroprevalence of tick-borne infections in endemic areas could be high. In this study, we investigated the seroprevalence of tick-borne pathogens (Anaplasma phagocytophilum, Borrelia burgdorferi, Babesia canis, Ehrlichia canis, Rickettsia conorii in hunting dogs, naturally infected with one or more pathogens. Serological test results of the investigated animals were compared to those from clinical examination, as well as from haematological and biochemical analyses. A total of 74.14% dogs were seropositive (R.conorii 44.83%, B. canis 32.76%, B. burgdorferi 25.86%, E. canis 13.79%, A. phagocytophilum 8.47%, with 25.86% of dogs seropositive to two pathogens, 15.52% seropositive to three pathogens, and 1.72% of dogs seropositive to four pathogens. Among all registered clinical signs, only pyrexia (p<0.05 and arrhythmia (p<0.05 were significant in seropositive dogs. There was no significant difference between seropositive and seronegative dogs regarding the majority of haematological and biochemical parameters. Statistically significant difference was registered for particular haematological (number of red blood cells and seroreactivity to B. burgdorferi and biochemical parameters (albumin concentration and seroreactivity to E. canis, and AST and seroreactivity to R. conorii but these values were not clinically significant. The high exposure to tick-borne pathogens suggests that ectoparasitic profilactic treatment is not adequate in examined population of hunting dogs. Clinical finding of pyrexia need to be further investigated and explained etiologically, which means that molecular diagnosis should be used in order to identify larger number of pathogens because of the possibility of coinfection. [Projekat Ministarstva nauke Republike Srbije, br. TR 31084

  5. Current Surveys of the Seroprevalence of Borrelia burgdorferi, Ehrlichia canis, Anaplasma phagocytophilum, Leishmania infantum, Babesia canis, Angiostrongylus vasorum and Dirofilaria immitis in Dogs in Bulgaria.

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    Pantchev, Nikola; Schnyder, Manuela; Vrhovec, Majda Globokar; Schaper, Roland; Tsachev, Ilia

    2015-08-01

    Canine vector-borne diseases (CVBDs) have increasingly become a focus of interest in recent years. Some of the CVBDs are zoonotic and may therefore also represent a risk for the human population. Different factors are in discussion to explain the expansion of vectors and pathogens into formerly unaffected areas. Knowledge of the prevalence and distribution of CVBDs in Bulgaria is scant overall and most data rely on single case descriptions. The aim of the present study was to determine the seroprevalence of important CVBDs in 167 dogs from central-southern Bulgaria (Stara Zagora), with special emphasis on hitherto uninvestigated babesiosis and angiostrongylosis, on poorly investigated Lyme borreliosis and canine granulocytic anaplasmosis, and on the potentially zoonotic dirofilariosis and leishmaniosis. Relatively high prevalence rates were documented for anti-Babesia canis antibodies, Dirofilaria immitis antigen (16.2 %; 27/167 each), anti-Ehrlichia canis (21 %; 35/167) and anti-Anaplasma phagocytophilum antibodies (30.5 - 46.1 %; 51 - 77/167), while Borrelia burgdorferi seroprevalence was low (2.4 %; 4/167). All samples were negative for Leishmania infantum antibodies and Angiostrongylus vasorum antigen and antibodies. In total, 64.7 % (108/167) of the samples indicated infection or exposure to at least one agent and a high proportion of dual infections (39.8 %; 43/108) was demonstrated. Multiple infections with up to four different organisms were also detected. Our data underline the importance of CVBDs and especially of co-infections which could influence the clinical outcome in dogs.

  6. Serological detection of Anaplasma phagocytophilum, Borrelia burgdorferi sensu lato and Ehrlichia canis antibodies and Dirofilaria immitis antigen in a countrywide survey in dogs in Poland.

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    Krämer, Friederike; Schaper, Roland; Schunack, Bettina; Połozowski, Andrzej; Piekarska, Jolanta; Szwedko, Aleksandra; Jodies, Robert; Kowalska, Dagmara; Schüpbach, Dörte; Pantchev, Nikola

    2014-09-01

    Canine vector-borne diseases (CVBDs) have increasingly become a focus of attention in the past few years. Nevertheless, in many parts of Europe information on their occurrence is still scarce. In a large study in Poland 3,094 serum samples taken from dogs throughout all 16 Polish provinces were tested using a commercial kit for the detection of circulating antibodies against Anaplasma phagocytophilum, Borrelia burgdorferi sensu lato and Ehrlichia canis and of Dirofilaria immitis antigen. A total of 12.31% (381/3,094; 95% confidence interval [CI]: 11.18-13.52%) and 3.75% (116/3,094; 95% CI: 3.11-4.48%) of the dogs were positive for A. phagocytophilum and B. burgdorferi s.l. antibodies, respectively. Furthermore, 0.26% (8/3,094; 95% CI: 0.11-0.51%) were positive for E. canis antibodies and 0.16% (5/3,094; 95% CI: 0.05-0.38%) for D. immitis antigen. The highest percentages of A. phagocytophilum-positive dogs were noted in Lesser Poland, Silesia and Łódź Provinces. For B. burgdorferi s.l., the highest prevalence was recorded in Łódź Province. Co-infections with A. phagocytophilum and B. burgdorferi s.l. were recorded in 1.71% of all examined dogs (53/3,094; 95% CI: 1.29-2.23%). One dog even had a triple infection, testing positive for E. canis too. Both A. phagocytophilum and B. burgdorferi s.l. have previously been reported in Poland and were confirmed in the present study by positive samples from all 16 provinces. Concerning E. canis and D. immitis travel history or importation cannot be excluded as factors which may have determined the occurrence of these pathogens in the relevant animals. Practitioners in Poland should be aware of the above mentioned CVBDs and of prophylactic measures to protect dogs and their owners.

  7. Seroprevalence and geographic distribution of Dirofilaria immitis and tick-borne infections (Anaplasma phagocytophilum, Borrelia burgdorferi sensu lato, and Ehrlichia canis) in dogs from Romania.

    Science.gov (United States)

    Mircean, Viorica; Dumitrache, Mirabela Oana; Györke, Adriana; Pantchev, Nikola; Jodies, Robert; Mihalca, Andrei Daniel; Cozma, Vasile

    2012-07-01

    Tick-borne diseases are of great concern worldwide. Despite this, in Romania there is only limited information regarding the prevalence of vector-borne pathogens in dogs. In all, 1146 serum samples were tested by SNAP(®) 4Dx(®) (IDEXX Laboratories, Inc., Westbrook, ME) for Anaplasma phagocytophilum, Borrelia burgdorferi, and Ehrlichia canis antibodies, and for Dirofilaria immitis antigen. The correlation between positive cases and their geographic distribution, as well as potential risk factors (age, sex, breed, type of dog, habitat, and prophylactic treatments) were evaluated. Overall, 129 dogs (11.3%) were serologically-positive to one or more of the tested pathogens. The seroprevalence for the four infectious agents were: A. phagocytophilum 5.5% (63/1146), D. immitis 3.3% (38/1146), E. canis 2.1% (24/1146), and B. burgdorferi 0.5% (6/1146). Co-infection with E. canis and A. phagocytophilum was registered in 2 dogs (0.2%). The geographical distribution of the seropositive cases suggests clustered foci in southern regions and in the western part of the country for D. immitis, and in the southeastern region (Constanţa County) for E. canis. A. phagocytophilum and B. burgdorferi showed a homogenous distribution, with a tendency for Lyme-positive samples to concentrate in central Romania. For D. immitis, A. phagocytophilum, and E. canis, administering prophylactic treatments was a risk factor associated with infection. Another associated risk factor was the type of dog (stray dogs were at risk being positive for D. immitis, shelter dogs for E. canis, and hunting dogs for B. burgdorferi). The prevalence of D. immitis was significantly higher in males and in dogs older than 2 years. This survey represents the first data detailing A. phagocytophilum and E. canis seroprevalence in Romanian dogs, and the most comprehensive epidemiological study on vector-borne infections in dogs from this country.

  8. Epitope-based vaccines with the Anaplasma marginale MSP1a functional motif induce a balanced humoral and cellular immune response in mice.

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    Paula S Santos

    Full Text Available Bovine anaplasmosis is a hemoparasitic disease that causes considerable economic loss to the dairy and beef industries. Cattle immunized with the Anaplasma marginale MSP1 outer membrane protein complex presents a protective humoral immune response; however, its efficacy is variable. Immunodominant epitopes seem to be a key-limiting factor for the adaptive immunity. We have successfully demonstrated that critical motifs of the MSP1a functional epitope are essential for antibody recognition of infected animal sera, but its protective immunity is yet to be tested. We have evaluated two synthetic vaccine formulations against A. marginale, using epitope-based approach in mice. Mice infection with bovine anaplasmosis was demonstrated by qPCR analysis of erythrocytes after 15-day exposure. A proof-of-concept was obtained in this murine model, in which peptides conjugated to bovine serum albumin were used for immunization in three 15-day intervals by intraperitoneal injections before challenging with live bacteria. Blood samples were analyzed for the presence of specific IgG2a and IgG1 antibodies, as well as for the rickettsemia analysis. A panel containing the cytokines' transcriptional profile for innate and adaptive immune responses was carried out through qPCR. Immunized BALB/c mice challenged with A. marginale presented stable body weight, reduced number of infected erythrocytes, and no mortality; and among control groups mortality rates ranged from 15% to 29%. Additionally, vaccines have significantly induced higher IgG2a than IgG1 response, followed by increased expression of pro-inflammatory cytokines. This is a successful demonstration of epitope-based vaccines, and protection against anaplasmosis may be associated with elicitation of effector functions of humoral and cellular immune responses in murine model.

  9. Are patients with erythema migrans who have leukopenia and/or thrombocytopenia coinfected with Anaplasma phagocytophilum or tick-borne encephalitis virus?

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    Franc Strle

    Full Text Available Lyme borreliosis (LB, tick-borne encephalitis (TBE and human granulocytic anaplasmosis (HGA are endemic in central part of Slovenia. We tested the hypothesis that patients with erythema migrans (EM from this region, who have leukopenia and/or thrombocytopenia (typical findings in HGA and in the initial phase of TBE but not in patients with LB are coinfected with Anaplasma phagocytophilum and/or with TBE virus, i.e. that cytopenia is a result of concomitant HGA or the initial phase of TBE. Comparison of clinical and laboratory findings for 67 patients with EM who disclosed leukopenia/thrombocytopenia with the corresponding results in sex- and age-matched patients with EM and normal blood cell counts revealed no differences. In addition, patients with typical EM and leukopenia and/or thrombocytopenia tested negative for the presence of IgM and IgG antibodies to TBE virus by ELISA as well as for the presence of specific IgG antibodies to A. phagocytophilum antigens by IFA in acute and convalescent serum samples. Thus, none of 67 patients (95% CI: 0 to 5.3% with typical EM (the presence of this skin lesion attests for early Lyme borreliosis and is the evidence for a recent tick bite was found to be coinfected with A. phagocytophilum or had a recent primary infection with TBE virus. The findings in the present study indicate that in Slovenia, and probably in other European countries endemic for LB, TBE and HGA, patients with early LB are rarely coinfected with the other tick-transmitted agents.

  10. Nanoparticle-Based Delivery of Anaplasma marginale Membrane Proteins; VirB9-1 and VirB10 Produced in the Pichia pastoris Expression System

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    Bing Zhang

    2016-11-01

    Full Text Available Bovine anaplasmosis or cattle-tick fever is a tick-borne haemolytic disease caused by the rickettsial haemoparasite Anaplasma marginale in tropical and subtropical areas of the world. While difficult to express, the proteins VirB9-1 and VirB10 are immunogenic components of the outer membrane type IV secretion system that have been identified as candidate antigens for vaccines targeting of A. marginale. Soluble VirB9-1 and VirB10 were successfully expressed using Pichia pastoris. When formulated with the self-adjuvanting silica vesicles, SV-100 (diameter: 50 nm, and pore entrance size: 6 nm, 200 µg of VirB9-1 and VirB10 were adsorbed per milligram of nanoparticle. The VirB9-1 and VirB10, SV-100 formulations were shown to induce higher antibody responses in mice compared to the QuilA formulations. Moreover, intracellular staining of selected cytokines demonstrated that both VirB9-1 and VirB10 formulations induced cell-mediated immune responses in mice. Importantly, the SV-100 VirB9-1 and VirB10 complexes were shown to specifically stimulate bovine T-cell linages derived from calves immunised with A. marginale outer membrane fractions, suggesting formulations will be useful for bovine immunisation and protection studies. Overall this study demonstrates the potential of self-adjuvanting silica vesicle formulations to address current deficiencies in vaccine delivery applications.

  11. Nuclease Tudor-SN Is Involved in Tick dsRNA-Mediated RNA Interference and Feeding but Not in Defense against Flaviviral or Anaplasma phagocytophilum Rickettsial Infection.

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    Nieves Ayllón

    Full Text Available Tudor staphylococcal nuclease (Tudor-SN and Argonaute (Ago are conserved components of the basic RNA interference (RNAi machinery with a variety of functions including immune response and gene regulation. The RNAi machinery has been characterized in tick vectors of human and animal diseases but information is not available on the role of Tudor-SN in tick RNAi and other cellular processes. Our hypothesis is that tick Tudor-SN is part of the RNAi machinery and may be involved in innate immune response and other cellular processes. To address this hypothesis, Ixodes scapularis and I. ricinus ticks and/or cell lines were used to annotate and characterize the role of Tudor-SN in dsRNA-mediated RNAi, immune response to infection with the rickettsia Anaplasma phagocytophilum and the flaviviruses TBEV or LGTV and tick feeding. The results showed that Tudor-SN is conserved in ticks and involved in dsRNA-mediated RNAi and tick feeding but not in defense against infection with the examined viral and rickettsial pathogens. The effect of Tudor-SN gene knockdown on tick feeding could be due to down-regulation of genes that are required for protein processing and blood digestion through a mechanism that may involve selective degradation of dsRNAs enriched in G:U pairs that form as a result of adenosine-to-inosine RNA editing. These results demonstrated that Tudor-SN plays a role in tick RNAi pathway and feeding but no strong evidence for a role in innate immune responses to pathogen infection was found.

  12. Anaplasma phagocytophilum increases the levels of histone modifying enzymes to inhibit cell apoptosis and facilitate pathogen infection in the tick vector Ixodes scapularis.

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    Cabezas-Cruz, Alejandro; Alberdi, Pilar; Ayllón, Nieves; Valdés, James J; Pierce, Raymond; Villar, Margarita; de la Fuente, José

    2016-04-01

    Epigenetic mechanisms have not been characterized in ticks despite their importance as vectors of human and animal diseases worldwide. The objective of this study was to characterize the histones and histone modifying enzymes (HMEs) of the tick vector Ixodes scapularis and their role during Anaplasma phagocytophilum infection. We first identified 5 histones and 34 HMEs in I. scapularis in comparison with similar proteins in model organisms. Then, we used transcriptomic and proteomic data to analyze the mRNA and protein levels of I. scapularis histones and HMEs in response to A. phagocytophilum infection of tick tissues and cultured cells. Finally, selected HMEs were functionally characterized by pharmacological studies in cultured tick cells. The results suggest that A. phagocytophilum manipulates tick cell epigenetics to increase I. scapularis p300/CBP, histone deacetylase, and Sirtuin levels, resulting in an inhibition of cell apoptosis that in turn facilitates pathogen infection and multiplication. These results also suggest that a compensatory mechanism might exist by which A. phagocytophilum manipulates tick HMEs to regulate transcription and apoptosis in a tissue-specific manner to facilitate infection, but preserving tick fitness to guarantee survival of both pathogens and ticks. Our study also indicates that the pathogen manipulates arthropod and vertebrate cell epigenetics in similar ways to inhibit the host response to infection. Epigenetic regulation of tick biological processes is an essential element of the infection by A. phagocytophilum and the study of the mechanisms and principal actors involved is likely to provide clues for the development of anti-tick drugs and vaccines.

  13. Babesia spp. and Anaplasma phagocytophilum in questing ticks, ticks parasitizing rodents and the parasitized rodents – Analyzing the host-pathogen-vector interface in a metropolitan area

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    Silaghi Cornelia

    2012-09-01

    Full Text Available Abstract Background The aims of this study were to evaluate the host-tick-pathogen interface of Babesia spp. and Anaplasma phagocytophilum in restored areas in both questing and host-attached Ixodes ricinus and Dermacentor reticulatus and their small mammalian hosts. Methods Questing ticks were collected from 5 sites within the city of Leipzig, Germany, in 2009. Small mammals were trapped at 3 of the 5 sites during 2010 and 2011. DNA extracts of questing and host-attached I. ricinus and D. reticulatus and of several tissue types of small mammals (the majority bank voles and yellow-necked mice, were investigated by PCR followed by sequencing for the occurrence of DNA of Babesia spp. and by real-time PCR for A. phagocytophilum. A selected number of samples positive for A. phagocytophilum were further investigated for variants of the partial 16S rRNA gene. Co-infection with Rickettsia spp. in the questing ticks was additionally investigated. Results 4.1% of questing I. ricinus ticks, but no D. reticulatus, were positive for Babesia sp. and 8.7% of I. ricinus for A. phagocytophilum. Sequencing revealed B. microti, B. capreoli and Babesia spp. EU1 in Leipzig and sequence analysis of the partial 16S RNA gene of A. phagocytophilum revealed variants either rarely reported in human cases or associated with cervid hosts. The statistical analysis revealed significantly less ticks infected with A. phagocytophilum in a city park in Leipzig as compared to the other sampling sites. A. phagocytophilum-DNA was detected in 2 bank voles, DNA of B. microti in 1 striped field-mouse and of Babesia sp. EU1 in the skin tissue of a mole. Co-infections were detected. Conclusion Our results show the involvement of small mammals in the natural endemic cycles of tick-borne pathogens. A more thorough understanding of the interactions of ticks, pathogens and hosts is the essential basis for effective preventive control measures.

  14. Anaplasma phagocytophilum APH0032 Is Exposed on the Cytosolic Face of the Pathogen-Occupied Vacuole and Co-opts Host Cell SUMOylation

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    Oki, Aminat T.; Huang, Bernice; Beyer, Andrea R.; May, Levi J.; Truchan, Hilary K.; Walker, Naomi J.; Galloway, Nathan L.; Borjesson, Dori L.; Carlyon, Jason A.

    2016-01-01

    Anaplasma phagocytophilum, a member of the family Anaplasmataceae and the obligate intracellular bacterium that causes granulocytic anaplasmosis, resides in a host cell-derived vacuole. Bacterial proteins that localize to the A. phagocytophilum-occupied vacuole membrane (AVM) are critical host-pathogen interfaces. Of the few bacterial AVM proteins that have been identified, the domains responsible for AVM localization and the host cell pathways that they co-opt are poorly defined. APH0032 is an effector that is expressed and localizes to the AVM late during the infection cycle. Herein, the APH0032 domain that is essential for associating with host cell membranes was mapped. Immunofluorescent labeling of infected cells that had been differentially permeabilized confirmed that APH0032 is exposed on the AVM's cytosolic face, signifying its potential to interface with host cell processes. SUMOylation is the covalent attachment of a member of the small ubiquitin-like modifier (SUMO) family of proteins to lysines in target substrates. Previous work from our laboratory determined that SUMOylation is important for A. phagocytophilum survival and that SUMOylated proteins decorate the AVM. Algorithmic prediction analyses identified APH0032 as a candidate for SUMOylation. Endogenous APH0032 was precipitated from infected cells using a SUMO affinity matrix, confirming that the effector co-opts SUMOylation during infection. APH0032 pronouncedly colocalized with SUMO1, but not SUMO2/3 moieties on the AVM. Ectopic expression of APH0032 in A. phagocytophilum infected host cells significantly boosted the bacterial load. This study delineates the first domain of any Anaplasmataceae protein that is essential for associating with the pathogen-occupied vacuole membrane, demonstrates the importance of APH0032 to infection, and identifies it as the second A. phagocytophilum effector that co-opts SUMOylation, thus underscoring the relevance of this post-translational modification to

  15. Prevalence and genetic characterization of Anaplasma marginale in zebu cattle (Bos indicus) and their ticks (Amblyomma variegatum, Rhipicephalus microplus) from Madagascar.

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    Pothmann, Daniela; Poppert, Sven; Rakotozandrindrainy, Raphael; Hogan, Benedikt; Mastropaolo, Mariano; Thiel, Claudia; Silaghi, Cornelia

    2016-10-01

    Tick-borne bovine anaplasmosis, caused by the obligate intracellular pathogen Anaplasma marginale (Rickettsiales: Anaplasmataceae), is a major constraint to cattle production in tropical and subtropical regions. From Madagascar, clinical cases were published but data based on molecular methods regarding the prevalence and genetic diversity of this pathogen on the island are lacking. The aims of this study were to investigate (1) the prevalence of A. marginale in Malagasy zebu cattle (Bos indicus) and their ticks with a species-specific real-time PCR, (2) the genetic diversity of A. marginale based on tandem repeats and microsatellites of the msp1α gene, and (3) the phylogenetic relationship between A. marginale isolates from Madagascar and strains found worldwide. Two hundred fourteen blood samples and 1822 ticks from 214 zebu cattle were collected. Rhipicephalus (R) microplus (40.2%) and Amblyomma (A) variegatum (59.8%) were identified on the cattle. A. marginale DNA was found in 89.7% of the examined zebu cattle and in 62.3% of the examined ticks. The tandem repeat and microsatellite analyses of the mspa1 gene showed high genetic diversity among the isolates between and within the different regions and high infection potential. Eighteen of the 25 tandem repeats identified have not been described before. Phylogenetic analysis revealed clustering of A. marginale strains from Madagascar with South Africa, America and Israel. A common ancestor may originate from South Africa and may have evolved due to phylogeographic characteristics or by a history of cattle movement. Its high prevalence in cattle and ticks, together with a low number of clinical manifestations and a high genetic heterogeneity among the investigated strains, confirms endemic stability of A. marginale in cattle from Madagascar.

  16. Serological survey of Borrelia burgdorferi sensu lato, Anaplasma phagocytophilum, and Ehrlichia canis infections in rural and urban dogs in Central Italy

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    Valentina Virginia Ebani

    2014-11-01

    Full Text Available [b]Introduction[i][/i][/b][i]. Borrelia burgdorferi [/i]sensu lato (s.l. and Anaplasma phagocytophilum are well known zoonotic pathogens, whereas[i] Ehrlichia canis[/i] is usually considered to be of veterinary concern, although on the basis of recent reports it also seems to be able to infect humans. [b]objective[/b]. The aim of the study was to determine the seroprevalence of [i]B. burgdorferi [/i]s.l., A. phagocytophilum and [i]E. canis[/i] in an Italian canine population, and to verify if there are differences between dogs living in urban areas and those from a rural environment. [b]materials and method.[/b] Blood sera of 1,965 dogs, 1,235 from cities and 730 from rural areas, were tested by indirect immunofluorescent assay (IFAT. [b]results[/b]. The overall seroprevalence was highest for E. canis (7.07%, followed by [i]A. phagocytophilum[/i] (4.68%, and [i]B. burgdorferi[/i] s.l. (1.47%. Rural dogs showed the highest seroprevalence to [i]B. burgdorferi[/i] s.l. and [i]A. phagocytophilum[/i]. No significant differences were observed between rural and urban [i]E. canis[/i]-positive dogs. A low percentage (1.32% of dogs with dual seropositivity was detected, and no triple positive reactions were observed. No significant differences were detected in the seroprevalence of the three agents in relationship to the age and gender of the dogs. Seroprevalence in the five years considered were not statistically different, except for the lowest rate for [i]E. canis[/i] observed in 2012. [b]conclusions[/b]. The results confirm the presence of [i]B. burgdorferi[/i] s.l., [i]A. phagocytophilum[/i] and [i]E. canis[/i] in Italian dogs in both urban and rural areas. Monitoring pet dogs, which share the same environment with their owners, is useful for identifying the presence of tick-borne disease agents of both veterinary and public health significance

  17. Tissue-Specific Signatures in the Transcriptional Response to Anaplasma phagocytophilum Infection of Ixodes scapularis and Ixodes ricinus Tick Cell Lines

    Science.gov (United States)

    Alberdi, Pilar; Mansfield, Karen L.; Manzano-Román, Raúl; Cook, Charlotte; Ayllón, Nieves; Villar, Margarita; Johnson, Nicholas; Fooks, Anthony R.; de la Fuente, José

    2016-01-01

    Anaplasma phagocytophilum are transmitted by Ixodes spp. ticks and have become one of the most common and relevant tick-borne pathogens due to their impact on human and animal health. Recent results have increased our understanding of the molecular interactions between Ixodes scapularis and A. phagocytophilum through the demonstration of tissue-specific molecular pathways that ensure pathogen infection, development and transmission by ticks. However, little is known about the Ixodes ricinus genes and proteins involved in the response to A. phagocytophilum infection. The tick species I. scapularis and I. ricinus are evolutionarily closely related and therefore similar responses are expected in A. phagocytophilum-infected cells. However, differences may exist between I. scapularis ISE6 and I. ricinus IRE/CTVM20 tick cells associated with tissue-specific signatures of these cell lines. To address this hypothesis, the transcriptional response to A. phagocytophilum infection was characterized by RNA sequencing and compared between I. scapularis ISE6 and I. ricinus IRE/CTVM20 tick cell lines. The transcriptional response to infection of I. scapularis ISE6 cells resembled that of tick hemocytes while the response in I. ricinus IRE/CTVM20 cells was more closely related to that reported previously in infected tick midguts. The inhibition of cell apoptosis by A. phagocytophilum appears to be a key adaptation mechanism to facilitate infection of both vertebrate and tick cells and was used to investigate further the tissue-specific response of tick cell lines to pathogen infection. The results supported a role for the intrinsic pathway in the inhibition of cell apoptosis by A. phagocytophilum infection of I. scapularis ISE6 cells. In contrast, the results in I. ricinus IRE/CTVM20 cells were similar to those obtained in tick midguts and suggested a role for the JAK/STAT pathway in the inhibition of apoptosis in tick cells infected with A. phagocytophilum. Nevertheless, tick

  18. Tissue-Specific Signatures in the Transcriptional Response to Anaplasma phagocytophilum Infection of Ixodes scapularis and Ixodes ricinus Tick Cell Lines.

    Science.gov (United States)

    Alberdi, Pilar; Mansfield, Karen L; Manzano-Román, Raúl; Cook, Charlotte; Ayllón, Nieves; Villar, Margarita; Johnson, Nicholas; Fooks, Anthony R; de la Fuente, José

    2016-01-01

    Anaplasma phagocytophilum are transmitted by Ixodes spp. ticks and have become one of the most common and relevant tick-borne pathogens due to their impact on human and animal health. Recent results have increased our understanding of the molecular interactions between Ixodes scapularis and A. phagocytophilum through the demonstration of tissue-specific molecular pathways that ensure pathogen infection, development and transmission by ticks. However, little is known about the Ixodes ricinus genes and proteins involved in the response to A. phagocytophilum infection. The tick species I. scapularis and I. ricinus are evolutionarily closely related and therefore similar responses are expected in A. phagocytophilum-infected cells. However, differences may exist between I. scapularis ISE6 and I. ricinus IRE/CTVM20 tick cells associated with tissue-specific signatures of these cell lines. To address this hypothesis, the transcriptional response to A. phagocytophilum infection was characterized by RNA sequencing and compared between I. scapularis ISE6 and I. ricinus IRE/CTVM20 tick cell lines. The transcriptional response to infection of I. scapularis ISE6 cells resembled that of tick hemocytes while the response in I. ricinus IRE/CTVM20 cells was more closely related to that reported previously in infected tick midguts. The inhibition of cell apoptosis by A. phagocytophilum appears to be a key adaptation mechanism to facilitate infection of both vertebrate and tick cells and was used to investigate further the tissue-specific response of tick cell lines to pathogen infection. The results supported a role for the intrinsic pathway in the inhibition of cell apoptosis by A. phagocytophilum infection of I. scapularis ISE6 cells. In contrast, the results in I. ricinus IRE/CTVM20 cells were similar to those obtained in tick midguts and suggested a role for the JAK/STAT pathway in the inhibition of apoptosis in tick cells infected with A. phagocytophilum. Nevertheless, tick

  19. Prevalence of Dirofilaria immitis, Ehrlichia canis, Borrelia burgdorferi sensu lato, Anaplasma spp. and Leishmania infantum in apparently healthy and CVBD-suspect dogs in Portugal - a national serological study

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    Cardoso Luís

    2012-03-01

    Full Text Available Abstract Background Canine vector-borne diseases (CVBDs are caused by a wide range of pathogens transmitted to dogs by arthropods including ticks and insects. Many CVBD-agents are of zoonotic concern, with dogs potentially serving as reservoirs and sentinels for human infections. The present study aimed at assessing the seroprevalence of infection with or exposure to Dirofilaria immitis, Ehrlichia canis, Borrelia burgdorferi sensu lato, Anaplasma spp. and Leishmania infantum in dogs in Portugal. Methods Based on 120 veterinary medical centres from all the regions of mainland and insular Portugal, 557 apparently healthy and 628 CVBD-suspect dogs were sampled. Serum, plasma or whole blood was tested for qualitative detection of D. immitis antigen and antibodies to E. canis, B. burgdorferi s. l., Anaplasma spp. and L. infantum with two commercial in-clinic enzyme-linked immunosorbent assay kits. Odds ratios (OR were calculated by logistic regression analysis to identify independent risk factors of exposure to the vector-borne agents. Results Total positivity levels to D. immitis, E. canis, B. burgdorferi, Anaplasma spp., L. infantum, one or more agents and mixed agents were 3.6%, 4.1%, 0.2%, 4.5%, 4.3%, 14.0% and 2.0% in the healthy group, and 8.9%, 16.4%, 0.5%, 9.2%, 25.2%, 46.3% and 11.6% in the clinically suspect group, respectively. Non-use of ectoparasiticides was a risk factor for positivity to one or more agents both in the apparently healthy (OR = 2.1 and CVBD-suspect (OR = 1.5 dogs. Seropositivity to L. infantum (OR = 7.6, E. canis (OR = 4.1 and D. immitis (OR = 2.4 were identified as risk factors for the presence of clinical signs compatible with CVBDs. Positivity to mixed agents was not found to be a risk factor for disease. Conclusions Dogs in Portugal are at risk of becoming infected with vector-borne pathogens, some of which are of zoonotic concern. CVBDs should be considered by practitioners and prophylactic measures must be put in

  20. Comparação de nested-PCR com o diagnóstico direto na detecção de Ehrlichia canis e Anaplasma platys em cães Comparison of nested-PCR with blood smear examination in detection of Ehrlichia canis and Anaplasma platys in dogs

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    Carlos A. N. Ramos

    2009-12-01

    Full Text Available Os sinais clínicos das infecções por Ehrlichia canis e Anaplasma platys são similares, e o diagnóstico desses patógenos feito por esfregaços sanguíneos corados é difícil devido à sensibilidade e especificidade. Por outro lado, os diagnósticos moleculares são altamente sensíveis e específicos, e nested-PCRs têm sido otimizadas para o diagnóstico preciso desses patógenos em cães. Em um Hospital Veterinário Escola, amostras de sangue total com EDTA foram obtidas de 100 cães, e esfregaços foram feitos das amostras de sangue para busca dos parasitos intracelulares. Para cada amostra, DNA foi extraído e submetido à nPCR para detecção de E. canis e A. platys. Os resultados dos esfregaços sanguíneos mostraram que 9% dos animais foram positivos para E. canis e 21% para A. platys. Com relação à nPCR, 57 e 55% dos cães foram positivos para E. canis e A. platys, respectivamente. Quando comparados com a nPCR, os esfregaços sanguíneos corados revelaram resultados falso-negativos para E. canis e A. platys. Os resultados indicam que a nPCR é altamente sensível e específica para detecção de ambos os patógenos, e os diagnósticos moleculares podem ser mais úteis nos Hospitais Veterinários.The clinical signs of Ehrlichia canis and Anaplasma platys infection are similar, and the diagnosis of these pathogens made by stained blood smears is poor due sensibility and specificity. On the other hand, the molecular diagnosis is highly sensitive and specific and nested-PCR have been optimized for accurate diagnosis these pathogens in dogs. At the veterinary teaching hospital, whole-blood samples with EDTA were obtained from 100 dogs and smears were made from blood samples for evaluation for intracellular parasites. For each sample, DNA was extracted and submitted to nPCR analysis for detection of E. canis and A. platys. The results of stained blood smears showed 9% of the animals were positive for E. canis and 21% for A. platys

  1. Frequency of antibodies to Babesia bigemina, B. bovis, Anaplasma marginale, Trypanosoma vivax and Borrelia burdgorferi in cattle from the northeastern region of the state of Pará, Brazil Freqüência de anticorpos para Babesia bigemina, B. bovis, Anaplasma marginale, Trypanosoma vivax e Borrelia burgdorferi em bovinos do nordeste do Estado do Pará, Brasil

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    Daniel S. Guedes Junior

    2008-06-01

    Full Text Available Babesiosis, anaplasmosis, and trypanosomosis are relevant diseases, potentially causing morbidity in cattle, leading to economic losses. Borreliosis is import as a potential zoonosis. The objective of this study was to determine, by indirect enzyme-linked immunosorbent assay (ELISA, the frequency of seropositive cattle to Babesia bigemina, B. bovis, Anaplasma marginale, Trypanosoma vivax and Borrelia burgdorferi in cattle from the Northeastern region of Pará, Brazil. Sera samples from 246 female adult cattle from municipalities of Castanhal and São Miguel do Guamá were used. Crude antigens ELISAs were used to detect antibodies to all agents, except to A. marginale, to which an indirect ELISA with recombinant major surface 1a protein (MSP1a antigen was used. Overall frequencies of seropositive animals were: B. bigemina - 99.2%; B. bovis - 98.8%; A. marginale - 68.3%; T. vivax - 93.1% and B. burgdorferi - 54.9%. The frequencies of seropositive cattle to B. bovis and B. bigemina suggest a high rate of transmission of these organisms by tick in the studied region, which can be classified as enzootically stable to these hemoprotozoans. The low frequency of seropositive cattle to A. marginale may be attributed to a lower sensitivity of the recombinant antigen ELISA utilized or a distinct rate of inoculation of this rickettsia by ticks, as compared with Babesia sp. transmission. The high frequency of seropositive cattle to T. vivax indicates that this hemoprotozoan is prevalent in herds from the Northeastern region of Pará. The rate of animal that showed homologues antibodies to B. burgdorferi indicates the presence of the tickborne spirochaetal agent in the cattle population in the studied region.A babesiose, a anaplasmose e a tripanossomose são enfermidades relevantes, potencialmente causadoras de morbidade em bovinos, levando a perdas econômicas. A borreliose assume importância como zoonose potencial. O objetivo desse estudo foi determinar

  2. 北京市房山区长角血蜱携带山羊无形体的调查∗%INVESTIGATION FOR ANAPLASMA CAPRA IN HAEMAPHYSALIS LONGICORNIS OF BEIJING AREA, CHINA

    Institute of Scientific and Technical Information of China (English)

    王亚伟; 曹务春; 江佳富; 贾娜; 郭东辉; 江瑞若; 常巧呈; 蒋宝贵; 刘洪波; 魏然

    2016-01-01

    为确认我国华北地区广泛分布的长角血蜱是否携带新近发现的山羊无形体Anaplasma capra,2012、2015年5~9月期间,在北京房山地区采集长角血蜱并进行检测。结果共采集游离的长角血蜱311只,其中成蜱95只,若蜱156只,幼蜱60只。提取蜱基因组DNA后对山羊无形体的gltA和16S rRNA两个基因片段进行PCR扩增。结果显示,共有3(3�2%)只成蜱阳性,若蜱和幼蜱中未检测到山羊无形体。遗传进化分析显示,扩增出的gltA和16S rRNA序列和早先报道的牡丹江人感染的A. capra的序列相一致。证实了在我国华北地区的媒介蜱携带该新发现的山羊无形体,该地区为山羊无形体潜在的自然疫源地。加强该地区蜱媒传染病的防控工作具有必要性和紧迫性。%Anaplasma capra, an emerged tick⁃borne agent, maintained its natural cycles via ticks and host reserviors. To survey for A. capra infection in Haemaphysalis longicornis in North China, a total of 311 H. longicornis ( 95 adults, 156 nymphs, 60 larvae) were collected in Beijing during May to Sept. in 2012 and 2015. A. capra was identified from 3 adults (3�2%) when amplified by PCR targeting gltA gene and rrs (16S rRNA). No infection was found in the nymphs and larvae. Phylogenetic analyses revealed that the partial gltA and 16S rRNA sequences obtained in the ticks are identical to A. capra harvseted from Mudanjiang′s patients in 2014. The finding of A. capra in ticks in North China indicates the natural foci may exist in North China and it is urgent and necessary to prevent and control the emerged tick borne disease in this area.

  3. Prevalence of Borrelia burgdorferi (Spirochaetales: Spirochaetaceae), Anaplasma phagocytophilum (Rickettsiales: Anaplasmataceae), and Babesia microti (Piroplasmida: Babesiidae) in Ixodes scapularis (Acari: Ixodidae) collected from recreational lands in the Hudson Valley Region, New York State.

    Science.gov (United States)

    Prusinski, M A; Kokas, J E; Hukey, K T; Kogut, S J; Lee, J; Backenson, P B

    2014-01-01

    ABSTRACT Blacklegged ticks, Ixodes scapularis Say, were collected from 27 sites in eight New York State counties from 2003 to 2006 to determine the prevalence and distribution of tick-borne pathogens in public-use areas over a 4-yr period. In total, 11,204 I. scapularis (3,300 nymphs and 7,904 adults) were individually analyzed using polymerase chain reaction to detect the presence of Borrelia burgdorferi (causative agent of Lyme disease), Anaplasma phagocytophilum (formerly Ehrlichia phagocytophila, causative agent of human granulocytic anaplasmosis), and Babesia microti (causative agent of human babesiosis). Overall prevalence of B. burgdorferi, A. phagocytophilum, and B. microti was 14.4, 6.5, and 2.7% in nymphs and 45.7, 12.3, and 2.5% in adult ticks, respectively. Rates varied geographically and temporally during the time period examined, and were related to measurements of tick density. Average rate ofpolymicrobial infection for nymphs and adults, respectively, was 1.5 and 8.5% overall, with 0.5 and 6.3% coinfection of B. burgdorferi and A. phagocytophilum, 1.0 and 1.5% B. burgdorferi and B. microti, and 0.05 and 0.6% A. phagocytophilum and B. microti. Thirty-three individual adult ticks from seven study sites in Westchester, Putnam, Dutchess, and Rockland counties tested positive for simultaneous infection with all three agents by multiplex polymerase chain reaction assay.

  4. Initial development and preliminary evaluation of a multiplex bead assay to detect antibodies to Ehrlichia canis, Anaplasma platys, and Ehrlichia chaffeensis outer membrane peptides in naturally infected dogs from Grenada, West Indies.

    Science.gov (United States)

    Wilkerson, Melinda J; Black, Kelley E; Lanza-Perea, Marta; Sharma, Bhumika; Gibson, Kathryn; Stone, Diana M; George, Anushka; Nair, Arathy D S; Ganta, Roman R

    2017-01-01

    Tick-borne bacteria, Ehrlichia canis, Anaplasma platys, and Ehrlichia chaffeensis are significant pathogens of dogs worldwide, and coinfections of E. canis and A. platys are common in dogs on the Caribbean islands. We developed and evaluated the performance of a multiplex bead-based assay to detect antibodies to E. canis, A. platys, and E. chaffeensis peptides in dogs from Grenada, West Indies, where E. canis and A. platys infections are endemic. Peptides from outer membrane proteins of P30 of E. canis, OMP-1X of A. platys, and P28-19/P28-14 of E. chaffeensis were coupled to magnetic beads. The multiplex peptide assay detected antibodies in dogs experimentally infected with E. canis and E. chaffeensis, but not in an A. platys experimentally infected dog. In contrast, the multiplex assay and an in-house enzyme-linked immunosorbent assay (ELISA) detected A. platys antibodies in naturally infected Grenadian dogs. Following testing of 104 Grenadian canine samples, multiplex assay results had good agreement with commercially available ELISA and immunofluorescent assay for E. canis antibody-positive dogs ( K values of 0.73 and 0.84), whereas A. platys multiplex results had poor agreement with these commercial assays ( K values of -0.02 and 0.01). Prevalence of seropositive E. canis and A. platys Grenadian dogs detected by the multiplex and commercial antibody assays were similar to previous reports. Although the multiplex peptide assay performed well in detecting the seropositive status of dogs to E. canis and had good agreement with commercial assays, better antigen targets are necessary for the antibody detection of A. platys.

  5. Investigação molecular de Ehrlichia spp. e Anaplasma platys em felinos domésticos: alterações clínicas, hematológicas e bioquímicas

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    Elisabete S Correa

    2011-10-01

    Full Text Available Ehrlichia sp. e Anaplasma platys são micro-organismos Gram negativos, parasitos intracelulares obrigatórios, residindo em vacúolos citoplasmáticos de leucócitos e plaquetas, encontrados no sangue periférico ou em tecidos. Poucos relatos têm sido feitos sobre erliquiose e anaplasmose em gatos no Brasil, os quais são baseados na presença de mórulas em leucócitos e plaquetas, ou pela detecção de anticorpos. O objetivo deste trabalho foi investigar a infecção natural por Ehrlichia sp. e A.platys em gatos no Município de Campos dos Goytacazes-RJ, através da hematoscopia e pela detecção do DNA desses agentes. Foram utilizadas amostras de sangue total e de soro de 91 gatos, independente de raça, sexo e idade. Realizaram-se hemograma, bioquímica sérica e PCR, utilizando oligonucleotídes para Ehrlichia sp. e A.platys. Os dados de hematoscopia mostraram que 9,89% dos gatos apresentaram mórulas em macroplaquetas. O DNA de A.platys foi detectado em 13,18% dos 91 animais e em 44,44% das amostras positivas à hematoscopia. O DNA de Ehrlichia sp. não foi detectado em nenhuma amostra. Nenhuma alteração foi observada nos sinais clínicos nem nos resultados laboratoriais nos animais estudados. Os dados sugerem que os felinos domésticos podem atuar como potenciais reservatórios para A. platys, como forma não sintomática das enfermidades relacionadas

  6. IgG and IgG2 antibodies from cattle naturally infected with Anaplasma marginale recognize the recombinant vaccine candidate antigens VirB9, VirB10, and elongation factor-Tu

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    Flábio R Araújo

    2008-03-01

    Full Text Available Anaplasma marginale is an important vector-borne rickettsia of ruminants in tropical and subtropical regions of the world. Immunization with purified outer membranes of this organism induces protection against acute anaplasmosis. Previous studies, with proteomic and genomic approach identified 21 proteins within the outer membrane immunogen in addition to previously characterized major surface protein1a-5 (MSP1a-5. Among the newly described proteins were VirB9, VirB10, and elongation factor-Tu (EF-Tu. VirB9, VirB10 are considered part of the type IV secretion system (TFSS, which mediates secretion or cell-to-cell transfer of macromolecules, proteins, or DNA-protein complexes in Gram-negative bacteria. EF-Tu can be located in the bacterial surface, mediating bacterial attachment to host cells, or in the bacterial cytoplasm for protein synthesis. However, the roles of VirB9, VirB10, and TFSS in A. marginale have not been defined. VirB9, VirB10, and EF-Tu have not been explored as vaccine antigens. In this study, we demonstrate that sera of cattle infected with A. marginale, with homologous or heterologous isolates recognize recombinant VirB9, VirB10, and EF-Tu. IgG2 from naturally infected cattle also reacts with these proteins. Recognition of epitopes by total IgG and by IgG2 from infected cattle with A. marginale support the inclusion of these proteins in recombinant vaccines against this rickettsia.

  7. Quantitation of Anaplasma marginale major surface protein (MSP)1a and MSP2 epitope-specific CD4+ T lymphocytes using bovine DRB3*1101 and DRB3*1201 tetramers.

    Science.gov (United States)

    Norimine, Junzo; Han, Sushan; Brown, Wendy C

    2006-09-01

    Antigen-specific CD4+ T cells play a critical role in protective immunity to many infectious pathogens. Although the antigen-specific CD4+ T cells can be measured by functional assays such as proliferation or cytokine enzyme-linked immunospot, such assays are limited to a specific function and cannot quantify anergic or suppressed T cells. In contrast, major histocompatiblity complex (MHC) class II tetramers can enumerate epitope-specific CD4+ T cells independent of function. In this paper, we report the construction of bovine leukocyte antigen MHC class II tetramers using a novel mammalian cell system to express soluble class II DRA/DRB3 molecules and defined immunodominant peptide epitopes of Anaplasma marginale major surface proteins (MSPs). Phycoerythrin-labeled tetramers were either loaded with exogenous peptide or constructed with the peptide epitope linked to the N terminus of the DRB3 chain. A DRB3*1101 tetramer loaded with MSP1a peptide F2-5B (ARSVLETLAGHVDALG) and DRB3*1201 tetramers loaded with MSP1a peptide F2-1-1b (GEGYATYLAQAFA) or MSP2 peptide P16-7 (NFAYFGGELGVRFAF) specifically stained antigen-specific CD4+ T cell lines and clones. Tetramers constructed with the T-cell epitope linked to the DRB3 chain were slightly better at labeling CD4+ T cells. In one cell line, the number of tetramer-positive T cells increased to approximately 94% of the CD4+ T cells after culture for 21 weeks with specific antigen. This novel technology should be useful to track the fate of antigen-specific CD4+ T-cell responses in cattle after immunization or infection with persistent pathogens, such as A. marginale, that modulate the host immune response.

  8. Molecular characteristics of emerging spotted fever group Rickettsiae and Anaplasma phagocytophilum in Hebei province, China%河北新发斑点热及人粒细胞无形体病实验室调查分析

    Institute of Scientific and Technical Information of China (English)

    孙印旗; 王勇; 姜霞; 姚娜; 钱振宇; 刘晓丽; 陈创夫; 张丽娟

    2015-01-01

    目的:对河北省新发蜱传斑点热群立克次体及人粒细胞无形体进行分子流行病学特征分析。方法对2009-2012年在河北省辛集、迁安及定州市收集的101份临床可疑立克次体病病例急性期血液DNA样本,采用2套巢氏PCR法分别扩增斑点热群立克次体热休克蛋白基因(groEL)和人粒细胞无形体16S rRNA基因并测序,序列采用NCBI网站进行Blast分析后,选择不同地区、不同宿主来源相应基因,使用DNAStar MegAlign同源分析。结果10.9%(11/101)的病例groEL扩增阳性,同源分析当地斑点热群立克次体groEL基因分为2个序列型,尽管有限片段无法区别立克次体种,但PCR阳性病例血清与我国常见黑龙江立克次体、西伯利亚立克次体、海南斑点热群立克次体及蚤传斑点热立克次体无抗原抗体反应,提示可能为新发斑点热。人粒细胞无形体16S rRNA基因扩增阳性率为8.9%(9/101),且病例急性期血清人粒细胞无形体IgM抗体全部阳性,其中3例急性期与恢复期血清发生IgG抗体转换。16S rRNA基因测序成功的9个序列(341 bp)100%同源,并且与该地区无形体病例分离株CZ-HGA-2100%同源。结论河北地区可能存在新发蜱传人粒细胞无形体病例及新发斑点热。加强立克次体实验室诊断及鉴别诊断、进一步开展病原学、相关媒介及宿主流行病学调查均具有重要的临床及公共卫生意义。%Objective To better understand the molecular epidemiological characteristics of emerging spotted fever group Rickettsiae and Anaplasma phagocytophilum identified in Hebei province. Methods One hundred and one clinical probable cases of typhus were collected from Xinji city, Qianan city and Dingzhou city, Hebei province during 2009-2012 and 101 acute stage and 17 convalescence blood samples were collected. Sera separated from blood were used for detecting the IgM and IgG antibodies against

  9. Identification of Anaplasma phagocytophilum in small mammals from Hengduan Mountains of Southwest China%中国西南横断山区小型兽类嗜吞噬细胞无形体基因的检测及序列测定

    Institute of Scientific and Technical Information of China (English)

    边长玲; 龚正达; 张丽云; 栗冬梅; 葛军旗; 李四全; 李璋鸿; 魏丽荣

    2009-01-01

    目的 了解中国西南横断山区小型兽类自然感染嗜吞噬细胞无形体的情况.方法 采集位于滇西北横断山区的高黎贡山山脉、香格里拉雪山等山地(海拔1000~4500m)林区的小型兽类脏器标本以低温保存和运输,所获标本在实验室应用聚合酶链反应(PCR)方法对嗜吞噬细胞无形体16S rRNA基因和Msp4基因片段进行扩增测序,并将所测序列与GenBank中注册的基因序列进行相似性比较.结果 共检测小型兽类5目18属35种共436只,从6属11种小型兽类中发现阳性标本32份,总阳性率为7.34%.其中,高黎贡山林区检测小型兽类标本25种301只,阳性26份,阳性率为8.64%(26/301);香格里拉雪山等林区检测小型兽类标本19种135只,阳性6份,阳性率为4.44%(6/135);阳性标本绝大部分发现于小型兽类中的啮齿类动物.序列比较分析表明:不同小型兽类间的16S rRNA基因序列相似性为99%~100%,且与吉林野鼠中检测的无形体相对应片段(GenBank:DQ449948)最相近,相似性达99%~100%.对其Msp4基因核苷酸序列进一步分析发现与GenBank中相应片段相似性为95%~97%,提示中国西南横断山区小型兽类所感染无形体株变异较大.结论 首次证实和发现中国西南横断山区6属11种小型兽类自然感染嗜吞噬细胞无形体,其中啮齿类动物可能是这一地区嗜吞噬细胞无形体的主要宿主.%Objective To investigate the prevalence of Anaplasma phagocytophilum in small mammals from the forest area of Hengduan Mountains in southwestern China.Methods Small mammals captured from Gaoligong and Xianggelila mountainous area of Yunnan province were detected by PCR amplification.The sequences of 16S rRNA and Msp4 gene fragments from positive samples were compared with corresponding sequences deposited in GenBank.Results A total number of 436 small animals,which belongs to 5 orders 18 genera 35 species were tested,32(7.34%)were positive in 6 genera 11 species

  10. Comparative Experimental Infection Study in Dogs with Ehrlichia canis, E. chaffeensis, Anaplasma platys and A. phagocytophilum.

    Science.gov (United States)

    Nair, Arathy D S; Cheng, Chuanmin; Ganta, Chanran K; Sanderson, Michael W; Alleman, Arthur R; Munderloh, Ulrike G; Ganta, Roman R

    2016-01-01

    Dogs acquire infections with the Anaplasmataceae family pathogens, E. canis, E. chaffeensis, E. ewingii, A. platys and A. phagocytophilum mostly during summer months when ticks are actively feeding on animals. These pathogens are also identified as causing diseases in people. Despite the long history of tick-borne diseases in dogs, much remains to be defined pertaining to the clinical and pathological outcomes of infections with these pathogens. In the current study, we performed experimental infections in dogs with E. canis, E. chaffeensis, A. platys and A. phagocytophilum. Animals were monitored for 42 days to evaluate infection-specific clinical, hematological and pathological differences. All four pathogens caused systemic persistent infections detectible throughout the 6 weeks of infection assessment. Fever was frequently detected in animals infected with E. canis, E. chaffeensis, and A. platys, but not in dogs infected with A. phagocytophilum. Hematological differences were evident in all four infected groups, although significant overlap existed between the groups. A marked reduction in packed cell volume that correlated with reduced erythrocytes and hemoglobin was observed only in E. canis infected animals. A decline in platelet numbers was common with E. canis, A. platys and A. phagocytophilum infections. Histopathological lesions in lung, liver and spleen were observed in all four groups of infected dogs; infection with E. canis had the highest pathological scores, followed by E. chaffeensis, then A. platys and A. phagocytophilum. All four pathogens induced IgG responses starting on day 7 post infection, which was predominantly comprised of IgG2 subclass antibodies. This is the first detailed investigation comparing the infection progression and host responses in dogs after inoculation with four pathogens belonging to the Anaplasmataceae family. The study revealed a significant overlap in clinical, hematological and pathological changes resulting from the infections.

  11. Comparative Experimental Infection Study in Dogs with Ehrlichia canis, E. chaffeensis, Anaplasma platys and A. phagocytophilum.

    Directory of Open Access Journals (Sweden)

    Arathy D S Nair

    Full Text Available Dogs acquire infections with the Anaplasmataceae family pathogens, E. canis, E. chaffeensis, E. ewingii, A. platys and A. phagocytophilum mostly during summer months when ticks are actively feeding on animals. These pathogens are also identified as causing diseases in people. Despite the long history of tick-borne diseases in dogs, much remains to be defined pertaining to the clinical and pathological outcomes of infections with these pathogens. In the current study, we performed experimental infections in dogs with E. canis, E. chaffeensis, A. platys and A. phagocytophilum. Animals were monitored for 42 days to evaluate infection-specific clinical, hematological and pathological differences. All four pathogens caused systemic persistent infections detectible throughout the 6 weeks of infection assessment. Fever was frequently detected in animals infected with E. canis, E. chaffeensis, and A. platys, but not in dogs infected with A. phagocytophilum. Hematological differences were evident in all four infected groups, although significant overlap existed between the groups. A marked reduction in packed cell volume that correlated with reduced erythrocytes and hemoglobin was observed only in E. canis infected animals. A decline in platelet numbers was common with E. canis, A. platys and A. phagocytophilum infections. Histopathological lesions in lung, liver and spleen were observed in all four groups of infected dogs; infection with E. canis had the highest pathological scores, followed by E. chaffeensis, then A. platys and A. phagocytophilum. All four pathogens induced IgG responses starting on day 7 post infection, which was predominantly comprised of IgG2 subclass antibodies. This is the first detailed investigation comparing the infection progression and host responses in dogs after inoculation with four pathogens belonging to the Anaplasmataceae family. The study revealed a significant overlap in clinical, hematological and pathological changes resulting from the infections.

  12. In vitro cultivation of Anaplasma marginale and A. phagocytophilum in tick cell lines: a review

    OpenAIRE

    Passos,Lygia Maria Friche

    2012-01-01

    Continuous cell lines have been established from several ixodid and argasid tick species, representing an excellent tool suitable for the isolation of pathogens and their subsequent propagation, which in turn allows the production of antigenic material for diagnostic tests, antibody and vaccine production, and also for studies on host-vector-pathogen relationships. This paper reviews the use of tick cells for culture initiation and maintenance of two obligate intracellular bacterial pathogens...

  13. Anaplasma marginale: Diversity, Virulence, and Vaccine Landscape through a Genomics Approach

    Science.gov (United States)

    Amaro-Estrada, Itzel; Rodríguez-Camarillo, Sergio Darío

    2016-01-01

    In order to understand the genetic diversity of A. marginale, several efforts have been made around the world. This rickettsia affects a significant number of ruminants, causing bovine anaplasmosis, so the interest in its virulence and how it is transmitted have drawn interest not only from a molecular point of view but also, recently, some genomics research have been performed to elucidate genes and proteins with potential as antigens. Unfortunately, so far, we still do not have a recombinant anaplasmosis vaccine. In this review, we present a landscape of the multiple approaches carried out from the genomic perspective to generate valuable information that could be used in a holistic way to finally develop an anaplasmosis vaccine. These approaches include the analysis of the genetic diversity of A. marginale and how this affects control measures for the disease. Anaplasmosis vaccine development is also reviewed from the conventional vaccinomics to genome-base vaccinology approach based on proteomics, metabolomics, and transcriptomics analyses reported. The use of these new omics approaches will undoubtedly reveal new targets of interest in the near future, comprising information of potential antigens and the immunogenic effect of A. marginale proteins. PMID:27610385

  14. Global transcriptional analysis reveals surface remodeling of anaplasma marginale in the tick vector

    Science.gov (United States)

    Background: Pathogens dependent upon vectors for transmission to new hosts undergo environment specific changes in gene transcription dependent on whether they are replicating in the vector or the mammalian host. Differential gene transcription, especially of potential vaccine candidates, is of inte...

  15. Anaplasma marginale: Diversity, Virulence, and Vaccine Landscape through a Genomics Approach

    Directory of Open Access Journals (Sweden)

    Rosa Estela Quiroz-Castañeda

    2016-01-01

    Full Text Available In order to understand the genetic diversity of A. marginale, several efforts have been made around the world. This rickettsia affects a significant number of ruminants, causing bovine anaplasmosis, so the interest in its virulence and how it is transmitted have drawn interest not only from a molecular point of view but also, recently, some genomics research have been performed to elucidate genes and proteins with potential as antigens. Unfortunately, so far, we still do not have a recombinant anaplasmosis vaccine. In this review, we present a landscape of the multiple approaches carried out from the genomic perspective to generate valuable information that could be used in a holistic way to finally develop an anaplasmosis vaccine. These approaches include the analysis of the genetic diversity of A. marginale and how this affects control measures for the disease. Anaplasmosis vaccine development is also reviewed from the conventional vaccinomics to genome-base vaccinology approach based on proteomics, metabolomics, and transcriptomics analyses reported. The use of these new omics approaches will undoubtedly reveal new targets of interest in the near future, comprising information of potential antigens and the immunogenic effect of A. marginale proteins.

  16. Seroprevalence against Borrelia burgdorferi sensu lato and occurence of antibody co-expression with Anaplasma phagocytophilum in dogs in Latvia

    OpenAIRE

    Berzina, Inese; Matise, Ilze

    2013-01-01

    Background Lyme disease is commonly diagnosed in humans in Latvia, but up to date no studies have been performed to investigate its prevalence in dogs. The aim of this study was to evaluate if seroprevalence against B. burgdorferi sensu lato (B. burgdorferi s.l.) and co-expression of antibodies against B.burgdorferi s.l. and A. phagocytophilum is higher in dogs with clinical suspicion of tick-borne diseases compared to healthy dogs. Findings Venous blood was taken from healthy dogs (n=441) an...

  17. Anaplasma phagocytophilum and Rickettsia spp. infections in hard ticks (Ixodes ricinus) in the city of Hanover (Germany): revisited.

    Science.gov (United States)

    Tappe, Julia; Strube, Christina

    2013-09-01

    The present study aimed to determine the prevalence of Rickettsiales (A. phagocytophilum and Rickettsia spp.) in 2100 I. ricinus ticks collected at 10 different sampling sites every month during the tick season 2010 in the city of Hanover, northern Germany. At the same time, the results served as a fifth-year-follow-up study to monitor whether changes or stagnation of tick infection rates - possibly due to climate change--were obvious or not. To detect infections with A. phagocytophilum and/or Rickettsia spp., tick samples were analysed by quantitative real-time PCR. Differentiation of Rickettsia species was accomplished using real-time pyrosequencing technology. Overall, 4.5% (94/2100) of the collected ticks were tested positive for A. phagocytophilum and 26.2% (551/2100) were positive for Rickettsia spp. infections. Species differentiation of Rickettsia-positive ticks via real-time pyrosequencing was possible in 48.6% (268/551) of samples, which were all identified as R. helvetica. Coinfections with both pathogens were found in 1.0% (20/2100) of ticks. Statistically significant seasonal fluctuations between sampling months as well as local differences between sampling sites were detected for Rickettsia spp. infection rates. For A. phagocytophilum infections, only significant seasonal variations were found. When comparing infection rates of Hanoverian ticks in 2010 to those in 2005, infection rates of A. phagocytophilum-infected nymphs increased statistically significant (P=0.008, power: 0.762) from 2.3% in 2005 (Schicht et al., 2011) to 4.5% in 2010. Rickettsia spp. infections in female ticks decreased significantly (P=0.049, power: 0.491) from 41.8% in 2005 (Schicht et al., 2012) to 32.4% in 2010. Comparison of the remaining tick stages showed no statistically significant differences.

  18. Rural Residents in China Are at Increased Risk of Exposure to Tick-Borne Pathogens Anaplasma phagocytophilum and Ehrlichia chaffeensis

    Directory of Open Access Journals (Sweden)

    Lijuan Zhang

    2014-01-01

    Full Text Available As emerging tick born rickettsial diseases caused by A. phagocytophilum and E. chaffeensis, anaplasmosis and ehrlichiosis have become a serious threat to human and animal health throughout the world. In particular, in China, an unusual transmission of nosocomial cases of human granulocytic anaplasmosis occurred in Anhui Province in 2006 and more recent coinfection case of A. phagocytophilum and E. chaffeensis was documented in Shandong Province. Although the seroprevalence of human granulocytic anaplasmosis (former human granulocytic ehrlichiosis, HGE has been documented in several studies, these data existed on local investigations, and also little data was reported on the seroprevalence of human monocytic ehrlichiosis (HME in China. In this cross-sectional epidemiological study, indirect immunofluorescence antibody assay (IFA proposed by WHO was used to detect A. phagocytophilum and E. chaffeensis IgG antibodies for 7,322 serum samples from agrarian residents from 9 provinces/cities and 819 urban residents from 2 provinces. Our data showed that farmers were at substantially increased risk of exposure. However, even among urban residents, risk was considerable. Seroprevalence of HGA and HME occurred in diverse regions of the country and tended to be the highest in young adults. Many species of ticks were confirmed carrying A. phagocytophilum organisms in China while several kinds of domestic animals including dog, goats, sheep, cattle, horse, wild rabbit, and some small wild rodents were proposed to be the reservoir hosts of A. phagocytophilum. The broad distribution of vector and hosts of the A. phagocytophilum and E. chaffeensis, especially the relationship between the generalized susceptibility of vectors and reservoirs and the severity of the disease’s clinical manifestations and the genetic variation of Chinese HGA isolates in China, is urgently needed to be further investigated.

  19. Superinfection exclusion of the ruminant pathogen anaplasma marginale in the tick vector is dependent on time between exposures to the strains

    Science.gov (United States)

    The remarkable genetic diversity of vector-borne pathogens allows for the establishment of superinfection in the mammalian host. To have a long-term impact on population strain structure, the introduced strains must also be transmitted by a vector population that has been exposed to the existing pri...

  20. Molecular detection of Anaplasma, Bartonella, and Borrelia species in ticks collected from migratory birds from Hong-do Island, Republic of Korea.

    Science.gov (United States)

    Kang, Jun-Gu; Kim, Heung-Chul; Choi, Chang-Yong; Nam, Hyun-Young; Chae, Hee-Young; Chong, Sung-Tae; Klein, Terry A; Ko, Sungjin; Chae, Joon-Seok

    2013-04-01

    Bird migration is a recurring annual and seasonal event undertaken by more than 100 species of birds in the southeast Asian and northeast Palearctic regions that pass through or remain for short periods from April to May and September to November at Hong-do Island, Republic of Korea (ROK). A total of 212 ticks (40 Haemaphysalis flava, 12 H. longicornis, 146 Ixodes turdus, 13 I. nipponensis, and 1 I. ornithophila) were collected from 65/2,161 (3.0%) migratory birds consisting of 21 species that were captured from January, 2008, through December, 2009, as part of the Migratory Birds Center, Hong-do bird banding program for studying bird migration patterns. Adult ticks were assayed individually while larvae and nymphs were pooled (1-22 and 1-6 ticks per pool, respectively) into 31 and 65 pools, respectively. Ticks were assayed for zoonotic pathogens by PCR using 16S rRNA, heat shock protein (groEL), and internal transcribed spacer (ITS) gene primers to amplify genera specific for Anapalsma, Bartonella, and Borrelia PCR amplicons. Using the 16S rRNA-based nested PCR, A. phagocytophilum (n=1) was detected in I. nipponensis collected from Zoothera sibirica and A. bovis (n=1) was detected in I. turdus collected from Emberiza chrysophrys. Borrelia turdi 16S rRNA genes (n=3) were detected in I. turdus and I. nipponensis collected from Turdus pallidus and Zoothera aurea. Borrelia spp. 16S rRNA genes (n=4) were detected in Ixodes ticks collected from Emberiza tristrami, T. pallidus, and Z. aurea. The Bartonella grahamii ITS gene (n=1) was detected by nested PCR assay in I. turdus collected from Z. aurea. These results provide insight into the potential role of migratory birds in the dispersal of ticks and associated tick-borne pathogens throughout their ranges in Asia.

  1. A study of the presence of B. burgdorferi, Anaplasma (previously Ehrlichia) phagocytophilum, Rickettsia, and Babesia in Ixodes ricinus collected within the territory of Belluno, Italy.

    Science.gov (United States)

    Piccolin, G; Benedetti, G; Doglioni, C; Lorenzato, C; Mancuso, S; Papa, N; Pitton, L; Ramon, M C; Zasio, C; Bertiato, G

    2006-01-01

    In the years 2000 and 2001, we sampled ticks in order to establish the distribution of Ixodes ricinus in the province of Belluno; 5987 tick samples from 244 sites throughout the province were gathered, by dragging for a 5-min period. In 40 sites, seasonal variations and cycle stages of the parasites were studied at monthly intervals from March to September. A polymerase chain reaction (PCR) technique was used to identify the tick-infected sites. Of 1931 individual ticks, 8.23% were positive for Borrelia burgdorferi, 4.4% were positive for Ehrlichia, 1.6% were positive for Rickettsia, and 1.6% were positive for Babesia. The co-presence of Borrelia and Ehrlichia (1.2%) and Babesia (0.5%), Borrelia, Ehrlichia and Rickettsia (0.1%) was also found.

  2. Canine and ovine tick-borne pathogens in camels, Nigeria

    OpenAIRE

    2016-01-01

    In April 2008, whole blood samples were collected from 36 dromedary camels in Sokoto, North-western Nigeria. Following PCR and reverse line blotting, twenty-two samples (61%) resulted positive for Ehrlichia/Anaplasma spp. and three (8%) for Theileria/Babesia spp., with three (8%) cases of co-infections being found. Both sequence and BLAST analyses identified Ehrlichia/Anaplasma spp. and Theileria/Babesia spp. positive cases as Anaplasma platys and Theileria ovis, respectively.This is the firs...

  3. Cloning and Sequence Analysis of the 16S rRNA Gene of Anaplasma bovis in Cattle%牛无浆体16S rRNA基因的克隆测序及分析

    Institute of Scientific and Technical Information of China (English)

    周作勇; 聂奎; 唐成; 胡世君; 周荣琼; 张泽

    2010-01-01

    从自然感染无浆体的重庆黄牛无菌采集血液,提取全血基因组,用血营养菌16S rRNA基因的通用引物进行PCR扩增,得到长约1500 bp的扩增片段,将其克隆到pMD18-T载体后进行测序,并与5条边缘无浆体、4条中央无浆体、4条牛无浆体、4条羊无浆体和3条嗜吞噬细胞无浆体16S rRNA基因序列进行系统发育分析.结果表明所克隆的基因片段长度为1412 bp,GenBank登录号为FJ169957.序列比较结果显示,所获得的序列与Kawa-hara公布的牛无浆体日本株(AB211163)同源性最高,达到99.0%,系统发育分析发现,该序列被聚类到牛无浆体群,并与嗜吞噬细胞无浆体群聚类到一个大的分支.本文从分子水平证实重庆地区存在牛无浆体,牛无浆体与嗜吞噬细胞无浆体的亲缘关系比其他3种无浆体更近.

  4. Investigation of tick-borne bacteria (Rickettsia spp., Anaplasma spp., Ehrlichia spp. and Borrelia spp.) in ticks collected from Andean tapirs, cattle and vegetation from a protected area in Ecuador

    OpenAIRE

    Pesquera, Cristina; Portillo, Aránzazu; Palomar, Ana M.; Oteo, José A.

    2015-01-01

    Background Ixodid ticks play an important role in the transmission and ecology of infectious diseases. Information about the circulation of tick-borne bacteria in ticks is lacking in Ecuador. Our aims were to investigate the tick species that parasitize Andean tapirs and cattle, and those present in the vegetation from the buffer zone of the Antisana Ecological Reserve and Cayambe-Coca National Park (Ecuador), and to investigate the presence of tick-borne bacteria. Methods Tick species were i...

  5. Seroepidemiological investigation on Anaplasma phagocytophilum in the population from northeast mountainous area of Beijing%北京东北部山区人群嗜吞噬细胞无形体血清流行病学调查

    Institute of Scientific and Technical Information of China (English)

    吕燕宁; 窦相峰; 陈丽娟; 孙玉兰; 张秀春; 关增智; 黎新宇; 王全意

    2016-01-01

    目的 了解北京东北部山区人群嗜吞噬细胞无形体的感染状况,为制定相应的防控策略提供依据.方法 在北京密云与怀柔区采集人群血清,采用间接免疫荧光法检测嗜吞噬细胞无形体IgG抗体,进行血清流行病学调查.结果 801份血清中嗜吞噬细胞无形体IgG抗体阳性者106份,阳性率13.23%,其中密云区为13.48%,怀柔区为12.70%,差异无统计学意义(P>0.05).结论 北京密云与怀柔区正常人群中均有嗜吞噬细胞无形体感染的存在,应加强人粒细胞无形体病的监测和防控工作.

  6. NCBI nr-aa BLAST: CBRC-ACAR-01-1026 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-ACAR-01-1026 ref|YP_153799.1| hypothetical protein AM518 [Anaplasma marginale str. St. Marie...s] gb|AAV86544.1| hypothetical protein AM518 [Anaplasma marginale str. St. Maries] YP_153799.1 3.7 24% ...

  7. NCBI nr-aa BLAST: CBRC-DNOV-01-3208 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-DNOV-01-3208 ref|YP_154339.1| cell division protein [Anaplasma marginale str. St. Marie...s] gb|AAV87084.1| cell division protein [Anaplasma marginale str. St. Maries] YP_154339.1 0.11 27% ...

  8. NCBI nr-aa BLAST: CBRC-LAFR-01-0905 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-LAFR-01-0905 ref|YP_153919.1| hypothetical protein AM688 [Anaplasma marginale str. St. Marie...s] gb|AAV86664.1| hypothetical protein AM688 [Anaplasma marginale str. St. Maries] YP_153919.1 5e-08 62% ...

  9. NCBI nr-aa BLAST: CBRC-ACAR-01-0693 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-ACAR-01-0693 ref|YP_153799.1| hypothetical protein AM518 [Anaplasma marginale str. St. Marie...s] gb|AAV86544.1| hypothetical protein AM518 [Anaplasma marginale str. St. Maries] YP_153799.1 0.24 26% ...

  10. NCBI nr-aa BLAST: CBRC-GGOR-01-1354 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-GGOR-01-1354 ref|YP_003328565.1| hypothetical protein ACIS_00677 [Anaplasma centrale str. Israel...] gb|ACZ49251.1| hypothetical protein ACIS_00677 [Anaplasma centrale str. Israel] YP_003328565.1 0.002 28% ...

  11. NCBI nr-aa BLAST: CBRC-GGOR-01-1354 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-GGOR-01-1354 ref|YP_003328481.1| hypothetical protein ACIS_00561 [Anaplasma centrale str. Israel...] gb|ACZ49167.1| hypothetical protein ACIS_00561 [Anaplasma centrale str. Israel] YP_003328481.1 0.002 28% ...

  12. Diagnosis of human granulocytic anaplasmosis in Belgium by combining molecular and serological methods

    Directory of Open Access Journals (Sweden)

    M. Hing

    2014-11-01

    Full Text Available We report here one new, hospitalized case of Anaplasma phagocytophilum in Belgium. The clinical presentation of anaplasmosis, its treatment and the molecular and serological relevant laboratory methods are briefly developed.

  13. Genetic identification and sequene analysis for high-risk group and animals of Anaplasma phagocytophilum infection in some areas of Fujian Province,China%福建部分地区人和动物嗜吞噬细胞无形体基因检测和序列分析

    Institute of Scientific and Technical Information of China (English)

    肖方震; 李丹萍; 徐国英; 陈阳; 邓艳琴

    2016-01-01

    目的 了解福建省人和动物嗜吞噬细胞无形体的感染状况以及基因特征.方法 收集永春县和上杭县高危人群、家畜的全血,采用巢式PCR扩增无形体16S rRNA片段,PCR产物纯化测序并与序列比对分析.结果 永春县牛的无形体感染率最高为53.85%,羊、狗无形体感染率分别为24.71%和12.50%.上杭县羊的无形体感染率为53.06%,高危人群的无形体感染率为8.33%.无形体基因序列与相应的参考序列同源性均达97%~100%.结论 福建省人和动物存在嗜吞噬细胞无形体感染.

  14. Prevalence of select vector-borne disease agents in owned dogs of Ghana

    Directory of Open Access Journals (Sweden)

    Lorelei L. Clarke

    2014-02-01

    Full Text Available Ticks, sera and ethylenediaminetetraacetic acid (EDTA blood were collected from dogs evaluated at the Amakom Veterinary Clinic in Kumasi, Ghana. Sera were evaluated for Dirofilaria immitis antigen and antibodies against Borrelia burgdorferi, Anaplasma phagocytophilum and Ehrlichia canis. Conventional polymerase chain reaction assays designed to amplify the deoxyribonucleic acid (DNA ofEhrlichia spp. or Anaplasma spp. or Neorickettsia spp. or Wolbachia spp., Babesia spp., Rickettsia spp., Hepatozoon spp., Bartonella spp. and the haemoplasmas were performed on DNA extracted from EDTA blood and all positive amplicons were sequenced. This small survey shows that the following vector-borne pathogens are present in urban Ghanian dogs: Ehrlichia canis, Hepatozoon canis,Dirofilaria immitis and Anaplasma platys. Bartonella henselae was isolated from ticks but not from the dogs.

  15. High-throughput screening of tick-borne pathogens in Europe

    DEFF Research Database (Denmark)

    Michelet, Lorraine; Delannoy, Sabine; Devillers, Elodie

    2014-01-01

    , Bartonella, Candidatus Neoehrlichia, Coxiella, Francisella, Babesia, and Theileria genus) across 94 samples. We successfully determined the prevalence of expected (Borrelia burgdorferi sensu lato, Anaplasma phagocytophilum, Rickettsia helvetica, Candidatus Neoehrlichia mikurensis, Babesia divergens, Babesia...... was conducted on 7050 Ixodes ricinus nymphs collected from France, Denmark, and the Netherlands using a powerful new high-throughput approach. This advanced methodology permitted the simultaneous detection of 25 bacterial, and 12 parasitic species (including; Borrelia, Anaplasma, Ehrlichia, Rickettsia...... venatorum), unexpected (Borrelia miyamotoi) and rare (Bartonella henselae) pathogens in the three European countries. Moreover we detected Borrelia spielmanii, Borrelia miyamotoi, Babesia divergens, and Babesia venatorum for the first time in Danish ticks. This surveillance method represents a major...

  16. Molecular detection of Neorickettsia risticii in Brazilian free-tailed bats (Tadarida brasiliensis from Buenos Aires , Argentina

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    Gabriel L. Cicuttin

    2013-05-01

    Full Text Available Neorickettsia risticii is the causative agent of Potomac Horse Fever, a severe febrile disease affecting horses, transmitted by trematodes species with a complex life cycle. A total of 30 insectivorous bats (Brazilian free-tailed bat Tadarida brasiliensis were analyzed by PCR for presence of genus Anaplasma, Ehrlichia, Neorickettsia and Rickettsia. Three samples showed positive reactions for genus Anaplasma, Ehrlichia and Neorickettsia, and the sequences were 99.67% identical to Neorickettsia risticii. The role of bats in the life cycle of N. risticii has yet to be elucidated; however bats may be reservoirs for this bacterium. To our knowledge, this is the first evidence of N. risticii in Argentina.

  17. Human granulocytic anaplasmosis acquired in Connecticut, USA, diagnosed in Vienna, Austria, 2015.

    Science.gov (United States)

    Markowicz, Mateusz; Schötta, Anna-Margarita; Wijnveld, Michiel; Stanek, Gerold

    2016-04-01

    Human granulocytic anaplasmosis (HGA) is caused by Anaplasma phagocytophilum, an intracellular pathogen transmitted by hard ticks. We report a patient who had acquired the infection in Connecticut, USA, and was diagnosed in Vienna, Austria, using PCR methods. Imported HGA from the United States to Austria is a rare event.

  18. The bacterial microbiome of dermacentor andersoni ticks influences pathogen susceptibility

    Science.gov (United States)

    Ticks are of medical and veterinary importance due to their ability to transmit pathogens to humans and animals. The Rocky Mountain wood tick, Dermacentor andersoni, is a vector of a number of pathogens, including Anaplasma marginale, which is the most widespread tick-borne pathogen of livestock. Al...

  19. Rickettsial Seroepidemiology among Farm Workers, Tianjin, People’s Republic of China

    Science.gov (United States)

    Shan, Ailan; Mathew, Bobby; Yin, Jieying; Fu, Xiuping; Lu, Jie; Xu, Jianguo; Dumler, J. Stephen

    2008-01-01

    High seroprevalence rates for Anaplasma phagocytophilum (8.8%), Coxiella burnetii (6.4%), Bartonella henselae (9.6%), and Rickettsia typhi (4.1%) in 365 farm workers near Tianjin, People’s Republic of China, suggest that human infections with these zoonotic bacteria are frequent and largely unrecognized. Demographic features of seropositive persons suggest distinct epidemiology, ecology, and risks. PMID:18507907

  20. Integrated tick and tick-borne disease control trials in crossbred dairy cattle in Malawi

    DEFF Research Database (Denmark)

    Tjørnehøj, Kirsten; Whiteland, A. P.; Mfitilodze, M. W.;

    1996-01-01

    Crossbred dairy heifers on a farm in an East Coast fever (ECF) endemic area in Malawi were immunised against Theileria parva, Anaplasma spp., Babesia bigemina, Babesia bovis and Cowdria ruminantium. They were treated at infrequent intervals with chlorfenvinphos to limit infestation with adult ticks......, without providing complete tick control. In one trial, which tested a threshold dipping regimen, 20 heifers were dipped only once in 6 months to control a flush of Boophilus microplus. Unimmunised controls showed serological evidence of exposure to T. parva and B. bigemina, and one died of ECF......, but there were no incidents of tick-borne disease in the immunised group. In a second trial, which tested a strategic dipping regimen, 107 animals were dipped 9 times over a 6 month period. Despite heavy challenge by B. bovis and moderate challenge by B. bigemina and Anaplasma spp, demonstrated serologically...

  1. Determining the repertoire of immunodominant proteins via whole-genome amplification of intracellular pathogens.

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    Michael J Dark

    Full Text Available Culturing many obligate intracellular bacteria is difficult or impossible. However, these organisms have numerous adaptations allowing for infection persistence and immune system evasion, making them some of the most interesting to study. Recent advancements in genome sequencing, pyrosequencing and Phi29 amplification, have allowed for examination of whole-genome sequences of intracellular bacteria without culture. We have applied both techniques to the model obligate intracellular pathogen Anaplasma marginale and the human pathogen Anaplasma phagocytophilum, in order to examine the ability of phi29 amplification to determine the sequence of genes allowing for immune system evasion and long-term persistence in the host. When compared to traditional pyrosequencing, phi29-mediated genome amplification had similar genome coverage, with no additional gaps in coverage. Additionally, all msp2 functional pseudogenes from two strains of A. marginale were detected and extracted from the phi29-amplified genomes, highlighting its utility in determining the full complement of genes involved in immune evasion.

  2. HEMOPARASITOSIS EN GANADERÍA DOBLE PROPÓSITO VENEZOLANA, DIAGNÓSTICO Y CONTROL: UNA REVISIÓN

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    Rita Tamasaukas

    2010-01-01

    Full Text Available Este trabajo de revisión de tesis doctoral incluye resultados de investigaciones realizadas en el periodo 1971 a 2009 sobre cuatro hemoparasitosis causados por protozoarios y rickettsiales en Venezuela y otros países, con énfasis en la infección mixta en rebaños bovinos, y su diagnóstico y control. En Venezuela, las afecciones causadas por agentes hemotrópicos parasitarios están distribuidas en todos los estados del país con vocación ganadera, en especial, en rebaños bovinos y bufalinos. Entre aquellos encontramos a especies del género Anaplasma (Anaplasma marginale, Babesia (Babesia bigemina y B. bovis, y Trypanosoma (Trypanosoma vivax, con una frecuencia endémica y con variaciones estacionales.

  3. Pathogen-mediated manipulation of arthropod microbiota to promote infection

    Science.gov (United States)

    Abraham, Nabil M.; Liu, Lei; Jutras, Brandon Lyon; Yadav, Akhilesh K.; Narasimhan, Sukanya; Gopalakrishnan, Vissagan; Ansari, Juliana M.; Jefferson, Kimberly K.; Cava, Felipe; Jacobs-Wagner, Christine; Fikrig, Erol

    2017-01-01

    Arthropods transmit diverse infectious agents; however, the ways microbes influence their vector to enhance colonization are poorly understood. Ixodes scapularis ticks harbor numerous human pathogens, including Anaplasma phagocytophilum, the agent of human granulocytic anaplasmosis. We now demonstrate that A. phagocytophilum modifies the I. scapularis microbiota to more efficiently infect the tick. A. phagocytophilum induces ticks to express Ixodes scapularis antifreeze glycoprotein (iafgp), which encodes a protein with several properties, including the ability to alter bacterial biofilm formation. IAFGP thereby perturbs the tick gut microbiota, which influences the integrity of the peritrophic matrix and gut barrier—critical obstacles for Anaplasma colonization. Mechanistically, IAFGP binds the terminal d-alanine residue of the pentapeptide chain of bacterial peptidoglycan, resulting in altered permeability and the capacity of bacteria to form biofilms. These data elucidate the molecular mechanisms by which a human pathogen appropriates an arthropod antibacterial protein to alter the gut microbiota and more effectively colonize the vector. PMID:28096373

  4. Comparative genomics of emerging human ehrlichiosis agents.

    Directory of Open Access Journals (Sweden)

    Julie C Dunning Hotopp

    2006-02-01

    Full Text Available Anaplasma (formerly Ehrlichia phagocytophilum, Ehrlichia chaffeensis, and Neorickettsia (formerly Ehrlichia sennetsu are intracellular vector-borne pathogens that cause human ehrlichiosis, an emerging infectious disease. We present the complete genome sequences of these organisms along with comparisons to other organisms in the Rickettsiales order. Ehrlichia spp. and Anaplasma spp. display a unique large expansion of immunodominant outer membrane proteins facilitating antigenic variation. All Rickettsiales have a diminished ability to synthesize amino acids compared to their closest free-living relatives. Unlike members of the Rickettsiaceae family, these pathogenic Anaplasmataceae are capable of making all major vitamins, cofactors, and nucleotides, which could confer a beneficial role in the invertebrate vector or the vertebrate host. Further analysis identified proteins potentially involved in vacuole confinement of the Anaplasmataceae, a life cycle involving a hematophagous vector, vertebrate pathogenesis, human pathogenesis, and lack of transovarial transmission. These discoveries provide significant insights into the biology of these obligate intracellular pathogens.

  5. FRECUENCIA DE HEMOPARÁSITOS EN BOVINOS DEL BAJO CAUCA Y ALTO SAN JORGE, 2000-2005

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    Mariana Herrera

    2008-12-01

    Full Text Available Objetivo. Determinar la frecuencia de hemoparásitos en los bovinos evaluados por microscopía directa. Materiales y métodos. Se aplicó un modelo epidemiológico cuyo diseño de investigación fue transeccional de tipo descriptivo retrospectivo transversal. Se analizó la totalidad de los datos de los últimos 5 años, proporcionados por el laboratorio Lavebac que presta su servicio a las regiones del Bajo Cauca Antioqueño y Córdoba. Se realizó un análisis de tendencias por el tipo de parásito, sistema de explotación, procedencia y época climática; seguido de un análisis comparativo de asociación utilizando la razón de disparidad (RD y el X2, mediante el programa EPI-INFO versión 3.3. Resultados. Se encontró una frecuencia hemoparasitaria del 22.5%, y de estos el 59.3% correspondió a Anaplasma sp, el 3.1% a Babesia sp y para Trypanosoma sp 30.9%. La región con mayor frecuencia hemoparasitaria fue Bajo Cauca (14%; los diagnósticos positivos fueron realizados con mayor frecuencia en época seca (14.9%, en la que se detectó mayor proporción de infección por Anaplasma sp. Conclusiones. Los resultados dan una aproximación a la frecuencia y distribución de Trypanosoma sp, Babesia sp y Anaplasma sp en dos zonas ganaderas de Colombia, en donde durante la época seca se encontró un mayor porcentaje de infección, y de los agentes hemoparasitarios Anaplasma sp es el más frecuente.

  6. Serosurveillance for livestock pathogens in free-ranging mule deer (Odocoileus hemionus.

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    Annette Roug

    Full Text Available Routine disease surveillance has been conducted for decades in mule deer (Odocoileus hemionus in California for pathogens shared between wildlife and domestic ruminants that may have implications for the animal production industry and wildlife health. Deer sampled from 1990 to 2007 (n = 2,619 were tested for exposure to six pathogens: bluetongue virus (BTV, epizootic hemorrhagic disease virus (EHDV, bovine viral diarrhea virus (BVDV, Leptospira spp., Anaplasma spp. and Brucella spp. We evaluated the relationship between exposure to these pathogens and demographic risk factors to identify broad patterns in seroprevalence across a large temporal and spatial scale. The overall seroprevalence for the entire study period was 13.4% for BTV, 16.8% for EHDV, 17.1% for BVDV, 6.5% for Leptospira spp., 0.2% for Brucella spp., and 17% for Anaplasma spp. Antibodies against BTV and EHDV were most prevalent in the deer populations of southern California. Antibodies against Leptospira spp. and Anaplasma spp. were most prevalent in coastal and central northern California whereas antibodies against BVDV were most prevalent in central-eastern and northeastern California. The overall seroprevalence for Anaplasma spp. was slightly lower than detected in previous studies. North and central eastern California contains large tracts of federal land grazed by livestock; therefore, possible contact between deer and livestock could explain the high BVDV seroprevalence found in these areas. Findings from this study will help to establish baseline values for future comparisons of pathogen exposure in deer, inform on long-term trends in deer population health and provide relevant information on the distribution of diseases that are shared between wildlife and livestock.

  7. Invertebrate vectors, parasites, and rickettsial agents in Guam

    Directory of Open Access Journals (Sweden)

    Johnson, J.

    2012-12-01

    Full Text Available We conducted a 3-week field study of ectoparasites of humans and domestic animals throughout Guam. Thirteen species of ectoparasitic arthropods were collected. Ectoparasites of medical or veterinary significance included the ticks Rhipicephalus sanguineus and Rhipicephalus microplus, fleas Ctenocephalides felis and Xenopsylla cheopsis, and the head louse Pediculus humanus capitis. Polymerase chain reaction based screening for rickettsial and protozoan pathogens detected pathogens in eight arthropods. These included Anaplasma platys, Coxiella burnetii, Babesia canis vogeli, and Hepatozoon canis.

  8. The search for true sources of TBEV and related pathogens : the danger of professed beliefs on pathogens, host and vector interplays

    OpenAIRE

    Gurung, Deepa

    2016-01-01

    Tick-borne disease in general is among the most important vector-borne diseases that are emerging as a threat to humans and is currently identified as a major health problem in many countries. Tick-borne encephalitis and lyme borreliosis are the main diseases transmitted by Ixodes ricinus ticks in Europe. Besides TBEV (Tick-borne encephalitis virus) and Borrelia burgdorferi sensu lato, I. ricinus is known to transmit other pathogenic microorganisms like- louping ill virus, Anaplasma phagocyto...

  9. Detection of Rickettsia amblyommii in association with a tick bite rash.

    Science.gov (United States)

    Billeter, Sarah A; Blanton, Hunter L; Little, Susan E; Levy, Michael G; Breitschwerdt, Edward B

    2007-01-01

    In the summer of 2006, an Amblyomma americanum tick was removed from a woman in central North Carolina, who subsequently developed a rash at the site of tick attachment. When examined by polymerase chain reaction (PCR) for Borrelia, Anaplasma, Ehrlichia, Babesia, Rickettsia, and Bartonella DNA, only the Rickettsia primers generated an amplicon, which was identified as "R. amblyommii" by sequencing. To our knowledge, this is the first case in which R. amblyommii was temporally associated with a rash.

  10. Prevalence of haemoparasitic infections in dairy cattle (Friesian breeds at nagari integrated dairy farms, Gauta-Nike village, Keffi local government area, Nasarawa state, north central of Nigeria

    Directory of Open Access Journals (Sweden)

    S.M. Abdullahi

    2014-02-01

    Full Text Available The of prevalence ofhaemoparasites of cattle located in Nagari Integrated Farms, Gauta-NikeVillage, Keffi Local Government Area, Nasarawa State, Nigeria was conducted inOctober 2012 where 50 Friesian cattle (male and females are kept on intensivesystem of management were randomly selected. Blood samples were collected in ananticoagulant sample bottle and submitted to the Parasitology Laboratory ofFaculty of Veterinary Medicine, Ahmadu Bello University Zaria, Kaduna state ofNigeria for parasitological examination. Giemsa stained thin blood smears wereexamined for hemoparasites and Hematocrit Centrifuge Technique (HCT was usedto determine the presence of motile parasites. An overall prevalence of 90%(82% female and 8% male was recorded for all samples examined, 21 (42% wereinfected with Anaplasma marginale, Theileria mutans shows 20 (40% prevalenceand 4 (8% were infected by Babesia bigemina. Mixed infection between Anaplasmamarginale and Babesia bigemina revealed 2 (4% while Anaplasma marginale andTheileria mutans was 7 (14%. There was a significant difference (P > 0.05in infections caused by Anaplasma marginale, Babesia bigemina and Theileriamutans (Table 1 and also between sexes(Table 3,  but there was no significant difference  (P<0.05 between any of the mixedinfections observed (Table 2. The result of this study shows thesehemoparasites are endemic in the cattle under study which may result in seriousdisease conditions when such animals are subjected to stressful condition.

  11. Tick-borne bacteria in Ixodes ricinus collected in southern Norway evaluated by a commercial kit and established real-time PCR protocols.

    Science.gov (United States)

    Quarsten, H; Skarpaas, T; Fajs, L; Noraas, S; Kjelland, V

    2015-06-01

    Ticks are important vectors of human pathogens. The knowledge of disease causing agents harboured by ticks in Norway is limited. The focus of this study was (a) to detect the bacteria of medical importance in ticks collected from the vegetation at locations in the southern part of the country and (b) to evaluate a novel commercially available multiplex PCR based method by comparing results with conventional established real-time PCR protocols. Borrelia burgdorferi sensu lato was confirmed to be the most prevalent pathogen detected (31%) among one hundred individually analysed adult ticks. Borrelia miyamotoi, a spirochete associated with relapsing fever, was detected in one sample. Anaplasma phagocytophilum was found in 4% of the ticks, followed by Rickettsia helvetica which was detected in one sample. Similar pathogen prevalence was also detected in 500 ticks analysed in pools. This is the first report of the spotted fever group Rickettsia in Norway. Francisella tularensis, Bartonella species or Coxiella burnetti was not detected. However, due to the low number of ticks analysed, the possible presence of these pathogens in the region cannot be ruled out. All isolates were screened by at least two different molecular methods for each bacterial target; one commercially available multiplex PCR based tick-borne bacteria flow chip system (Master Diagnostica) and corresponding real-time PCR protocols. The comparison of methods verified that most findings were detected by both methods (71 Borrelia, 15 Anaplasma and 2 Rickettsia), whereas two additional Borrelia and Anaplasma infected samples were detected by the real-time protocols.

  12. Prevalence of selected infectious disease agents in stray cats in Catalonia, Spain

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    Sara Ravicini

    2016-02-01

    Full Text Available Objectives The objective of the current study was to investigate the prevalence rates of the following infectious agents in 116 stray cats in the Barcelona area of Spain: Anaplasma phagocytophilum, Bartonella species, Borrelia burgdorferi, Chlamydia felis, Dirofilaria immitis, Ehrlichia species, feline calicivirus (FCV, feline herpesvirus-1 (FHV-1, feline leukaemia virus (FeLV, feline immunodeficiency virus (FIV, haemoplasmas, Mycoplasma species and Rickettsia species. Methods Serum antibodies were used to estimate the prevalence of exposure to A phagocytophilum, Bartonella species, B burgdorferi, Ehrlichia species and FIV; serum antigens were used to assess for infection by D immitis and FeLV; and molecular assays were used to amplify nucleic acids of Anaplasma species, Bartonella species, C felis, D immitis, Ehrlichia species, FCV, FHV-1, haemoplasmas, Mycoplasma species and Rickettsia species from blood and nasal or oral swabs. Results Of the 116 cats, 63 (54.3% had evidence of infection by Bartonella species, FeLV, FIV or a haemoplasma. Anaplasma species, Ehrlichia species or Rickettsia species DNA was not amplified from these cats. A total of 18/116 cats (15.5% were positive for FCV RNA (six cats, Mycoplasma species DNA (six cats, FHV-1 DNA (three cats or C felis DNA (three cats. Conclusions and relevance This study documents that shelter cats in Catalonia are exposed to many infectious agents with clinical and zoonotic significance, and that flea control is indicated for cats in the region.

  13. A survey of tick-borne pathogens in dogs and their ticks in the Pantanal biome, Brazil.

    Science.gov (United States)

    Melo, A L T; Witter, R; Martins, T F; Pacheco, T A; Alves, A S; Chitarra, C S; Dutra, V; Nakazato, L; Pacheco, R C; Labruna, M B; Aguiar, D M

    2016-03-01

    Tick and blood samples collected from domestic dogs in the Brazilian Pantanal were tested by molecular methods for the presence of tick-borne protozoa and bacteria. Among 320 sampled dogs, 3.13% were infected by Babesia vogeli (Piroplasmida: Babesiidae), 8.75% by Hepatozoon canis (Eucoccidiorida: Hepatozoidae), 7.19% by Anaplasma platys (Rickettsiales: Anaplasmataceae), and 0.94% by an unclassified Anaplasma sp. In three tick species collected from dogs, the following tick-borne agents were detected: (a) B. vogeli, An. platys and Ehrlichia canis (Rickettsiales: Anaplasmataceae), infecting Rhipicephalus sanguineus sensu lato (Ixodida: Ixodidae) ticks; (b) H. canis, an unclassified Anaplasma sp. and Rickettsia amblyommii (Rickettsiales: Rickettsiaceae), infecting Amblyomma cajennense sensu lato (Ixodida: Ixodidae) ticks, and (c) Rickettsia sp. strain Atlantic rainforest, an emerging human pathogen, infecting Amblyomma ovale ticks. Molecular analysis, based on a mitochondrial gene, revealed that the Am. cajennense s.l. ticks of the present study corresponded to Amblyomma sculptum, a member of the Am. cajennense species complex, and that Rh. sanguineus s.l. belonged to the tropical lineage. Whereas dogs are exposed to a number of tick-borne bacterial and protozoan agents in the Pantanal biome, humans are potentially exposed to infection by spotted fever group rickettsiae (e.g. R. amblyommii and Rickettsia sp. strain Atlantic rainforest) because both Am. sculptum and Am. ovale are among the most important human-biting ticks in Brazil.

  14. Detection of Babesia caballi in Amblyomma variegatum ticks (Acari: Ixodidae) collected from cattle in the Republic of Guinea.

    Science.gov (United States)

    Tomassone, L; Pagani, P; De Meneghi, D

    2005-06-01

    A reverse line blot hybridisation (RLB) assay was applied to screen Amblyomma variegatum adult ticks (n = 504) collected from N'Dama cattle in the Republic of Guinea. In a PCR, the V1 hypervariable region of the 16S ribosomal RNA (rRNA) gene was amplified with a set of primers unique for species of the genera Anaplasma and Ehrlichia, and the V4 hypervariable region of the 18S rRNA gene was amplified with primers specific for members of the genera Theileria and Babesia. Amplified PCR products from A. variegatum ticks were hybridised onto a membrane, to which oligonucleotide probes species-specific for Ehrlichia/Anaplasma and Theileria/Babesia parasites were covalently linked. No pathogens belonging to Ehrlichia/Anaplasma species were found, while 10 DNA samples resulted positive for Babesia caballi and 5 samples for Theileria velifera. This is the first report of B. caballi in A. variegatum ticks. One of the B. caballi positive samples was sequenced. This new strain (BcabGuinea) showed a 97% similarity to the Z15104 B. caballi GenBank sequence.

  15. Persistence mechanisms in tick-borne diseases : tick-borne diseases

    Directory of Open Access Journals (Sweden)

    A.F. Barbet

    2009-09-01

    Full Text Available The use of new, highly sensitive diagnostic methods has revealed persistent infections to be a common feature of different tick-borne diseases, such as babesiosis, anaplasmosis and heartwater. Antigenic variation can contribute to disease persistence through the continual elaboration of new surface structures, and we know in several instances how this is achieved. Known or suspected mechanisms of persistence in babesial parasites include cytoadhesion and rapid variation of the adhesive ligand in Babesia bovis and genetic diversity in several merozoite stage proteins of different Babesia spp. In Anaplasma, extensive variation in the pfam01617 gene family accompanies cycling of organism levels in chronic infection. One result from the pioneering research at Onderstepoort is the definition of a related polymorphic gene family that is likely involved in immunity against heartwater disease. We are beginning to understand the sizes of the antigenic repertoires and full definition is close, with the possibility of applying simultaneous high-throughput sequencing to the order of 1 000 small genomes. We also, for the first time, can consider modifying these genomes and looking at effects on persistence and virulence. However, important biological questions remain unanswered; for example, why we are seeing a new emerging Anaplasma infection of humans and is infection of endothelial cells by Anaplasma significant to persistence in vivo.

  16. The Microbiome of Ehrlichia-Infected and Uninfected Lone Star Ticks (Amblyomma americanum)

    Science.gov (United States)

    Trout Fryxell, R. T.; DeBruyn, J. M.

    2016-01-01

    The Lone Star tick, Amblyomma americanum, transmits several bacterial pathogens including species of Anaplasma and Ehrlichia. Amblyomma americanum also hosts a number of non-pathogenic bacterial endosymbionts. Recent studies of other arthropod and insect vectors have documented that commensal microflora can influence transmission of vector-borne pathogens; however, little is known about tick microbiomes and their possible influence on tick-borne diseases. Our objective was to compare bacterial communities associated with A. americanum, comparing Anaplasma/Ehrlichia -infected and uninfected ticks. Field-collected questing specimens (n = 50) were used in the analyses, of which 17 were identified as Anaplasma/Ehrlichia infected based on PCR amplification and sequencing of groEL genes. Bacterial communities from each specimen were characterized using Illumina sequencing of 16S rRNA gene amplicon libraries. There was a broad range in diversity between samples, with inverse Simpson’s Diversity indices ranging from 1.28–89.5. There were no statistical differences in the overall microbial community structure between PCR diagnosed Anaplasma/Ehrlichia-positive and negative ticks, but there were differences based on collection method (P < 0.05), collection site (P < 0.05), and sex (P < 0.1) suggesting that environmental factors may structure A. americanum microbiomes. Interestingly, there was not always agreement between Illumina sequencing and PCR diagnostics: Ehrlichia was identified in 16S rRNA gene libraries from three PCR-negative specimens; conversely, Ehrlichia was not found in libraries of six PCR-positive ticks. Illumina sequencing also helped identify co-infections, for example, one specimen had both Ehrlichia and Anaplasma. Other taxa of interest in these specimens included Coxiella, Borrelia, and Rickettsia. Identification of bacterial community differences between specimens of a single tick species from a single geographical site indicates that intra

  17. The Microbiome of Ehrlichia-Infected and Uninfected Lone Star Ticks (Amblyomma americanum.

    Directory of Open Access Journals (Sweden)

    R T Trout Fryxell

    Full Text Available The Lone Star tick, Amblyomma americanum, transmits several bacterial pathogens including species of Anaplasma and Ehrlichia. Amblyomma americanum also hosts a number of non-pathogenic bacterial endosymbionts. Recent studies of other arthropod and insect vectors have documented that commensal microflora can influence transmission of vector-borne pathogens; however, little is known about tick microbiomes and their possible influence on tick-borne diseases. Our objective was to compare bacterial communities associated with A. americanum, comparing Anaplasma/Ehrlichia -infected and uninfected ticks. Field-collected questing specimens (n = 50 were used in the analyses, of which 17 were identified as Anaplasma/Ehrlichia infected based on PCR amplification and sequencing of groEL genes. Bacterial communities from each specimen were characterized using Illumina sequencing of 16S rRNA gene amplicon libraries. There was a broad range in diversity between samples, with inverse Simpson's Diversity indices ranging from 1.28-89.5. There were no statistical differences in the overall microbial community structure between PCR diagnosed Anaplasma/Ehrlichia-positive and negative ticks, but there were differences based on collection method (P < 0.05, collection site (P < 0.05, and sex (P < 0.1 suggesting that environmental factors may structure A. americanum microbiomes. Interestingly, there was not always agreement between Illumina sequencing and PCR diagnostics: Ehrlichia was identified in 16S rRNA gene libraries from three PCR-negative specimens; conversely, Ehrlichia was not found in libraries of six PCR-positive ticks. Illumina sequencing also helped identify co-infections, for example, one specimen had both Ehrlichia and Anaplasma. Other taxa of interest in these specimens included Coxiella, Borrelia, and Rickettsia. Identification of bacterial community differences between specimens of a single tick species from a single geographical site indicates that

  18. The impact of 2 dipping systems on endemic stability to bovine babesiosis and anaplasmosis in cattle in 4 communally grazed areas in Limpopo Province, South Africa

    Directory of Open Access Journals (Sweden)

    B.O. Rikhotso

    2005-06-01

    Full Text Available A 12-month study was conducted in 4 communal grazing areas in the Bushbuckridge region, Limpopo Province, South Africa. The main objective was to investigate the impact of reduced acaricide application on endemic stability to bovine babesiosis (Babesia bigemina and Babesia bovis and anaplasmosis (Anaplasma marginale in the local cattle population. To this end 60 cattle in each communal grazing area were bled at the beginning and the conclusion of the experimental period and their sera were assayed for B. bovis, B. bigemina and Anaplasma antibodies. Cattle in the intensively dipped group were dipped 26 times and maintained on a 14-day dipping interval throughout the study, whereas cattle in the strategically dipped group were dipped only 13 times. Three cattle, from which adult ticks were collected, were selected from each village, while immature ticks were collected by drag-sampling the surrounding vegetation. During the dipping process, a questionnaire aimed at assessing the prevalence of clinical cases of tick-borne disease, abscesses and mortalities was completed by an Animal Health Technician at each diptank. An increase in seroprevalence to B. bovis and B. bigemina and a decrease in seroprevalence to Anaplasma was detected in the strategically dipped group while in the intensively dipped group the converse was true. Amblyomma hebraeum was the most numerous tick species on the cattle, and Rhipicephalus (Boophilus microplus was more plentiful than Rhipicephalus (Boophilus decoloratus. Drag samples yielded more immature stages of A. hebraeum than of Rhipicephalus (Boophilus spp. The incidence of clinical cases of tick-borne disease and of abscesses increased in the strategically dipped group at the start of the survey.

  19. Novel Babesia and Hepatozoon agents infecting non-volant small mammals in the Brazilian Pantanal, with the first record of the tick Ornithodoros guaporensis in Brazil.

    Science.gov (United States)

    Wolf, Rafael William; Aragona, Mônica; Muñoz-Leal, Sebastián; Pinto, Leticia Borges; Melo, Andréia Lima Tomé; Braga, Isis Assis; Costa, Jackeliny dos Santos; Martins, Thiago Fernandes; Marcili, Arlei; Pacheco, Richard de Campos; Labruna, Marcelo B; Aguiar, Daniel Moura

    2016-04-01

    Taking into account the diversity of small terrestrial mammals of the Pantanal, the present study aimed to verify the occurrence of infection by Ehrlichia spp., Anaplasma spp., Rickettsia spp., Hepatozoon spp., Babesia spp. and parasitism by ticks in non-volant small mammals collected in the Brazilian Pantanal. Samples of blood, liver and spleen were collected from 64 captured animals, 22 marsupials and 42 rodents. Pathogen detection was performed by the use of genus-specific Polymerase Chain Reaction (PCR) assays. Ticks collected from the animals consisted of Amblyomma sculptum and Amblyomma triste nymphs, and Ornithodoros guaporensis larvae. None of the vertebrate samples (blood, liver, or spleen) yielded detectable DNA of Rickettsia spp. or Ehrlichia spp. The blood of the rodent Hylaeamys megacephalus yielded an Anaplasma sp. genotype (partial 16S rRNA gene) 99% similar to multiple Anaplasma spp. genotypes around the world. The blood of three rodents of the species Calomys callosus were positive for a novel Hepatozoon sp. agent, phylogenetically related (18S rDNA gene) to distinct Hepatozoon genotypes that have been detected in rodents from different parts of the world. One marsupial (Monodelphis domestica) and three rodents (Thrichomys pachyurus) were positive to novel piroplasmid genotypes, phylogenetically (18S rDNA gene) related to Theileria bicornis, Cytauxzoon manul, and Cytauxzoon felis. The present study provides the first molecular detection of Hepatozoon sp. and piroplasmids in small mammals in Brazil. Additionally, we expanded the distribution of O. guaporensis to Brazil, since this tick species was previously known to occur only in Bolivia.

  20. Wildlife reservoirs for vector-borne canine, feline and zoonotic infections in Austria

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    Georg G. Duscher

    2015-04-01

    The role of wild ungulates, especially ruminants, as reservoirs for zoonotic disease on the other hand seems to be negligible, although the deer filaroid Onchocerca jakutensis has been described to infect humans. Deer may also harbour certain Anaplasma phagocytophilum strains with so far unclear potential to infect humans. The major role of deer as reservoirs is for ticks, mainly adults, thus maintaining the life cycle of these vectors and their distribution. Wild boar seem to be an exception among the ungulates as, in their interaction with the fox, they can introduce food-borne zoonotic agents such as Trichinella britovi and Alaria alata into the human food chain.

  1. Alguns ectoparasitas e protozoários em bovinos da República da Guiné-Bissau

    OpenAIRE

    ROSA, FERNANDA; Crespo, Maria Virgínia; Travassos Dias, J. A.

    1998-01-01

    Os autores assinalam a presença de algumas espécies de ixodídeos (Amblyomma variegatum, Boophilus annulatus, B. geigy e Hyalomma sp.) localizados em diferentes regiões do corpo de bovinos autóctones da República da Guiné-Bissau, colhidos entre Novembro de 1990 e Março de 1991. A pesquisa de protozoários, efectuada principalmente em esfregaços de sangue revelou a presença de Anaplasma marginale, Theileria mutans, Babesia bigemina e Sarcocystis sp. Não foram encontradas lesões relacionada...

  2. Human Granulocytic Anaplasmosis: First Reported Case in Canada

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    Michael D Parkins

    2009-01-01

    Full Text Available Human granulocytic anaplasmosis (HGA is a tick-borne rickettsial infection of peripheral blood neutrophils caused by Anaplasma phagocytophilum. While this infection is increasingly recognized as endemic throughout much of the United States, no Canadian cases have been previously described, despite the agent being identified in Canadian ticks. Herein we present a case of HGA acquired in an urban Alberta centre. Canadian physicians must be aware of the possibility of tick-borne rickettsial diseases as etiology of fever in individuals presenting with leukopenia/lymphopenia, thrombocytopenia and elevated transaminases during periods of tick activity. Prompt recognition and treatment are important in minimizing resultant morbidity and mortality.

  3. Human granulocytic anaplasmosis: First reported case in Canada.

    Science.gov (United States)

    Parkins, Michael D; Church, Deirdre L; Jiang, Xiu Yan; Gregson, Daniel B

    2009-01-01

    Human granulocytic anaplasmosis (HGA) is a tick-borne rickettsial infection of peripheral blood neutrophils caused by Anaplasma phagocytophilum. While this infection is increasingly recognized as endemic throughout much of the United States, no Canadian cases have been previously described, despite the agent being identified in Canadian ticks. Herein we present a case of HGA acquired in an urban Alberta centre. Canadian physicians must be aware of the possibility of tick-borne rickettsial diseases as etiology of fever in individuals presenting with leukopenia/lymphopenia, thrombocytopenia and elevated transaminases during periods of tick activity. Prompt recognition and treatment are important in minimizing resultant morbidity and mortality.

  4. Hematologic, serum chemistry and serologic values of Dall's sheep (Ovis dalli dalli) in Alaska.

    Science.gov (United States)

    Foreyt, W J; Smith, T C; Evermann, J F; Heimer, W E

    1983-04-01

    In June 1979, 73 Dall's sheep were captured near Tok, Alaska to determine selected hematologic and serum metabolite parameters and to determine the presence of antibodies to selected pathogens. Hematology and serum metabolite values were compared with values for domestic sheep and bighorn sheep (Ovis canadensis). Antibodies were detected against Brucella sp. (4%), Campylobacter feti (30%), contagious ecthyma virus (23%) and bovine parainfluenza type 3 virus (1%). Antibodies were not detected against Anaplasma sp., Leptospira sp., bovine virus diarrhea virus, bluetongue virus, infectious bovine rhinotracheitis virus, ovine progressive pneumonia, and Toxoplasma sp.

  5. Molecular and serological detection of tick-borne pathogens in dogs from an area endemic for Leishmania infantumin Mato Grosso do Sul, Brazil

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    Keyla Carstens Marques de Sousa

    Full Text Available Tick-borne pathogens affect a wide range of vertebrate hosts. To identify tick-borne pathogens among dogs from Campo Grande, MS, Brazil testing seropositive for Leishmania infantum (syn. L. chagasi, a serological and molecular study was conducted to detectEhrlichia canis, Anaplasma platys and Babesia vogeli in 60 serum and spleen samples. A confirmatory diagnosis ofL. infantum based on serological and molecular assays was also performed, as was sequence alignment and phylogenetic analysis to assess the identity of the parasite species infecting these animals. IgG antibodies toEhrlichia spp., B. vogeli and L. infantum were found, respectively, in 39 (65%, 49 (81.6% and 60 (100% of the sampled dogs. Twenty-seven (45%, fifty-four (90%, fifty-three (88.3%, two (3.3% and one (1.6% dog were positive, respectively, forE. canis, Leishmania spp., Leishmania donovani complex, Babesia sp. and Anaplasma sp. in PCR assays. After sequencing, the amplicons showed 99% of identity with E. canis, B. vogeli, A. platys andLeishmania chagasi isolates. The findings of this study indicate that L. infantum-seropositive dogs from Campo Grande are exposed to multiple tick-borne pathogens, which should therefore be included in the differential diagnosis of dogs with clinical suspicion of leishmaniasis.

  6. An Assessment of Whole Blood and Fractions by Nested PCR as a DNA Source for Diagnosing Canine Ehrlichiosis and Anaplasmosis

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    Tereza Emmanuelle de Farias Rotondano

    2012-01-01

    Full Text Available Ehrlichiosis and anaplasmosis are tick-borne diseases. Ehrlichia canis and Anaplasma platys infect mainly white cells and platelets, respectively. The main DNA source for PCR is peripheral blood, but the potential of blood cell fractions has not been extensively investigated. This study aims at assessment of whole blood (WB and blood fractions potential in nested PCR (nPCR to diagnose canine ehrlichiosis and anaplasmosis. The 16S rRNA gene was amplified in 71.4, 17.8, 31.57, and 30% of the WB, granulocyte (G, mononuclear cells (M, and buffy coat (BC samples. Compared to the WB, the sensitivity of the PCR was 42.86% for the M, and BC fractions, 21.43% for the G, and 33.33% for the blood clot (C. There was fair agreement between the WB and M, BC and C, and slight with the G. Fair agreement occurred between the nPCR and morulae in the blood smear. One animal was coinfected with A. platys and E. canis. This study provided the first evidence of A. platys infection in dogs in Paraíba, Brazil, and demonstrated that WB is a better DNA source than blood fractions to detect Ehrlichia and Anaplasma by nPCR, probably because of the plasma bacterial concentration following host cell lysis.

  7. An Alternative and Rapid Method for the Extraction of Nucleic Acids from Ixodid Ticks by Potassium Acetate Procedure

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    Islay Rodríguez

    2014-08-01

    Full Text Available Four variants of the potassium acetate procedure for DNA extraction from ixodid ticks at different stage of their life cycles were evaluated and compared with phenol-chloroform and ammonium hydroxide methods. The most rapid and most efficient variant was validated in the DNA extraction procedure from the engorged ticks collected from bovine, canine as well as from house ticks for the screening of Borrelia burgdorferi sensu lato, Anaplasma spp. and Babesia spp. The ammonium hydroxide procedure was used for non-engorged ticks. All the variants were efficient and allowed obtaining PCR-quality material according to the specific amplification of 16S rRNA gene fragment of the original tick. DNA extracted from the ticks under the study was tested by multiplex PCR for the screening of tick-borne pathogens. Anaplasma spp. and Babesia spp. amplification products were obtained from 29/48 extracts. Ammonium hydroxide protocol was not efficient for two extracts. Detection of amplification products from the PCR indicated that DNA had been successfully extracted. The potassium acetate procedure could be an alternative, rapid, and reliable method for DNA extraction from the ixodid ticks, mainly for poorly-resourced laboratories.

  8. 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.

  9. Ehrlichiosis y anaplasmosis en Costa Rica

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    Gaby Dolz

    2013-07-01

    Full Text Available La ehrlichiosis y la anaplasmosis son enfermedades infecciosas producidas por bacterias de la familia Anaplasmataceae y transmitidas por garrapatas. Ambas afectan, entre otras especies, al hombre, ocasionando sintomatología que puede ser asociada a un resfriado común o con signos clínicos compatibles con el dengue hemorrágico, patología que se presenta frecuentemente en Costa Rica. Tanto la ehrlichiosis como la anaplasmosis son consideradas también enfermedades de importancia en Medicina Veterinaria. A continuación se brinda una revisión sobre los hallazgos obtenidos en investigaciones realizadas en el país para determinar la presencia y distribución de Ehrlichia y Anaplasma en Costa Rica. Ehrlichia canis se encuentra ampliamente distribuida en el país y es la especie predominante en perros y garrapatas (Rhipicephalus sanguineus. Adicionalmente, se ha detectado, aunque en menor porcentaje, la presencia de Anaplasma platys y Anaplsma phagocytophilum en perros y sus garrapatas. También se ha determinado la presencia de A. phagocytophilum en un venado cola blanca, y de E. canis en humanos donadores de bancos de sangre mediante técnica serológica y molecular.

  10. A transversal study on antibodies against selected pathogens in dromedary camels in the Canary Islands, Spain.

    Science.gov (United States)

    Mentaberre, Gregorio; Gutiérrez, Carlos; Rodríguez, Noé F; Joseph, Sunitha; González-Barrio, David; Cabezón, Oscar; de la Fuente, José; Gortazar, Christian; Boadella, Mariana

    2013-12-27

    The Canary Islands contain the most important dromedary camel (Camelus dromedarius) population in the European Union and are the main export point of dromedaries to continental Europe and Latin America. We investigated the presence of antibodies against relevant disease agents in 100 Canarian camel sera. Selected blood samples of the same animals were also tested by PCR. Sera were tested for antibodies against Bluetongue virus (BTV; 0%), Bovine Viral Diarrhoea virus (BVDV; 0%), Camelpox virus (CPV; 8% by serum neutralization, 16% by ELISA), Peste des Petits Ruminants virus (PPRV, 0%), Rift Valley Fever virus (RVFV; 0%) and West Nile Fever virus (WNV; 3%), the bacterial pathogens Anaplasma sp. (3%), Brucella sp. (1%), Coxiella burnetii (19%), Mycobacterium avium paratuberculosis (MAP; 22%), Mycobacterium tuberculosis complex (MTC; 10%) and Rickettsia sp. (83%), and the parasites Toxoplasma gondii (36%) and Neospora caninum (86%). The most remarkable findings were the detection of antibodies against CPV and the high antibody prevalence against C. burnetii, Rickettsia sp., T. gondii and N. caninum. By PCR, we found no C. burnetii, N. caninum and Anaplasma sp. DNA in the tested samples. However, Rickettsia sp. DNA was detected in six antibody positive tested samples. These results should be taken into consideration in order to implement adequate control measures and avoid a potential dissemination of infections to other territories.

  11. High-throughput screening of tick-borne pathogens in Europe

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    Lorraine eMichelet

    2014-07-01

    Full Text Available Due to increased travel, climatic, and environmental changes, the incidence of tick-borne disease in both humans and animals is increasing throughout Europe. Therefore, extended surveillance tools are desirable. To accurately screen tick-borne pathogens, a large scale epidemiological study was conducted on 7050 Ixodes ricinus nymphs collected from France, Denmark, and the Netherlands using a powerful new high-throughput approach. This advanced methodology permitted the simultaneous detection of 25 bacterial, and 12 parasitic species (including; Borrelia, Anaplasma, Ehrlichia, Rickettsia, Bartonella, Candidatus Neoehrlichia, Coxiella, Francisella, Babesia, and Theileria genus across 94 samples. We successfully determined the prevalence of expected (Borrelia burgdorferi sensu lato, Anaplasma phagocytophilum, Rickettsia helvetica, Candidatus Neoehrlichia mikurensis, Babesia divergens, Babesia venatorum, unexpected (Borrelia miyamotoi and rare (Bartonella henselae pathogens in the three European countries. Moreover we detected Borrelia spielmanii, Borrelia miyamotoi, Babesia divergens, and Babesia venatorum for the first time in Danish ticks. This surveillance method represents a major improvement in epidemiological studies, able to facilitate comprehensive testing of tick-borne pathogens, and which can also be customized to monitor emerging diseases.

  12. Evidence of co-infection with Mycobacterium bovis and tick-borne pathogens in a naturally infected sheep flock.

    Science.gov (United States)

    López, Vladimir; Alberdi, Pilar; Fernández de Mera, Isabel G; Barasona, José Angel; Vicente, Joaquín; Garrido, Joseba M; Torina, Alessandra; Caracappa, Santo; Lelli, Rossella Colomba; Gortázar, Christian; de la Fuente, José

    2016-03-01

    Ticks are responsible for the transmission of pathogens of veterinary importance, including those affecting sheep. The current study was designed to investigate co-infections with tick-borne and other pathogens in a naturally infected sheep flock with poor health condition using serology and PCR. Infection with Anaplasma ovis was detected by serology and PCR in 56% of the animals. The presence of Rickettsia spp. of the Spotted Fever Group (SFG) was detected by PCR and sequence analysis in 31% of the animals. All the animals were negative for Anaplasma phagocytophilum either by serology or PCR. Twelve sheep were randomly selected for anatomopathological studies. Five of these animals presented lesions consistent with Mycobacterium tuberculosis complex (MTBC) infection and spoligotyping confirmed infection with Mycobacterium bovis spoligotype SB0339. Co-infection with tick-borne pathogens and MTBC could contribute to the poor health condition observed in these animals but other uncontrolled factors may also be responsible. The differential expression of immune response genes supported previous findings in ruminants and suggested that infection with tick-borne pathogens and M. bovis may results in unique gene expression patterns in sheep. The results underline the need for further research into the possible role of sheep in the epidemiology of animal tuberculosis.

  13. Tick survey for prevalent pathogens in peri-urban recreation sites in Saarland and Rhineland-Palatinate (Germany).

    Science.gov (United States)

    Mehlhorn, Heinz; Mehlhorn, Tim; Müller, Melanie; Vogt, Manfred; Rissland, Jürgen

    2016-03-01

    Ixodid ticks are important vectors of human pathogens in Central Europe. Despite this fact, prevalence studies are scarce, especially with regard to much-frequented peri-urban recreation sites. In this pilot study, 4.014 larvae, nymphs and adult ticks sampled monthly during the active seasons in 2011 and 2012 from 14 distinct collection sites in two German states (Saarland and Rhineland-Palatinate) were screened for Borrelia spp., Anaplasma spp. and tick-borne encephalitis virus. Mean prevalence rates were 19.8 % for Borrelia spp., 1.9 % for Anaplasma spp. and 0.1 % for tick-borne encephalitis virus (TBEV), which are in accordance with those reported from other regions in Germany and neighbouring countries. Nevertheless, the detection of TBEV-infected ticks is the first positive result after several unsuccessful efforts over the previous years in official "TBE-risk" zones of Saarland and Rhineland-Palatinate which supports the presumption of the origin of observed local infection. Besides ixodid ticks a non-engorged adult female tick of the invading species Dermacentor reticulatus has been found reflecting the appearance of another vector eventually jeopardising the health of host animals as well as humans.

  14. Molecular evidence for bacterial and protozoan pathogens in hard ticks from Romania.

    Science.gov (United States)

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

    2013-09-01

    The aim of the present study was to provide a preliminary insight into the diversity of tick-borne pathogens circulating at the domestic host-tick interface in Romania. For this, feeding and questing ticks were analyzed by real-time polymerase chain reaction (PCR) for the presence of Anaplasma phagocytophilum, Anaplasma platys, Ehrlichia canis, Borrelia burgdorferi sensu latu, and by PCR and subsequent sequencing for Rickettsia spp., Babesia spp. and Theileria spp. A total of 382 ticks, encompassing 5 species from 4 genera, were collected in April-July 2010 from different areas of Romania; of them, 40 were questing ticks and the remainder was collected from naturally infested cattle, sheep, goats, horses or dogs. Tick species analyzed included Ixodes ricinus, Dermacentor marginatus, Hyalomma marginatum, Rhipicephalus bursa, and Rhipicephalus sanguineus. Four rickettsiae of the spotted fever group of zoonotic concern were identified for the first time in Romania: Rickettsia monacensis and Rickettsia helvetica in I. ricinus, and Rickettsia slovaca and Rickettsia raoultii in D. marginatus. Other zoonotic pathogens such as A. phagocytophilum, Borrelia afzelii, and Babesia microti were found in I. ricinus. Pathogens of veterinary importance were also identified, including Theileria equi in H. marginatum, Babesia occultans in D. marginatus and H. marginatum, Theileria orientalis/sergenti/buffeli-group in I. ricinus and in H. marginatum and E. canis in R. sanguineus. These findings show a wide distribution of very diverse bacterial and protozoan pathogens at the domestic host-tick interface in Romania, with the potential of causing both animal and human diseases.

  15. Analysis of risk factors and prevalence of haemoplasma infection in dogs.

    Science.gov (United States)

    Aquino, L C; Kamani, J; Haruna, A M; Paludo, G R; Hicks, C A; Helps, C R; Tasker, S

    2016-05-15

    Mycoplasma haemocanis (Mhc) and 'Candidatus Mycoplasma haematoparvum' (CMhp) are canine haemoplasma species that can induce anaemia in immunocompromised and/or splenectomised dogs. This study aimed to determine the prevalence and phylogeny of canine haemoplasma species in dogs from Nigeria and describe any risk factors for infection. Canine haemoplasma species-specific and generic haemoplasma qPCR assays were used. The species-specific qPCR assays found Mhc infection in 18 of 245 dogs (7.3%), and CMhp infection in only one dog (0.4%). The generic haemoplasma qPCR assays were positive in 44 of 245 (17.9%) dogs. Twenty-five dogs had discordant qPCR results in that they were generic haemoplasma qPCR positive but species-specific qPCR negative. Further evaluation of these dogs by 16S rDNA sequencing gave limited results but 5 were confirmed to be infected with non-haemoplasma species: 2 Anaplasma phagocytophilum, 1 Anaplasma ovis, 1 Serratia marcescens and 1 Aerococcus spp. The 16S rRNA gene sequences from Mhc species showed>99.8% identity with each other and>99.6% identity with GenBank sequences, and resided in a single clade with other global Mhc and Mycoplasma haemofelis sequences, indicating low 16S rRNA genetic variability amongst this canine haemoplasma species.

  16. Comparative analysis of wolbachia genomes reveals streamlining and divergence of minimalist two-component systems.

    Science.gov (United States)

    Christensen, Steen; Serbus, Laura Renee

    2015-03-24

    Two-component regulatory systems are commonly used by bacteria to coordinate intracellular responses with environmental cues. These systems are composed of functional protein pairs consisting of a sensor histidine kinase and cognate response regulator. In contrast to the well-studied Caulobacter crescentus system, which carries dozens of these pairs, the streamlined bacterial endosymbiont Wolbachia pipientis encodes only two pairs: CckA/CtrA and PleC/PleD. Here, we used bioinformatic tools to compare characterized two-component system relays from C. crescentus, the related Anaplasmataceae species Anaplasma phagocytophilum and Ehrlichia chaffeensis, and 12 sequenced Wolbachia strains. We found the core protein pairs and a subset of interacting partners to be highly conserved within Wolbachia and these other Anaplasmataceae. Genes involved in two-component signaling were positioned differently within the various Wolbachia genomes, whereas the local context of each gene was conserved. Unlike Anaplasma and Ehrlichia, Wolbachia two-component genes were more consistently found clustered with metabolic genes. The domain architecture and key functional residues standard for two-component system proteins were well-conserved in Wolbachia, although residues that specify cognate pairing diverged substantially from other Anaplasmataceae. These findings indicate that Wolbachia two-component signaling pairs share considerable functional overlap with other α-proteobacterial systems, whereas their divergence suggests the potential for regulatory differences and cross-talk.

  17. Study on co-infection of tick-borne pathogens in Ixodes persulcatus in Charles Hilary, Xinjiang Uygur autonomous region%新疆维吾尔自治区夏尔西里自然保护区全沟硬蜱复合感染蜱媒病原研究

    Institute of Scientific and Technical Information of China (English)

    刘晓明; 张桂林; 刘然; 孙响; 郑重; 邱尔臣; 马晓玲

    2015-01-01

    目的 了解新疆维吾尔自治区(新疆)夏尔西里自然保护区全沟硬蜱蜱媒病原复合感染情况.方法 2012年6月至2013年6月在新疆夏尔西里自然保护区采用布旗法采集全沟硬蜱,通过巢式PCR方法对蜱体内莱姆病伯氏疏螺旋体[Borrelia(B.) burgdorferi] 5S~ 23S rRNA基因间隔区、无形体(Anaplasma)16S rRNA基因、斑点热立克次体(Rickettsia) ompA基因、Q热贝氏柯克斯体Coxiella(C.)burneti的com1基因、巴贝西原虫(Babesia)的Nss-rRNA基因进行扩增.结果 204只全沟硬蜱中104只蜱病原检测阳性,阳性率为50.98%,其中伯氏疏螺旋体阳性率为34.31%(n=70),斑点热立克次体阳性率为28.92%(n=59),嗜吞噬细胞无形体阳性率为9.31%(n=19),未检出Q热贝氏柯克斯体和巴贝西原虫.x2检验显示,不同病原感染率差异有统计学意义(x2=38.517,P=0.000).204只蜱中有39只存在复合感染,复合感染率为19.12%(39/204),在病原检测阳性蜱中复合感染率为37.50%(39/104).伯氏疏螺旋体与斑点热立克次体复合感染率为16.18%(n=33);伯氏疏螺旋体与嗜吞噬细胞无形体复合感染率为4.90%(n=10),斑点热立克次体与嗜吞噬细胞无形体复合感染率为2.94%(n=6),3种病原体复合感染率为2.45%(n=5).经x2检验,3种病原混合感染率差异有统计学意义(x2=41.328,P=0.000).结论 新疆夏尔西里自然保护区全沟硬蜱存在伯氏疏螺旋体、嗜吞噬细胞无形体、斑点热立克次体的自然感染,并存在2~3种病原的复合感染.%Objective To study the co-infection of tick-borne pathogens in Ixodes persulcatus collected in Charles Hilary,Xinjiang Uygur autonomous region (Xinjiang).Methods Ticks were collected by flagging in grassy areas in Charles Hilary,Xinjiang from June 2012 to June 2013.The 5S-23S rRNA intergenic region from Borrelia (B.) burgdorferi,16S rRNA gene from Anaplasma,ompA gene from spotted fever group Rickettsia,comI gene from Coxiella (C

  18. Parasites and vector-borne pathogens in client-owned dogs in Albania. Blood pathogens and seroprevalences of parasitic and other infectious agents.

    Science.gov (United States)

    Hamel, Dietmar; Shukullari, Enstela; Rapti, Dhimitër; Silaghi, Cornelia; Pfister, Kurt; Rehbein, Steffen

    2016-02-01

    Knowledge on the epidemiology of parasitic and vector-borne infections is still very limited for Albania, a country located in the Balkan Peninsula in southeast Europe. Recent publications indicated prevalence rates of up to 52% for vector-borne infections in less-cared dogs in Albania. To provide data on the epidemiological situation in dogs under veterinary care, a total of 602 client-owned dogs presented to four small animal clinics between March 2010 and April 2011 in Tirana, Albania, were screened by examination of Giemsa-stained blood smears, PCR, and serological methods for the presence of arthropod-borne infections, as well as Neospora caninum and Toxoplasma gondii. Eight different pathogens, namely Babesia vogeli, Hepatozoon canis, Leishmania infantum, Dirofilaria immitis, Anaplasma phagocytophilum, Anaplasma platys, Ehrlichia canis, and Mycoplasma haemocanis, were detected by direct methods with prevalence rates ranging from 1 to 9%. Seroprevalence for Babesia spp., L. infantum, Anaplasma spp., and E. canis were 6.6, 5.1, 24.1, and 20.8%, respectively. Dogs >1 year of age were positive for vector-borne infections significantly more often than younger dogs (p = 0.003). More than half (51.7%) of the dogs were seroreactive to T. gondii and 18.3% to N. caninum. This is the first report on the detection of A. phagocytophilum, A. platys, E. canis, and M. haemocanis by PCR as well as the serological confirmation of exposure of dogs to N. caninum and T. gondii in Albania. The spectrum of pathogens and the seroprevalences for N. caninum and T. gondii in client-owned dogs from Tirana, Albania, are comparable to that reported in other countries in the Mediterranean Basin. The prevalence rates of vector-borne pathogens are at the lower range of that reported in studies from this geographical region. This is probably due to increased awareness of the owners of pet dogs, including better husbandry conditions and ectoparasiticidal treatment, thus limiting exposure

  19. First molecular detection of tick-borne pathogens in dogs from Jiangxi, China

    Science.gov (United States)

    ZHENG, Weiqing; LIU, Mingming; MOUMOUNI, Paul Franck Adjou; LIU, Xiaoqing; EFSTRATIOU, Artemis; LIU, Zhanbin; LIU, Yangqing; TAO, Huiying; GUO, Huanping; WANG, Guanbo; GAO, Yang; LI, Zifen; RINGO, Aaron Edmund; JIRAPATTHARASATE, Charoonluk; CHEN, Haiying; XUAN, Xuenan

    2016-01-01

    In this study, blood samples obtained from 162 dogs in Jiangxi, China, were employed in molecular screening of canine tick-borne pathogens by PCR and sequencing. Babesia spp. gene fragment was detected in 12 (7.41%) dogs. All samples were negative for Hepatozoon spp., Ehrlichia canis, Coxiella spp., Borrelia spp., Rickettsia spp. and Anaplasma platys. Species-specific PCR analysis further confirmed that 8 (4.94%) and 4 (2.47%) dogs were infected by Babesia canis vogeli and Babesia gibsoni, respectively. Based on our analyses, Babesia spp. infection in Jiangxi appeared not related to age, gender, breed, usage, activity and health status or tick infestation history of the dogs. This is the first molecular report of Babesia canis vogeli and Babesia gibsoni in dogs from Jiangxi, China. PMID:27890889

  20. Study on coinfecting vector-borne pathogens in dogs and ticks in Rio Grande do Norte, Brazil

    Directory of Open Access Journals (Sweden)

    Luiz Ricardo Gonçalves

    Full Text Available Since dogs presenting several vector borne diseases can show none or nonspecific clinical signs depending on the phase of infection, the assessment of the particular agents involved is mandatory. The present study aimed to investigate the presence of Babesia spp., Ehrlichia spp., Anaplasma spp., Hepatozoon spp. and Leishmania spp. in blood samples and ticks, collected from two dogs from Rio Grande do Norte showing suggestive tick-borne disease by using molecular techniques. DNA of E. canis, H. canis and L. infantum were detected in blood samples and R. sanguineus ticks collected from dogs. Among all samples analyzed, two showed the presence of multiple infections with E. canis, H. canis and L. infantum chagasi. Here we highlighted the need for molecular differential diagnosis in dogs showing nonspecific clinical signs.

  1. Tick-borne pathogens of potential zoonotic importance in the southern African Region

    Directory of Open Access Journals (Sweden)

    Simbarashe Chitanga

    2014-02-01

    Full Text Available The aim of this communication is to provide preliminary information on the tick-borne pathogens of potential zoonotic importance present in southern Africa, mainly focusing on their geographical distribution and host range, and to identify research gaps. The following tick-borne zoonoses have been reported to occur in southern Africa based mainly on case reports: Crimean–Congo haemorrhagic fever caused by Crimean–Congo haemorrhagic fever virus; ehrlichiosis caused by Ehrlichia ruminantium, Ehrlichia canis and Anaplasma phagocytophilum; babesiosis caused by Babesia microti; relapsing fever caused by Borrelia duttonii and rickettsioses caused by Rickettsia africae, Rickettsia aeschlimannii and Rickettsia conorii. The epidemiological factors influencing their occurrence are briefly reviewed.

  2. Canine vector-borne diseases in Brazil

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    Dantas-Torres Filipe

    2008-08-01

    Full Text Available Abstract Canine vector-borne diseases (CVBDs are highly prevalent in Brazil and represent a challenge to veterinarians and public health workers, since some diseases are of great zoonotic potential. Dogs are affected by many protozoa (e.g., Babesia vogeli, Leishmania infantum, and Trypanosoma cruzi, bacteria (e.g., Anaplasma platys and Ehrlichia canis, and helminths (e.g., Dirofilaria immitis and Dipylidium caninum that are transmitted by a diverse range of arthropod vectors, including ticks, fleas, lice, triatomines, mosquitoes, tabanids, and phlebotomine sand flies. This article focuses on several aspects (etiology, transmission, distribution, prevalence, risk factors, diagnosis, control, prevention, and public health significance of CVBDs in Brazil and discusses research gaps to be addressed in future studies.

  3. Human granulocytic anaplasmosis in Austria: epidemiological, clinical, and laboratory findings in five consecutive patients from Tyrol, Austria.

    Science.gov (United States)

    Walder, Gernot; Fuchs, Dietmar; Sarcletti, Mario; Berek, Klaus; Falkensammer, Barbara; Huber, Klaus; Petrovec, Miro; Dierich, Manfred P; Würzner, Reinhard

    2006-05-01

    We report five consecutive cases of Anaplasma (A.) phagocytophilum infection (the causative agent of human granulocytic anaplasmosis (HGA)) from western Austria. All infections were acquired between June and August in 2003 and 2004 in the Inn valley (Tyrol, Austria). Four patients required hospitalisation, one patient was treated as an outpatient. During the acute stage of illness, laboratory findings included thrombocytopenia (5/5), elevated C-reactive protein (5/5), elevated neopterin (5/5), elevated lactate dehydrogenase (4/5), and elevation of liver enzymes (4/5). Leukopenia (3/5) and elevated procalcitonin (2/5) were less frequently observed. All patients were treated with tetracyclines, which led to prompt improvement of the clinical conditions. Anti-platelet antibodies were observed in one of four patients, but remained unchanged after complete covalescence.

  4. Problem of ticks and tick-borne diseases in India with special emphasis on progress in tick control research: a review.

    Science.gov (United States)

    Ghosh, Srikant; Nagar, Gaurav

    2014-12-01

    Ticks, as vectors of several zoonotic diseases, are ranked second only to mosquitoes as vectors. The diseases spread by ticks are a major constraint to animal productivity while causing morbidity and mortality in both animals and humans. A number of tick species have been recognised since long as vectors of lethal pathogens, viz. Crimean-Congo haemorrhagic fever virus (CCHFV), Kyasanur forest disease virus (KFDV), Babesia spp, Theileria, Rickettsia conorii, Anaplasma marginale, etc. and the damages caused by them are well-recognised. There is a need to reassess the renewed threat posed by the tick vectors and to prioritize the tick control research programme. This review is focused on the major tick-borne human and animal diseases in India and the progress in vector control research with emphasis on acaricide resistance, tick vaccine and the development of potential phytoacaricides as an integral part of integrated tick control programme.

  5. Wild birds and urban ecology of ticks and tick-borne pathogens, Chicago, Illinois, USA, 2005-2010.

    Science.gov (United States)

    Hamer, Sarah A; Goldberg, Tony L; Kitron, Uriel D; Brawn, Jeffrey D; Anderson, Tavis K; Loss, Scott R; Walker, Edward D; Hamer, Gabriel L

    2012-10-01

    Bird-facilitated introduction of ticks and associated pathogens is postulated to promote invasion of tick-borne zoonotic diseases into urban areas. Results of a longitudinal study conducted in suburban Chicago, Illinois, USA, during 2005-2010 show that 1.6% of 6,180 wild birds captured in mist nets harbored ticks. Tick species in order of abundance were Haemaphysalis leporispalustris, Ixodes dentatus, and I. scapularis, but 2 neotropical tick species of the genus Amblyomma were sampled during the spring migration. I. scapularis ticks were absent at the beginning of the study but constituted the majority of ticks by study end and were found predominantly on birds captured in areas designated as urban green spaces. Of 120 ticks, 5 were infected with Borrelia burgdorferi, spanning 3 ribotypes, but none were infected with Anaplasma phagocytophilum. Results allow inferences about propagule pressure for introduction of tick-borne diseases and emphasize the large sample sizes required to estimate this pressure.

  6. Metabolismo oxidativo dos neutrófilos de bezerros das raças Nelore e Limousin nos primeiros quatro meses de vida Neutrophil oxidative burst in Nelore and Limousin calves in the first four months of life

    Directory of Open Access Journals (Sweden)

    Márcio Carvalho da Costa

    2008-09-01

    Full Text Available Com o objetivo de investigar alguns mecanismos de defesa em bezerros de raças de corte criadas a campo, foram utilizados 90 animais recém-nascidos aparentemente sadios, 45 da raça Nelore e 45 da raça Limousin. Amostras de sangue foram colhidas de cada bezerro entre 24 e 36 horas de vida e aos 15, 30, 60, 90 e 120 dias. Determinaram-se o leucograma e o metabolismo oxidativo dos neutrófilos por meio do teste da redução do tetrazólio de nitroazul (NBT, provas não estimulada (NBT-NE e estimulada (NBT-E. Investigou-se a taxa de parasitemia determinada por Anaplasma marginale, Babesia bigemina e B. bovis. Utilizou-se a análise de variância de medidas repetidas para estudar o comportamento das variáveis hematológicas com o avançar da idade. O teste de Krushkal-Wallis foi empregado para caracterizar a variação da porcentagem de neutrófilos reativos relacionada à idade. Comparações entre as raças foram realizadas em cada idade por meio do teste de Mann-Whithney. A contagem total de leucócitos aumentou com a idade nas duas raças estudadas. Observou-se diminuição do número de neutrófilos e aumento do de linfócitos, ocorrendo sua inversão antes dos 15 dias de idade. A capacidade oxidativa dos neutrófilos foi menor nos bezerros recém-nascidos e aumentou com o avançar da idade. Os neutrófilos dos bezerros Limousin apresentaram maior capacidade de redução do NBT do que os dos bezerros Nelore, notadamente após o segundo mês de vida. Essa diferença não provocou reflexos sobre a saúde dos animais e pode ser resultado da infecção natural e assintomática com o Anaplasma marginale.To study some defence mechanisms of beef calves kept on range conditions, 90 healthy calves, 45 Nelore and 45 Limousin, were bled at 24 to 36 hours of life, 15, 30, 60, 90 and, 120 days. Leukogram and neutrophil oxidative burst through the non-stimulated and the stimulated NBT tests were determined. The parasitemia with Anaplasma marginale

  7. New criteria for selecting the origin of DNA replication in Wolbachia and closely related bacteria

    DEFF Research Database (Denmark)

    Ioannidis, Panagiotis; Dunning Hotopp, Julie C; Sapountzis, Panagiotis;

    2007-01-01

    , the origin of DNA replication (ori) regions were identified in silico for Wolbachia strains and eleven other related bacteria belonging to Ehrlichia, Anaplasma, and Rickettsia genera. These features include DnaA-, CtrA- and IHF-binding sites as well as the flanking genes in C. crescentus. The Wolbachia ori...... bacteria while fundamental characteristics like presence of DnaA and IHF binding sites as well as the boundary genes are more widely conserved. The relative paucity of CtrA binding sites in the ori regions, as well as the absence of key enzymes associated with DNA replication in the respective genomes......BACKGROUND: The annotated genomes of two closely related strains of the intracellular bacterium Wolbachia pipientis have been reported without the identifications of the putative origin of replication (ori). Identifying the ori of these bacteria and related alpha-Proteobacteria as well...

  8. Fatal parasitosis in blackbucks (Antilope cervicapra: a possible factor risk in hunting units

    Directory of Open Access Journals (Sweden)

    Ned de la Cruz-Hernández

    2015-11-01

    Full Text Available In February 2012, a reproductive group of 60 adult blackbucks (Antilope cervicapra from Veracruz, Mexico was relocated to hunting units in eastern and northeastern Mexico. Seven individuals died due to hemorrhagic parasitic, abomasitis and enteritis caused by Haemonchus spp., Setaria spp., and Trichostrongylids. Deaths were associated with hepatic necrosis, bilateral congestive distention of heart and fibrinonecrotic bronchopneumonia. Also Anaplasma marginale was identified. The blackbucks’ population displayed a general mortality rate of 11.67%, where 25% of total male and 9.62% of total female died. The mortality was controlled by segregation of all remaining blackbucks and the treatment for internal and external parasites (biting flies and ticks. After the treatment, no fatality cases related to parasitosis were recorded. The results presented here exhibit the high relevance of parasitosis as possible factor risk in the survival of tis specie.

  9. 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.

  10. Fatal human anaplasmosis associated with macrophage activation syndrome in Greece and the Public Health response.

    Science.gov (United States)

    Tsiodras, Sotirios; Spanakis, Nikos; Spanakos, Gregory; Pervanidou, Danai; Georgakopoulou, Theano; Campos, Elsa; Petra, Theofania; Kanellopoulos, Petros; Georgiadis, George; Antalis, Emmanouil; Kontos, Vassileios; Giannopoulos, Lambros A; Tselentis, Yiannis; Papa, Anna; Tsakris, Athanassios; Saroglou, George

    2017-02-08

    Human granulocytic anaplasmosis (HGA) is a tick-borne disease caused by Anaplasma phagocytophilum that has the potential to spread in new geographical areas. The first fatal case of HGA in Greece is presented. Fever of unknown origin, renal and respiratory insufficiency and development of macrophage activation syndrome characterized the clinical presentation. Amplification and sequencing of a fragment of the groEL gene revealed the presence of A. phagocytophilum. The epidemiological and clinical features were collected during an epidemiological investigation. Public health measures were instituted by the Hellenic Centre for Disease Control and Prevention. The Public Health intervention required the collaboration of epidemiologists, veterinarians and microbiologists. Emphasis was given to communication activities and misconceptions concerning canines and their role in the disease. The emergence of human anaplasmosis in a new geographical area highlights the importance of disease awareness and of the need for continued support for tick and tick-borne disease surveillance networks.

  11. Antigenic Variation in Bacterial Pathogens.

    Science.gov (United States)

    Palmer, Guy H; Bankhead, Troy; Seifert, H Steven

    2016-02-01

    Antigenic variation is a strategy used by a broad diversity of microbial pathogens to persist within the mammalian host. Whereas viruses make use of a minimal proofreading capacity combined with large amounts of progeny to use random mutation for variant generation, antigenically variant bacteria have evolved mechanisms which use a stable genome, which aids in protecting the fitness of the progeny. Here, three well-characterized and highly antigenically variant bacterial pathogens are discussed: Anaplasma, Borrelia, and Neisseria. These three pathogens display a variety of mechanisms used to create the structural and antigenic variation needed for immune escape and long-term persistence. Intrahost antigenic variation is the focus; however, the role of these immune escape mechanisms at the population level is also presented.

  12. Influence of dipping practices on the seroprevalence of babesiosis and anaplasmosis in the foot-and-mouth disease buffer zone adjoining the Kruger National Park in South Africa.

    Science.gov (United States)

    Stevens, K B; Spickett, A M; Vosloo, W; Pfeiffer, D U; Dyason, E; Du Plessis, B

    2007-06-01

    A serological survey of bovine babesiosis and anaplasmosis was conducted in the foot-and-mouth disease buffer zone surrounding the Kruger National Park in South Africa between 2001 and 2003 to determine whether the withdrawal of government-subsidized dipping in certain regions had affected the seroprevalence of these tick-borne diseases. Seroprevalence of Anaplasma marginale and Babesia bovis increased during the study period. This increase was greater in Limpopo Province where farmers had to supply their own acaricide than in Mpumalanga Province where dipping materials were provided by the local Veterinary Services. The number of animals testing positive for B. bigemina decreased in both provinces during the study period, which was attributed to possible vector displacement rather than more effective tick control measures. Responses to a questionnaire on ticks and tick-borne diseases revealed local knowledge on the subject to be highly variable and sometimes incorrect.

  13. Rickettsial pathogens and arthropod vectors of medical and veterinary significance on Kwajalein Atoll and Wake Island

    Directory of Open Access Journals (Sweden)

    Durden, L.

    2012-05-01

    Full Text Available Modern surveys of ectoparasites and potential vector-borne pathogens in the Republic of the Marshall Islands and Wake Island are poorly documented. We report on field surveys of ectoparasites from 2010 with collections from dogs, cats, and rats. Five ectoparasites were identified: the cat flea Ctenocephalides felis, a sucking louse Hoplopleura pacifica, the mites Laelaps nuttalli and Radfordia ensifera, and the brown dog tickRhipicephalus sanguineus. Ectoparasites were screened for rickettsial pathogens. DNA from Anaplasma platys, a Coxiella symbiont of Rhipicephalus sanguineus, anda Rickettsia sp. were identified by PCR and DNA sequencing from ticks and fleas on Kwajalein Atoll. An unidentified spotted fever group Rickettsia was detected in a pool of Laelaps nuttalli and Hoplopleura pacifica from Wake Island. The records of Hoplopleura pacifica, Laelaps nuttalli, and Radfordia ensifera and the pathogens are new for Kwajalein Atoll and Wake Island.

  14. Ehrlichia sp. infection in carthorses of low-income owners, Southern Brazil.

    Science.gov (United States)

    Vieira, Thállitha S; Vieira, Rafael F; Krawczak, Felipe S; Soares, Herbert S; Guimarães, Ana M; Barros-Filho, Ivan R; Marcondes, Mary; Labruna, Marcelo B; Biondo, Alexander W; Vidotto, Odilon

    2016-10-01

    Although well established in dogs, Ehrlichia sp. infection has been scarcely reported in horses. The aim was to perform a comprehensive serological and molecular survey for the detection of Ehrlichia spp. in carthorses from Southern Brazil. Blood samples from 190 carthorses from Paraná State were sampled. Horses were also tested for Borrelia burgdorferi and Anaplasma phagocytophilum. Anti-Ehrlichia sp. antibodies were detected by a commercial rapid ELISA, and immunofluorescence antibody assays (IFA) with E. chaffeensis and E. canis as crude antigens. The molecular and phylogenetic analysis of Ehrlichia sp. was based on 16S rRNA and dsb genes. A total of 52 (27.4%), 4 (2.1%), and 3 (1.6%) horses were positive for Ehrlichia spp., Anaplasma spp. and Borrelia burgdorferi, respectively, by the commercial rapid ELISA. Thirty-eight (20.0%) and 37 (19.5%) horses showed anti-E. chaffeensis and anti-E. canis antibodies by IFA, respectively. One blood sample that also showed anti-E. chaffeensis antibodies was PCR positive for the 16S rRNA and dsb genes of Ehrlichia spp., showing an identity of>98.0% to the uncultured Ehrlichia sp. previously detected in Brazilian jaguars (Panthera onca). Anti-Ehrlichia sp. antibodies and Ehrlichia DNA were detected in carthorses from Southern Brazil, which may post public health concerns due to intimate contact with low-income owners. This is the first report of a natural infection of this bacteria in horses from South America. Clinical signs and the tick vector remain unknown.

  15. Tick-borne rickettsial pathogens in ticks and small mammals in Korea.

    Science.gov (United States)

    Kim, Chul-Min; Yi, Ying-Hua; Yu, Do-Hyeon; Lee, Mi-Jin; Cho, Mae-Rim; Desai, Atul R; Shringi, Smriti; Klein, Terry A; Kim, Heung-Chul; Song, Jin-Won; Baek, Luck-Ju; Chong, Sung-Tae; O'guinn, Monica L; Lee, John S; Lee, In-Yong; Park, Jin-Ho; Foley, Janet; Chae, Joon-Seok

    2006-09-01

    In order to investigate the prevalence of tick-borne infectious agents among ticks, ticks comprising five species from two genera (Hemaphysalis spp. and Ixodes spp.) were screened using molecular techniques. Ticks (3,135) were collected from small wild-caught mammals or by dragging/flagging in the Republic of Korea (ROK) and were pooled into a total of 1,638 samples (1 to 27 ticks per pool). From the 1,638 tick samples, species-specific fragments of Anaplasma phagocytophilum (1 sample), Anaplasma platys (52 samples), Ehrlichia chaffeensis (29 samples), Ehrlichia ewingii (2 samples), Ehrlichia canis (18 samples), and Rickettsia rickettsii (28 samples) were amplified by PCR assay. Twenty-one pooled and individual tick samples had mixed infections of two (15 samples) or three (6 samples) pathogens. In addition, 424 spleen samples from small captured mammals (389 rodents, 33 insectivores, and 2 weasels) were screened for selected zoonotic pathogens. Species-specific DNA fragments of A. phagocytophilum (110 samples), A. platys (68 samples), E. chaffeensis (8 samples), E. ewingii (26 samples), E. canis (51 samples), and Rickettsia sp. (22 samples) were amplified by PCR assay. One hundred thirty small mammals had single infections, while 4, 14, and 21 striped field mice (Apodemus agrarius) had mixed infections of four, three, and two pathogens, respectively. Phylogenetic analysis based on nucleotide sequence comparison also revealed that Korean strains of E. chaffeensis clustered closely with those from China and the United States, while the Rickettsia (rOmpA) sequences clustered within a clade together with a Chinese strain. These results suggest that these agents should be considered in differential diagnosis while examining cases of acute febrile illnesses in humans as well as animals in the ROK.

  16. Tick-borne Diseases (Borreliosis, Anaplasmosis, Babesiosis) in German and Austrian Dogs: Status quo and Review of Distribution, Transmission, Clinical Findings, Diagnostics and Prophylaxis.

    Science.gov (United States)

    Pantchev, Nikola; Pluta, Silvia; Huisinga, Elke; Nather, Stephanie; Scheufelen, Miriam; Vrhovec, Majda Globokar; Schweinitz, Andrea; Hampel, Herwig; Straubinger, Reinhard K

    2015-08-01

    Tick-borne diseases (TBD) in dogs have gained in significance in German and Austrian veterinary practices. The widespread European tick species Ixodes ricinus represents an important vector for spirochaetes of the Borrelia burgdorferi sensu lato group and Rickettsiales such as Anaplasma phagocytophilum. The meadow or ornate dog tick (Dermacentor reticulatus) is an important vector for Babesia canis, as is the brown dog tick (Rhipicephalus sanguineus) for Babesia vogeli in the Mediterranean region. The present work covers pathogen transmission by tick vectors, including the mechanisms and the minimum intervals required, in conjunction with possible non-vector-borne transmission routes. It also addresses the incubation periods, pathogenicity and clinical findings associated with each pathogen and genospecies and presents case examples. Current data on prevalence, annual fluctuations and distribution in various pre-selected dog populations (symptomatic versus asymptomatic) in both countries are depicted in maps. Reasons for changes in prevalence (especially of Borrelia) are discussed. Criteria and algorithms for clinical diagnosis and monitoring in dogs, including case history, direct detection (blood smears, molecular detection by species-specific PCR and sequencing) and indirect methods (whole-cell and peptide-based antibody tests), are presented, together with laboratory abnormalities (haematology, clinical chemistry, urine). The role of anti-C6 antibody concentration (ACAC) and its correlation with proteinuria and Lyme nephritis are assessed on the basis of new data. Consideration is also given to the importance of blood smears, PCR and serology in the case of anaplasmosis and babesiosis, and the diagnostic value of combining these methods. The relevance of molecular differentiation of Anaplasma species (A. phagocytophilum versus A. platys) and Babesia spp. (large versus small forms) in cases of serological cross-reaction is emphasized. A summary is given of

  17. Prevalence of tick-borne pathogens in Ixodes ricinus and Dermacentor reticulatus ticks from different geographical locations in Belarus.

    Science.gov (United States)

    Reye, Anna L; Stegniy, Valentina; Mishaeva, Nina P; Velhin, Sviataslau; Hübschen, Judith M; Ignatyev, George; Muller, Claude P

    2013-01-01

    Worldwide, ticks are important vectors of human and animal pathogens. Besides Lyme Borreliosis, a variety of other bacterial and protozoal tick-borne infections are of medical interest in Europe. In this study, 553 questing and feeding Ixodes ricinus (n = 327) and Dermacentor reticulatus ticks (n = 226) were analysed by PCR for Borrelia, Rickettsia, Anaplasma, Coxiella, Francisella and Babesia species. Overall, the pathogen prevalence in ticks was 30.6% for I. ricinus and 45.6% for D. reticulatus. The majority of infections were caused by members of the spotted-fever group rickettsiae (24.4%), 9.4% of ticks were positive for Borrelia burgdorferi sensu lato, with Borrelia afzelii being the most frequently detected species (40.4%). Pathogens with low prevalence rates in ticks were Anaplasma phagocytophilum (2.2%), Coxiella burnetii (0.9%), Francisella tularensis subspecies (0.7%), Bartonella henselae (0.7%), Babesia microti (0.5%) and Babesia venatorum (0.4%). On a regional level, hotspots of pathogens were identified for A. phagocytophilum (12.5-17.2%), F. tularensis ssp. (5.5%) and C. burnetii (9.1%), suggesting established zoonotic cycles of these pathogens at least at these sites. Our survey revealed a high burden of tick-borne pathogens in questing and feeding I. ricinus and D. reticulatus ticks collected in different regions in Belarus, indicating a potential risk for humans and animals. Identified hotspots of infected ticks should be included in future surveillance studies, especially when F. tularensis ssp. and C. burnetii are involved.

  18. 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.

  19. Prevalence of Tick-Borne Pathogens in Ixodes ricinus and Dermacentor reticulatus Ticks from Different Geographical Locations in Belarus

    Science.gov (United States)

    Reye, Anna L.; Stegniy, Valentina; Mishaeva, Nina P.; Velhin, Sviataslau; Hübschen, Judith M.; Ignatyev, George; Muller, Claude P.

    2013-01-01

    Worldwide, ticks are important vectors of human and animal pathogens. Besides Lyme Borreliosis, a variety of other bacterial and protozoal tick-borne infections are of medical interest in Europe. In this study, 553 questing and feeding Ixodes ricinus (n = 327) and Dermacentor reticulatus ticks (n = 226) were analysed by PCR for Borrelia, Rickettsia, Anaplasma, Coxiella, Francisella and Babesia species. Overall, the pathogen prevalence in ticks was 30.6% for I. ricinus and 45.6% for D. reticulatus. The majority of infections were caused by members of the spotted-fever group rickettsiae (24.4%), 9.4% of ticks were positive for Borrelia burgdorferi sensu lato, with Borrelia afzelii being the most frequently detected species (40.4%). Pathogens with low prevalence rates in ticks were Anaplasma phagocytophilum (2.2%), Coxiella burnetii (0.9%), Francisella tularensis subspecies (0.7%), Bartonella henselae (0.7%), Babesia microti (0.5%) and Babesia venatorum (0.4%). On a regional level, hotspots of pathogens were identified for A. phagocytophilum (12.5–17.2%), F. tularensis ssp. (5.5%) and C. burnetii (9.1%), suggesting established zoonotic cycles of these pathogens at least at these sites. Our survey revealed a high burden of tick-borne pathogens in questing and feeding I. ricinus and D. reticulatus ticks collected in different regions in Belarus, indicating a potential risk for humans and animals. Identified hotspots of infected ticks should be included in future surveillance studies, especially when F. tularensis ssp. and C. burnetii are involved. PMID:23349900

  20. Choque séptico por ehrliquiosis

    Directory of Open Access Journals (Sweden)

    Viviana Hernández-de Mezerville

    2007-04-01

    Full Text Available La ehrliquiosis humana es una infección zoonótica transmitida por "garrapatas duras" causada por Anaplasma sp y Ehrlichia sp. Puede presentarse como un cuadro pseudogripal con fiebre, malestar general, mialgias y cefalea o hasta un choque séptico con falla multiorgánica y muerte. La mitad de los pacientes requieren atención hospitalaria en algún momento y se reporta una mortalidad de hasta el 5% secundaria a enfermedades oportunistas o a hemorragias. El diagnóstico se puede sospechar visualizando las mórulas intracitoplasmáticas en las células afectadas en frotis de sangre periférica, y se confirma con serología, con cultivos o con detección molecular. El tratamiento de elección es la doxiciclina con buena respuesta clínica. Se reporta un caso probable de ehrliquiosis humana asociado a choque séptico y falla multiorgánica tratado exitosamente en el Hospital Dr. Rafael Angel Calderón Guardia.Human ehrlichiosis is a tick-borne infectious disease caused by Anaplasma sp and Ehrlichia sp. The presentation can range from a flu-like syndrome with fever, malaise, myalgia and headache to a septic shock state with multiorganic dysfunction. Half the patients require in- hospital care and the mortality rate is 5% as a result of opportunistic infections or hemorrhage. The diagnosis can be made by the visualization of intracytoplasmatic morulae in the blood smear, serology, cultives or polymerase chain reaction. The treatment of choice is doxycicline and generally carries a good response. We report a case of human ehrlichiosis associated with septic shock and multiorganic dysfunction successfully treated at the Calderon Guardia Hospital.

  1. Molecular detection of bacterial and parasitic pathogens in hard ticks from Portugal.

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    Maia, Carla; Ferreira, Andreia; Nunes, Mónica; Vieira, Maria Luísa; Campino, Lenea; Cardoso, Luís

    2014-06-01

    Ticks are important vector arthropods of human and animal pathogens. As information about agents of disease circulating in vectors in Portugal is limited, the aim of the present study was to detect bacteria and parasites with veterinary and zoonotic importance in ticks collected from dogs, cats, and field vegetation. A total of 925 ticks, comprising 888 (96.0%) adults, 8 (0.9%) nymphs, and 29 (3.1%) larvae, were collected in 4 geographic areas (districts) of Portugal. Among those, 620 (67.0%) were removed from naturally infested dogs, 42 (4.5%) from cats, and 263 (28.4%) were questing ticks obtained from field vegetation. Rhipicephalus sanguineus was the predominant tick species, and the only one collected from dogs and vegetation, while all Ixodes ricinus specimens (n=6) were recovered from cats. Rickettsia massiliae and Rickettsia conorii were identified in 35 ticks collected from cats and dogs and in 3 ticks collected from dogs. Among ticks collected from cats or dogs, 4 Rh. sanguineus specimens were detected with Hepatozoon felis, 3 with Anaplasma platys, 2 with Hepatozoon canis, one with Anaplasma phagocytophilum, one with Babesia vogeli, one with Borrelia burgdorferi sensu lato and one with Cercopithifilaria spp. Rickettsia helvetica was detected in one I. ricinus tick collected from a cat. To the best of our knowledge, this was the first time that Cercopithifilaria spp., Ba. vogeli, H. canis, and H. felis have been detected in ticks from Portugal. The wide range of tick-borne pathogens identified, some of zoonotic concern, suggests a risk for the emergence of tick-borne diseases in domestic animals and humans in Portugal. Further studies on these and other tick-borne agents should be performed to better understand their epidemiological and clinical importance, and to support the implementation of effective control measures.

  2. Survey of Ixodes pacificus Ticks in California Reveals a Diversity of Microorganisms and a Novel and Widespread Anaplasmataceae Species.

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    Mark W Eshoo

    Full Text Available Ixodes pacificus ticks can harbor a wide range of human and animal pathogens. To survey the prevalence of tick-borne known and putative pathogens, we tested 982 individual adult and nymphal I. pacificus ticks collected throughout California between 2007 and 2009 using a broad-range PCR and electrospray ionization mass spectrometry (PCR/ESI-MS assay designed to detect a wide range of tick-borne microorganisms. Overall, 1.4% of the ticks were found to be infected with Borrelia burgdorferi, 2.0% were infected with Borrelia miyamotoi and 0.3% were infected with Anaplasma phagocytophilum. In addition, 3.0% were infected with Babesia odocoilei. About 1.2% of the ticks were co-infected with more than one pathogen or putative pathogen. In addition, we identified a novel Anaplasmataceae species that we characterized by sequencing of its 16S rRNA, groEL, gltA, and rpoB genes. Sequence analysis indicated that this organism is phylogenetically distinct from known Anaplasma species with its closest genetic near neighbors coming from Asia. The prevalence of this novel Anaplasmataceae species was as high as 21% at one site, and it was detected in 4.9% of ticks tested statewide. Based upon this genetic characterization we propose that this organism be called 'Candidatus Cryptoplasma californiense'. Knowledge of this novel microbe will provide awareness for the community about the breadth of the I. pacificus microbiome, the concept that this bacterium could be more widely spread; and an opportunity to explore whether this bacterium also contributes to human or animal disease burden.

  3. Small risk of developing symptomatic tick-borne diseases following a tick bite in the Netherlands

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    Hofhuis Agnetha

    2011-02-01

    Full Text Available Abstract Background In The Netherlands, the incidence of Lyme borreliosis is on the rise. Besides its causative agent, Borrelia burgdorferi s.l., other potential pathogens like Rickettsia, Babesia and Ehrlichia species are present in Ixodes ricinus ticks. The risk of disease associated with these microorganisms after tick-bites remains, however, largely unclear. A prospective study was performed to investigate how many persons with tick-bites develop localized or systemic symptoms and whether these are associated with tick-borne microorganisms. Results In total, 297 Ixodes ricinus ticks were collected from 246 study participants who consulted a general practitioner on the island of Ameland for tick bites. Ticks were subjected to PCR to detect DNA of Borrelia burgdorferi s.l., Rickettsia spp., Babesia spp. or Ehrlichia/Anaplasma spp.. Sixteen percent of the collected ticks were positive for Borrelia burgdorferi s.l., 19% for Rickettsia spp., 12% for Ehrlichia/Anaplasma spp. and 10% for Babesia spp.. At least six months after the tick bite, study participants were interviewed on symptoms by means of a standard questionnaire. 14 out of 193 participants (8.3% reported reddening at the bite site and 6 participants (4.1% reported systemic symptoms. No association between symptoms and tick-borne microorganisms was found. Attachment duration ≥24 h was positively associated with reddening at the bite site and systemic symptoms. Using logistic regression techniques, reddening was positively correlated with presence of Borrelia afzelii, and having 'any symptoms' was positively associated with attachment duration. Conclusion The risk of contracting acute Lyme borreliosis, rickettsiosis, babesiosis or ehrlichiosis from a single tick bite was

  4. Prevalence of tick-borne pathogens in Ixodes ricinus and Dermacentor reticulatus ticks from different geographical locations in Belarus.

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    Anna L Reye

    Full Text Available Worldwide, ticks are important vectors of human and animal pathogens. Besides Lyme Borreliosis, a variety of other bacterial and protozoal tick-borne infections are of medical interest in Europe. In this study, 553 questing and feeding Ixodes ricinus (n = 327 and Dermacentor reticulatus ticks (n = 226 were analysed by PCR for Borrelia, Rickettsia, Anaplasma, Coxiella, Francisella and Babesia species. Overall, the pathogen prevalence in ticks was 30.6% for I. ricinus and 45.6% for D. reticulatus. The majority of infections were caused by members of the spotted-fever group rickettsiae (24.4%, 9.4% of ticks were positive for Borrelia burgdorferi sensu lato, with Borrelia afzelii being the most frequently detected species (40.4%. Pathogens with low prevalence rates in ticks were Anaplasma phagocytophilum (2.2%, Coxiella burnetii (0.9%, Francisella tularensis subspecies (0.7%, Bartonella henselae (0.7%, Babesia microti (0.5% and Babesia venatorum (0.4%. On a regional level, hotspots of pathogens were identified for A. phagocytophilum (12.5-17.2%, F. tularensis ssp. (5.5% and C. burnetii (9.1%, suggesting established zoonotic cycles of these pathogens at least at these sites. Our survey revealed a high burden of tick-borne pathogens in questing and feeding I. ricinus and D. reticulatus ticks collected in different regions in Belarus, indicating a potential risk for humans and animals. Identified hotspots of infected ticks should be included in future surveillance studies, especially when F. tularensis ssp. and C. burnetii are involved.

  5. Prevalence of Toxoplasma gondii, Bartonella species and haemoplasma infection in cats in South Africa.

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    Lobetti, Remo; Lappin, Michael R

    2012-12-01

    Vector-borne agents and Toxoplasma gondii are common in cats, with many being zoonotic. The current study investigated the prevalence of selected infectious agents in cats from Johannesburg, South Africa, for which no published data exists. Whole blood and sera were obtained from 102 cats with a variety of disease conditions. Total DNA was extracted from the blood and assayed using PCR techniques for Mycoplasma haemofelis, Candidatus M haemominutum, Candidatus M turicensis, Bartonella species, Ehrlichia species and Anaplasma species. Enzyme-linked immunosorbent assays were used to detect IgG and IgM serum antibodies to T gondii and IgG serum antibodies to Bartonella species. Associations between test results, patient characteristics and haematological values were also evaluated. Overall, 56 cats (55%) were positive in one or more of the assays. Haemoplasma DNA was amplified from 26 cats [M haemofelis: four cats (3.9%); Candidatus M haemominutum from 22 cats (21.6%)] and Bartonella species DNA was amplified from eight cats [Bartonella henselae: five cats (4.9%); Bartonella clarridgeieae: three cats (2.9%)]; DNA of Ehrlichia species or Anaplasma species were not amplified. Of the cats, 24 (23.5%) were seropositive for Bartonella IgG and 18 (17.6%) were positive for T gondii IgM (12 cats), IgG (eight cats), or both (two cats). The study concluded that Bartonella species haemoplasmas and T gondii are common in client-owned cats in the region and the diagnosis of feline vector-borne agents and T gondii is difficult without the use of specific diagnostic tests, as there are minimal patient characteristics or haematological changes that indicate infection.

  6. Arthropod-borne pathogens circulating in free-roaming domestic cats in a zoo environment in Brazil.

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    André, Marcos Rogério; Baccarim Denardi, Nathani Cristina; Marques de Sousa, Keyla Carstens; Gonçalves, Luiz Ricardo; Henrique, Paloma Canedo; Grosse Rossi Ontivero, Claudia Regina; Lima Gonzalez, Irys Hany; Cabral Nery, Carolina Vaz; Fernandes Chagas, Carolina Romeiro; Monticelli, Cauê; Alexandre de Santis, Ana Cláudia Gabriela; Machado, Rosangela Zacarias

    2014-09-01

    Recently, tick and flea-borne pathogens have been detected in wild carnivores maintained in captivity in Brazilian zoos. Since free-roaming cats are frequently found in Brazilian zoos, they could act as reservoirs for arthropod-borne pathogens, which could be transmitted to endangered wild carnivores maintained in captivity in these institutions. On the other hand, stray cats in zoos may play a role as sentinels to pathogens that circulate among wild animals in captivity. The present work aimed to detect the presence of Anaplasmataceae agents, hemoplasmas, Bartonella species, piroplasmas, and Hepatozoon sp. DNA in blood samples of 37 free-roaming cats in a Brazilian zoo. Three (8%) cats were positive for Anaplasma spp. closed related to Anaplasma phagocytophilum; 12 (32%) cats were positive for hemoplasmas [two (5%) for Mycoplasma haemofelis, five (13.5%) for Candidatus Mycoplasma haemominutum, and five (13.5%) for Candidatus Mycoplasma turicensis]; 11 (30%) were positive for Bartonella spp., six (16%) were positive Babesia vogeli and one (3%) for Theileria sp. Coinfection with multiple arthropod-borne agentes was observed in sampled cats. None of sampled cats were positive for Ehrlichia spp., Cytauxzoon spp., or Hepatozoon spp. in PCR. This is the first molecular detection of Babesia vogeli and Theileria sp. in domestic cats in Brazil. The control of the population of free-roaming cats in these conservation institutions is much needed aiming to prevent the potential transmission to endangered wild animals maintained in captivity, such as wild neotropical wild felids, as well as to human beings visiting zoos.

  7. DIAGNÓSTICO EM ANAPLASMOSE BOVINA DIAGNOSIS IN BOVINE ANAPLASMOSIS

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    Odilon Vidotto

    2001-04-01

    Full Text Available Anaplasma marginale (THEILER 1910 é uma rickettsia intra-eritrocitária obrigatória de ruminantes susceptíveis, transmitida biológica e mecanicamente por carrapatos e insetos hematófagos. Ela determina o aparecimento de formas clínicas aguda, superaguda, leve e/ou crônica, com um período pré-patente de 20 a 40 dias seguido por parasitemia e intensa anemia, provocando perdas com um custo estimado de 40 milhões de dólares anuais; está amplamente distribuída nas regiões tropicais, subtropicais e temperadas do mundo. A. marginale confere imunidade de origem humoral e celular que não é dependente de infecção persistente. O diagnóstico é baseado em sinais clínicos, detecção do microrganismo no sangue, anticorpos no soro ou alterações patológicas post mortem. O objetivo deste trabalho é reunir informações sobre aspectos de diagnóstico da anaplasmose bovina.Anaplasma marginale (THEILER, 1910, a rickettsial hemoparasite of ruminants, is transmited biologically and mecanically by ticks and haematofagous insects. It has a pre-patent period of 20 to 40 days, followed by high parasitemia and severe anemia. Infected animals can develop a mild, aguda, hiperaguda or cronic clinical forms of the disease. A. marginale has an worldwide distribuition with high incidence in tropical and subtropical regions. A. marginale promove an immunity not dependent of a persistent infection, involving both, humural and celular immunological mechanisms. Diagnosis is based on clinical signs, detection of the microrganisms on the blood, antibodies on the serum or post-mortem pathological changes. Informations about the diagnostic aspects of anaplasmosis are reviewed.

  8. Molecular detection of vector-borne bacteria and protozoa in healthy hunting dogs from Central Italy

    Institute of Scientific and Technical Information of China (English)

    Valentina; Virginia; Ebani; Simona; Nardoni; Giulia; Fognani; Linda; Mugnaini; Fabrizio; Bertelloni; Guido; Rocchigiani; Roberto; Amerigo; Papini; Francesco; Stefani; Francesca; Mancianti

    2015-01-01

    Objective:To determine the pi’evalence of vector-bome bacteria and protozoa in hunting dogs living in Central Italy.Methods:Molecular testing was executed on DNA which was extracted from blood specimens collected from 117 asymptomatic dogs to detect Anaplasma phagocytophilum,Babesia canis(B.canis),Bartonella spp..Coxiella burnetii(C.burnetii).Ehrlichia canis.Hepatozoon canis.and Leislnnania infantum.Results:A total of 48 dogs(41.0%) were infested by Ixodes ricinus and Rhipicephalus sanguineus ticks.Tick-borne infections were observed in 64(54.7%) animals.More in detail.38 dogs(32.5%) screened positive for Hepatozoon canis,24(20.5%) for Bartonella rinsonii subsp.berkhoffii.20(17.1%) for Leishmania infantum,6(5.1%) for C.burnetii,5(4.3%) for B.canis(3 B.canis vogeli and 2 B.canis canis),3(2.5%) for Anaplasma phagocytophilum,and 2(1.7%) for Ehrlichia canis.Mixed infection by 2 agents occurred in 17(14.5%) subjects,by 3 agents in 7(6.0%) dogs,and by 4 agents in 1(0.9%) animal.Conclusions:The results demonstrated that several vector-borne pathogens were circulating in this region and dogs infected by these agents were usually asymptomatic.A relevant finding was the presence of DNA of C.burnetii,a severe zoonotic agent,in the 5.1% of tested dogs,which can be source of infection for their owners not only through tick bites,but also directly with urine,feces and birth products.

  9. Type-specific PCR assays for Babesia bovis msa-1 genotypes in Asia: Revisiting the genetic diversity in Sri Lanka, Mongolia, and Vietnam.

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    Liyanagunawardena, Nilukshi; Sivakumar, Thillaiampalam; Kothalawala, Hemal; Silva, Seekkuge Susil Priyantha; Battsetseg, Badgar; Lan, Dinh Thi Bich; Inoue, Noboru; Igarashi, Ikuo; Yokoyama, Naoaki

    2016-01-01

    Babesia bovis is the most virulent Babesia organism, resulting in a high mortality rate in cattle. The genetic diversity of B. bovis merozoite surface antigens (MSAs), such as MSA-1, MSA-2b, and MSA-2c, might be linked to altered immune profiles in the host animals. The present study aimed to develop type-specific PCR assays for Asian msa-1 genotypes, thereby re-analyzing the genetic diversity of msa-1 in Sri Lanka, Mongolia, and Vietnam. Specific primers were designed for nine Asian msa-1 genotypes, which had been detected based on the phylogeny constructed using msa-1 gene sequences retrieved from the GenBank database. Specificity of the type-specific PCR assays was confirmed using plasmids containing the inserts of msa-1 gene fragments that represent Asian genotypes. Furthermore, no amplicons were observed by these PCR assays when DNA samples of Babesia bigemina, Babesia ovata, Theileria annulata, Theileria orientalis, Trypanosoma evansi, Trypanosoma theileri, Anaplasma marginale, and Anaplasma bovis, and non-infected bovine blood were analyzed. In total, 109 B. bovis-positive blood DNA samples sourced from Sri Lanka (44 cattle), Mongolia (26 cattle), and Vietnam (23 cattle and 16 water buffaloes) were then screened by the type-specific PCR assays. The sequences derived from all of the PCR amplicons were phylogenetically analyzed. Out of 109 DNA samples, 23 (20 from cattle and 3 from water buffaloes) were positive for at least one genotype. In agreement with previous studies, five and four different genotypes were detected among the DNA samples from Sri Lanka and Vietnam, respectively. In contrast, four genotypes, including three novel genotypes, were detected from Mongolia. Five DNA samples were found to be co-infected with multiple genotypes. The sequences of the PCR amplicons clustered phylogenetically within the corresponding clades. These findings indicated that the type-specific PCR assays described herein are useful for the determination of genotypic

  10. Infecção natural por hemoparasitos em bezerros submetidos à quimio-profilaxia aos 30 dias de idade Natural infection by hemoparasites in calves submitted to chemo -prophylaxis wonder 30 days of age

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    Rosângela A. Da Silva

    2007-09-01

    Full Text Available O complexo Tristeza Parasitária acarreta grandes prejuízos à pecuária bovina nacional. Os principais agentes etiológicos são Babesia bigemina, B. bovis e Anaplasma marginale, sendo o carrapato Boophilus microplus o principal vetor. Este trabalho relata a ocorrência de infecção natural por hemoparasitos da tristeza parasitária bovina em 36 bezerros com alta infestação natural por carrapatos e submetidos à quimioprofilaxia aos 30 dias de idade. Babesia bigemina (33,3%, B. bovis (11,1% e A. marginale (13,9% foram detectados em esfregaços sangüíneos de 16 animais (44,4% de diferentes idades. Seis bezerros apresentaram sintomas (16,7% e um morreu (2,8%. O número de casos clínicos foi decorrente de uma associação de fatores, destacando-se a alta infestação precoce por carrapatos e a baixa imunidade passiva em período em que os bezerros ainda não haviam desenvolvido imunidade ativa suficiente.The tick-borne disease (TBD brings great damages to cattle breeding. The most important etiologic agents are Babesia bigemina, B. bovis and Anaplasma marginale, being the tick Boophilus microplus the main vector. This work reports the occurrence of natural infection by hemoparasites of TBD in 36 calves with high ticks natural infestation submitted to chemoprophylaxis with 30 days year-old. The blood smears from animals of different ages were analized and were found B. bigemina (33.3%, B. bovis (11.1% and A. marginale (13.9%. Six animals had clinical symptoms (16.7% and one dead (2.8%. The number of clinical cases ocurred in consequence of an association of factors as high infestation of ticks and low passive immunity in period that calves had not developed enough active immunity.

  11. Animal and human Ehrlichiosis/ Erliquiose nos animais e no homem

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    Odilon Vidotto

    2001-05-01

    Full Text Available Ehrlichiosis is a disease caused by gram negative obligate intracellular bacterial organisms belonging to the Rickettsiales Order, Anaplasmataceae Family, Genus Ehrlichia and Anaplasma. These organisms may parasite leukocytes, erythrocytes or platelets, leading to abnormalities in many organs. Clinical signs are variable depending on the severity of the infection, host immune response, affected organs, the specific Ehrlichia species involved, and the presence of coinfection with other Ehrlichiae or tickborne disease. The incidence of ehrlichiosis has increased over the last years in both animals and men. The etiological diagnosis is important for adequate epidemiological monitoring. Most tests currently being used have limitations. The recent use of molecular biology diagnostic techniques allow the characterization of the specie or species that are infecting a given patient.A erliquiose é uma doença causada por bactérias gram negativas estritamente intracelulares, pertencentes a Ordem Rickettsiales, Família Anaplasmataceae, Gêneros Ehrlichia e Anaplasma. As diferentes erlíquias podem parasitar leucócitos, eritrócitos e plaquetas levando a alterações em vários órgãos. Os sinais clínicos variam com a severidade da infecção, a resposta imunológica do hospedeiro, os órgãos atingidos, a espécie de erlíquia envolvida e a presenca de co-infecção com outras erlíquias ou outros microrganismos transmitidos pelo mesmo vetor. A incidência de erliquiose vem aumentando nos últimos anos tanto nos animais como no homem. O diagnóstico etiológico é importante para o monitoramento epidemiológico, porém a maioria dos testes usados rotineiramente apresenta limitações. A recente introdução de técnicas diagnósticas que empregam biologia molecular permitem caracterizar quais espécies de erlíquia estão infectando o paciente.

  12. 黑龙江口岸新发蜱传疾病的调查%Investigation of emerging tick-borne diseases at Heilongjiang ports

    Institute of Scientific and Technical Information of China (English)

    程成; 鞠文东; 焦丹; 付维明; 王艳梅; 王延禄; 呼满霞; 杨丽炜; 崔永民

    2015-01-01

    目的 调查黑龙江口岸地区蜱携带新发蜱传病原体的情况.方法 针对黑龙江省5个中俄边境口岸,2014年4-6月采集蜱类样本188份,利用PCR方法分别扩增5种新发蜱传病原体的特异性核酸片段,通过基因测序进一步鉴定并分型.结果 共检测出莱姆病螺旋体(Borrelia burgdorferi)阳性样本11例,巴贝西原虫(Babesia)1例,斑点热群立克次体(spotted fever group rickettsia) 101例,巴尔通体(Bartonella)6例,嗜吞噬细胞无形体(Anaplasma phagocytophilum)10例,感染率分别为5.85%、0.53%、53.7%、3.72%、5.32%.共发现8种病原体,分别是嘎氏疏螺旋体(Borrelia afzelii)、伽氏疏螺旋体(Borrelia garinii、巴贝西原虫属(Babesia sp.)、劳氏立克次体(Rickettsia raoultii)、新塔拉塞维奇立克次体(Candidatus Rickettsia tarasevichiae)、黑龙江立克次体(Rickettsia heilongjiangensis)、巴尔通体属(Bartonella sp.)、嗜吞噬细胞无形体(Anaplasma phagocytophilum).结论 黑龙江中俄边境存在新发蜱传疾病多样性的特征,提示应重点关注斑点热群立克次体感染,并开展蜱及蜱传疾病的常态监测.

  13. 黑龙江密山口岸三种蜱媒传染病在媒介蜱中复合感染的调查

    Institute of Scientific and Technical Information of China (English)

    程成; 鞠文东; 付维明; 徐宁; 张剑锋; 王艳梅; 王旭; 耿聪; 杨丽炜

    2015-01-01

    目的 调查黑龙江密山口岸三种蟀媒传染病在媒介蜱中复合感染的情况.方法 采集蜱类标本44份,建立伯氏疏螺旋体(Borrelia burgdorferi)、斑点热群立克次体(spotted fever group rickettsia)、嗜吞噬细胞无形体(anaplasma phagocytophilum)三种新发蜱传疾病病原体特异性通用引物,利用PCR方法对所有标本进行特异性片段扩增,对阳性标本测序并进行序列分析.结果 在所检测的44份标本中,共检测出伯氏疏螺旋体阳性标本4份,斑点热群立克次体阳性标本6份,嗜吞噬细胞无形体阳性标本2份,感染率分别为13.63%、9.10%和4.55%.在全沟硬蜱检测到两份分别是莱姆病螺旋体与斑点热群立克次体及嗜吞噬细胞无形体和斑点热群立克次体的复合感染,复合感染率为4.55%.结论 黑龙江密山地区至少存在两种莱姆病螺旋体(Borrelia garinii,Borrelia afzelii),三种斑点热群立克次体(Rickettsia raoultii,Candidatus Rickettsia tarasevichiae,Rickettsia heilongjiangensis),和嗜吞噬细胞无形体(anaplasma phagocytophilum);全沟硬蜱存在复合感染.

  14. Emerging incidence of Lyme borreliosis, babesiosis, bartonellosis, and granulocytic ehrlichiosis in Australia

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    Mayne PJ

    2011-12-01

    Full Text Available Peter J MayneInternational Lyme and Associated Diseases Society, Bethesda, MD, USABackground: Borrelia burgdorferi, the causative agent of Lyme disease (LD, and Babesia, Bartonella, and Ehrlichia species (spp. are recognized tick-borne pathogens in humans worldwide. Using serology and molecular testing, the incidence of these pathogens was investigated in symptomatic patients from Australia.Methods: Sera were analyzed by an immunofluorescent antibody assay (IFA followed by immunoglobulin (IgG and IgM Western blot (WB assays. Both whole blood and sera were analyzed for detection of specific Borrelia spp. DNA using multiplex polymerase chain reaction (PCR testing. Simultaneously, patients were tested for Babesia microti, Babesia duncani, Anaplasma phagocytophilum, Ehrlichia chaffeensis, and Bartonella henselae infection by IgG and IgM IFA serology, PCR, and fluorescent in situ hybridization (FISH.Results: Most patients reported symptom onset in Australia without recent overseas travel. 28 of 51 (55% tested positive for LD. Of 41 patients tested for tick-borne coinfections, 13 (32% were positive for Babesia spp. and nine (22% were positive for Bartonella spp. Twenty-five patients were tested for Ehrlichia spp. and (16% were positive for Anaplasma phagocytophilum while none were positive for Ehrlichia chaffeensis. Among the 51 patients tested for LD, 21 (41% had evidence of more than one tick-borne infection. Positive tests for LD, Babesia duncani, Babesia microti, and Bartonella henselae were demonstrated in an individual who had never left the state of Queensland. Positive testing for these pathogens was found in three others whose movements were restricted to the east coast of Australia.Conclusion: The study identified a much larger tick-borne disease (TBD burden within the Australian community than hitherto reported. In particular, the first cases of endemic human Babesia and Bartonella disease in Australia with coexisting Borrelia infection are

  15. A survey of canine tick-borne diseases in India

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    Coleman Glen T

    2011-07-01

    Full Text Available Abstract Background There are few published reports on canine Babesia, Ehrlichia, Anaplasma, Hepatozoon and haemotropic Mycoplasma infections in India and most describe clinical disease in individual dogs, diagnosed by morphological observation of the microorganisms in stained blood smears. This study investigated the occurrence and distribution of canine tick-borne disease (TBD pathogens using a combination of conventional and molecular diagnostic techniques in four cities in India. Results On microscopy examination, only Hepatozoon gamonts were observed in twelve out of 525 (2.3%; 95% CI: 1.2, 4 blood smears. Using polymerase chain reaction (PCR, a total of 261 from 525 dogs (49.7%; 95% CI: 45.4, 54.1 in this study were infected with one or more canine tick-borne pathogen. Hepatozoon canis (30%; 95% CI: 26.0, 34.0 was the most common TBD pathogen found infecting dogs in India followed by Ehrlichia canis (20.6%; 95% CI: 17.2, 24.3, Mycoplasma haemocanis (12.2%; 95% CI: 9.5, 15.3, Anaplasma platys (6.5%; 95% CI: 4.5, 8.9, Babesia vogeli (5.5%, 95% CI: 3.7, 7.8 and Babesia gibsoni (0.2%, 95% CI: 0.01, 1.06. Concurrent infection with more than one TBD pathogen occurred in 39% of cases. Potential tick vectors, Rhipicephalus (most commonly and/or Haemaphysalis ticks were found on 278 (53% of dogs examined. Conclusions At least 6 species of canine tick-borne pathogens are present in India. Hepatozoon canis was the most common pathogen and ticks belonging to the genus Rhipicephalus were encountered most frequently. Polymerase chain reaction was more sensitive in detecting circulating pathogens compared with peripheral blood smear examination. As co-infections with canine TBD pathogens were common, Indian veterinary practitioners should be cognisant that the discovery of one such pathogen raises the potential for multiple infections which may warrant different clinical management strategies.

  16. Anaplasmosis bovina

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    Belkis Corona, Majela Rodríguez y Siomara Martínez.

    2005-04-01

    Full Text Available Anaplasma marginale es una rickettsia del genogrupo II de las Ehrlichias, que parasita los eritrocitos maduros del ganado bovino y causa severas pérdidas económicas fundamentalmente en las zonas tropicales y subtropicales (Palmer y col., 1999. Este microorganismo presenta múltiple variabilidad antigénica, de morfología, virulencia, transmisibilidad por garrapatas y habilidad para inducir protección cruzada contra aislamientos heterólogos (Palmer y McElwain, 1995. Se han caracterizado seis proteínas de superficie de membrana de los cuerpos iniciales de este organismo, portadoras de epitopes B y T, denominadas proteínas mayoritarias de superficie (MSPs y designadas 1a, 1b, 2, 3, 4 y 5. Estas proteínas son reconocidas por anticuerpos neutralizantes y se encuentran en una estrecha relación intermolecular en la superficie de la membrana de los cuerpos iniciales. Algunas de estas proteínas inducen una protección total o parcial en animales vacunados, aunque el nivel y la uniformidad de la misma, es variable (Palmer y McElwain, 1995. A pesar de las cuantiosas pérdidas económicas producidas todos los años, a nivel mundial hasta el momento no se cuenta con un método de control eficaz contra la enfermedad, por lo que resulta de gran importancia desarrollar una vacuna capaz de prevenir la infección con este patógeno y contar con técnicas de diagnóstico más sensibles y específicas que permitan la detección de animales portadores para ser utilizadas en estudios epizootiológicos y para el control de la enfermedad (Echaide y col., 1998. Por lo que a partir de estas premisas nos proponemos como objetivo realizar una exhaustiva revisión bibliográfica acerca de la anaplasmosis bovina y su agente causal, Anaplasma marginale.

  17. Molecular Detection of Tick-Borne Pathogens in Humans with Tick Bites and Erythema Migrans, in the Netherlands

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    Jahfari, Setareh; Hofhuis, Agnetha; Fonville, Manoj; van der Giessen, Joke; van Pelt, Wilfrid; Sprong, Hein

    2016-01-01

    Background Tick-borne diseases are the most prevalent vector-borne diseases in Europe. Knowledge on the incidence and clinical presentation of other tick-borne diseases than Lyme borreliosis and tick-borne encephalitis is minimal, despite the high human exposure to these pathogens through tick bites. Using molecular detection techniques, the frequency of tick-borne infections after exposure through tick bites was estimated. Methods Ticks, blood samples and questionnaires on health status were collected from patients that visited their general practitioner with a tick bite or erythema migrans in 2007 and 2008. The presence of several tick-borne pathogens in 314 ticks and 626 blood samples of this cohort were analyzed using PCR-based methods. Using multivariate logistic regression, associations were explored between pathogens detected in blood and self-reported symptoms at enrolment and during a three-month follow-up period. Results Half of the ticks removed from humans tested positive for Borrelia burgdorferi sensu lato, Anaplasma phagocytophilum, Candidatus Neoehrlichia mikurensis, Rickettsia helvetica, Rickettsia monacensis, Borrelia miyamotoi and several Babesia species. Among 92 Borrelia burgdorferi s. l. positive ticks, 33% carried another pathogen from a different genus. In blood of sixteen out of 626 persons with tick bites or erythema migrans, DNA was detected from Candidatus Neoehrlichia mikurensis (n = 7), Anaplasma phagocytophilum (n = 5), Babesia divergens (n = 3), Borrelia miyamotoi (n = 1) and Borrelia burgdorferi s. l. (n = 1). None of these sixteen individuals reported any overt symptoms that would indicate a corresponding illness during the three-month follow-up period. No associations were found between the presence of pathogen DNA in blood and; self-reported symptoms, with pathogen DNA in the corresponding ticks (n = 8), reported tick attachment duration, tick engorgement, or antibiotic treatment at enrolment. Conclusions Based on molecular

  18. Tick fever in cattle in the region of Botucatu - SP: a retrospective study of 1986-2007/ Tristeza Parasitária em bovinos na região de Botucatu – SP: estudo retrospectivo de 1986 – 2007

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    Alexandre Secorum Borges

    Full Text Available A retrospective study of Cattle Tick Fever was made with animals up to one year old, which occurred from 1986-2007 in Botucatu-SP in the influence area of the Veterinary Hospital of the School of Veterinary Medicine and Animal Science – UNESP. There were 232 cases and 57 deaths. The main etiological agent causing the disease on this region is Anaplasma marginale identified as a single agent in 31.5% of the cases. Most of the cases occurred on autumn in mixed-blood animals two to six months old. Predominant clinical signs included apathy, hyporexia or anorexia, dehydration, weight loss and pale mucosa. The high incidence and mortality observed justify prophylactic actions to have the balance between host, agent, vector, and environment, thus avoiding the clinical form of disease and death of livestock herds in the region.Foi realizado um estudo retrospectivo dos casos de tristeza parasitária bovina (TPB em bovinos com até um ano de idade ocorridos na região de Botucatu-SP, área de influência do Hospital Veterinário da Faculdade de Medicina Veterinária e Zootecnia de Botucatu – UNESP, entre os anos de 1986 e 2007. Ocorreram, nesse período, 232 casos da doença e 57 mortes. O principal agente etiológico causador da doença nesta região foi o Anaplasma marginale, identificado como um único agente em 31,5% dos episódios da doença. A maioria dos casos ocorreu no Outono, em animais mestiços e com dois a seis meses de idade. Os sinais clínicos predominantes foram: apatia, hiporexia ou anorexia, desidratação, perda de peso e palidez das mucosas. A elevada incidência e mortalidade observados justificam ações profiláticas visando o equilíbrio entre hospedeiro, agente, vetor, e meio ambiente, evitando, assim, a forma clínica da doença e morte dos animais em rebanhos da região.

  19. Surveillance of syndrome of fever with thrombocytopenia and etiological detection in Beijing%北京市发热伴血小板减少综合征监测和病原检测分析

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    窦相峰; 林晖; 张秀春; 吕燕宁; 王全意; 王小梅; 田丽丽; 黎新宇; 何战英; 孙玉兰; 关增智

    2011-01-01

    目的 分析北京市发热伴血小板或白细胞减少综合征监测和严重发热伴血小板减少综合征病毒(SFTSV)感染情况.方法 对符合监测对象定义的病例(体温≥37.5℃伴血小板<80×109/L或白细胞<3.0×109/L)进行流行病学调查并采集乙二胺四乙酸抗凝和非抗凝血液标本分别检测嗜吞噬细胞无形体和SFTSV.结果 2011年4-7月共监测到45例具有发热伴血小板或白细胞减少症状病例,其中男性27例,年龄在13~81岁之间.其中2例经实验室检测为新型布尼亚病毒感染病例,1例死亡,均为外地输入病例,未检测到嗜吞噬细胞无形体感染病例.结论 北京市监测系统尚未发现本地感染新型布尼亚病毒病例.%Objective To evaluate the surveillance of syndrome of fever with thrombocytopenia or leucopenia and infection status of severe fever with thrombocytopenia syndrome virus (SFTSV) in Beijing. Methods The cases of fever ( ≥37. 5℃) with thrombocytopenia ( < 80 × 109/L) or leucopenia ( < 3.0 ×109/L) were investigated with standard questionnaire and their blood samples were collected with non-anticoagulation tube and EDTA tube. RNA of SFTSV and DNA of Anaplasma phagocytophilum were detected by real-time RT-PCR and nested PCR respectively. Results During April-July in 2011, forty-five suspected cases of fever with thrombocytopenia or leucopenia were detected, including 27 in males and 18 in females aged 13-81 years. Two cases of SFTSV infection were laboratory-confirmed by real-time RT-PCR, which were from Henan and Anhui provinces, and 1 was fatal. No Anaplasma phagocytophilum were detected. Conclusion No local SFTSV infection cases had been found through the surveillance system in Beijing.

  20. A field survey on parasites and antibodies against selected pathogens in owned dogs in Lilongwe, Malawi

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    Karin Alvåsen

    2016-02-01

    Full Text Available The aim of this study was to screen for selected parasites and antibody levels against vectorborne pathogens in owned dogs in Lilongwe, Malawi. The study population consisted of 100 dogs; 80 participating in vaccination–spaying campaigns and 20 visiting a veterinary clinic as paying clients. All dogs went through a general physical examination including visual examination for signs of ectoparasites. A total of 100 blood samples were analysed using commercial snap tests and 40 faecal samples by egg flotation in saturated sodium chloride. The sampled dogs had a seroprevalence of 12% for Anaplasma spp., 22% for Ehrlichia spp., 4% for Dirofilaria immitis and 1% for Leishmania spp. Eggs from Ancylostoma spp. were found in 80% of the faecal samples, whereas eggs of Trichuris vulpis, Toxocara canis and Toxascaris leonina were only present in 3%, 8% and 13% of the samples, respectively. Ectoparasites such as Ctenocephalides sp., Trichodectes sp. and ticks were present on 98%, 25% and 11%, respectively, of the campaign dogs. Among client dogs, 35% had Ctenocephalides fleas, 10% had Trichodectes lice and none had ticks. Public education and prophylactic treatment could be used to improve the animal welfare of dogs; this would most likely also have positive impact on public health.

  1. Tick-borne rickettsial pathogens in questing ticks, removed from humans and animals in Mexico

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    Vargas-Sandoval, Margarita; Torres, Javier

    2016-01-01

    Tick-borne rickettsial diseases (TBRD) are commonly encountered in medical and veterinary clinical settings. The control of these diseases is difficult, requiring disruption of a complex transmission chain involving a vertebrate host and ticks. The geographical distribution of the diseases is related to distribution of the vector, which is an indicator of risk for the population. A total of 1,107 ticks were collected by tick dragging from forests, ecotourism parks and hosts at 101 sites in 22 of the 32 states of Mexico. Collected ticks were placed in 1.5 mL cryovials containing 70% ethanol and were identified to species. Ticks were pooled according to location/host of collection, date of collection, sex, and stage of development. A total of 51 ticks were assayed by polymerase chain reaction (PCR) to confirm species identification using morphological methods. A total of 477 pools of ticks were assayed using PCR techniques for selected tick-borne pathogens. Anaplasma phagocytophilum was the most commonly detected pathogen (45 pools), followed by, Ehrlichia (E.) canis (42), Rickettsia (R.) rickettsii (11), E. chaffeensis (8), and R. amblyommii (1). Rhipicephalus sanguineus was the tick most frequently positive for selected pathogens. Overall, our results indicate that potential tick vectors positive for rickettsial pathogens are distributed throughout the area surveyed in Mexico. PMID:26726019

  2. Prevalence of antibodies to spotted fever group Rickettsia spp. and Ehrlichia spp. in coyotes (Canis latrans) in Oklahoma and Texas, USA.

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    Starkey, Lindsay A; West, Misti D; Barrett, Anne W; Saucier, Jill M; O'Connor, Tom P; Paras, Kelsey L; Reiskind, Michael H; Reichard, Mason V; Little, Susan E

    2013-07-01

    Coyotes (Canis latrans) are commonly infested with ticks, including Amblyomma americanum, the predominant vector of Ehrlichia chaffeensis and Ehrlichia ewingii; Dermacentor variabilis, an important vector of Rickettsia rickettsii; and Amblyomma maculatum, a major vector of Rickettsia parkeri, a spotted fever group (SFG) Rickettsia. To determine the degree to which coyotes are infected with or exposed to tick-borne bacterial disease agents, serum samples collected from coyotes in Oklahoma and Texas were tested for antibodies reactive to R. rickettsii, Ehrlichia canis, E. chaffeensis, E. ewingii, Borrelia burgdorferi, and Anaplasma phagocytophilum by indirect fluorescent antibody (IFA) testing or enzyme-linked immunosorbent assay (ELISA). Of the coyotes tested, 60% (46/77) and 64% (47/74) had antibodies reactive to R. rickettsii and E. chaffeensis, respectively, on IFA. Additionally, 5% (4/77) had antibodies reactive to E. canis, but not B. burgdorferi or A. phagocytophilum, on SNAP(®) 4Dx(®) ELISA; subsequent serologic analysis by plate ELISA using species-specific peptides revealed antibodies to E. ewingii, E. canis, and E. chaffeensis in 46% (23/50), 18% (9/50), and 4% (2/50) of serum samples, respectively. Taken together, these data indicate that coyotes in this region are commonly exposed to SFG Rickettsia and E. ewingii and that further consideration of coyotes as a component of the maintenance cycle for these pathogens may be warranted.

  3. Comparison of selected canine vector-borne diseases between urban animal shelter and rural hunting dogs in Korea

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    Ahn KyuSung

    2010-04-01

    Full Text Available Abstract A serological survey for Dirofilaria immitis, Anaplasma phagocytophilum, Ehrlichia canis, and Borrelia burgdorferi infections in rural hunting and urban shelter dogs mainly from southwestern regions of the Republic of Korea (South Korea was conducted. From a total of 229 wild boar or pheasant hunting dogs, the number of serologically positive dogs for any of the four pathogens was 93 (40.6%. The highest prevalence observed was D. immitis (22.3%, followed by A. phagocytophilum (18.8%, E. canis (6.1% and the lowest prevalence was B. burgdorferi (2.2%. In contrast, stray dogs found within the city limits of Gwangju showed seropositivity only to D. immitis (14.6%, and none of the 692 dogs responded positive for A. phagocytophilum, E. canis or B. burgdorferi antibodies. This study indicates that the risk of exposure to vector-borne diseases in rural hunting dogs can be quite high in Korea, while the urban environment may not be suitable for tick infestation on dogs, as evidenced by the low infection status of tick-borne pathogens in stray dogs.

  4. Prevalence of canine monocitic ehrlichiosis and canine thrombocytic anaplasmosis in dogs suspected of hemoparasitosis in Cuiabá Mato Grosso

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    Rute Witter

    2014-02-01

    Full Text Available The present study evaluated Canine Monocytic Ehrlichiosis and Canine Thrombocytic Anaplasmosis in 77 dogs treated at the Veterinary Hospital (HOVET of the Federal University of Mato Grosso (UFMT in 2009. An association among clinical and hematological, serological and molecular data was investigated. Dogs were evaluated by partial amplification of dsb and 16S rRNA of Ehrlichia canis and Anaplasma platys and by Indirect Fluorescence Antigen Test (IFAT for E. canis. DNA of E. canis and A. platys were detected in 18 (23.3% and 07 (9.1% dogs respectively. Fifty four (70.1% dogs were positive by IFAT with titers ranging from 40 to 327,680. There were higher frequencies of E. canis positive PCR in dogs less than 12 months old, with anemia, lymphopenia and thrombocytopenia (P?0.05. Leukopenic dogs presented tendency to be A. platys PCR positive (P=0.07. Thrombocytopenia and hyperproteinemia of dogs were associated with high anti-E. canis antibody titers (P?0.05. Lymphadenopathy and pulmonary changes were observed in 15 (30.6% and 4 (57.1% E. canis PCR positive dogs (P?0.05. No other clinical features were associated between E. canis and A. platys infected dogs. E. canis was the only specie of Ehrlichia detected in dogs attended at the HOVET with a higher infection rate in young dogs, and statistically associated with anemic and thrombocytopenic dogs. On the other side, A. platys presented low occurrence among evaluated dogs.

  5. First record of autochthonous canine ehrlichiosis caused by Ehrlichia canis in Romania.

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    Morar, Doru; Dărăbuş, Gheorghe; Imre, Mirela; Ilie, Marius Stelian; Imre, Kálmán

    2015-06-01

    This case study describes the first genetically confirmed and clinically manifested autochthonous Ehrlichia canis infection in a 9-year-old female mixed-breed dog from Romania. Health screening of the dog included clinical examination, evaluation of stained peripheral blood smear and hematologic variables, as well as serologic testing and molecular analysis. Clinical signs included fever, apathy, dehydration, pale mucous membranes, and weakness. The microscopic examination of the blood smear and immunologic assays for Borrelia burgdorferi, Anaplasma phagocytophilum, and E canis antibodies, and for Dirofilaria immitis antigen yielded negative results. Hematologic abnormalities included moderate nonregenerative anemia, leucopenia with neutropenia, and moderate thrombocytopenia. The biochemical abnormalities identified were hypoalbuminemia, and mildly increased serum enzyme activities of AST and ALT. In addition, increased urea and creatinine levels associated with low urine specific gravity and proteinuria were also present. Nested PCR amplification of the partial E canis 16S rRNA gene demonstrated the presence of this rickettsial pathogen in the dog's blood, which subsequently was confirmed through sequencing based on the 100% homology with GenBank deposited E canis isolates. After specific treatment with doxycycline (10 mg/kg, orally, SID) for one month, the proteinuria, and hematologic and serum biochemical abnormalities with the exception of mild azotemia resolved. This report supports the geographical expansion of canine ehrlichiosis caused by E canis in nonendemic regions of Europe.

  6. A comparison of DNA extraction protocols from blood spotted on FTA cards for the detection of tick-borne pathogens by Reverse Line Blot hybridization.

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    Hailemariam, Zerihun; Ahmed, Jabbar Sabir; Clausen, Peter-Henning; Nijhof, Ard Menzo

    2017-01-01

    An essential step in the molecular detection of tick-borne pathogens (TBPs) in blood is the extraction of DNA. When cooled storage of blood under field conditions prior to DNA extraction in a dedicated laboratory is not possible, the storage of blood on filter paper forms a promising alternative. We evaluated six DNA extraction methods from blood spotted on FTA Classic(®) cards (FTA cards), to determine the optimal protocol for the subsequent molecular detection of TBPs by PCR and the Reverse Line Blot hybridization assay (RLB). Ten-fold serial dilutions of bovine blood infected with Babesia bovis, Theileria mutans, Anaplasma marginale or Ehrlichia ruminantium were made by dilution with uninfected blood and spotted on FTA cards. Subsequently, DNA was extracted from FTA cards using six different DNA extraction protocols. DNA was also isolated from whole blood dilutions using a commercial kit. PCR/RLB results showed that washing of 3mm discs punched from FTA cards with FTA purification reagent followed by DNA extraction using Chelex(®) resin was the most sensitive procedure. The detection limit could be improved when more discs were used as starting material for the DNA extraction, whereby the use of sixteen 3mm discs proved to be most practical. The presented best practice method for the extraction of DNA from blood spotted on FTA cards will facilitate epidemiological studies on TBPs. It may be particularly useful for field studies where a cold chain is absent.

  7. Prevalence of vector-borne bacterial pathogens in riparian brush rabbits (Sylvilagus bachmani riparius) and their ticks.

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    Schmitz, Kelly M; Foley, Janet E; Kasten, Rickie W; Chomel, Bruno B; Larsen, R Scott

    2014-04-01

    From June to October 2010, 48 endangered riparian brush rabbits (Sylvilagus bachmani riparius) were trapped at a captive propagation site in central California with the intention of release into re-established habitats. During prerelease examinations, ticks and blood samples were collected for surveillance for Rickettsia spp., Anaplasma phagocytophilum, Borrelia burgdorferi, and Bartonella spp. Ticks were identified, and DNA was extracted for PCR analysis. Serology was performed to detect exposure to Rickettsia spp., B. burgdorferi, and A. phagocytophilum. DNA was extracted from blood samples and analyzed for A. phagocytophilum using PCR assays. Rabbit blood samples were also cultured for Bartonella spp. Haemaphysalis leporispalustris ticks were detected on all rabbits except one. A total of 375 ticks were collected, with 48% of the rabbits (23 rabbits) having a burden ranging from 0 to 5 ticks, 15% (seven rabbits) from 6 to 10 ticks, 25% (12 rabbits) from 11 to 15 ticks, and 12% (six rabbits) with >15 ticks. There was no evidence of B. burgdorferi or R. rickettsii in tick or rabbit samples. There was also no evidence of Bartonella spp. in the rabbit samples. Four tick samples and 14 rabbits were weakly PCR positive for A. phagocytophilum, and six rabbits were antibody positive for A. phagocytophilum. These results suggest that there may be little risk of these tick-borne diseases in riparian brush rabbits or to the people in contact with them.

  8. Prevalence of pathogenic bacteria in Ixodes ricinus ticks in Central Bohemia.

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    Klubal, Radek; Kopecky, Jan; Nesvorna, Marta; Sparagano, Olivier A E; Thomayerova, Jana; Hubert, Jan

    2016-01-01

    Bacteria associated with the tick Ixodes ricinus were assessed in specimens unattached or attached to the skin of cats, dogs and humans, collected in the Czech Republic. The bacteria were detected by PCR in 97 of 142 pooled samples including 204 ticks, i.e. 1-7 ticks per sample, collected at the same time from one host. A fragment of the bacterial 16S rRNA gene was amplified, cloned and sequenced from 32 randomly selected samples. The most frequent sequences were those related to Candidatus Midichloria midichlori (71% of cloned sequences), followed by Diplorickettsia (13%), Spiroplasma (3%), Rickettsia (3%), Pasteurella (3%), Morganella (3%), Pseudomonas (2%), Bacillus (1%), Methylobacterium (1%) and Phyllobacterium (1%). The phylogenetic analysis of Spiroplasma 16S rRNA gene sequences showed two groups related to Spiroplasma eriocheiris and Spiroplasma melliferum, respectively. Using group-specific primers, the following potentially pathogenic bacteria were detected: Borellia (in 20% of the 142 samples), Rickettsia (12%), Spiroplasma (5%), Diplorickettsia (5%) and Anaplasma (2%). In total, 68% of I. ricinus samples (97/142) contained detectable bacteria and 13% contained two or more putative pathogenic groups. The prevalence of tick-borne bacteria was similar to the observations in other European countries.

  9. Effect of Imidocarb dipropionate on the immune response to Foot and Mouth Disease vaccine in healthy and anaplasmosis-infected calves

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    N. A. Afifi

    2014-03-01

    Full Text Available Aim: This work was performed to investigate the effect of a potent anti-protozoan drug, Imidocarb on the cell mediated and humoral immune response to foot and mouth disease vaccine (FMDV, O1 strain in normal and Anaplasmosis-infected calves. Materials and Methods: A total of 55 male mixed bred calves were used and divided into two main groups of 25 calves each. The first group was healthy and the second was Anaplasma - infected calves. FMDV was administered in both groups. Calves of the first and second groups were subdivided into equal five subgroups of 5 calves each. The first subgroup was vaccinated control. The treated subgroups were each given 3 mg / kg body weight Imidocarb dipropionate in a single intramuscular dose at one week before vaccination, at time of vaccination, one week and two weeks post vaccination with FMDV (O1, respectively. The cellular immune response in the different groups was evaluated weekly, however antibody titers were measured by ELISA and serum neutralization test Results: Imidocarb increased rate of erythrocyte rosette forming lymphocytes when it was administered one week before vaccination, at time of vaccination and one week post vaccination. Imidocarb increased antibody titre of FMDV in both normal and anaplasmosis-infected calves. The protection rate due to challenge with virulent FMDV was high in treated calves as compared with the vaccinated control. Conclusion: The best immunopotentiating effect of Imidocarb is achieved by dosing one week before vaccinating calves with FMD vaccine.

  10. Molecular evidence of spotted fever group rickettsiae and Anaplasmataceae from ticks and stray dogs in Bangladesh.

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    Qiu, Yongjin; Nakao, Ryo; Thu, May June; Akter, Shirin; Alam, Mohammad Zahangir; Kato, Satomi; Katakura, Ken; Sugimoto, Chihiro

    2016-03-01

    Emerging tick-borne diseases (TBDs) are important foci for human and animal health worldwide. However, these diseases are sometimes over looked, especially in countries with limited resources to perform molecular-based surveys. The aim of this study was to detect and characterize spotted fever group (SFG) rickettsiae and Anaplasmataceae in Bangladesh, which are important tick-borne pathogens for humans and animals worldwide. A total of 50 canine blood samples, 15 ticks collected from dogs, and 154 ticks collected from cattle were screened for the presence of SFG rickettsiae and Anaplasmataceae using molecular-based methods such as PCR and real-time PCR. The sequence analysis of the amplified products detected two different genotypes of SFG rickettsiae in ticks from cattle. The genotype detected in Rhipicephalus microplus was closely related to Rickettsia monacensis, while the genotype detected in Haemaphysalis bispinosa was closely related to Rickettsia sp. found in Korea and Japan. Anaplasma bovis was detected in canine blood and ticks (Rhipicephalus sanguineus and H. bispinosa). Unexpectedly, the partial genome sequence of Wolbachia sp., presumably associated with the nematode Dirofilaria immitis, was identified in canine blood. The present study provides the first molecular evidence of SFG rickettsiae and A. bovis in Bangladesh, indicating the possible emergence of previously unrecognized TBDs in this country.

  11. Genetic Diversity of Tick-Borne Rickettsial Pathogens; Insights Gained from Distant Strains

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    Sebastián Aguilar Pierlé

    2014-01-01

    Full Text Available The ability to capture genetic variation with unprecedented resolution improves our understanding of bacterial populations and their ability to cause disease. The goal of the pathogenomics era is to define genetic diversity that results in disease. Despite the economic losses caused by vector-borne bacteria in the Order Rickettsiales, little is known about the genetic variants responsible for observed phenotypes. The tick-transmitted rickettsial pathogen Anaplasma marginale infects cattle in tropical and subtropical regions worldwide, including Australia. Genomic analysis of North American A. marginale strains reveals a closed core genome defined by high levels of Single Nucleotide Polymorphisms (SNPs. Here we report the first genome sequences and comparative analysis for Australian strains that differ in virulence and transmissibility. A list of genetic differences that segregate with phenotype was evaluated for the ability to distinguish the attenuated strain from virulent field strains. Phylogenetic analyses of the Australian strains revealed a marked evolutionary distance from all previously sequenced strains. SNP analysis showed a strikingly reduced genetic diversity between these strains, with the smallest number of SNPs detected between any two A. marginale strains. The low diversity between these phenotypically distinct bacteria presents a unique opportunity to identify the genetic determinants of virulence and transmission.

  12. Flying ticks: anciently evolved associations that constitute a risk of infectious disease spread.

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    de la Fuente, José; Estrada-Peña, Agustín; Cabezas-Cruz, Alejandro; Brey, Ricardo

    2015-10-15

    Ticks are important vectors of emerging zoonotic diseases affecting human and animal health worldwide. Ticks are often found on wild birds, which have been long recognized as a potential risk factor for dissemination of ticks and tick-borne pathogens (TBP), thus raising societal concerns and prompting research into their biology and ecology. To fully understand the role of birds in disseminating some ticks species and TBP, it is important to consider the evolutionary relationships between birds, ticks and transmitted pathogens. In this paper we reviewed the possible role of birds in the dissemination of TBP as a result of the evolution of host-tick-pathogen associations. Birds are central elements in the ecological networks of ticks, hosts and TBP. The study of host-tick-pathogen associations reveals a prominent role for birds in the dissemination of Borrelia spp. and Anaplasma phagocytophilum, with little contribution to the possible dissemination of other TBP. Birds have played a major role during tick evolution, which explains why they are by far the most important hosts supporting the ecological networks of ticks and several TBP. The immune response of birds to ticks and TBP has been largely overlooked. To implement effective measures for the control of tick-borne diseases, it is necessary to study bird-tick and bird-pathogen molecular interactions including the immune response of birds to tick infestation and pathogen infection.

  13. Chronic coinfections in patients diagnosed with chronic Lyme disease: a systematic literature review

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    Lantos, Paul M.; Wormser, Gary P.

    2014-01-01

    Purpose The controversial diagnosis of chronic Lyme disease is often given to patients with prolonged, medically unexplained physical symptoms. Many such patients are also treated for chronic co-infections with Babesia, Anaplasma, or Bartonella in the absence of typical presentations, objective clinical findings, or laboratory confirmation of active infection. We have undertaken a systematic review of the literature to evaluate several aspects of this practice. Methods Five systematic literature searches were performed using Boolean operators and the PubMed search engine. Results The literature searches did not demonstrate convincing evidence of 1) chronic anaplasmosis infection, 2) treatment responsive symptomatic chronic babesiosis in immunocompetent persons in the absence of fever, laboratory abnormalities and detectable parasitemia, 3) either geographically widespread or treatment responsive symptomatic chronic infection with Babesia duncani in the absence of fever, laboratory abnormalities and detectable parasitemia, 4) tick-borne transmission of Bartonella species, or 5) simultaneous Lyme disease and Bartonella infection. Conclusions The medical literature does not support the diagnosis of chronic, atypical tick-borne coinfections in patients with chronic, nonspecific illnesses. PMID:24929022

  14. Tabanids: neglected subjects of research, but important vectors of disease agents!

    Science.gov (United States)

    Baldacchino, Frédéric; Desquesnes, Marc; Mihok, Steve; Foil, Lane D; Duvallet, Gérard; Jittapalapong, Sathaporn

    2014-12-01

    Tabanids are nuisance pests for people and livestock because of their painful and irritating bite, persistent biting behavior, and blood ingestion. About 4400 tabanid species have been described; they are seasonally present in all kinds of landscapes, latitudes, and altitudes. High populations have a significant economic impact on outdoor activities, tourism, and livestock production. Tabanids are also vectors of animal disease agents, including viruses, bacteria and parasites. However, tabanids have received little attention in comparison with other hematophagous Diptera. Here, we highlight the many direct and indirect impacts of tabanids and provide a brief summary of tabanid morphology, biology, and life cycle. Impacts include pathogen transmission, parasite transportation (Dermatobia hominis), biological transmission (Loa loa), and mechanical transmission of viruses, such as equine infectious anemia virus, protozoa, such as Trypanosoma evansi and Besnotia besnoiti, and bacteria, such as Bacillus anthracis and Anaplasma marginale. We discuss parameters of mechanical transmission and its mathematical modeling. Control methods for tabanid populations are also summarized; these include trapping, the use of insecticides, repellents, and livestock protection. Lastly recommendations are provided for the direction of future research.

  15. Prevalence Survey of Selected Bovine Pathogens in Water Buffaloes in the North Region of Brazil

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    Jenevaldo Barbosa da Silva

    2014-01-01

    Full Text Available Although the largest buffalo herd in the occident is in the north region of Brazil, few studies have been conducted to assess the prevalence of selected parasitic diseases in buffalo herd. The present study was therefore conducted to investigate the epidemiological of Toxoplasma gondii, Neospora caninum, Anaplasma marginale, Babesia bigemina, and Babesia bovis in water buffaloes in the north region of Brazil. A total of 4796 buffalo blood samples were randomly collected from five provinces and simultaneously analyzed by the IFAT and ELISA. The serological prevalence of T. gondii and N. caninum was 41.3% and 55.5% in ELISA and 35.7% and 48.8% in IFAT, respectively. The overall prevalence of A. marginale, B. bovis, and B. bigemina was 63%, 25%, and 21% by ELISA and 50.0%, 22.5%, and 18.8% by IFAT, respectively. This study shows valuable information regarding the serological survey of selected bovine pathogens in water buffaloes in the north region of Brazil which will likely be very beneficial for the management and control programs of this disease.

  16. Enzootic instability for bovine anaplasmosis on family farms located in southwestern Paraná, Brazil

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    Tatiane Sott

    Full Text Available Abstract The objective of this study was to assess the occurence of animals seropositive for Anaplasma marginale in the municipality of Realeza, Paraná State, Brazil. Blood samples were collected from 344 cows on 18 small farms in the municipality of Realeza-PR. The animals’serum samples were forwarded to the Federal University of Fronteira do Sul, in order to investigate the occurrence of anti-A. marginale IgG antibodies by an enzyme-linked immunosorbent assay commercial kit. IgG antibodies to A. marginale were detected in cattle from 77.7% of the farms. To the best author's knowledge, this is the first report of occurrence of A. marginale in cattle in southwestern Paraná. The serological assay showed that 24.4% of the animals were seropositive, thus characterizing the location investigated as an area of enzootic instability for the disease. The family farms located in the municipality of Realeza-PR showed enzootic instability for bovine anaplasmosis. It is necessary to conduct disease monitoring programs in association with preventive measures in order to ensure the sanitary quality of the herds and to reduce economic losses for the farmers. In addition, it is essential to implement educational extension actions that allow farmers to acquire knowledge, attitudes and perceptions regarding the risk factors that contribute towards herd A. marginale-infection.

  17. Expansion of Variant Diversity Associated with a High Prevalence of Pathogen Strain Superinfection under Conditions of Natural Transmission

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    Ueti, Massaro W.; Tan, Yunbing; Broschat, Shira L.; Castañeda Ortiz, Elizabeth J.; Camacho-Nuez, Minerva; Mosqueda, Juan J.; Scoles, Glen A.; Grimes, Matthew; Brayton, Kelly A.

    2012-01-01

    Superinfection occurs when a second, genetically distinct pathogen strain infects a host that has already mounted an immune response to a primary strain. For antigenically variant pathogens, the primary strain itself expresses a broad diversity of variants over time. Thus, successful superinfection would require that the secondary strain express a unique set of variants. We tested this hypothesis under conditions of natural transmission in both temperate and tropical regions where, respectively, single-strain infections and strain superinfections of the tick-borne pathogen Anaplasma marginale predominate. Our conclusion that strain superinfection is associated with a significant increase in variant diversity is supported by progressive analysis of variant composition: (i) animals with naturally acquired superinfection had a statistically significantly greater number of unique variant sequences than animals either experimentally infected with single strains or infected with a single strain naturally, (ii) the greater number of unique sequences reflected a statistically significant increase in primary structural diversity in the superinfected animals, and (iii) the increase in primary structural diversity reflected increased combinations of the newly identified hypervariable microdomains. The role of population immunity in establishing temporal and spatial patterns of infection and disease has been well established. The results of the present study, which examined strain structure under conditions of natural transmission and population immunity, support that high levels of endemicity also drive pathogen divergence toward greater strain diversity. PMID:22585962

  18. Choque séptico por ehrliquiosis

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    Viviana Hernández-de Mezerville

    2007-04-01

    Full Text Available La ehrliquiosis humana es una infección zoonótica transmitida por "garrapatas duras" causada por Anaplasma sp y Ehrlichia sp. Puede presentarse como un cuadro pseudogripal con fiebre, malestar general, mialgias y cefalea o hasta un choque séptico con falla multiorgánica y muerte. La mitad de los pacientes requieren atención hospitalaria en algún momento y se reporta una mortalidad de hasta el 5% secundaria a enfermedades oportunistas o a hemorragias. El diagnóstico se puede sospechar visualizando las mórulas intracitoplasmáticas en las células afectadas en frotis de sangre periférica, y se confirma con serología, con cultivos o con detección molecular. El tratamiento de elección es la doxiciclina con buena respuesta clínica. Se reporta un caso probable de ehrliquiosis humana asociado a choque séptico y falla multiorgánica tratado exitosamente en el Hospital Dr. Rafael Angel Calderón Guardia.

  19. Transport of ixodid ticks and tick-borne pathogens by migratory birds.

    Science.gov (United States)

    Hasle, Gunnar

    2013-01-01

    Birds, particularly passerines, can be parasitized by Ixodid ticks, which may be infected with tick-borne pathogens, like Borrelia spp., Babesia spp., Anaplasma, Rickettsia/Coxiella, and tick-borne encephalitis virus. The prevalence of ticks on birds varies over years, season, locality and different bird species. The prevalence of ticks on different species depends mainly on the degree of feeding on the ground. In Europe, the Turdus spp., especially the blackbird, Turdus merula, appears to be most important for harboring ticks. Birds can easily cross barriers, like fences, mountains, glaciers, desserts and oceans, which would stop mammals, and they can move much faster than the wingless hosts. Birds can potentially transport tick-borne pathogens by transporting infected ticks, by being infected with tick-borne pathogens and transmit the pathogens to the ticks, and possibly act as hosts for transfer of pathogens between ticks through co-feeding. Knowledge of the bird migration routes and of the spatial distribution of tick species and tick-borne pathogens is crucial for understanding the possible impact of birds as spreaders of ticks and tick-borne pathogens. Successful colonization of new tick species or introduction of new tick-borne pathogens will depend on suitable climate, vegetation and hosts. Although it has never been demonstrated that a new tick species, or a new tick pathogen, actually has been established in a new locality after being seeded there by birds, evidence strongly suggests that this could occur.

  20. Rapid identification and differentiation of Theileria sergenti and Theileria sinensis using a loop-mediated isothermal amplification (LAMP) assay.

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    Liu, Aihong; Guan, Guiquan; Du, Pengfei; Gou, Huitian; Zhang, Jun; Liu, Zhijie; Ma, Milin; Ren, Qiaoyun; Liu, Junlong; Yang, Jifei; Li, Youquan; Niu, Qinli; Bai, Qi; Yin, Hong; Luo, Jianxun

    2013-01-16

    The present study developed and validated a species-specific loop-mediated isothermal amplification (LAMP) assay for the rapid detection and discrimination of two benign bovine Theileria species -T. sergenti and T. sinensis. The LAMP assay is inexpensive and easy to perform and involves a rapid reaction-the amplification can be performed in 55 min or 50 min under isothermal conditions of 61°C or 63°C, respectively, by employing a set of four species-specific primer mixtures. The results can be checked using agarose gels. The optimal assay conditions, under which the assay exhibited with no cross-reaction with other closely related tick-borne parasites (T. annulata, Babesia bovis, B. bigemina, B. major, B. ovata, B. U. sp., Anaplasma marginale) or between the two Theileria species of interest, was established. The assay is approximately 10-fold more sensitive than the conventional specific PCR assay. The LAMP assay was validated using DNA from 6 standard stocks in the laboratory and was evaluated for its diagnostic utility using blood samples collected from experimentally and naturally infection cattle or yaks in China. These findings indicate that this Theileria species-specific LAMP assay may have potential clinical applications for the detection and differentiation of two benign bovine Theileria species -T. sergenti and T. sinensis, especially in endemic countries.

  1. Detection of naturally infected vector ticks (acari: ixodidae by different species of babesia and theileria agents from three different enzootic parts of iran.

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    Mohammad Abdigoudarzi

    2013-12-01

    Full Text Available Diagnostic study of vector ticks for different pathogens transmitted specifically have been done by Iranian old scientists working on the basis of biological transmission of pathogens. In this study we decided to confirm natural infection of different collected ticks from three different provinces of Iran.Ticks were collected from livestock (sheep, goats and cattle during favorable seasons (April to September 2007 and 2008. Slide preparations were stained by Giemsa and Feulgen and were studied searching for any trace of infection. Positive DNA from infected blood or tissue samples was provided and was used as positive control. First, PCR optimization for positive DNA was done, and then tick samples were subjected to specific PCR.Eleven pairs of primers were designed for detection of Theileria, Babesia and Anaplasma spp. Totally 21 tick samples were detected to be infected with protozoa. Hyalomma anatolicum anatolicum and Rhipicephalus turanicus from Fars Province were infected with T. lestoquardi at two different places. Hyalomma detritum was infected with T. lestoquardi in Lorestan Province and Rh. turanicus was infected to Ba. ovis from Fars Province.Totally 21 tick samples were detected to be infected with protozoa. Every sample is regarded with host-environment related factors. Since there are complex relations of vectors and their relevant protozoa, different procedures are presented for future studies.

  2. Investigation and control of blood protozoonoses in sheep%张家川县绵羊血液原虫病的调查与防治

    Institute of Scientific and Technical Information of China (English)

    豆晓峰; 付福利; 王耀平; 马学文

    2004-01-01

    对甘肃省张家川县绵羊蜱媒血液原虫病进行了流行病学调查.结果发现,绵羊体表寄生蜱类5种,即青海血蜱(Haemaphysalis qinghaiensis)、长角血蜱(H.longicornis)、革蜱(Dermacentor spp.)、草原革蜱(D.nuttalli)和微小牛蜱(Boophilus microplus),其中青海血蜱为传播媒介优势种.绵羊泰勒虫(Theileria ovis)感染率为62.2%,无形体(Anaplasma ovis)感染率为16.7%,且有双重感染的病例.体表喷洒药物灭蜱试验表明,50mg/L倍特对蜱类的半数致死时间(LT50)为4.91h,用药后3d体表残留活蜱数为3.4±2.2只,14d后再次染蜱数为49.7±12.0只,羊只血液原虫病感染率由38.1%降至8.8%(P0.01).

  3. Detecting and differentiating Theileria sergenti and Theileria sinensis in cattle and yaks by PCR based on major piroplasm surface protein (MPSP).

    Science.gov (United States)

    Liu, Aihong; Guan, Guiquan; Liu, Zhijie; Liu, Junlong; Leblanc, Neil; Li, Youquan; Gao, Jinliang; Ma, Milin; Niu, Qinli; Ren, Qiaoyun; Bai, Qi; Yin, Hong; Luo, Jianxun

    2010-12-01

    Theileria sergenti and Theileria sinensis are closely related members of benign Theileria species found in cattle and yaks in China. They are morphologically indistinguishable. A polymerase chain reaction (PCR) targeting major piroplasm surface protein of T. sergenti and T. sinensis was developed in this study. The newly developed oligonucleotide primer set was able to specifically amplify the DNA of T. sinensis and in conjunction with primers for T. sergenti and these two species could be detected and distinguished. Specificity testing also revealed that there was no cross-reaction with the other tick-borne diseases Theileria annulata, Babesia ovata, Anaplasma marginale as well as bovine white blood cells. Phylogenetic analysis based on the MPSP gene sequences confirmed the specificity of PCR assays. The sensitivity of the methods was 0.1pg DNA for the T. sergenti PCR and 1pg DNA for T. sinensis PCR. Two hundred and thirty-six field blood samples from of cattle and yaks were collected from five different geographical regions in China where benign Theileria species have been found. T. sergenti was found in all five provinces but was absent from one county in Gansu Province. T. sinensis was only found in Gansu Province. In both counties in Gansu where the parasites co-existed, mixed infections were detected. Our results indicate that the PCR methods developed in this study are suitable for the detection and differentiation of T. sergenti and T. sinensis.

  4. Occurrence of haemoparasites in cattle in Monduli district, northern Tanzania

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    Isihaka J. Haji

    2014-02-01

    Full Text Available Haemoparasite infections are among the most economically important cattle diseases in sub-Saharan Africa. The present study investigated the occurrence of haemoparasites in 295 indigenous cattle from five villages (Mswakini, Lake Manyara, Naitolia, Makuyuni and Nanja of the Monduli district, a wildlife-domestic animal-human interface area in northern Tanzania. The data showed that the overall occurrence of haemoparasites in the sampled cattle was 12.5% (95% CI: 8.7% – 16.3%, involving single and mixed infections with Theileria parva, Anaplasma marginale, Babesia bovis, Trypanosoma vivax and Trypanosoma brucei. The highest haemoparasite occurrence was recorded in Lake Manyara (18.3%; 95% CI: 8.5% – 28.1%, and the lowest was recorded in Nanja (6.5%; 95% CI: 0.4% – 12.6%. This preliminary study, furthermore, provided evidence of the possible arthropod vectors (ticks and tsetse flies that may be involved in the transmission of haemoparasites to cattle in the Monduli district. It is envisaged that this survey will stimulate more studies to determine the prevalence of haemoparasites in livestock by using more sensitive molecular techniques.

  5. Lyme disease: the next decade

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    Raphael B Stricker

    2011-01-01

    Full Text Available Raphael B Stricker, Lorraine JohnsonInternational Lyme and Associated Diseases Society, Bethesda, MD, USAAbstract: Although Lyme disease remains a controversial illness, recent events have created an unprecedented opportunity to make progress against this serious tick-borne infection. Evidence presented during the legally mandated review of the restrictive Lyme guidelines of the Infectious Diseases Society of America (IDSA has confirmed the potential for persistent infection with the Lyme spirochete, Borrelia burgdorferi, as well as the complicating role of tick-borne coinfections such as Babesia, Anaplasma, Ehrlichia, and Bartonella species associated with failure of short-course antibiotic therapy. Furthermore, renewed interest in the role of cell wall-deficient (CWD forms in chronic bacterial infection and progress in understanding the molecular mechanisms of biofilms has focused attention on these processes in chronic Lyme disease. Recognition of the importance of CWD forms and biofilms in persistent B. burgdorferi infection should stimulate pharmaceutical research into new antimicrobial agents that target these mechanisms of chronic infection with the Lyme spirochete. Concurrent clinical implementation of proteomic screening offers a chance to correct significant deficiencies in Lyme testing. Advances in these areas have the potential to revolutionize the diagnosis and treatment of Lyme disease in the coming decade.Keywords: Lyme disease, Borrelia burgdorferi, L-forms, cysts, biofilms, proteomics

  6. Serological investigation of vector-borne disease in dogs from rural areas of China

    Institute of Scientific and Technical Information of China (English)

    Shiwen Wang; Jing He; Lijuan Zhang

    2012-01-01

    Objective:To evaluate the Anaplasma phagocytophilum (A. phagocytophilum), Ehrlichia canis (E. canis), Dirofilaria immitis (D. immitis) (canine heartworm), Borrelia burgdorferi (B. burgdorferi) infections in countryside dogs from Yunnan, Hainan and Anhui provinces. Methods: Serum samples were collected from 26 dogs in Yunnan, Hainan and Anhui provinces. The samples were tested using a commercial ELISA rapid diagnostic assay kit (SNAP® 4Dx®; IDEXX Laboratories, Inc. U.S.A.). Meanwhile, indirect immunofluorescence assay (IFA) recommended by WHO was conducted to detect IgG to A. phagocytophilum. Two methods were analyzed and compared. Results: The number of serologically positive dogs for IgG to A. phagocytophilum was only 2 which was from Hainan province and none of the 26 dogs responded positive for E. canis, D. immitis (canine heartworm), and B. burgdorferi by ELISA rapid diagnostic method. The number of serologically positive dogs for IgG to A. phagocytophilum was 13 (50%) by IFA method. Data of the two methods were analyzed by statistical software and the difference was statistically significant (P=0.002). Conclusions: It can be concluded that IFA method was more sensitive than ELISA rapid diagnostic method. However, we need conduct further and intensive epidemiology survey on tick-born diseases pathogens including A. phagocytophilum, E. canis, D. immitis (canine heartworm), and B. burgdorferi which have public health significance.

  7. Spatial distribution of vector borne disease agents in dogs in Aegean region, Turkey

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    Kerem Ural

    2014-06-01

    Full Text Available Objective. Assess the spatial distribution of seroprevalence of infection with or exposure to 4 vector-borne pathogens Ehrlichia canis, Anaplasma phagocytophilum, Borrelia burgdorferi and Dirofilaria immitis, across the coastal states of the Aegean region with special reference to clinical signs and haematological variances related to disease condition. Materials and methods. A convenience sample, targeting blood from at least 10 pet dogs from İzmir, Aydin, Denizli, Mugla and Manisa cities involved was evaluated using a canine point-of-care ELISA kit. Results. Out of 307 dogs tested the overall seroprevalence was highest for E. canis (24.42%, followed by E. canis + A. phagocytophilum co-infection (10.42%, A. phagocytophilum (7.49% and D. immitis (2.28%. Only 2 cases were seropositive to B. burgdorferi albeit 10 dogs were co-infected with more than 2 agents. For both dogs infected with E. canis and co-infected with E. canis and A. phagocytophilum, anemia, thrombocytopenia and leukocytosis, were more commonly detected, whereas thrombocytopenia and leukocytosis were significant finding in dogs infected with A. phagocytophilum or D. immitis, respectively. Variance analysis showed significant differences for mean RBC, Hb, PCV and PLT values (p<0.01 among control group and other groups. Conclusions. Seropositivity for vector-borne pathogens other than B. burgdorferi, is moderately to widely distributed in dogs residing in the Aegean region in Turkey.

  8. No Observed Effect of Landscape Fragmentation on Pathogen Infection Prevalence in Blacklegged Ticks (Ixodes scapularis in the Northeastern United States.

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    Christine P Zolnik

    Full Text Available Pathogen prevalence within blacklegged ticks (Ixodes scapularis Say, 1821 tends to vary across sites and geographic regions, but the underlying causes of this variation are not well understood. Efforts to understand the ecology of Lyme disease have led to the proposition that sites with higher host diversity will result in lower disease risk due to an increase in the abundance of inefficient reservoir species relative to the abundance of species that are highly competent reservoirs. Although the Lyme disease transmission cycle is often cited as a model for this "dilution effect hypothesis", little empirical evidence exists to support that claim. Here we tested the dilution effect hypothesis for two pathogens transmitted by the blacklegged tick along an urban-to-rural gradient in the northeastern United States using landscape fragmentation as a proxy for host biodiversity. Percent impervious surface and habitat fragment size around each site were determined to assess the effect of landscape fragmentation on nymphal blacklegged tick infection with Borrelia burgdorferi and Anaplasma phagocytophilum. Our results do not support the dilution effect hypothesis for either pathogen and are in agreement with the few studies to date that have tested this idea using either a landscape proxy or direct measures of host biodiversity.

  9. Investigation of anaplasmosis in Yiyuan County, Shandong Province, China

    Institute of Scientific and Technical Information of China (English)

    Lijuan Zhang; Feng Cui; Lingling Wang; Lingling Zhang; Jingshan Zhang; Shiwen Wang; Shuxia Yang

    2011-01-01

    Objective:To investigate the situation of anaplasmosis in Yiyuan county, Shandong Province. Methods:A total of 26 blood samples from febrile patients suspected of anaplasmosis,48 blood samples from healthy farmers,8 from dogs, and10 from goats and170 ticks were collected in the same area during2005-2007, and detected by serological and molecular methods.Results:Eight confirmed cases and6probable cases were determined using serologic and molecular methods. The seroprevalence ofAnaplasma phagocytophilum (A. phagocytophilum) was26.7%in healthy cases. Nine out of10sheep samples and7 out of8 dog samples reacted positively to theA. phagocytophilumantigen.PCR amplification and sequencing of the16SrRNA ofA. phagocytophilum gene showed that some samples from patients, goats and ticks were100% identical. The seroprevalence ofRickettsia typhi was22.9%,Orientia tsutsugamushi6.3%, Rickettsia sibirica27.1%,Coxiella burnetii18.8%,Bartonella henselae31.3%, andBorrelia burgdorferi41.6%.Conclusions: It is important to make differential diagnosis of febrile patients and to apply treatment with specific antibiotics. It is needed to enforce essential prevention and control measures including tick control and to improve sanitation conditions.

  10. Prevalence of Bartonella species, haemoplasmas and Toxoplasma gondii in cats in Scotland.

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    Bennett, Alexander D; Gunn-Moore, Danielle A; Brewer, Melissa; Lappin, Michael R

    2011-08-01

    The objective of this study was to determine the prevalence rates for select infectious agents of cats presented to the Royal (Dick) School of Veterinary Studies at the University of Edinburgh, Scotland. Whole blood, serum, and oral mucosal and nail bed swabs were collected. While Ehrlichia species, Anaplasma species or Rickettsia felis DNA were not amplified from any cat, 44.2% of the cats had evidence of infection or exposure to either a Bartonella species (15.3% were seropositive and 5.8% polymerase chain reaction (PCR) positive), a haemoplasma (28.6% PCR positive), and/or Toxoplasma gondii (19.2% seropositive). No Bartonella species DNA was amplified from the nail or oral mucosal swabs despite a 5.8% amplification rate from the blood samples. This finding likely reflects the absence of Ctenocephalides felis infection from our study population, as this organism is a key component for Bartonella species translocation in cats. The results from this study support the use of flea control products to lessen exposure of cats (and people) to Bartonella species and support discouraging the feeding of raw meat to cats and preventing them from hunting to lessen T gondii infection.

  11. The development of an enzyme-linked immunosorbent assay for Trypanosoma vivax antibodies and its use in epidemiological surveys

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    Claudio R Madruga

    2006-11-01

    Full Text Available There are data indicating that the distribution of Trypanosoma vivax in the Brazilian territory is expanding with potential to reach other areas, where the vectors are present. The detection of anti-trypanosomal antibodies in serum provides important information of the trypanosomal status in cattle herds. For this reason, an enzyme-linked immunosorbent assay (Tv-ELISA-Ab with crude antigen from one Brazilian isolate of T. vivax was developed and evaluated. The sensitivity and specificity were respectively 97.6 and 96.9%. In the evaluation of cross-reactions, three calves inoculated with T. evansi trypimastigotes blood forms showed optical densities (OD under the cut-off during the whole experimental period, except one at 45 days post-inoculation. With relation to Babesia bovis, B. bigemina, and Anaplasma marginale, which are endemic hemoparasites in the studied area, the cross-reactions were shown to be 5.7, 5.3, and 1.1%, respectively. The first serological survey of Pantanal and state of Pará showed that T. vivax is widespread, although regions within both areas had significantly different prevalences. Therefore, this Tv-ELISA-Ab may be a more appropriate test for epidemiological studies in developing countries because the diagnostic laboratories in most countries may be able to perform an ELISA, which is not true for polymerase chain reaction.

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

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    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.

  13. Serosurvey for selected disease agents in white-tailed deer from Mexico.

    Science.gov (United States)

    Martinez, A; Salinas, A; Martinez, F; Cantu, A; Miller, D K

    1999-10-01

    Serum samples from 350 white-tailed deer (Odocoileus virginianus texanus) collected in March 1994 from northeastern Mexico were tested for the prevalence of antibody activity against five infectious diseases of ruminants. The prevalence rate was 81% for bluetongue virus (BTV) of all serotypes, 72% for epizootic hemorrhagic disease virus (EHDV), 3% for Borrelia burgdorferi, 69% for Anaplasma marginale, and 0% for Brucella abortus, B. melitensis, and B. ovis. These are diseases that affect domestic ruminants, and deer may act as a reservoir of infection. In addition, if deer are translocated, they may introduce pathogens to formerly disease-free areas. The high seroprevalence of BTV and EHDV cannot be related to the presence of hemorrhagic disease in the deer in this region. This is the first report to indicate the presence of B. burgdorferi infection of deer in Mexico. Despite the high prevalence of A. marginale titers, it is uncertain that deer play a role in the epizootiology of cattle anaplasmosis in the region. Apparently, white-tailed deer are unimportant in the epizootiology of brucellosis of both cattle and goats in northeastern Mexico.

  14. Detection of Parasites and Parasitic Infections of Free-Ranging Wildlife on a Game Ranch in Zambia: A Challenge for Disease Control

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    Hetron Mweemba Munang'andu

    2012-01-01

    Full Text Available Ex-situ conservancies are expanding alternatives to livestock production in Zambia albeit the lack of information on circulating infectious parasites from wildlife. Therefore, 12 wildlife species were examined on a game ranch were all species were found to be infected by Rhipecephalus spp. Haemoparasite infections were estimated at 7.37% (n=95 with Babesia spp. detected in bushbuck (Tragelaphus scriptus; Anaplasma marginale in impala (Aepyceros melampus and puku (Kobus vardonii for the first time in Zambia. The majority of worm species isolated from bovids were not detected in equids and, vice versa. Our findings intimate ecological and behavioural patterns of some animals as deterministic to exposure. Kafue lechwe (Kobus leche kafuensis had the widest range of worm species with more infected organs than other animals suggesting their semi aquatic nature contributory to prolonged worm exposure compared to other animals. On the other hand, Kafue lechwe had the least tick infections attributable more to shorter attachment periods as they spend prolonged periods submerged in water. Our findings indicate the vital role that wildlife plays in the epidemiology of parasitic diseases. To reduce the infection burden, control measures should be focused on reducing transmission to highly susceptible animal species as described herein.

  15. 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.

  16. Recent study on canine vector-borne zoonoses in southern Slovakia - serologic survey.

    Science.gov (United States)

    Čabanová, Viktória; Pantchev, Nikola; Hurníková, Zuzana; Miterpáková, Martina

    2015-12-01

    Over the last decade a significant spread of Canine Vector Borne Diseases has been recorded in Central Europe. The aim of the study described here, was to collect current data on the occurrence and distribution of three major canine vector-borne pathogens in the veterinary clinical practice by a newly-developed commercial ELISA test for the detection of Dirofilaria immitis antigen as well as specific circulating antibodies to Anaplasma phagocytophilum and Borrelia burgdorferi sensu lato. Circulating D. immitis antigen was detected in five of 180 investigated sera samples. Two of D. immitis seropositive dogs revealed also microfilariae of D. repens in the blood and three of them were negative for the presence of microfilariae in the Knott's test. From the practical point of view, the finding of D. immitis occult infections might influence existing knowledge about distribution of this species among dogs in Central European countries. In 11.7% of the tested dogs the presence of specific antibodies against A. phagocytophilum was confirmed. Antibodies against B. burgdorferi s.l. were detected in 2.8% of tested sera samples. Coinfection with A. phagocytophilum and B. burgdorferi s.l. was observed in two dogs from Košice district in south-eastern Slovakia. Our data point toward the presence of Canine Vector Borne Diseases in the studied area. Therefore, veterinarians should include these diseases in their differential diagnosis and higher awareness should be focused also on prophylactic measures to prevent the pathogens transmission by arthropod vectors.

  17. [Toxinology of bovine paraplegic syndrome].

    Science.gov (United States)

    Sevcik, C; Brito, J C; D'Suze, G; Mijares, A J; Domínguez, M G

    1993-01-01

    A clinical entity named "Bovine Paraplegic Syndrome" ("Síndrome Parapléjico de los Bovinos") has spread alarmingly, in the cattle growing areas of the central and eastern plains of Venezuela. Approximately four million cattle are bread in the area were the disease occurs. The mortality index due to the disease ranges 5 to 25% of the animals at risk, mostly cows, pregnant or lactating. The principal characteristic of the bovine paraplegic syndrome is decubitus, ventral or sternal, in animals that make vane efforts to stand when stimulated. The diagnosis is established ruling out, clinically and with laboratory findings, that the animals are suffering known diseases with similar symptoms such as paralytic rabies, botulism and blood parasites such Trypanosoma sp., Babesia sp., and Anaplasma sp.. Death occurs always, usually after few days, and to this date there is no known treatment able to save the sick cows. In this work, we describe results that suggest the presence of a toxin in the cattle suffering and prone to suffer the syndrome; it is a natural toxin produced by ruminal bacteria. In squid giant axons under voltage clamp conditions, this toxin is very specific to block sodium current during nerve electrical activity.

  18. Toxicology of a bovine paraplegic syndrome.

    Science.gov (United States)

    Sevcik, C; Brito, J C; D'Suze, G; Domínguez-Bello, M G; Lovera, M; Mijares, A J; Bónoli, S

    1993-12-01

    A clinical entity named 'bovine paraplegic syndrome' ('síndrome parapléjico de los bovinos') has spread alarmingly in the cattle-growing areas of the central and eastern plains of Venezuela. It is estimated that four million cattle are bred in the area where the disease occurs. The mortality ranges from 5 to 25% of the animals at risk, mostly pregnant or lactating cows. The principal characteristic of the bovine paraplegic syndrome is ventral or sternal decubitus, in animals that make vain efforts to stand when stimulated. The diagnosis is established when all other possible causes (e.g. paralytic rabies, botulism and blood parasites such as Anaplasma marginal, Babesia bovis, B. bigemina, and Trypanosoma vivax) have been ruled out clinically and by laboratory tests. Death always occurs, usually after a few days, and there is no known treatment. In this work, we describe results that show the presence of a toxin in the cattle suffering from, or liable to suffer from the syndrome. The toxin is produced by ruminal bacteria. In squid giant axons under voltage clamp conditions, the toxin blocks the sodium current. We detected the toxin analytically by absorbance measurements at 340 nm after reacting with picrylsulfonic acid. We obtained a good separation of the toxin with isocratic high pressure liquid chromatography, using 40% methanol in water on phenylborasil columns.

  19. Anaplasmataceae and Borrelia burgdorferi sensu lato in the sand lizard Lacerta agilis and co-infection of these bacteria in hosted Ixodes ricinus ticks

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    Ekner Anna

    2011-09-01

    Full Text Available Abstract Background Anaplasmataceae and Borrelia burgdorferi s.l. are important tick-borne bacteria maintained in nature by transmission between ticks and vertebrate hosts. However, the potential role of lizards as hosts has not been sufficiently studied. Results The current study showed that 23 of 171 examined sand lizards Lacerta agilis were PCR positive for Anaplasmataceae. The nucleotide sequences of the several selected PCR products showed 100% homology with Anaplasma spp. found in Ixodes ricinus collected in Tunisia and Morocco (AY672415 - AY672420. 1.2% of lizard collar scale samples were PCR positive for B. lusitaniae. In addition, 12 of 290 examined I. ricinus were PCR positive for B. burgdorferi s.l. and 82 were PCR positive for Anaplasmatacea. The number of ticks per lizard and the number of ticks PCR positive for both microorganisms per lizard were strongly correlated. Moreover, we found a significant correlation between numbers of ticks infected with Anaplasmataceae and with B. burgdorferi s.l. living on the same lizard. However, there was no significant correlation between detection of both bacteria in the same tick. Conclusions To the best of our knowledge, this is the first report of Anaplasmataceae DNA and additionally the second report of B. burgdorferi s.l DNA detection in the sand lizard.

  20. Ixodes ricinus and its transmitted pathogens in urban and peri-urban areas in Europe: new hazards and relevance for public health

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    Annapaola eRizzoli

    2014-12-01

    Full Text Available Tick-borne diseases represent major public and animal health issues worldwide. Ixodes ricinus, primarily associated with deciduous and mixed forests, is the principal vector of causative agents of viral, bacterial and protozoan zoonotic diseases in Europe. Recently, abundant tick populations have been observed in European urban green areas, which are of public health relevance due to exposure of humans and domesticated animals to potentially infected ticks. In urban habitats, small and medium sized mammals, birds, companion animals (dogs, cats and larger mammals (roe deer, wild boar play a role in maintenance of tick populations and as reservoirs of tick-borne pathogens. Presence of ticks infected with tick-borne encephalitis virus and high prevalence of ticks infected with Borrelia burgdorferi s.l., causing Lyme borreliosis, have been reported from urbanized areas in Europe. Emerging pathogens, including bacteria of the order Rickettsiales (Anaplasma phagocytophilum, 'Candidatus Neoehrlichia mikurensis', Rickettsia helvetica, R. monacensis, Borrelia miyamatoi and protozoans (Babesia divergens, B. venatorum and B. microti have also been detected in urban tick populations. Understanding the ecology of ticks and their associations with hosts in a European urbanized environment is crucial to quantify parameters necessary for risk pre-assessment and identification of public health strategies for control and prevention of tick-borne diseases.

  1. Interaction of the tick immune system with transmitted pathogens

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    Ondrej eHajdusek

    2013-07-01

    Full Text Available Ticks are hematophagous arachnids transmitting a wide variety of pathogens including viruses, bacteria, and protozoans to their vertebrate hosts. The tick vector competence has to be intimately linked to the ability of transmitted pathogens to evade tick defense mechanisms encountered on their route through the tick body comprising midgut, hemolymph, salivary glands or ovaries. Tick innate immunity is, like in other invertebrates, based on an orchestrated action of humoral and cellular immune responses. The direct antimicrobial defense in ticks is accomplished by a variety of small molecules such as defensins, lysozymes or by tick-specific antimicrobial compounds such as microplusin/hebraein or 5.3-kDa family proteins. Phagocytosis of the invading microbes by tick hemocytes seems to be mediated by the primordial complement-like system composed of thioester-containing proteins, fibrinogen-related lectins and convertase-like factors. Moreover, an important role in survival of the ingested microbes seems to be played by host proteins and redox balance maintenance in the tick midgut. Here, we summarize recent knowledge about the major components of tick immune system and focus on their interaction with the relevant tick-transmitted pathogens, represented by spirochetes (Borrelia, rickettsiae (Anaplasma, and protozoans (Babesia. Availability of the tick genomic database and feasibility of functional genomics based on RNA interference greatly contribute to the understanding of molecular and cellular interplay at the tick-pathogen interface and may provide new targets for blocking the transmission of tick pathogens.

  2. A proline racemase based PCR for identification of Trypanosoma vivax in cattle blood.

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    Regassa Fikru

    Full Text Available A study was conducted to develop a Trypanosoma vivax (T. vivax specific PCR based on the T. vivax proline racemase (TvPRAC gene. Forward and reverse primers were designed that bind at 764-783 bp and 983-1002 bp of the gene. To assess its specificity, TvPRAC PCR was conducted on DNA extracted from different haemotropic pathogens: T. vivax from Nigeria, Ethiopia and Venezuela, T. congolense Savannah type, T. brucei brucei, T. evansi, T. equiperdum, T. theileri, Theileria parva, Anaplasma marginale, Babesia bovis and Babesia bigemina and from bovine, goat, mouse, camel and human blood. The analytical sensitivity of the TvPRAC PCR was compared with that of the ITS-1 PCR and the 18S PCR-RFLP on a dilution series of T. vivax DNA in water. The diagnostic performance of the three PCRs was compared on 411 Ethiopian bovine blood specimens collected in a former study. TvPRAC PCR proved to be fully specific for T. vivax, irrespective of its geographical origin. Its analytical sensitivity was lower than that of ITS-1 PCR. On these bovine specimens, TvPRAC PCR detected 8.3% T. vivax infections while ITS-1 PCR and 18S PCR-RFLP detected respectively 22.6 and 6.1% T. vivax infections. The study demonstrates that a proline racemase based PCR could be used, preferably in combination with ITS-1 PCR, as a species-specific diagnostic test for T. vivax infections worldwide.

  3. Tick-borne rickettsial pathogens in questing ticks, removed from humans and animals in Mexico.

    Science.gov (United States)

    Sosa-Gutierrez, Carolina G; Vargas-Sandoval, Margarita; Torres, Javier; Gordillo-Pérez, Guadalupe

    2016-09-30

    Tick-borne rickettsial diseases (TBRD) are commonly encountered in medical and veterinary clinical settings. The control of these diseases is difficult, requiring disruption of a complex transmission chain involving a vertebrate host and ticks. The geographical distribution of the diseases is related to distribution of the vector, which is an indicator of risk for the population. A total of 1107 were collected by tick drag from forests, ecotourism parks and hosts at 101 sites in 22 of the 32 states of Mexico. Collected ticks were placed in 1.5 mL cryovials containing 70% ethanol and were identified to species. Ticks were pooled according to location/host of collection, date of collection, sex, and stage of development. A total of 51 ticks were assayed by polymerase chain reaction (PCR) to confirm species identification using morphological methods. A total of 477 pools of ticks were assayed using PCR techniques for selected tick-borne pathogens. Anaplasma phagocytophilum was the most commonly detected pathogen (45 pools), followed by, Ehrlichia (E.) canis (42), Rickettsia (R.) rickettsii (11), E. chaffeensis (8), and R. amblyommii (1). Rhipicephalus sanguineus was the tick most frequently positive for selected pathogens. Overall, our results indicate that potential tick vectors positive for rickettsial pathogens are distributed throughout the area surveyed in Mexico.

  4. Parasitism and mutualism in Wolbachia

    DEFF Research Database (Denmark)

    Bordenstein, Seth R; Paraskevopoulos, Charalampos; Dunning Hotopp, Julie C;

    2009-01-01

    Ecological and evolutionary theories predict that parasitism and mutualism are not fixed endpoints of the symbiotic spectrum. Rather, parasitism and mutualism may be host or environment dependent, induced by the same genetic machinery, and shifted due to selection. These models presume the existe......Ecological and evolutionary theories predict that parasitism and mutualism are not fixed endpoints of the symbiotic spectrum. Rather, parasitism and mutualism may be host or environment dependent, induced by the same genetic machinery, and shifted due to selection. These models presume...... the existence of genetic or environmental variation that can spur incipient changes in symbiotic lifestyle. However, for obligate intracellular bacteria whose genomes are highly reduced, studies specify that discrete symbiotic associations can be evolutionarily stable for hundreds of millions of years...... in symbiotic lifestyle with a comprehensive, phylogenomic analysis. Contrary to previous claims, we show unequivocally that the transition in lifestyle cannot be reconstructed with current methods due to long-branch attraction (LBA) artifacts of the distant Anaplasma and Ehrlichia outgroups. Despite the use...

  5. Coexistence of emerging bacterial pathogens in Ixodes ricinus ticks in Serbia*

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    Tomanović S.

    2010-09-01

    Full Text Available The list of tick-borne pathogens is long, varied and includes viruses, bacteria, protozoa and nematodes. As all of these agents can exist in ticks, their co-infections have been previously reported. We studied co-infections of emerging bacterial pathogens (Borrelia burgdorferi sensu lato, Anaplasma phagocytophilum and Francisella tularensis in Ixodes ricinus ticks in Serbia. Using PCR technique, we detected species-specific sequences, rrf-rrl rDNA intergenic spacer for B. burgdorferi s.l., p44/msp2 paralogs for A. phagocytophilum, and the 17 kDa lipoprotein gene, TUL4, for F. tularensis, respectively, in total DNA extracted from the ticks. Common infections with more than one pathogen were detected in 42 (28.8 % of 146 infected I. ricinus ticks. Co-infections with two pathogens were present in 39 (26.7 % of infected ticks. Simultaneous presence of A. phagocytophilum and different genospecies of B. burgdorferi s.l. complex was recorded in 16 ticks, co-infection with different B. burgdorferi s. l. genospecies was found in 15 ticks and eight ticks harbored mixed infections with F. tularensis and B. burgdorferi s.l. genospecies. Less common were triple pathogen species infections, detected in three ticks, one infected with A. phagocytophilum / B. burgdorferi s.s. / B. lusitaniae and two infected with F. tularensis / B. burgdorferi s.s. / B. lusitaniae. No mixed infections of A. phagocytophilum and F. tularensis were detected.

  6. Neorickettsia sennetsu as a Neglected Cause of Fever in South-East Asia.

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    Sabine Dittrich

    Full Text Available Neorickettsia sennetsu infection is rarely recognized, with less than 100 globally reported patients over the last 50 years. The disease is thought to be contracted by eating raw fish, a staple of many South-East Asian cuisines. In 2009, the first patient with sennetsu was identified in the Lao PDR (Laos, raising the question as to how common this organism and related species are in patients presenting with fever. We investigated the frequency of N. sennetsu infection at hospitals in diverse areas of Laos. Consenting febrile hospital inpatients from central (Vientiane: n = 1,013, northern (Luang Namtha: n = 453 and southern (Salavan: n = 171 Laos were screened by PCR for N. sennetsu, if no previous positive direct diagnostic test was available. A PCR-restriction fragment length polymorphism assay was developed to differentiate between N. sennetsu, Ehrlichia chaffeensis and Anaplasma phagocytophilum. To allow more detailed studies of N. sennetsu, culture was successfully established using a reference strain (ATCC VR-367, identifying a canine-macrophage cell line (DH82 to be most suitable to visually identify infection. After screening, N. sennetsu was identified and sequence confirmed in four (4/1,637; 0.2% Lao patients. Despite the previously identified high seroprevalence of N. sennetsu antibodies in the Lao population (~17%, acute N. sennetsu infection with sufficient clinical signs to prompt hospitalization appears to be rare. The reservoir, zoonotic cycle and pathogenicity of N. sennetsu remain unclear and require further investigations.

  7. Transport of Ixodid ticks and tick-borne pathogens by migratory birds.

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    Gunnar eHasle

    2013-09-01

    Full Text Available Birds, particularly passerines, can be parasitized by Ixodid ticks, which may be infected with tick-borne pathogens, like Borrelia spp., Babesia spp., Anaplasma, Rickettsia/Coxiella, and tick-borne encephalitis virus. The prevalence of ticks on birds varies over years, season, locality and different bird species. The prevalence of ticks on different species depends mainly on the degree of feeding on the ground. In Europe, the Turdus spp., especially the blackbird, Turdus merula, appears to be most important for harboring ticks. Birds can easily cross barriers, like fences, mountains, glaciers, desserts and oceans, which would stop mammals, and they can move much faster than the wingless hosts. Birds can potentially transport tick-borne pathogens by transporting infected ticks, by being infected with tick-borne pathogens and transmit the pathogens to the ticks, and possibly act as hosts for transfer of pathogens between ticks through co-feeding. Knowledge of the bird migration routes and of the spatial distribution of tick species and tick-borne pathogens is crucial for understanding the possible impact of birds as spreaders of ticks and tick-borne pathogens. Successful colonization of new tick species or introduction of new tick-borne pathogens will depend on suitable climate, vegetation and hosts. Although it has never been demonstrated that a new tick species, or a new tick pathogen, actually has been established in a new locality after being seeded there by birds, evidence strongly suggests that this could occur.

  8. “Nothing is permanent but change”* -- Antigenic variation in persistent bacterial pathogens

    Science.gov (United States)

    Palmer, Guy H.; Bankhead, Troy; Lukehart, Sheila A.

    2012-01-01

    Summary Pathogens persist in immunocompetent mammalian hosts using various strategies, including evasion of immune effectors by antigenic variation. Among highly antigenically variant bacteria, gene conversion is used to generate novel expressed variants from otherwise silent donor sequences. Recombination using oligonucleotide segments from multiple donors is a combinatorial mechanism that tremendously expands the variant repertoire, allowing thousands of variants to be generated from a relatively small donor pool. Three bacterial pathogens, each encoded by a small genome (Borrelia burgdorferi VlsE diversity is encoded and expressed on a linear plasmid required for persistence and recent experiments have demonstrated that VlsE recombination is necessary for persistence in the immunocompetent host. In contrast, both Treponema pallidum TprK and Anaplasma marginale Msp2 expression sites and donors are chromosomally encoded. Both T. pallidum and A. marginale generate antigenic variants in vivo in individual hosts and studies at the population level reveal marked strain diversity in the variant repertoire that may underlie pathogen strain structure and the capacity for re-infection and heterologous strain superinfection. Here, we review gene conversion in bacterial antigenic variation and discuss the short- and long-term selective pressures that shape the variant repertoire. PMID:19709057

  9. 'Nothing is permanent but change'- antigenic variation in persistent bacterial pathogens.

    Science.gov (United States)

    Palmer, Guy H; Bankhead, Troy; Lukehart, Sheila A

    2009-12-01

    Pathogens persist in immunocompetent mammalian hosts using various strategies, including evasion of immune effectors by antigenic variation. Among highly antigenically variant bacteria, gene conversion is used to generate novel expressed variants from otherwise silent donor sequences. Recombination using oligonucleotide segments from multiple donors is a combinatorial mechanism that tremendously expands the variant repertoire, allowing thousands of variants to be generated from a relatively small donor pool. Three bacterial pathogens, each encoded by a small genome (Borrelia burgdorferi VlsE diversity is encoded and expressed on a linear plasmid required for persistence and recent experiments have demonstrated that VlsE recombination is necessary for persistence in the immunocompetent host. In contrast, both Treponema pallidum TprK and Anaplasma marginale Msp2 expression sites and donors are chromosomally encoded. Both T. pallidum and A. marginale generate antigenic variants in vivo in individual hosts and studies at the population level reveal marked strain diversity in the variant repertoire that may underlie pathogen strain structure and the capacity for re-infection and heterologous strain superinfection. Here, we review gene conversion in bacterial antigenic variation and discuss the short- and long-term selective pressures that shape the variant repertoire.

  10. Tick-borne ehrlichiosis infection in human beings

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    S. Ganguly

    2008-11-01

    Full Text Available Human monocytic ehrlichiosis is a tick-borne infectious disease transmitted by several tick species, especially Amblyomma spp caused by Ehrlichia chaffeensis. E. chaffeensis is an obligatory intracellular, tick-transmitted bacterium that is maintained in nature in a cycle involving at least one and perhaps several vertebrate reservoir hosts. Two additional Ehrlichia spp, Anaplasma (formerly Ehrlichia phagocytophila (the agent of human granulocytic ehrlichiosis [HGE] and E. ewingii (a cause of granulocytic ehrlichiosis in dogs act as human pathogens. Human E. chaffeensis infections have generally been reported in North America, Asia and Europe, but recently human cases have been reported in Brazil only. Human monocytic ehrlichiosis is diagnosed by demonstration of a four-fold or greater change in antibody titer to E. chaffeensis antigen by IFA in paired serum samples, or a positive PCR assay and confirmation of E. chaffeensis DNA, or identification of morulae in leukocytes and a positive IFA titer to E. chaffeensis antigen, or immunostaining of E. chaffeensis antigen in a biopsy or autopsy sample, or culture of E. chaffeensis from a clinical specimen.

  11. Molecular detection of tick-borne pathogens in Ixodes ricinus from Moldova collected in 1960.

    Science.gov (United States)

    Movila, Alexandru; Toderas, Ion; Uspenskaia, Inga; Conovalov, Jurii

    2013-06-01

    This study is the first report about the prevalence of tick-borne pathogens, as well as their (co-)infection rates, in the museum-archived I. ricinus female ticks collected in Moldova in 1960. A total of 16.7% (21/126) ticks was mono-infected. Borrelia burgdorferi sensu stricto was revealed as the most abundant species (4.8%) followed by B. garinii (1.6%), B. afzelii (0.8%), B. valaisiana (0.8%), and B. lusitaniae (0.8%). DNA of Rickettsia helvetica (2.4%), R. monacensis (2.4%), Anaplasma phagocytophilum (2.4%), 'Candidatus Neoehrlichia mikurensis' (0.8%), and Babesia microti (0.8%) were also detected, indicating the occurrence of these emerging tick-borne microorganisms in Moldova since 1960 at least. In this study, we detected a co-infection (0.8%; 1/126 tested ticks) between B. microti and R. helvetica. Additional investigations are warranted to further characterize a historical snapshot of the distribution of tick-borne pathogens in Europe.

  12. Advances in genetic manipulation of obligate intracellular bacterial pathogens

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    Paul eBeare

    2011-05-01

    Full Text Available Infections by obligate intracellular bacterial pathogens result in significant morbidity and mortality worldwide. These bacteria include Chlamydia spp., which causes millions of cases of sexually transmitted disease and blinding trachoma annually, and members of the α-proteobacterial genera Anaplasma, Ehrlichia, Orientia and Rickettsia, agents of serious human illnesses including epidemic typhus. Coxiella burnetii, the agent of human Q fever, has also been considered a prototypical obligate intracellular bacterium, but recent host cell-free (axenic growth has rescued it from obligatism. The historic genetic intractability of obligate intracellular bacteria has severely limited molecular dissection of their unique lifestyles and virulence factors involved in pathogenesis. Host cell restricted growth is a significant barrier to genetic transformation that can make simple procedures for free-living bacteria, such as cloning, exceedingly difficult. Low transformation efficiency requiring long term culture in host cells to expand small transformant populations is another obstacle. Despite numerous technical limitations, the last decade has witnessed significant gains in genetic manipulation of obligate intracellular bacteria including allelic exchange. Continued development of genetic tools should soon enable routine mutation and complementation strategies for virulence factor discovery and stimulate renewed interest in these refractory pathogens. In this review, we discuss the technical challenges associated with genetic transformation of obligate intracellular bacteria and highlight advances made with individual genera.

  13. Human ehrlichiosis

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    Đokić Milomir

    2006-01-01

    Full Text Available Background. Human ehrlichiosis is a newly recognized disease. It is a tick-borne disease caused by several bacterial species of the genhus Erlichia. These are small gram-negative pleomorphic cocci, that are obligatory intracellular bacteria. Tick Ixodes is the principle vector in Europe, and Amblyomma americanum in the United States. Bacterial organisms replicate in a tick, and are transmited from infected cells in a vector to the blood cells of animals or humans. Human ehrlichiosis is a name for a group of diseases caused by different species of Ehrlichia. One of them is the disease named human monocytic ehrlichiosis, caused by Ehrlichia chaffeensis, and the other is a human granulocytic ehrlichiosis caused by Anaplasma phagocytophilia. Case report. We reported a 23-year-old patient admitted for the clinical treatment with the symptoms of high febrility (above 40 °C, headache, vomiting, general weakness and exhaustion, but without data on a tick bite. The patient was treated with trimetoprim-sulfamethoxazole for a week when Ehrlichia chaffeensis was confirmed by the immunofluoroscence test, and the therapy contimed with doxacyclin. Conclusion. Human ehrlichiosis is also present in our country, so this disease should be considered everyday, especially in infectology practice.

  14. Molecular evidence of tick-borne hemoprotozoan-parasites (Theileria ovis and Babesia ovis) and bacteria in ticks and blood from small ruminants in Northern Algeria.

    Science.gov (United States)

    Aouadi, Atef; Leulmi, Hamza; Boucheikhchoukh, Mehdi; Benakhla, Ahmed; Raoult, Didier; Parola, Philippe

    2017-02-01

    Using qPCR, standard PCR and/or sequencing, we investigated the presence of tick-associated microorganisms in ticks and blood from sheep and goats from Souk Ahras, Algeria. Borrelia theileri, was detected in (7/120, 5.8%) blood from sheep and (13/120, 10.8%) goats. Anaplasma ovis was screened in (38/73, 52%) Rhipicephalus bursa and (5/22, 22.7%) R. turanicus and in (74/120, 61.7%), (65/120, 54.2%) blood of sheep and goats respectively. Coxiella burnetii tested positive in R. bursa (4/73, 5.5%) and (7/120, 5.8%) blood of sheep and (2/120, 1.7%) goats. Theileria ovis was detected in (50/147, 34%) R. bursa and (3/22, 13.6%) R. turanicus and in (64/120, 53.3%) blood of sheep and (25/120, 20.8%) goats. Babesia ovis was screened positive in (23/147, 15.6%) R. bursa and (7/48, 14.6%) R. turanicus. Our findings expand knowledge about the repertoire of tick-borne microorganisms present in ectoparasites and/or the blood of small ruminants in Algeria.

  15. Kekerabatan Genetik Caplak Rhiphicephalus (Boophilus microplus Asal IndonesiaBerdasarkan Sekuen Internal Transcribed Spacer-2 (GENETIC RELATIONSHIP INDONESIAN RHIPHICEPHALUS (BOOPHILUS MICROPLUS TICK BASED ON INTERNAL TRANSCRIBED SPACER-2 SEQUENSE

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    Ana Sahara

    2015-10-01

    Full Text Available Rhiphicephalus (Boophilus microplus is important obligatory blood feeding ectoparasites transmittingmany different viral, bacterial and protozoan and plays a role as a vector of Babesia sp., The leria sp. andAnaplasma sp. in cattle. The accuracy in identifying and distinguishing interspecies and intraspeciesdiversity among parasites is needed to understand the epidemiology, biology and capacity as a vector.Variations in the DNA base sequence of the internal transcribed spacer region2 (ITS 2 has been used asa molecular marker for identification in an effort to determine phylogenetic relationships. The aim of thisstudy was to determine the ITS 2 gene nucleotide sequence of R. microplus, which was expected to beuseful for accurate identification the parasite diversity and phylogenetic relationship among many differentspecies. DNA amplification was conducted using BOO2 forward dan BOO2 reverse primers. The DNAsamples containing ITS2 region fragment of 1099 nt were derived from the nucleotide sequence multiplealignments of R.microplus and other ticks genes obtained from Gene bank using Clustal W software, andthen analyzed using the MEGA program version 6. Genetic distances based on nucleotide sequence weredetermined with Kimura 2-parameter method producing the smallest genetic distance of 0 % and 1.2 %.Construction of phylogenetic trees using the Neighbor joining method showed that ticks from variousregions in Indonesia was species complex which have a closer with R.microplus isolates from India, Laos,South Africa, China and Australia R.australis origin.

  16. Zoonotic vector-borne bacterial pathogens in California mountain lions (Puma concolor), 1987-2010.

    Science.gov (United States)

    Girard, Yvette A; Swift, Pamela; Chomel, Bruno B; Kasten, Rickie W; Fleer, Katryna; Foley, Janet E; Torres, Steven G; Johnson, Christine K

    2012-11-01

    Sera collected from 442 mountain lions in 48 California counties between the years of 1987 and 2010 were tested using immunofluorescence assays and agglutination tests for the presence of antibodies reactive to Yersinia pestis, Francisella tularensis, Bartonella henselae, Borrelia burgdorferi, and Anaplasma phagocytophilum antigens. Data were analyzed for spatial and temporal trends in seropositivity. Seroprevalences for B. burgdorferi (19.9%) and B. henselae (37.1%) were relatively high, with the highest exposure in the Central Coast region for B. henselae. B. henselae DNA amplified in mountain lion samples was genetically similar to human-derived Houston-1 and domestic cat-derived U4 B. henselae strains at the gltA and ftsZ loci. The statewide seroprevalences of Y. pestis (1.4%), F. tularensis (1.4%), and A. phagocytophilum (5.9%), were comparatively low. Sera from Y. pestis- and F. tularensis-seropositive mountain lions were primarily collected in the Eastern and Western Sierra Nevada, and samples reactive to Y. pestis antigen were collected exclusively from adult females. Adult age (≥ 2 years) was a risk factor for B. burgdorferi exposure. Over 70% of tested animals were killed on depredation permits, and therefore were active near areas with livestock and human residential communities. Surveillance of mountain lions for these bacterial vector-borne and zoonotic agents may be informative to public health authorities, and the data are useful for detecting enzootic and peridomestic pathogen transmission patterns, particularly in combination with molecular characterization of the infecting organisms.

  17. Study on ticks and tick-borne zoonoses in public parks in Italy.

    Science.gov (United States)

    Corrain, R; Drigo, M; Fenati, M; Menandro, M L; Mondin, A; Pasotto, D; Martini, M

    2012-11-01

    A survey on tick density and on tick-borne zoonoses was carried out in four public parks in the outskirts of Imola (northern Italy) from June to October 2006. All stages of Ixodes ricinus and only larvae of Riphicephalus sanguineus were recovered by dragging, performed on 100-m transects. Almost all ticks (99%) were harvested in one park. I. ricinus density (nymphs/100 m(2) ) ranged from 0 in park L to 6.3 in park F. Nymphs and adults of I. ricinus were subjected to PCR for Anaplasma phagocytophilum, Bartonella spp., Borrelia burgdorferi s. l. and Rickettsia spp. The observed prevalences were 38.3% for Bartonella henselae, 5.2% for Bartonella clarridgeiae, 10.4% for B. burgdorferi s. l., 2.6% for Rickettsia helvetica and 13% for Rickettsia monacensis, respectively. No DNA of A. phagocytophilum was found. Acarological risks (AR) were calculated as probabilities of collecting at least one infected nymph per transect. The AR values calculated for the various zoonotic agents were 11.4% for R. helvetica, 27.7% for B. clarridgeiae, 49.7% for B. burgdorferi s. l., 57.2% for R. monacensis and 90.4% for B. henselae, respectively. In this study, B. clarridgeiae was for the first time identified in I. ricinus ticks.

  18. Serologic survey for selected arboviruses and other potential pathogens in wildlife from Mexico.

    Science.gov (United States)

    Aguirre, A A; McLean, R G; Cook, R S; Quan, T J

    1992-07-01

    During 1988 and 1989, a serologic survey of wildlife was conducted in northeastern Mexico to determine the presence, prevalence, and distribution of arboviruses and other selected disease agents. Eighty mammal specimens were tested. Antibodies to vesicular stomatitis-Indiana, Venezuelan equine encephalitis-Mena II, Rio Grande virus, and vesicular stomatitis-New Jersey were detected predominantly in small mammals. Deer and mouflon (Ovis musimon) had antibodies to bluetongue and epizootic hemorrhagic disease. Two species had serologic evidence of recent exposure to Francisella tularensis. A white-tailed deer (Odocoileus virginianus) had antibodies to Anaplasma marginale. All specimens tested for antibodies against Yersinia pestis and Brucella abortus were negative. Sera from 315 birds were tested for antibody against five equine encephalitis viruses and six avian pathogens. During 1988, antibodies to Venezuelan equine encephalitis-Mena II, Venezuelan equine encephalitis-TC83, St. Louis encephalitis, eastern equine encephalitis, and western equine encephalitis were detected in birds of several species. Antibodies to Pasteurella multocida and Newcastle disease virus were also detected. Birds from five species presented antibodies to Mycoplasma meleagridis. Specimens tested for M. gallisepticum, M. synoviae, and Chlamydia psittaci were negative. To the best of our knowledge, this survey represents the first serologic evidence of bluetongue, Cache Valley virus, epizootic hemorrhagic disease, Jamestown Canyon virus, vesicular stomatitis-Indiana, vesicular stomatitis-New Jersey, Rio Grande virus, and tularemia reported among wildlife in Mexico.

  19. Canine granulocytic anaplasmosis: a review.

    Science.gov (United States)

    Carrade, D D; Foley, J E; Borjesson, D L; Sykes, J E

    2009-01-01

    Anaplasma phagocytophilum is an emerging pathogen of humans, horses, and dogs worldwide that is transmitted by Ixodid ticks and maintained in a variety of small wild mammal species. Recent studies suggest that multiple strains of A. phagocytophilum may be circulating in wild and domestic animal populations, and these strains may have differential host tropisms and pathogenicity. The organism infects and survives within neutrophils by disabling key neutrophil functions, including neutrophil motility, phagocytosis, the oxidative burst mechanism, and neutrophil-endothelial cell interactions, as well as interfering with neutrophil apoptosis. Coinfections with other tick-borne pathogens may occur, especially Borrelia burgdorferi. A. phagocytophilum causes an acute febrile illness in dogs with lethargy and inappetence. Less frequent signs include lameness, coughing, polydipsia, intermittent vomiting, and hemorrhages. Diagnosis is based on finding morulae within granulocytes in the peripheral blood, the combination of acute and convalescent serology using immunofluorescent antibody techniques, and detection of the DNA of A. phagocytophilum using specific polymerase chain reaction assays. Whether persistent infection or reinfection with A. phagocytophilum occurs after natural infection requires additional study, with most reports suggesting that anaplasmosis is a self-limiting disease in dogs that responds well to a 2-week course of doxycycline therapy.

  20. Occurrence of haemoparasites in cattle in Monduli district, northern Tanzania.

    Science.gov (United States)

    Haji, Isihaka J; Malele, Imna; Namangala, Boniface

    2014-11-13

    Haemoparasite infections are among the most economically important cattle diseases in sub-Saharan Africa. The present study investigated the occurrence of haemoparasites in 295 indigenous cattle from five villages (Mswakini, Lake Manyara, Naitolia, Makuyuni and Nanja) of the Monduli district, a wildlife-domestic animal-human interface area in northern Tanzania. The data showed that the overall occurrence of haemoparasites in the sampled cattle was 12.5% (95% CI: 8.7% - 16.3%), involving single and mixed infections with Theileria parva, Anaplasma marginale, Babesia bovis, Trypanosoma vivax and Trypanosoma brucei. The highest haemoparasite occurrence was recorded in Lake Manyara (18.3%; 95% CI: 8.5% - 28.1%), and the lowest was recorded in Nanja (6.5%; 95% CI: 0.4% - 12.6%). This preliminary study, furthermore, provided evidence of the possible arthropod vectors (ticks and tsetse flies) that may be involved in the transmission of haemoparasites to cattle in the Monduli district. It is envisaged that this survey will stimulate more studies to determine the prevalence of haemoparasites in livestock by using more sensitive molecular techniques.

  1. Molecular diagnosis of Anaplasmataceae organisms in dogs with clinical and microscopical signs of ehrlichiosis Diagnóstico molecular de agentes da família Anaplasmataceae em cães com sinais clínicos e microscópios de erliquiose

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    Ana Sílvia Dagnone

    2009-12-01

    Full Text Available Ehrlichioses are important emerging zoonotic tick-borne diseases that can affect both animals and humans. Clinical manifestations of ehrlichiosis caused by different members of Anaplasmataceae in dogs are similar to each other and to other diseases showing systemic manifestation. The observation of inclusions in white blood cells and in platelets cannot be used to confirm the Anaplasmataceae etiologic agent of the disease. In this work we assessed the presence of Anaplasmataceae agents in 51 dogs from two different cities (Jaboticabal and Campo Grande showing clinical and microscopical diagnosis of ehrlichiosis, by using molecular techniques. Anaplasmataceae DNA were amplified in 46/51 (90.2% of the blood samples; 22 (40% samples from Jaboticabal and 10 (18.2% from Campo Grande were positive for E. canis nPCR. Anaplasma platys DNA was amplified in 2 samples from Jaboticabal and in 11 from Campo Grande. Phylogenetic analysis of E. canis and A. platys DNA confirmed the infection agent and showed that PCR is the most reliable method to diagnose ehrlichial infection.Erliquioses são importantes enfermidades emergentes transmitidas por carrapatos que podem afetar os animais e o homem. Em cães, as manifestações clínicas da erliquiose causada por diferentes membros da Família Anaplasmataceae são similares entre si e entre outras enfermidades de manifestação sistêmica. A observação de inclusões em leucócitos e plaquetas não pode ser utilizada para diagnosticar o agente etiológico pertencente à Família Anaplasmataceae. O presente trabalho objetivou detectar, por meio de técnicas moleculares, a presença de agentes da Família Anaplasmataceae em 51 cães de duas diferentes cidades (Jaboticabal, SP e Campo Grande, MS apresentando sinais clínicos e microscópios sugestivos de erliquiose. DNA de agentes da Família Anaplasmataceae foi amplificado em 46/51 (90,2% das amostras de sangue; 22 (40% amostras de Jaboticabal e 10 (18,2% amostras

  2. Apoptose na infecção experimental de cães domésticos com Ehrlichia canis Apoptosis in experimental infection with Ehrlichia canis in domestic dogs

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    Irma Ximena Barbosa Sanchez

    2012-08-01

    Full Text Available A Erliquiose canina é uma zoonose causada pela Ehrlichia canis, bactéria Gram negativa de distribuição mundial. Alguns cães com erliquiose se tornam portadores assintomáticos enquanto outros desenvolvem uma doença aguda com morte rápida. A apoptose pode ser importante na eliminação de patógenos intracelulares, podendo, nas infecções por Ehrlichia sp. e Anaplasma sp., ocorrer modulação da apoptose celular para prolongar a sobrevivência desses organismos. Para avaliação do papel da apoptose na erliquiose canina, sete cães foram distribuídos em dois grupos. No Grupo inoculado, realizou-se a infecção por via intravenosa com sangue infectado com E. canis (isolado Jaboticabal, sendo realizada a inoculação com PBS estéril nos animais pertencentes ao Grupo Controle. Semanalmente e até 35 dias pós-inoculação, amostras de sangue foram coletadas e submetidas a n-PCR e reação de imunofluorescência (RIFI para confirmação da infecção. No 36° dia pós-inoculação, os animais foram eutanasiados, sendo as amostras de baço, linfonodo, rim e fígado coletadas e processadas para as técnicas de TUNEL e imunohistoquímica (Bcl-2, Bax. Verificou-se pela n-PCR que os animais inoculados se tornaram positivos para E. canis a partir da segunda semana. Pela RIFI, verificou-se animais com sorologia positiva a partir da terceira semana pós-inoculação. No grupo controle, todos os testes realizados foram negativos para E. canis. Apesar da reação de TUNEL mostrar maior incidência de apoptose no Grupo Inoculado, sendo o baço e os linfonodos os órgãos que apresentaram maior marcação, os resultados da imunohistoquímica para Bcl-2 e Bax indicam que a via intrínseca de apoptose não é importante nos órgãos analisados.Some dogs infected with Ehrlichia canis become asymptomatic while others develop an acute illness followed by quick death. Apoptosis may be an important mechanism for elimination of intracellular pathogens. Also

  3. Survey on tick-borne anaplasmataceae in the south edge of Gurbantunggut desert%古尔班通古特沙漠南缘蜱携带无形体的调查

    Institute of Scientific and Technical Information of China (English)

    孟庆玲; 乔军; 盛金良; 王俊伟; 王为升; 姚娜; 陈创夫; 张丽娟

    2012-01-01

    To understand the tick-borne anaplasmataceae in the south edge of Gurbantunggut desert, ticks specimens from 11 sites in three different habitats areas were collected and identified. Nested PCR was carried to detect positive ticks carring anaplasmataceae,and then 5% end variable region of 16S rRNA gene of anaplasmataceae was amplified by semi-nested PCR for cloning and sequencing. The obtained DNA sequences were compared with the GenBank sequence and phylogenetic tree was constructed with the Mega 5.0 software. The 708 ticks collected from 11 sites were identified as 4 genera and 8 species. 25 were detected positive in the 236 tick samples, with a positive rate of 10.59 %. By sequence analyses, Ehrlichia cha ff eensis , Ehrlichia eanis , Ana plasrna marginale and Ana plasrna ovis were positive in Hyalomrna asiaticum ,Rhipicephalus sanguineeus , Hyalornma detritum and Haemaphysalis longi- cornis,respectively. The results showed that there exists anaplasmataceae in ticks parasited in livestock from the south edge of Gurbantunggut desert,suggesting that the region may be one of natural loci of anaplasmosis.%为了解古尔班通古特沙漠南缘蜱携带无形体的状况,在该地域3个不同生境区域11个地点采集家畜寄生蜱标本,进行分类鉴定;运用套式PCR筛选无形体阳性蜱样本,并用半套式PCR扩增无形体科16SrRNA基因5′端高变区,进行克隆测序;将所获得的DNA序列与GenBank收录的序列作比对,并用Mega 5.0软件构建系统发育树,鉴定蜱携带无形体的种类。结果共采集708只蜱,鉴定为4个属8个种。在分类后的236份蜱样本中,检测出无形体阳性25份,阳性率为10.59%。序列比较发现,在亚洲璃眼蜱、血红扇头蜱、残缘璃眼蜱、长角血蜱中分别存在查菲埃立克体(Ehrlichia chaffeensis)、犬埃立克体(Ehrlichia canis)、边缘无形体(Anaplasma marginale)和绵羊无形体(Anaplasma ovis)的16SrRNA基因片段。研究结果证实

  4. Serosurvey of tick-borne pathogens in dogs from urban and rural areas from Parana State, Brazil Avaliação sorológica de patógenos transmitidos por carrapatos em cães urbanos e rurais do estado do Paraná, Brasil

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    Thállitha Samih Wischral Jayme Vieira

    2013-03-01

    Full Text Available Considering the zoonotic potential of tick-borne disease (TBD agents and the fact that dogs may act as sentinels for human infection, the aim of the present study was to determine the seroprevalence of TBD agents and risk factors for exposure in two different canine populations from Parana State, Southern Brazil. A total of 138 dog serum samples from urban (UA (n=68 and rural (RA (n=70 areas were tested with commercial ELISA rapid test for Anaplasma phagocytophilum, Ehrlichia canis and Borrelia burgdorferi antibodies and indirect immunofluorescence assay (IFAT for Babesia vogeli. An overall of 92∕138 (66.7% dogs, being 62∕68 (91.2% from UA and 30∕70 (42.9% from RA, were seropositive for at least one TBD agent. From the total number of dogs, sixty-two were positive for E. canis (44.9%, 19 (13.8% for A. phagocytophilum, and 64 (46.4% for B. vogeli. Anti-B. burgdorferi antibodies were not detected. Dogs from UA showed a higher percentage of tick infestation (p = 0.0135 and were highly associated with seropositivity to E. canis (p = 0.000005, A. phagocytophilum (p = 0.0001, and B. vogeli (p = 0.0012. In summary, the findings indicate that dogs from urban areas present higher potential risk exposure to TBD pathogens than those from rural areas.Considerando o potencial zoonótico das doenças transmitidas por carrapatos (DTCs e que os cães podem atuar como sentinelas para infecções em humanos, os objetivos deste estudo foram determinar a soroprevalência de agentes das DTCs e fatores de risco para a exposição em duas diferentes populações caninas do Estado do Paraná, região Sul do Brasil. Um total de 138 amostras de soro de cães de área urbana (AU (n = 68 e rural (AR (n = 70 foram testadas utilizando um teste de ELISA comercial rápido para detecção de anticorpos contra Anaplasma phagocytophilum, Ehrlichia canis e Borrelia burgdorferi e imunofluorescência indireta (IFI para Babesia vogeli. Um total de 92∕138 (66,7% cães, sendo

  5. Detection of co-infection with Lyme spirochetes and Spotted fever group rickettsiae in a group of Haemaphysalis longicornis%一组长角血蜱中检出莱姆病螺旋体和斑点热群立克次体复合感染

    Institute of Scientific and Technical Information of China (English)

    孟真; 姜理平; 陆群英; 程苏云; 叶菊莲; 占利

    2008-01-01

    目的 了解浙江省山区野生动物和蜱中莱姆病、斑点热、埃立克体病(无形体病)的感染情况.方法 采用巢式PCR对采集的鼠、蜱标本进行莱姆病伯氏疏螺旋体、斑点热群立克次体、埃立克体(无形体)特异性核酸片段检测分析.结果 从121份鼠标本和105组蜱标本中检出阳性结果 14份.鼠标本中检出伯氏疏螺旋体5S~23S rDNA间隔区片段1份和埃立克体(无形体)16SrDNA 5'端片段2份.蜱标本中检出阳性11份,包括伯氏疏螺旋体5S~23S rDNA间隔区片段3份和斑点热群立克次体外膜蛋白OmpA基因5'端片段8份.其中1组长角血蜱成虫标本为伯氏疏螺旋体和斑点热群立克次体复合感染,5S~23S rRNA基因间隔区和ompA基因片段均阳性,分别与伯氏疏螺旋体法雷氏基因型和马赛立克次体株等关系较近.结论 在同一组长角血蜱成虫中同时检出莱姆病疏螺旋体和斑点热群立克次体复合感染.%Objective The present study was conducted to investigate the infection of Lyme disease, Spotted fever, Ehrlichiosis (anaplasmosisin) in wild animals and ticks in the mountain areas of Zhejiang province. Methods Nested polymerase chain reaction was used to amplify specific DNA sequences of Lyme spirochetes, Spotted fever group rickettsiae, Ehrlichia (anaplasma) from samples of mice and ticks. Results 14 positive samples were identified from 121 mice and 105 groups of ticks. Among mice samples, one positive 5S-23S rDNA intergenic spacer of Borreia burgdorferi and two 5' fragments of Ehrlichia (anaplasma) 16S rDNA were obtained. 11 positive results were detected from tick samples including three 5S-23S rDNA intergenic spacer regions of Borreia burgdorferi and eight 5' fragments of Spotted fever group rickettsiae outer member protein A gene. One group of adult ticks, Haemaphysalis longicornis, which had been collected from eastern mountain area were detected to have co-infected with Lyme spirochetes and

  6. Molecular characterization of Ehrlichia canis infecting dogs, Buenos Aires.

    Science.gov (United States)

    Cicuttin, Gabriel L; De Salvo, María N; Gury Dohmen, Federico E

    2016-07-01

    Canine monocytic ehrlichiosis (CME) is a worldwide potentially fatal tick-borne rickettsial disease of dogs caused by Ehrlichia canis and transmitted by Rhipicephalus sanguineus sensu lato. CME diagnosis includes indirect (serology) and direct (e.g. blood smears and PCR) methods. PCR is more sensitive and specific than direct microscopic examination and positive PCR results confirm infection, whereas positive serologic test results only confirm exposure. The aim of the present study was to perform a molecular characterization of E. canis from canine samples of the Metropolitan Area of Buenos Aires. We studied 223 blood samples of dogs submitted to our institute for CME diagnoses. The samples were initially screened for Anaplasmataceae family by PCR, resulting in 30 positive dogs (13.4%). Subsequently, positive DNAs were analyzed by nested PCR 16S rRNA specific for E. canis or Anaplasma platys, resulting in 15 (6.7%) and 16 (7.2%) positive dogs, respectively. For molecular characterization, samples positive for E. canis were subjected to amplification of a fragment of the dsb and p28 genes. The nucleotide sequences obtained for the dsb fragment resulted in 100% identity with others E. canis found in dogs from different regions of worldwide. The nucleotide sequences obtained for p28 gene resulted in 100% of identity with each other and closely with E. canis str. Jaboticabal (Brazil). Identity with others sequences of E. canis ranged from 76.9 to 79.7%. The occurrence of canine cases molecularly confirmed in Metropolitan Area of Buenos Aires highlights the need for more studies in order to understand epidemiological factors associated with CME, especially the disease transmission dynamic in South America given the existence of two lineages of R. sanguineus sensu lato with different vectorial capacity for transmission of E. canis.

  7. The pCS20 PCR assay for Ehrlichia ruminantium does not cross-react with the novel deer ehrlichial agent found in white-tailed deer in the United States of America

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    S.M. Mahan

    2004-11-01

    Full Text Available White-tailed deer are susceptible to heartwater (Ehrlichia [Cowdria] ruminantium infection and are likely to suffer high mortality if the disease spreads to the United States. It is vital, therefore, to validate a highly specific and sensitive detection method for E. ruminantium infection that can be reliably used in testing white-tailed deer, which are reservoirs of antigenically or genetically related agents such as Ehrlichia chaffeensis, Anaplasma (Ehrlichia phagocytophilum (HGE agent and Ehrlichia ewingii. Recently, a novel but as yet unnamed ehrlichial species, the white-tailed deer ehrlichia (WTDE, has been discovered in deer populations in the United States. Although the significance of WTDE as a pathogen is unknown at present, it can be distinguished from other Ehrlichia spp. based on 16S rRNA gene sequence analysis. In this study it was differentiated from E. ruminantium by the use of the pCS20 PCR assay which has high specificity and sensitivity for the detection of E. ruminantium. This assay did not amplify DNA from the WTDE DNA samples isolated from deer resident in Florida, Georgia and Missouri, but amplified the specific 279 bp fragment from E. ruminantium DNA. The specificity of the pCS20 PCR assay for E. ruminantium was confirmed by Southern hybridization. Similarly, the 16S PCR primers (nested that amplify a specific 405-412 bp fragment from the WTDE DNA samples, did not amplify any product from E. ruminantium DNA. This result demonstrates that it would be possible to differentiate between E. ruminantium and the novel WTDE agent found in white tailed deer by applying the two respective PCR assays followed by Southern hybridizations. Since the pCS20 PCR assay also does not amplify any DNA products from E. chaffeensis or Ehrlichia canis DNA, it is therefore the method of choice for the detection of E. ruminantium in these deer and other animal hosts.

  8. Ehrlichiosis, babesiosis, anaplasmosis and hepatozoonosis in dogs from St. Kitts, West Indies.

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    Patrick J Kelly

    Full Text Available BACKGROUND: Although tick-borne diseases are important causes of morbidity and mortality in dogs in tropical areas, there is little information on the agents causing these infections in the Caribbean. METHODOLOGY: We used PCRs to test blood from a cross-section of dogs on St Kitts for Ehrlichia (E. canis, Babesia (B. spp., Anaplasma (A. spp. and Hepatozoon (H. spp. Antibodies against E. canis and A. phagocytophilum/platys were detected using commercial immunochromatography tests. Records of the dogs were examined retrospectively to obtain clinical and laboratory data. PRINCIPAL FINDINGS: There was serological and/or PCR evidence of infections of dogs with E. canis (27%; 46/170, Babesia spp. (24%; 90/372 including B. canis vogeli (12%; 43/372 and B. gibsoni (10%; 36/372, A. platys (11%; 17/157 and H. canis (6%; 15/266. We could not identify the Babesia sp. detected in nine dogs. There was evidence of multiple infections with dual infections with E. canis and B. canis vogeli (8%; 14/179 or B. gibsoni (7%; 11/170 being the most common. There was agreement between immunochromatography and PCR test results for E. canis for 87% of dogs. Only 13% of exposed dogs had signs of a tick-borne disease and 38% had laboratory abnormalities. All 10 dogs presenting for a recheck after treatment of E. canis with doxycycline were apparently healthy although all remained seropositive and six still had laboratory abnormalities despite an average of two treatments with the most recent being around 12 months previously. Infections with Babesia spp. were also mainly subclinical with only 6% (4/67 showing clinical signs and 13% (9/67 having laboratory abnormalities. Similarly, animals with evidence of infections with A. platys and H. canis were largely apparently healthy with only occasional laboratory abnormalities. CONCLUSIONS: Dogs are commonly infected with tick-borne pathogens in the Caribbean with most having no clinical signs or laboratory abnormalities.

  9. Application of highly sensitive saturation labeling to the analysis of differential protein expression in infected ticks from limited samples

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    Villar Margarita

    2010-08-01

    Full Text Available Abstract Background Ticks are vectors of pathogens that affect human and animal health worldwide. Proteomics and genomics studies of infected ticks are required to understand tick-pathogen interactions and identify potential vaccine antigens to control pathogen transmission. One of the limitations for proteomics research in ticks is the amount of protein that can be obtained from these organisms. In the work reported here, individual naturally-infected and uninfected Rhipicephalus spp. ticks were processed using a method that permits simultaneous extraction of DNA, RNA and proteins. This approach allowed using DNA to determine pathogen infection, protein for proteomics studies and RNA to characterize mRNA levels for some of the differentially expressed proteins. Differential protein expression in response to natural infection with different pathogens was characterized by two-dimensional (2-D differential in gel electrophoresis (DIGE saturation labeling in combination with mass spectrometry analysis. To our knowledge, this is the first report of the application of DIGE saturation labeling to study tick proteins. Results Questing and feeding Rhipicephalus spp. adult ticks were collected in 27 farms located in different Sicilian regions. From 300 collected ticks, only 16 were found to be infected: R. sanguineus with Rickettsia conorii and Ehrlichia canis; R. bursa with Theileria annulata; and R. turanicus with Anaplasma ovis. The proteomic analysis conducted from a limited amount of proteins allowed the identification of host, pathogen and tick proteins differentially expressed as a consequence of infection. Conclusion These results showed that DIGE saturation labeling is a powerful technology for proteomics studies in small number of ticks and provided new information about the effect of pathogen infection in ticks.

  10. Prevalence and Diversity of Tick-Borne Pathogens in Nymphal Ixodes scapularis (Acari: Ixodidae) in Eastern National Parks.

    Science.gov (United States)

    Johnson, Tammi L; Graham, Christine B; Boegler, Karen A; Cherry, Cara C; Maes, Sarah E; Pilgard, Mark A; Hojgaard, Andrias; Buttke, Danielle E; Eisen, Rebecca J

    2016-12-27

    Tick-borne pathogens transmitted by Ixodes scapularis Say (Acari: Ixodidae), also known as the deer tick or blacklegged tick, are increasing in incidence and geographic distribution in the United States. We examined the risk of tick-borne disease exposure in 9 national parks across six Northeastern and Mid-Atlantic States and the District of Columbia in 2014 and 2015. To assess the recreational risk to park visitors, we sampled for ticks along frequently used trails and calculated the density of I. scapularis nymphs (DON) and the density of infected nymphs (DIN). We determined the nymphal infection prevalence of I. scapularis with a suite of tick-borne pathogens including Borrelia burgdorferi, Borrelia miyamotoi, Anaplasma phagocytophilum, and Babesia microti Ixodes scapularis nymphs were found in all national park units; DON ranged from 0.40 to 13.73 nymphs per 100 m(2) Borrelia burgdorferi, the causative agent of Lyme disease, was found at all sites where I. scapularis was documented; DIN with B. burgdorferi ranged from 0.06 to 5.71 nymphs per 100 m(2) Borrelia miyamotoi and A. phagocytophilum were documented at 60% and 70% of the parks, respectively, while Ba. microti occurred at just 20% of the parks. Ixodes scapularis is well established across much of the Northeastern and Mid-Atlantic States, and our results are generally consistent with previous studies conducted near the areas we sampled. Newly established I. scapularis populations were documented in two locations: Washington, D.C. (Rock Creek Park) and Greene County, Virginia (Shenandoah National Park). This research demonstrates the potential risk of tick-borne pathogen exposure in national parks and can be used to educate park visitors about the importance of preventative actions to minimize tick exposure.

  11. Zoonotic infections among employees from Great Smoky Mountains and Rocky Mountain National Parks, 2008-2009.

    Science.gov (United States)

    Adjemian, Jennifer; Weber, Ingrid B; McQuiston, Jennifer; Griffith, Kevin S; Mead, Paul S; Nicholson, William; Roche, Aubree; Schriefer, Martin; Fischer, Marc; Kosoy, Olga; Laven, Janeen J; Stoddard, Robyn A; Hoffmaster, Alex R; Smith, Theresa; Bui, Duy; Wilkins, Patricia P; Jones, Jeffery L; Gupton, Paige N; Quinn, Conrad P; Messonnier, Nancy; Higgins, Charles; Wong, David

    2012-11-01

    U.S. National Park Service employees may have prolonged exposure to wildlife and arthropods, placing them at increased risk of infection with endemic zoonoses. To evaluate possible zoonotic risks present at both Great Smoky Mountains (GRSM) and Rocky Mountain (ROMO) National Parks, we assessed park employees for baseline seroprevalence to specific zoonotic pathogens, followed by evaluation of incident infections over a 1-year study period. Park personnel showed evidence of prior infection with a variety of zoonotic agents, including California serogroup bunyaviruses (31.9%), Bartonella henselae (26.7%), spotted fever group rickettsiae (22.2%), Toxoplasma gondii (11.1%), Anaplasma phagocytophilum (8.1%), Brucella spp. (8.9%), flaviviruses (2.2%), and Bacillus anthracis (1.5%). Over a 1-year study period, we detected incident infections with leptospirosis (5.7%), B. henselae (5.7%), spotted fever group rickettsiae (1.5%), T. gondii (1.5%), B. anthracis (1.5%), and La Crosse virus (1.5%) in staff members at GRSM, and with spotted fever group rickettsiae (8.5%) and B. henselae (4.3%) in staff at ROMO. The risk of any incident infection was greater for employees who worked as resource managers (OR 7.4; 95% CI 1.4,37.5; p=0.02), and as law enforcement rangers/rescue crew (OR 6.5; 95% CI 1.1,36.5; p=0.03), relative to those who worked primarily in administration or management. The results of this study increase our understanding of the pathogens circulating within both parks, and can be used to inform the development of effective guidelines and interventions to increase visitor and staff awareness and help prevent exposure to zoonotic agents.

  12. Molecular detection of Rickettsia conorii and other zoonotic spotted fever group rickettsiae in ticks, Romania.

    Science.gov (United States)

    Ionita, Mariana; Silaghi, Cornelia; Mitrea, Ioan Liviu; Edouard, Sophie; Parola, Philippe; Pfister, Kurt

    2016-02-01

    The diverse tick fauna as well as the abundance of tick populations in Romania represent potential risks for both human and animal health. Spotted fever group (SFG) rickettsiae are recognized as important agents of emerging human tick-borne diseases worldwide. However, the epidemiology of rickettsial diseases has been poorly investigated in Romania. In urban habitats, companion animals which are frequently exposed to tick infestation, play a role in maintenance of tick populations and as reservoirs of tick-borne pathogens. Therefore, the aim of the present study was to investigate the occurrence of SFG rickettsiae in ticks infesting dogs in a greater urban area in South-eastern Romania. Adult ixodid ticks (n=205), including Rhipicephalus sanguineus sensu lato (n=120), Dermacentor reticulatus (n=76) and Ixodes ricinus (n=9) were collected from naturally infested dogs and were screened for SFG rickettsiae using conventional PCR followed by sequencing. Additionally, ticks were screened for DNA of Babesia spp., Hepatozoon spp., Ehrlichia canis, and Anaplasma platys. Four zoonotic SFG rickettsiae were identified: Rickettsia raoultii (16%) and Rickettsia slovaca (3%) in D. reticulatus, Rickettsia monacensis (11%) in I. ricinus, and Rickettsia conorii (0.8%) in Rh. sanguineus s.l. Moreover, pathogens of veterinary importance, such as B. canis (21%) in D. reticulatus and E. canis (7.5%) in Rh. sanguineus s.l. were identified. The findings expand the knowledge on distribution of SFG rickettsiae as well as canine pathogens in Romania. Additionally, this is the first report describing the molecular detection of R. conorii in ticks from Romania.

  13. Role of sand lizards in the ecology of Lyme and other tick-borne diseases in the Netherlands

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    Spitzen - van der Sluijs Annemarieke

    2010-05-01

    Full Text Available Abstract Background Lizards are considered zooprophylactic for almost all Borrelia burgdorferi species, and act as dilution hosts in parts of North America. Whether European lizards significantly reduce the ability of B. burgdorferi to maintain itself in enzootic cycles, and consequently decrease the infection rate of Ixodes ricinus ticks for B. burgdorferi and other tick-borne pathogens in Western Europe is not clear. Results Ticks were collected from sand lizards, their habitat (heath and from the adjacent forest. DNA of tick-borne pathogens was detected by PCR followed by reverse line blotting. Tick densities were measured at all four locations by blanket dragging. Nymphs and adult ticks collected from lizards had a significantly lower (1.4% prevalence of B. burgdorferi sensu lato, compared to questing ticks in heath (24% or forest (19%. The prevalence of Rickettsia helvetica was significantly higher in ticks from lizards (19% than those from woodland (10% whereas neither was significantly different from the prevalence in ticks from heather (15%. The prevalence of Anaplasma and Ehrlichia spp in heather (12% and forest (14% were comparable, but significantly lower in ticks from sand lizards (5.4%. The prevalence of Babesia spp in ticks varied between 0 and 5.3%. Tick load of lizards ranged from 1 - 16. Tick densities were ~ 5-fold lower in the heather areas than in woodlands at all four sites. Conclusions Despite their apparent low reservoir competence, the presence of sand lizards had insignificant impact on the B. burgdorferi s.l. infection rate of questing ticks. In contrast, sand lizards might act as reservoir hosts for R. helvetica. Remarkably, the public health risk from tick-borne diseases is approximately five times lower in heather than in woodland, due to the low tick densities in heather.

  14. Novel Ehrlichia and Hepatozoon agents infecting the crab-eating fox (Cerdocyon thous) in southeastern Brazil.

    Science.gov (United States)

    Almeida, Aliny P; Souza, Tayse D; Marcili, Arlei; Labruna, Marcelo B

    2013-05-01

    This study evaluated infection by vector-borne agents in 58 crab-eating fox (Cerdocyon thous L.) that were road-killed in an Atlantic rainforest reserve in the state of Espírito Santo, southeastern Brazil. Spleen, lung, or blood samples collected from the foxes were tested in the laboratory by a battery of polymerase chain reaction (PCR) assays targeting bacteria of the genera Rickettsia, Borrelia, Coxiella, Anaplasma, and Ehrlichia; and protozoa of the genera Babesia, Hepatozoon, and Leishmania. Of the targeted organisms, evidence of infection in the foxes was detected for Ehrlichia and Hepatozoon organisms only. Overall, six (10.3%) foxes were infected by an ehrlichial agent closely related to an ehrlichial agent recently detected in free-ranging Jaguars [(Panthera onca (L.)] in central-western Brazil, and to Ehrlichia ruminantium. For Hepatozoon, 28 (48.3%) foxes were infected by an agent closely related to Hepatozoon sp. Curupira 2 and H. americanum; and one (1.7%) fox was infected by an organism closely related to reptile-associated Hepatozoon agents. Finally, 11 (19.0%) foxes were found infested by Amblyomma cajennense (F.) nymphs, which were all PCR negative for the range of vector-borne agents cited above. Because the haplotypes found in free-ranging foxes are genetically closely related to pathogens of great veterinary importance, namely E. ruminantium and H. americanum, it is highly desirable to know if these novel organisms have any important role as agents of diseases in domestic animals and wildlife in Brazil.

  15. Disaggregating Tropical Disease Prevalence by Climatic and Vegetative Zones within Tropical West Africa.

    Science.gov (United States)

    Beckley, Carl S; Shaban, Salisu; Palmer, Guy H; Hudak, Andrew T; Noh, Susan M; Futse, James E

    2016-01-01

    Tropical infectious disease prevalence is dependent on many socio-cultural determinants. However, rainfall and temperature frequently underlie overall prevalence, particularly for vector-borne diseases. As a result these diseases have increased prevalence in tropical as compared to temperate regions. Specific to tropical Africa, the tendency to incorrectly infer that tropical diseases are uniformly prevalent has been partially overcome with solid epidemiologic data. This finer resolution data is important in multiple contexts, including understanding risk, predictive value in disease diagnosis, and population immunity. We hypothesized that within the context of a tropical climate, vector-borne pathogen prevalence would significantly differ according to zonal differences in rainfall, temperature, relative humidity and vegetation condition. We then determined if these environmental data were predictive of pathogen prevalence. First we determined the prevalence of three major pathogens of cattle, Anaplasma marginale, Babesia bigemina and Theileria spp, in the three vegetation zones where cattle are predominantly raised in Ghana: Guinea savannah, semi-deciduous forest, and coastal savannah. The prevalence of A. marginale was 63%, 26% for Theileria spp and 2% for B. bigemina. A. marginale and Theileria spp. were significantly more prevalent in the coastal savannah as compared to either the Guinea savanna or the semi-deciduous forest, supporting acceptance of the first hypothesis. To test the predictive power of environmental variables, the data over a three year period were considered in best subsets multiple linear regression models predicting prevalence of each pathogen. Corrected Akaike Information Criteria (AICc) were assigned to the alternative models to compare their utility. Competitive models for each response were averaged using AICc weights. Rainfall was most predictive of pathogen prevalence, and EVI also contributed to A. marginale and B. bigemina prevalence

  16. Co-infection and genetic diversity of tick-borne pathogens in roe deer from Poland.

    Science.gov (United States)

    Welc-Falęciak, Renata; Werszko, Joanna; Cydzik, Krystian; Bajer, Anna; Michalik, Jerzy; Behnke, Jerzy M

    2013-05-01

    Wild species are essential hosts for maintaining Ixodes ticks and the tick-borne diseases. The aim of our study was to estimate the prevalence, the rate of co-infection with Babesia, Bartonella, and Anaplasma phagocytophilum, and the molecular diversity of tick-borne pathogens in roe deer in Poland. Almost half of the tested samples provided evidence of infection with at least 1 species. A. phagocytophilum (37.3%) was the most common and Bartonella (13.4%) the rarest infection. A total of 18.3% of all positive samples from roe deer were infected with at least 2 pathogens, and one-third of those were co-infected with A. phagocytophilum, Bartonella, and Babesia species. On the basis of multilocus molecular studies we conclude that: (1) Two different genetic variants of A. phagocytophilum, zoonotic and nonzoonotic, are widely distributed in Polish roe deer population; (2) the roe deer is the host for zoonotic Babesia (Bab. venatorum, Bab. divergens), closely related or identical with strains/species found in humans; (3) our Bab. capreoli and Bab. divergens isolates differed from reported genotypes at 2 conserved base positions, i.e., positions 631 and 663; and (4) this is the first description of Bart. schoenbuchensis infections in roe deer in Poland. We present 1 of the first complex epidemiological studies on the prevalence of Babesia, Bartonella, and A. phagocytophilum in naturally infected populations of roe deer. These game animals clearly have an important role as reservoir hosts of tick-borne pathogens, but the pathogenicity and zoonotic potential of the parasite genotypes hosted by roe deer requires further detailed investigation.

  17. Association of BoLA-DRB3 alleles with tick-borne disease tolerance in dairy cattle in a tropical environment.

    Science.gov (United States)

    Duangjinda, M; Jindatajak, Y; Tipvong, W; Sriwarothai, J; Pattarajinda, V; Katawatin, S; Boonkum, W

    2013-09-23

    Tick-borne disease is one of the most harmful tropical diseases in dairy production. Selection of dairy cows for tolerance to tick-borne disease is a challenging concept for dairy breeders in the tropics. The objectives of this study were (1) to detect specific tick-borne pathogen in cattle of different genetics and (2) to examine the polymorphisms of DRB3.2 alleles in Thai dairy cattle and find the allelic association with tick-borne disease tolerance. Specific primers to Anaplasma marginale (AM), Babesia bigemina (BG) and Babesia bovis (BB) were used to detect the infections by PCR. The results showed that the high proportion of infections were found in Bos indicus (Sahiwal, n=95) and crossbred Holstein × Zebu (75:25 Holstein:Zebu, n=101), compared to high Holstein fraction crossbreed (≥ 87.5% Holstein, n=187). The proportion of triple infections was also highly found in high Holstein fractions crossbreed. This study confirmed that Zebuine (Bos indicus) had a higher degree of tolerance, even when infected by tick-borne pathogens, compared to high Holstein fraction crossbred. The associated alleles of DRB3.2 for tick-borne pathogen infection tolerance were found: DRB3*14 and *41 were found to be tolerant to A. marginale; *14 to B. bovis; and *10 and *51 to B. bigemina. These tolerance alleles could be used as potential markers for selection in dairy genetic evaluation. The associated alleles for susceptibility were also found: *2 was found to be susceptible to A. marginale; *3 and *16 to B. bovis; and *20 to B. bigemina. These susceptibility alleles could be used as markers for culling, and selection favoring susceptibility alleles should be considered to maintain heterozygote advantage and pathogen-specific memories in the herd.

  18. Molecular detection and characterization of tick-borne pathogens in dogs and ticks from Nigeria.

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    Joshua Kamani

    Full Text Available BACKGROUND: Only limited information is currently available on the prevalence of vector borne and zoonotic pathogens in dogs and ticks in Nigeria. The aim of this study was to use molecular techniques to detect and characterize vector borne pathogens in dogs and ticks from Nigeria. METHODOLOGY/PRINCIPAL FINDINGS: Blood samples and ticks (Rhipicephalus sanguineus, Rhipicephalus turanicus and Heamaphysalis leachi collected from 181 dogs from Nigeria were molecularly screened for human and animal vector-borne pathogens by PCR and sequencing. DNA of Hepatozoon canis (41.4%, Ehrlichia canis (12.7%, Rickettsia spp. (8.8%, Babesia rossi (6.6%, Anaplasma platys (6.6%, Babesia vogeli (0.6% and Theileria sp. (0.6% was detected in the blood samples. DNA of E. canis (23.7%, H. canis (21.1%, Rickettsia spp. (10.5%, Candidatus Neoehrlichia mikurensis (5.3% and A. platys (1.9% was detected in 258 ticks collected from 42 of the 181 dogs. Co- infections with two pathogens were present in 37% of the dogs examined and one dog was co-infected with 3 pathogens. DNA of Rickettsia conorii israelensis was detected in one dog and Rhipicephalus sanguineus tick. DNA of another human pathogen, Candidatus N. mikurensis was detected in Rhipicephalus sanguineus and Heamaphysalis leachi ticks, and is the first description of Candidatus N. mikurensis in Africa. The Theileria sp. DNA detected in a local dog in this study had 98% sequence identity to Theileria ovis from sheep. CONCLUSIONS/SIGNIFICANCE: The results of this study indicate that human and animal pathogens are abundant in dogs and their ticks in Nigeria and portray the potential high risk of human exposure to infection with these agents.

  19. Serologic screening for 13 infectious agents in roe deer (Capreolus capreolus in Flanders

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    Paul Tavernier

    2015-11-01

    Full Text Available Introduction: In order to investigate the role of roe deer in the maintenance and transmission of infectious animal and human diseases in Flanders, we conducted a serologic screening in 12 hunting areas. Materials and methods: Roe deer sera collected between 2008 and 2013 (n=190 were examined for antibodies against 13 infectious agents, using indirect enzyme-linked immunosorbent assay, virus neutralisation, immunofluorescence, or microagglutination test, depending on the agent. Results and discussion: High numbers of seropositives were found for Anaplasma phagocytophilum (45.8%, Toxoplasma gondii (43.2% and Schmallenberg virus (27.9%, the latter with a distinct temporal distribution pattern following the outbreak in domestic ruminants. Lower antibody prevalence was found for Chlamydia abortus (6.7%, tick-borne encephalitis virus (5.1%, Neospora caninum (4.8%, and Mycobacterium avium subsp paratuberculosis (4.1%. The lowest prevalences were found for Leptospira (1.7%, bovine viral diarrhoea virus 1 (1.3%, and Coxiella burnetii (1.2%. No antibodies were found against Brucella sp., bovine herpesvirus 1, and bluetongue virus. A significant difference in seroprevalence between ages (higher in adults >1 year was found for N. caninum. Four doubtful reacting sera accounted for a significant difference in seroprevalence between sexes for C. abortus (higher in females. Conclusions: Despite the more intensive landscape use in Flanders, the results are consistent with other European studies. Apart from maintaining C. abortus and MAP, roe deer do not seem to play an important role in the epidemiology of the examined zoonotic and domestic animal pathogens. Nevertheless, their meaning as sentinels should not be neglected in the absence of other wild cervid species.

  20. A comparison of nonlinear mixed models and response to selection of tick-infestation on lambs

    Science.gov (United States)

    2017-01-01

    Tick-borne fever (TBF) is stated as one of the main disease challenges in Norwegian sheep farming during the grazing season. TBF is caused by the bacterium Anaplasma phagocytophilum that is transmitted by the tick Ixodes ricinus. A sustainable strategy to control tick-infestation is to breed for genetically robust animals. In order to use selection to genetically improve traits we need reliable estimates of genetic parameters. The standard procedures for estimating variance components assume a Gaussian distribution of the data. However, tick-count data is a discrete variable and, thus, standard procedures using linear models may not be appropriate. Thus, the objectives of this study were twofold: 1) to compare four alternative non-linear models: Poisson, negative binomial, zero-inflated Poisson and zero-inflated negative binomial based on their goodness of fit for quantifying genetic variation, as well as heritability for tick-count and 2) to investigate potential response to selection against tick-count based on truncation selection given the estimated genetic parameters from the best fit model. Our results showed that zero-inflated Poisson was the most parsimonious model for the analysis of tick count data. The resulting estimates of variance components and high heritability (0.32) led us to conclude that genetic determinism is relevant on tick count. A reduction of the breeding values for tick-count by one sire-dam genetic standard deviation on the liability scale will reduce the number of tick counts below an average of 1. An appropriate breeding scheme could control tick-count and, as a consequence, probably reduce TBF in sheep. PMID:28257433

  1. Disaggregating Tropical Disease Prevalence by Climatic and Vegetative Zones within Tropical West Africa.

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    Carl S Beckley

    Full Text Available Tropical infectious disease prevalence is dependent on many socio-cultural determinants. However, rainfall and temperature frequently underlie overall prevalence, particularly for vector-borne diseases. As a result these diseases have increased prevalence in tropical as compared to temperate regions. Specific to tropical Africa, the tendency to incorrectly infer that tropical diseases are uniformly prevalent has been partially overcome with solid epidemiologic data. This finer resolution data is important in multiple contexts, including understanding risk, predictive value in disease diagnosis, and population immunity. We hypothesized that within the context of a tropical climate, vector-borne pathogen prevalence would significantly differ according to zonal differences in rainfall, temperature, relative humidity and vegetation condition. We then determined if these environmental data were predictive of pathogen prevalence. First we determined the prevalence of three major pathogens of cattle, Anaplasma marginale, Babesia bigemina and Theileria spp, in the three vegetation zones where cattle are predominantly raised in Ghana: Guinea savannah, semi-deciduous forest, and coastal savannah. The prevalence of A. marginale was 63%, 26% for Theileria spp and 2% for B. bigemina. A. marginale and Theileria spp. were significantly more prevalent in the coastal savannah as compared to either the Guinea savanna or the semi-deciduous forest, supporting acceptance of the first hypothesis. To test the predictive power of environmental variables, the data over a three year period were considered in best subsets multiple linear regression models predicting prevalence of each pathogen. Corrected Akaike Information Criteria (AICc were assigned to the alternative models to compare their utility. Competitive models for each response were averaged using AICc weights. Rainfall was most predictive of pathogen prevalence, and EVI also contributed to A. marginale and B

  2. Theileriosis in six dogs in South Africa and its potential clinical significance

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    Chantal T. Rosa

    2014-02-01

    Full Text Available Theileriosis is a tick-borne disease caused by a piroplasma of the genus Theileria that can causeanaemia and thrombocytopenia. Its clinical importance for dogs’ remains poorly understood,as only some develop clinical signs. In this study, physical and laboratory findings, treatment and outcomes of six client-owned diseased dogs presented at the Onderstepoort Veterinary Academic Hospital are described retrospectively. In the dogs, Theileria species (n = 4and Theileria equi (n = 2 were detected by a polymerase chain reaction (PCR-reverse blothybridisation assay in blood samples, whilst PCR for Babesia, Anaplasma and Ehrlichia were negative. The most common physical findings were pale mucous membranes (five out of six dogs, bleeding tendencies (five out of six dogs and lethargy (three out of six dogs. All dogs were thrombocytopenic [median 59.5 x 109/L (range 13–199] and five out of six dogs were anaemic [median haematocrit 18% (range 5–32]. Bone marrow core biopsies performed in two dogs showed myelofibrosis. Theileriosis was treated with imidocarb dipropionate and the suspected secondary immune-mediated haematological disorders with prednisolone and azathioprine. Five dogs achieved clinical cure and post-treatment PCR performed in three out of five dogs confirmed absence of circulating parasitaemia. An immune-mediated response to Theileria species is thought to result in anaemia and/or thrombocytopenia in diseased dogs with theileriosis. A bleeding tendency, most likely secondary to thrombocytopenia and/or thrombocytopathy, was the most significant clinical finding in these cases. The link between thrombocytopenia, anaemia and myelofibrosis in theileriosis requires further investigation and theileriosis should be considered a differential diagnosis for dogs presenting with anaemia and/or thrombocytopenia in endemic tick-borne disease areas.

  3. Disaggregating Tropical Disease Prevalence by Climatic and Vegetative Zones within Tropical West Africa

    Science.gov (United States)

    Beckley, Carl S.; Shaban, Salisu; Palmer, Guy H.; Hudak, Andrew T.; Noh, Susan M.; Futse, James E.

    2016-01-01

    Tropical infectious disease prevalence is dependent on many socio-cultural determinants. However, rainfall and temperature frequently underlie overall prevalence, particularly for vector-borne diseases. As a result these diseases have increased prevalence in tropical as compared to temperate regions. Specific to tropical Africa, the tendency to incorrectly infer that tropical diseases are uniformly prevalent has been partially overcome with solid epidemiologic data. This finer resolution data is important in multiple contexts, including understanding risk, predictive value in disease diagnosis, and population immunity. We hypothesized that within the context of a tropical climate, vector-borne pathogen prevalence would significantly differ according to zonal differences in rainfall, temperature, relative humidity and vegetation condition. We then determined if these environmental data were predictive of pathogen prevalence. First we determined the prevalence of three major pathogens of cattle, Anaplasma marginale, Babesia bigemina and Theileria spp, in the three vegetation zones where cattle are predominantly raised in Ghana: Guinea savannah, semi-deciduous forest, and coastal savannah. The prevalence of A. marginale was 63%, 26% for Theileria spp and 2% for B. bigemina. A. marginale and Theileria spp. were significantly more prevalent in the coastal savannah as compared to either the Guinea savanna or the semi-deciduous forest, supporting acceptance of the first hypothesis. To test the predictive power of environmental variables, the data over a three year period were considered in best subsets multiple linear regression models predicting prevalence of each pathogen. Corrected Akaike Information Criteria (AICc) were assigned to the alternative models to compare their utility. Competitive models for each response were averaged using AICc weights. Rainfall was most predictive of pathogen prevalence, and EVI also contributed to A. marginale and B. bigemina prevalence

  4. New criteria for selecting the origin of DNA replication in Wolbachia and closely related bacteria

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    Baldo Laura

    2007-06-01

    Full Text Available Abstract Background The annotated genomes of two closely related strains of the intracellular bacterium Wolbachia pipientis have been reported without the identifications of the putative origin of replication (ori. Identifying the ori of these bacteria and related alpha-Proteobacteria as well as their patterns of sequence evolution will aid studies of cell replication and cell density, as well as the potential genetic manipulation of these widespread intracellular bacteria. Results Using features that have been previously experimentally verified in the alpha-Proteobacterium Caulobacter crescentus, the origin of DNA replication (ori regions were identified in silico for Wolbachia strains and eleven other related bacteria belonging to Ehrlichia, Anaplasma, and Rickettsia genera. These features include DnaA-, CtrA- and IHF-binding sites as well as the flanking genes in C. crescentus. The Wolbachia ori boundary genes were found to be hemE and COG1253 protein (CBS domain protein. Comparisons of the putative ori region among related Wolbachia strains showed higher conservation of bases within binding sites. Conclusion The sequences of the ori regions described here are only similar among closely related bacteria while fundamental characteristics like presence of DnaA and IHF binding sites as well as the boundary genes are more widely conserved. The relative paucity of CtrA binding sites in the ori regions, as well as the absence of key enzymes associated with DNA replication in the respective genomes, suggest that several of these obligate intracellular bacteria may have altered replication mechanisms. Based on these analyses, criteria are set forth for identifying the ori region in genome sequencing projects.

  5. Seasonal activity and tick-borne pathogen infection rates of Ixodes ricinus ticks in Hungary.

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    Egyed, László; Elő, Péter; Sréter-Lancz, Zsuzsanna; Széll, Zoltán; Balogh, Zsuzsanna; Sréter, Tamás

    2012-04-01

    Ixodes ricinus is the most important tick species in Europe as it is most widely distributed and transmits the majority of tick-borne zoonotic pathogens. As limited data are available for Hungary, the aim of the present study was to investigate the seasonal timing of questing by I. ricinus and the infection rate of this tick species with all major tick-borne zoonotic pathogens. Monthly collections of I. ricinus were carried out over 3 consecutive years by dragging a blanket in 6 biotopes representing different areas of Hungary. Altogether, 1800 nymphs (300 per collection point) were screened as pooled samples (each of 5 specimens) by PCR-based methods for tick-borne pathogens. I. ricinus larvae, nymphs, and adults had bimodal activity patterns with a major peak in the spring. As newly moulted ticks of all stages are thought to emerge in the autumn of each year, it appears that most newly emerged ticks delayed their questing until the following spring. The minimum prevalence of Borrelia burgdorferi sensu lato was 2.5%. Borr. afzelii, Borr. burgdorferi sensu stricto, Borr. garinii, Borr. lusitaniae, and Borr. valaisiana were identified by hybridization. The minimum infection rate with spotted fever group rickettsiae was 1.9%. Rickettsia helvetica was identified in all biotopes. The minimum prevalence of Anaplasma phagocytophilum, Babesia divergens and Bab. microti was low (0.3-0.5%). Bartonella spp.-, Francisella tularensis-, and TBE virus-specific amplification products were not detected. Relative to the results of comparable studies carried out in the Carpathian Basin, the prevalence of tick-borne pathogens was low in Hungary. This might be attributed to the climatic difference between the lowland areas of Hungary and submountain areas of the surrounding countries involved in the studies.

  6. Serological surveillance of vector-borne and zoonotic diseases among hunters in eastern Poland

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    Małgorzata Tokarska-Rodak

    2016-01-01

    Full Text Available Background & objectives: Many etiological agents of zoonoses are considered as significant biological hazard to people visiting forested areas frequently, for instance, hunters. They may be exposed to ticks, rodents, and birds as well as excreta/secretions of wild animals or contaminated water and soil. Hence, this population is at risk of contracting infection with pathogens such as Borrelia burgdorferi sensu lato (s.l., Anaplasma phagocytophilum, Babesia spp., tick-borne encephalitis virus, Bartonella spp., Francisella tularensis, Echinococcus spp., or hantaviruses. The aim of the study was to assess the seroprevalence of zoonotic agents, viz. A. phagocytophilum, hantaviruses, and Echinococcus spp., with special regard to B. burgdorferi s.l., among hunters in Lubelskie Voivodeship (eastern Poland. Methods: Serum samples collected from 134 hunters from Lubelskie Voivodeship were analyzed with the use of immunological techniques (enzyme-linked immunosorbent assay, line immunoblot test, and indirect fluorescence assay for the presence of antibodies against the agents. Results: Specific antibodies were detected in 66% of the tested individuals. Antibodies against B. burgdorferi s.l. (39%, A. phagocytophilum (30%, hantaviruses (9%, and Echinococcus spp. (8% were detected individually or as mixed results. Interpretation & conclusion: The results confirm that there is a risk of exposure to different pathogens in the forested areas in eastern Poland and that hunters are highly vulnerable to infection with the examined zoonotic agents. A significant proportion of co-occurring antibodies against different pathogens was noticed. Thus, hunters have to take special care of their health status evaluation and mitigate the exposure risk by using adequate prophylaxis measures.

  7. Life history and demographic drivers of reservoir competence for three tick-borne zoonotic pathogens.

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    Richard S Ostfeld

    Full Text Available Animal and plant species differ dramatically in their quality as hosts for multi-host pathogens, but the causes of this variation are poorly understood. A group of small mammals, including small rodents and shrews, are among the most competent natural reservoirs for three tick-borne zoonotic pathogens, Borrelia burgdorferi, Babesia microti, and Anaplasma phagocytophilum, in eastern North America. For a group of nine commonly-infected mammals spanning >2 orders of magnitude in body mass, we asked whether life history features or surrogates for (unknown encounter rates with ticks, predicted reservoir competence for each pathogen. Life history features associated with a fast pace of life generally were positively correlated with reservoir competence. However, a model comparison approach revealed that host population density, as a proxy for encounter rates between hosts and pathogens, generally received more support than did life history features. The specific life history features and the importance of host population density differed somewhat between the different pathogens. We interpret these results as supporting two alternative but non-exclusive hypotheses for why ecologically widespread, synanthropic species are often the most competent reservoirs for multi-host pathogens. First, multi-host pathogens might adapt to those hosts they are most likely to experience, which are likely to be the most abundant and/or frequently bitten by tick vectors. Second, species with fast life histories might allocate less to certain immune defenses, which could increase their reservoir competence. Results suggest that of the host species that might potentially be exposed, those with comparatively high population densities, small bodies, and fast pace of life will often be keystone reservoirs that should be targeted for surveillance or management.

  8. Seroprevalence of canine dirofilariosis, granulocytic anaplasmosis and lyme borreliosis of public health importance in dogs from India’s North East

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    S. K. Borthakur

    2014-09-01

    Full Text Available Aim: Vector-borne infections namely dirofilariosis, ehrlichiosis, anaplasmosis and lyme borreliosis are being recognized as emerging and/or re-emerging problems in dogs and man due to rapid extension of zoogeographical ranges of many causative agents through international tourism and increase mobility of dogs at national and international level towards meeting the demand for companion animals in the present day society. Anticipating such situation, a serological study was conducted in dogs from North East India to estimate the prevalence of zoonotically important Dirofilaria immitis, Anaplasma phagocytophilum and Borrelia burgdorferi along with Ehrlichia canis. Materials and Methods: Serological study was carried out using enzyme immunoassay in commercial SNAP 4DX® test kit (Idexx Laboratories, USA. The study was conducted in 191 dogs comprising 82 pets, 57 stray and 52 working dogs owned by defence organizations. Results: The study revealed seroprevalence of mosquito-borne D. immitis (17.80%, tick-borne E. canis (22.51% and A. phagocytophilum (4.71% with an overall 41.88% prevalence of pathogens in single or co-infection. Serological evidence of tick-borne lyme borreliosis due to B. burgdorferi could not be established in dogs in the present study. Of the zoonotic species, highest prevalence of D. immitis was found in the stray dogs (22.80% and that of A. phagocytophilum in pet dogs (6.09%. Conclusion: The results of the present serological study serve as baseline information on the prevalence of A. phagocytophilum in dogs reported for the first time in India and reaffirmation on the high prevalence of D. immitis and E. canis in the North East India.

  9. Subacute transverse myelitis with Lyme profile dissociation

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    Ajjan, Mohammed

    2008-06-01

    Full Text Available Introduction: Transverse myelitis is a very rare neurologic syndrome with an incidence per year of 1-5 per million population. We are presenting an interesting case of subacute transverse myelitis with its MRI (magnetic resonance imaging and CSF (cerebrospinal fluid findings. Case: A 46-year-old African-American woman presented with decreased sensation in the lower extremities which started three weeks ago when she had a 36-hour episode of sore throat. She reported numbness up to the level just below the breasts. Lyme disease antibodies total IgG (immunoglobulin G and IgM (immunoglobulin M in the blood was positive. Antinuclear antibody profile was within normal limits. MRI of the cervical spine showed swelling in the lower cervical cord with contrast enhancement. Cerebrospinal fluid was clear with negative Borrelia Burgdorferi IgG and IgM. Herpes simplex, mycoplasma, coxiella, anaplasma, cryptococcus and hepatitis B were all negative. No oligoclonal bands were detected. Quick improvement ensued after she was given IV Ceftriaxone for 7 days. The patient was discharged on the 8th day in stable condition. She continued on doxycycline for 21 days. Conclusions: Transverse myelitis should be included in the differential diagnosis of any patient presenting with acute or subacute myelopathy in association with localized contrast enhancement in the spinal cord especially if flu-like prodromal symptoms were reported. Lyme disease serology is indicated in patients with neurological symptoms keeping in mind that dissociation in Lyme antibody titers between the blood and the CSF is possible.

  10. 16S rRNA gene clone library analysis of bacterial communities of the tick with infection of 4 species of pathogens%4种病原菌特异基因片段阳性蜱的16S rRNA基因克隆文库分析

    Institute of Scientific and Technical Information of China (English)

    张守印; 俞东征; 孙继民; 贺金荣; 付秀萍; 张景山; 张建华; 蔡虹; 马凤琴; 海荣

    2009-01-01

    Objective To develop the method of 16S rRNA gene clone library for tick bacterial flora analysis, and to analyze the detection effective of pathogens in tick and capacity of bacterial flora diversity. Methods Primers were designed according to the specific gene of Borrelia burgdorferi, Bartonella henselae, Anaplasma phagocytophilum, Ehrlichia chaffeensis and templates were choosen by positive PCR result to amplify the DNA extracted from the ticks. One set of primers targeting 16S rRNA gene conserved region were chosen to amplify certain fragments, DNA extraction, PCR reaction, cloning and sequencing. Nucleotide sequences were compared with GenBank database. Calculated Coverage values of clone library and Shannon-Wiener diversity index. Results Sixteen defined genus-or species-bacteria were detected in 103 valid sequences. Eight species were edge type (Clone No. > 5). Three kinds of pathogens were identified (Borrelia burgdorferi, Bartonella henselae and Rickettsia sp). Three kinds of pathogens were not edge type(Clone No. 5个);检测到伯氏疏螺旋体、汉赛巴通体和立克次体3种病原菌,但这3种病原菌均不是优势类型(克隆子数均<5个).Coverage值为96.11%,Shannon-Wiener多样性指数为2.40.克隆序列分析结果表明,蜱寄生细菌主要为α、γ变形菌纲,占56.25%(9/16).结论 16S rRNA基因序列分析可以对蜱标本进行菌群相对定量研究,可以同时检出多种病原菌,是一种较好的细菌菌群多样性分析和病原菌筛检方法.

  11. Passive Surveillance of Ixodes scapularis (Say), Their Biting Activity, and Associated Pathogens in Massachusetts

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    Xu, Guang; Mather, Thomas N.; Hollingsworth, Craig S.

    2016-01-01

    Abstract A passive surveillance of tick-borne pathogens was conducted over a 7-year period (2006–2012), in which a total of 3551 ticks were submitted to the University of Massachusetts for PCR testing. The vast majority of these ticks were Ixodes scapularis from Massachusetts (N = 2088) and hence were the focus of further analysis. Two TaqMan duplex qPCR assays were developed to test I. scapularis ticks for the presence of three human pathogens: Borrelia burgdorferi, Anaplasma phagocytophilum, and Babesia microti. I. scapularis submissions were concentrated from Cape Cod, the eastern half of the state outside of the Boston metropolitan area, parts of Franklin and Hampshire counties along the Quabbin Reservoir watershed, and southwestern Berkshire county. Differences in seasonal activity pattern were observed for different developmental stages of I. scapularis. The largest proportion of tick bite victims were age 9 years and under. Nymphal ticks were found more often on lower extremities of their hosts, while more adult ticks were found on the head. Overall infection rate of B. burgdorferi, A. phagocytophilum, and B. microti in human-biting ticks was 29.6%, 4.6%, and 1.8%, respectively. B. burgdorferi-infected ticks were widely distributed, but A. phagocytophilum- and B. microti-infected I. scapularis were found mainly in the eastern half of the state. We found that 1.8%, 1.0%, and 0.4% of ticks were coinfected by B. burgdorferi and A. phagocytophilum, B. burgdorferi and B. microti, and A. phagocytophilum and B. microti, respectively, and 0.3% of ticks had triple coinfection. PMID:27248292

  12. Harvested white-tailed deer as sentinel hosts for early establishing Ixodes scapularis populations and risk from vector-borne zoonoses in southeastern Canada.

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    Bouchard, C; Leighton, P A; Beauchamp, G; Nguon, S; Trudel, L; Milord, F; Lindsay, L R; Bélanger, D; Ogden, N H

    2013-03-01

    Due to recent establishment of the blacklegged tick, Ixodes scapularis Say, in southeastern Canada, tick-borne zoonoses (Lyme disease, human granulocytotropic anaplasmosis, and babesiosis) are of growing concern for public health. Using white-tailed deer (Odocoileus virginianus) culled in southwestern Quebec during 2007-2008, we investigated whether hunter-killed deer could act as sentinels for early establishing tick populations and for tick-borne pathogens. Accounting for environmental characteristics of culling sites, and age and sex of deer, we investigated whether their tick infestation levels could identify locations of known tick populations detected in active surveillance, presumed tick populations detected by passive surveillance, or both. We also used spatial cluster analyses to identify spatial patterns of tick infestation and occurrence of tick-borne zoonoses infection in ticks collected from the deer. Adult ticks were found on 15% of the 583 deer examined. Adult male deer had the greatest number (approximately 90%) of adult ticks. Overall, 3, 15, and 0% of the ticks collected were polymerase chain reaction (PCR)-positive for Borrelia burgdorferi, Anaplasma phagocytophilum, and Babesia microti, respectively. Our statistical analyses suggest that sex and age of deer, temperature, precipitation, and an index of tick dispersion by migratory birds were significantly associated with tick infestation levels. Cluster analysis identified significant clusters of deer carrying ticks PCR-positive for A. phagocytophilum, and for deer carrying two or more I. scapularis. Our study suggests that hunter-killed deer may be effective as sentinels for emerging areas of tick-borne anaplasmosis. They may have limited use as sentinels for early emerging I. scapularis tick populations and emerging Lyme disease risk.

  13. Ixodes ricinus and Its Endosymbiont Midichloria mitochondrii: A Comparative Proteomic Analysis of Salivary Glands and Ovaries.

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    Monica Di Venere

    Full Text Available Hard ticks are hematophagous arthropods that act as vectors of numerous pathogenic microorganisms of high relevance in human and veterinary medicine. Ixodes ricinus is one of the most important tick species in Europe, due to its role of vector of pathogenic bacteria such as Borrelia burgdorferi and Anaplasma phagocytophilum, of viruses such as tick borne encephalitis virus and of protozoans as Babesia spp. In addition to these pathogens, I. ricinus harbors a symbiotic bacterium, Midichloria mitochondrii. This is the dominant bacteria associated to I. ricinus, but its biological role is not yet understood. Most M. mitochondrii symbionts are localized in the tick ovaries, and they are transmitted to the progeny. M. mitochondrii bacteria have however also been detected in the salivary glands and saliva of I. ricinus, as well as in the blood of vertebrate hosts of the tick, prompting the hypothesis of an infectious role of this bacterium. To investigate, from a proteomic point of view, the tick I. ricinus and its symbiont, we generated the protein profile of the ovary tissue (OT and of salivary glands (SG of adult females of this tick species. To compare the OT and SG profiles, 2-DE profiling followed by LC-MS/MS protein identification were performed. We detected 21 spots showing significant differences in the relative abundance between the OT and SG, ten of which showed 4- to 18-fold increase/decrease in density. This work allowed to establish a method to characterize the proteome of I. ricinus, and to detect multiple proteins that exhibit a differential expression profile in OT and SG. Additionally, we were able to use an immunoproteomic approach to detect a protein from the symbiont. Finally, the method here developed will pave the way for future studies on the proteomics of I. ricinus, with the goals of better understanding the biology of this vector and of its symbiont M. mitochondrii.

  14. Detection of A. phagocytophilum and E. chaffeensis in patient and mouse blood and ticks by a duplex real-time PCR assay.

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    Dong, Tuo; Qu, Zhangyi; Zhang, Lijuan

    2013-01-01

    Human granulocytic anaplasmosis (HGA) and human monocytic ehrlichiosis (HME) are emerging, tick-borne, zoonotic infectious diseases caused by Anaplasma phagocytophilum and Ehrlichia chaffeensis, respectively. Early diagnosis is essential for rapid clinical treatment to avoid misdiagnosis and severe patient outcomes. Simple, sensitive and reliable diagnostic methods are urgently needed. In this study, we developed a duplex real-time PCR assay targeting the A. phagocytophilum ankA gene and the E. chaffeensis TRP120 gene, respectively. The lowest limit of detection of the duplex real-time PCR assay was 100 copies of the targeted A. phagocytophilum ankA gene and the E. chaffeensis TRP120 gene per reaction, and the specificity was 100%. Detection in blood DNA samples from the acute stage of illness for 22 HGA cases and 8 HME cases indicated that the duplex real-time PCR assay was more sensitive than the nested PCR assay. The infection of Citellusundulatus Pallas with A. phagocytophilum and E. chaffeensis was first confirmed in Xinjiang Province and the positive rate was 3.1% for A. phagocytophilum, 6.3% for E. chaffeensis and 3.1% for co-infection with both pathogens. The rates of A. phagocytophilum and E. chaffeensis infection of D. silvarum ticks collected from Shanxi Province were 8.2% and 14.8%, respectively, and the co-infection rate was 3.3%. The rates of A. phagocytophilum and E. chaffeensis infection in H. longicornis ticks collected from Shandong Province were 1.6% and 6.3%, respectively, and the co-infection rate was 1.6%.

  15. Directory of Open Access Journals (Sweden)

    Fredrick M. Kivaria

    2012-09-01

    Full Text Available A cross-sectional study was conducted between September and October 2010 in five states of South Sudan that were selected on the basis of the perceived risk of tick-borne diseases. The purpose was to investigate epidemiological parameters of tick-borne diseases in South Sudan and their uses in future control strategies. A total of 805 calves were assessed by clinical, microscopic and serological examination and tick counts. The indirect Enzyme-Linked Immuno-Sorbent Assay (ELISA was used to detect antibodies to Theileria parva, Theileria mutans, Anaplasma marginale and Babesian bigemina. Sero-conversion risks for T. parva and T. mutans were 27.3% and 31.3% respectively, whilst the risk was 57.6% and 52.8% for A. marginale and B. bigemina, respectively. Major tick species identified include Rhipicephalus appendiculatus, Rhipicephalus decoloratus, Rhipicephalus microplus, Amblyomma variegatum, and Rhipicephalus evertsi. There was great variation (P ≤ 0.001 in the number of all these ticks, both between herds in a state and between calves in an individual herd. The low and intermediate sero-conversion risks observed in the study states suggest that immunisation against East Coast fever (ECF is justified. Fortunately, three major genotypes that were identified by applying Polymerase Chain Reaction Restriction Fragment Length Polymorphism (PCRRFLP analysis on the p104 to the blood samples and T. parva Muguga, matched very well with T. parva Kiambu 5 and T. parva Muguga; therefore the Muguga cocktail can be used for the immunisation of cattle in South Sudan. However, prospective studies are required to develop optimal control measures for tick-borne diseases under different ecological and husbandry practices in South Sudan.

  16. Dog overpopulation and burden of exposure to canine distemper virus and other pathogens on Santa Cruz Island, Galapagos.

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    Diaz, Nicole M; Mendez, Gabriella S; Grijalva, C Jaime; Walden, Heather S; Cruz, Marilyn; Aragon, Eduardo; Hernandez, Jorge A

    2016-01-01

    Dog overpopulation and diseases are hazards to native island species and humans on the Galapagos. Vaccination and importation of dogs are prohibited on the Galapagos. Risk management of these hazards requires the use of science-based risk assessment and risk communication. The objectives of the study reported here were (i) to estimate the human:dog ratio and (ii) the prevalence of and identify exposure factors associated with positive antibody titers to canine distemper virus (CDV) and other pathogens, as well as infection with intestinal parasites in owned dogs on Santa Cruz Island, Galapagos in September 2014. The observed human:dog ratio was 6.148:1 which extrapolates to 2503 dogs (two times more than a recent dog count conducted by Galapagos Biosecurity Agency in March 2014). The proportion of spayed female dogs (50%) was higher, compared to neutered male dogs (30%) (p=0.04). Prevalence of dogs with positive antibody titers to CDV was 36% (95% CI=26, 46%), to canine parvovirus was 89% (95% CI=82, 95%), and to canine adenovirus was 40% (95% CI=30, 51%). The frequency of seropositive dogs to CDV was lower in urban dogs (26%), compared to rural dogs (53%) (p<0.05). A positive interaction effect between rural residence and spay/neuter status on seropositivity to CDV was observed, which we discuss in this report. Because vaccination is prohibited, the dog population on Santa Cruz is susceptible to an outbreak of CDV (particularly among urban dogs) with potential spill over to marine mammals. Dog's age (1-2 or 3-14 years old, compared to younger dogs), and residence (rural, urban) were associated with positive antibody titers to parvovirus, adenovirus, Ehrlichia spp., or Anaplasma spp., as well as infection with Ancylostoma spp., an intestinal parasite in dogs that can be transmitted to humans, particularly children. These results provide the most comprehensive assessment of dog overpopulation and exposure to CDV and other pathogens on the Galapagos to date.

  17. Ehrlichiae and ehrlichial diseases in china.

    Science.gov (United States)

    Wen, Bohai; Cao, Wuchun; Pan, Hua

    2003-06-01

    The various ticks collected from different areas of China were examined for the existence of ehrlichial agents by polymerase chain reaction (PCR) with genus- or species-specific primers designed on the basis of ehrlichial 16S rRNA genes and sequence analyses. In southern China, E. chaffeensis was detected in Amblyomma testudinarium ticks from infested cattle, Haemaphysalis yeni ticks from hare, and Ixodes ovatus ticks from Muntiacus reevesi. E. canis was identified in Rhipicephalus sanguineus ticks from dogs and Boophilus microplus ticks from goats. A new species of the genus Ehrlichia, closely related to E. chaffeensis, and Anaplasma marginale were found in B. microplus ticks from cattle in Tibet. In northern China, E. chaffeensis was detected in Dermacentor silvarum and I. persulcatus ticks; the granulocytic ehrlichial agents were detected in I. persulcatus ticks from an area where Lyme disease is endemic. Canine ehrlichiosis was found in southern China and E. canis and E. platys were identified in dogs; human ehrlichioses were demonstrated by amplifying the 16S rRNA genes of E. chaffeensis and granulocytic ehrlichial agents from patients' blood specimens. In comparison of 16S rRNA gene sequences, the sequences of E. chaffeensis, E. canis, and E. platys in China were found to be different from that in other countries at certain nucleotide positions. These results reveal that a variety of tick-borne ehrlichial agents and diseases exist in China, and the ehrlichial agents and their tick-vectors are same as or different from that in other countries at species or strain levels.

  18. The differential interaction of Brucella and ochrobactrum with innate immunity reveals traits related to the evolution of stealthy pathogens.

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    Elías Barquero-Calvo

    Full Text Available BACKGROUND: During evolution, innate immunity has been tuned to recognize pathogen-associated molecular patterns. However, some alpha-Proteobacteria are stealthy intracellular pathogens not readily detected by this system. Brucella members follow this strategy and are highly virulent, but other Brucellaceae like Ochrobactrum are rhizosphere inhabitants and only opportunistic pathogens. To gain insight into the emergence of the stealthy strategy, we compared these two phylogenetically close but biologically divergent bacteria. METHODOLOGY/PRINCIPAL FINDINGS: In contrast to Brucella abortus, Ochrobactrum anthropi did not replicate within professional and non-professional phagocytes and, whereas neutrophils had a limited action on B. abortus, they were essential to control O. anthropi infections. O. anthropi triggered proinflammatory responses markedly lower than Salmonella enterica but higher than B. abortus. In macrophages and dendritic cells, the corresponding lipopolysaccharides reproduced these grades of activation, and binding of O. anthropi lipopolysaccharide to the TLR4 co-receptor MD-2 and NF-kappaB induction laid between those of B. abortus and enteric bacteria lipopolysaccharides. These differences correlate with reported variations in lipopolysaccharide core sugars, sensitivity to bactericidal peptides and outer membrane permeability. CONCLUSIONS/SIGNIFICANCE: The results suggest that Brucellaceae ancestors carried molecules not readily recognized by innate immunity, so that non-drastic variations led to the emergence of stealthy intracellular parasites. They also suggest that some critical envelope properties, like selective permeability, are profoundly altered upon modification of pathogen-associated molecular patterns, and that this represents a further adaptation to the host. It is proposed that this adaptive trend is relevant in other intracellular alpha-Proteobacteria like Bartonella, Rickettsia, Anaplasma, Ehrlichia and Wolbachia.

  19. Fatal pyogranulomatous myocarditis in 10 Boxer puppies.

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    Detmer, Susan E; Bouljihad, Mostafa; Hayden, David W; Schefers, Jeremy M; Armien, Anibal; Wünschmann, Arno

    2016-03-01

    Over a period of 5 years, 10 pure-bred Boxer puppies, 9-16 weeks old, were presented with a history of sudden death and were diagnosed with pyogranulomatous myocarditis. The myocarditis was characterized by a mixed infiltrate composed predominantly of neutrophils and macrophages. In our retrospective study, original case records and archived materials were examined. All dogs were positive for Borrelia burgdorferi on immunohistochemistry (IHC). There was no evidence of infectious agents in formalin-fixed, paraffin-embedded (FFPE) heart tissue sections stained with hematoxylin and eosin, Ziehl-Neelsen, Gram, Grocott methenamine silver, Warthin-Starry, Von Kossa, and Steiner-Chapman stains. IHC for Chlamydia sp., Toxoplasma gondii, Neospora caninum, West Nile virus, and canine parvovirus also yielded a negative result in all dogs. Polymerase chain reaction testing for vector-borne pathogens on heart tissue from 9 of the dogs (1 frozen and 8 FFPE samples) yielded positive results for 1 dog with B. burgdorferi as well as Anaplasma phagocytophilum in another dog. Subsequently, 2 additional cases were found in a French Bulldog and a French Bulldog-Beagle mix that had identical morphology, test results, age, and seasonality to these 10 Boxer dogs. The similarities in the seasonality, signalment of the affected dogs, and the gross and microscopic lesions suggest a common etiology. Positive IHC and morphologic similarities to human Lyme carditis indicate that B. burgdorferi is likely the agent involved. An additional consideration for these cases is the possibility of a breed-specific autoimmune myocarditis or potential predisposition for cardiopathogenic agents in young Boxers.

  20. To Be or Not to Be Associated: Power study of four statistical modeling approaches to identify parasite associations in cross-sectional studies

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    Elise eVaumourin

    2014-05-01

    Full Text Available A growing number of studies are reporting simultaneous infections by parasites in many different hosts. The detection of whether these parasites are significantly associated is important in medicine and epidemiology. Numerous approaches to detect associations are available, but only a few provide statistical tests. Furthermore, they generally test for an overall detection of association and do not identify which parasite is associated with which other one. Here, we developed a new approach, the association screening approach, to detect the overall and the detail of multi-parasite associations. We studied the power of this new approach and of three other known ones (i.e. the generalized chi-square, the network and the multinomial GLM approaches to identify parasite associations either due to parasite interactions or to confounding factors. We applied these four approaches to detect associations within two populations of multi-infected hosts: 1 rodents infected with Bartonella sp., Babesia microti and Anaplasma phagocytophilum and 2 bovine population infected with Theileria sp. and Babesia sp.. We found that the best power is obtained with the screening model and the generalized chi-square test. The differentiation between associations, which are due to confounding factors and parasite interactions was not possible. The screening approach significantly identified associations between Bartonella doshiae and B. microti, and between T. parva, T. mutans and T. velifera. Thus, the screening approach was relevant to test the overall presence of parasite associations and identify the parasite combinations that are significantly over- or under-represented. Unravelling whether the associations are due to real biological interactions or confounding factors should be further investigated. Nevertheless, in the age of genomics and the advent of new technologies, it is a considerable asset to speed up researches focusing on the mechanisms driving interactions

  1. Understanding Anaplasmataceae pathogenesis using ‘Omics’ approaches

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    Ludovic ePruneau

    2014-07-01

    Full Text Available This paper examines how Omics approaches improve our understanding of Anaplasmataceae pathogenesis, through a global and integrative strategy to identify genes and proteins involved in biochemical pathways key for pathogen-host-vector interactions.The Anaplasmataceae family comprises obligate intracellular bacteria mainly transmitted by arthropods. These bacteria are responsible for major human and animal endemic and emerging infectious diseases with important economic and public health impacts. In order to improve disease control strategies, it is essential to better understand their pathogenesis. Our work focused on four Anaplasmataceae, which cause important animal, human and zoonotic diseases: Anaplasma marginale, A. phagocytophilum, Ehrlichia chaffeensis and E. ruminantium. Wolbachia spp. an endosymbiont of arthropods was also included in this review as a model of a non-pathogenic Anaplasmataceae.A gap analysis on Omics approaches on Anaplasmataceae was performed, which highlighted a lack of studies on the genes and proteins involved in the infection of hosts and vectors. Furthermore, most of the studies have been done on the pathogen itself, mainly on infectious free-living forms and rarely on intracellular forms. In order to perform a transcriptomic analysis of the intracellular stage of development, researchers developed methods to enrich bacterial transcripts from infected cells. These methods are described in this paper. Bacterial genes encoding outer membrane proteins, post-translational modifications, eukaryotic repeated motif proteins, proteins involved in osmotic and oxidative stress and hypothetical proteins have been identified to play a key role in Anaplasmataceae pathogenesis. Further investigations on the function of these outer membrane proteins and hypothetical proteins will be essential to confirm their role in the pathogenesis. Our work underlines the need for further studies in this domain and on host and vector responses

  2. Identification of 24h Ixodes scapularis immunogenic tick saliva proteins.

    Science.gov (United States)

    Lewis, Lauren A; Radulović, Željko M; Kim, Tae K; Porter, Lindsay M; Mulenga, Albert

    2015-04-01

    Ixodes scapularis is arguably the most medically important tick species in the United States. This tick transmits 5 of the 14 human tick-borne disease (TBD) agents in the USA: Borrelia burgdorferi, Anaplasma phagocytophilum, B. miyamotoi, Babesia microti, and Powassan virus disease. Except for the Powassan virus disease, I. scapularis-vectored TBD agents require more than 24h post attachment to be transmitted. This study describes identification of 24h immunogenic I. scapularis tick saliva proteins, which could provide opportunities to develop strategies to stop tick feeding before transmission of the majority of pathogens. A 24h fed female I. scapularis phage display cDNA expression library was biopanned using rabbit antibodies to 24h fed I. scapularis female tick saliva proteins, subjected to next generation sequencing, de novo assembly, and bioinformatic analyses. A total of 182 contigs were assembled, of which ∼19% (35/182) are novel and did not show identity to any known proteins in GenBank. The remaining ∼81% (147/182) of contigs were provisionally identified based on matches in GenBank including ∼18% (27/147) that matched protein sequences previously annotated as hypothetical and putative tick saliva proteins. Others include proteases and protease inhibitors (∼3%, 5/147), transporters and/or ligand binding proteins (∼6%, 9/147), immunogenic tick saliva housekeeping enzyme-like (17%, 25/147), ribosomal protein-like (∼31%, 46/147), and those classified as miscellaneous (∼24%, 35/147). Notable among the miscellaneous class include antimicrobial peptides (microplusin and ricinusin), myosin-like proteins that have been previously found in tick saliva, and heat shock tick saliva protein. Data in this study provides the foundation for in-depth analysis of I. scapularis feeding during the first 24h, before the majority of TBD agents can be transmitted.

  3. PCR diagnosis of tick-borne pathogens in Maharashtra state, India indicates fitness cost associated with carrier infections is greater for crossbreed than native cattle breeds

    Science.gov (United States)

    Kolte, Sunil W.; Larcombe, Stephen D.; Jadhao, Suresh G.; Magar, Swapnil P.; Warthi, Ganesh; Kurkure, Nitin V.; Glass, Elizabeth J.; Shiels, Brian R.

    2017-01-01

    Tick-borne pathogens (TBP) are responsible for significant economic losses to cattle production, globally. This is particularly true in countries like India where TBP constrain rearing of high yielding Bos taurus, as they show susceptibility to acute tick borne disease (TBD), most notably tropical theileriosis caused by Theileria annulata. This has led to a programme of cross breeding Bos taurus (Holstein-Friesian or Jersey) with native Bos indicus (numerous) breeds to generate cattle that are more resistant to disease. However, the cost to fitness of subclinical carrier infection in crossbreeds relative to native breeds is unknown, but could represent a significant hidden economic cost. In this study, a total of 1052 bovine blood samples, together with associated data on host type, sex and body score, were collected from apparently healthy animals in four different agro-climatic zones of Maharashtra state. Samples were screened by PCR for detection of five major TBPs: T. annulata, T. orientalis, B. bigemina, B. bovis and Anaplasma spp.. The results demonstrated that single and co-infection with TBP are common, and although differences in pathogen spp. prevalence across the climatic zones were detected, simplistic regression models predicted that host type, sex and location are all likely to impact on prevalence of TBP. In order to remove issues with autocorrelation between variables, a subset of the dataset was modelled to assess any impact of TBP infection on body score of crossbreed versus native breed cattle (breed type). The model showed significant association between infection with TBP (particularly apicomplexan parasites) and poorer body condition for crossbreed animals. These findings indicate potential cost of TBP carrier infection on crossbreed productivity. Thus, there is a case for development of strategies for targeted breeding to combine productivity traits with disease resistance, or to prevent transmission of TBP in India for economic benefit. PMID

  4. Emergence of oriental theileriosis in cattle and its transmission through Rhipicephalus (Boophilus microplus in Assam, India

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    Parikshit Kakati

    2015-09-01

    Full Text Available Aim: The aim of the present study was to investigate the presence of Theileria in blood samples of crossbred and indigenous adult cows raised under unorganized small scale farming system in a Babesia and Anaplasma endemic geographical area from Assam, India and to see its transmission through Rhipicephalus (Boophilus microplus ticks. Materials and Methods: For the present study, 57 clinical cases of cattle suspected to be of hemoparasitic infections were taken into consideration. The parasites were identified based on morphology in giemsa stained blood smear followed by polymerase chain reaction (PCR. Sera samples were tested for T. annulata antibodies in plate and Dot-ELISA. PCR was also conducted in eggs of Rhipicephalus (Boophilus microplus tick collected from a Theileria orientalis positive animal. Results: PCR amplified 1124, 776, and 160 bp DNA fragments of B. bigemina (64.91%,T. orientalis(21.05% and A. marginale (14.03%, respectively. This assay further conducted in 12 T. orientalis positive blood samples with primers of Buffeli, Chitose, and Ikeda variants of T. orientalis showed 3 samples positive to Ikeda type and none for Buffeli and Chitose. Babesia bovis and Theileria annulata specific primers also did not amplify any fragment during the PCR assay of the blood samples. Further, all sera samples tested negative to T. annulata antibodies in Plate and Dot-ELISA. PCR conducted in eggs of R (B.microplus tick collected from a T. orientalis positive animal revealed presence of the parasite DNA. Gradual improvement in physical condition leading to complete recovery in 10 out of 12 T. orientalis infected clinical cases treated with buparvaquone(at 2.5mg/kg.b.wt I/M was the feedback obtained from field veterinarians and the cattle owners. Conclusion: The present investigation represents the first report of occurrence of T. orientalis in cattle of Assam with involvement of pathogenic Ikeda strain in clinical outbreaks and its possible natural

  5. Population-based passive tick surveillance and detection of expanding foci of blacklegged ticks Ixodes scapularis and the Lyme disease agent Borrelia burgdorferi in Ontario, Canada.

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    Nelder, Mark P; Russell, Curtis; Lindsay, L Robbin; Dhar, Badal; Patel, Samir N; Johnson, Steven; Moore, Stephen; Kristjanson, Erik; Li, Ye; Ralevski, Filip

    2014-01-01

    We identified ticks submitted by the public from 2008 through 2012 in Ontario, Canada, and tested blacklegged ticks Ixodes scapularis for Borrelia burgdorferi and Anaplasma phagocytophilum. Among the 18 species of ticks identified, I. scapularis, Dermacentor variabilis, Ixodes cookei and Amblyomma americanum represented 98.1% of the 14,369 ticks submitted. Rates of blacklegged tick submission per 100,000 population were highest in Ontario's Eastern region; D. variabilis in Central West and Eastern regions; I. cookei in Eastern and South West regions; and A. americanum had a scattered distribution. Rates of blacklegged tick submission per 100,000 population were highest from children (0-9 years old) and older adults (55-74 years old). In two health units in the Eastern region (i.e., Leeds, Grenville & Lanark District and Kingston-Frontenac and Lennox & Addington), the rate of submission for engorged and B. burgdorferi-positive blacklegged ticks was 47× higher than the rest of Ontario. Rate of spread for blacklegged ticks was relatively faster and across a larger geographic area along the northern shore of Lake Ontario/St. Lawrence River, compared with slower spread from isolated populations along the northern shore of Lake Erie. The infection prevalence of B. burgdorferi in blacklegged ticks increased in Ontario over the study period from 8.4% in 2008 to 19.1% in 2012. The prevalence of B. burgdorferi-positive blacklegged ticks increased yearly during the surveillance period and, while increases were not uniform across all regions, increases were greatest in the Central West region, followed by Eastern and South West regions. The overall infection prevalence of A. phagocytophilum in blacklegged ticks was 0.3%. This study provides essential information on ticks of medical importance in Ontario, and identifies demographic and geographic areas for focused public education on the prevention of tick bites and tick-borne diseases.

  6. Anemia and mechanism of erythrocyte destruction in ducks with acute Leucocytozoon infections

    Science.gov (United States)

    Kocan, R.M.

    1968-01-01

    In the anemia which accompanies infection by Leucocytozoon simondi in Pekin ducks there was a far greater loss of erythrocytes than could be accounted for as a result of direct physical rupture by the parasite. Erythrocyte loss began at the same time the 1st parasites appeared in the blood and was severest just prior to maximum parasitemia. Blood replacement and parasite loss occurred simultaneously. Examination of the spleen and bone marrow revealed that erythrophagocytosis was not the cause of anemia as reported for infections of Plasmodium, Babesia and Anaplasma. An anti-erythrocyte (A-E) factor was found in the serum of acutely infected ducks which agglutinated and hemolyzed normal untreated duck erythrocytes as well as infected cells. This A-E factor appeared when the 1st red cell loss was detected and reached its maximum titer just prior to the greatest red cell loss. Titers of the A-E factor were determined using normal uninfected erythrocytes at temperatures between 4 and 42 C. Cells agglutinated below 25 C and hemolyzed at 37 and 42 C. These results indicated that the A-E factor could be responsible for loss of cells other than those which were infected and could thus produce an excess loss of red cells. Attempts to implicate the A-E factor as an autoantibody were all negative. The A-E factor was present in the gamma fraction of acute serum but no anamnestic response could be detected when recovered ducks were reinfected. Anemia was never as severe in reinfections as in primary infections. The A-E factor also never reached as high a titer and was removed from the circulation very rapidly in reinfected ducks. It is concluded that red cell loss in ducks with acute Leucocytozoon disease results from intravascular hemolysis rather than erythrophagocytosis. The A-E factor responsible for hemolysis is more likely a parasite product rather than autoantibody.

  7. 浙江省鼠形动物蜱源立克次体感染状况调查%A Cross-sectional study on tick-borne Rickettsiae infections among murine-like animals

    Institute of Scientific and Technical Information of China (English)

    柴程良; 陈恩富; 孙继民; 陆群英; 凌锋; 姜理平; 葛君华; 顾时平; 叶晓东; 刘社兰

    2014-01-01

    目的:了解浙江省不同类型地理环境中鼠形动物蜱源立克次体的感染状况。方法2009-2011年在安吉县、金东区和天台县用夹夜法捕获鼠形动物,进行分类鉴定、无菌采集肝脾标本,并采用巢式PCR方法检测无形体属与埃立克体属16SrRNA 以及立克次体属(包括斑疹伤寒群和斑点热群)与东方体属热休克蛋白基因groEL。结果3个调查点共捕获鼠形动物14种851头,社鼠(30.32%)、黑线姬鼠(18.80%)和青毛鼠(11.75%)为主要优势种,其中金东区、安吉县和天台县的优势鼠种分别为社鼠(39.20%)、青毛鼠(32.05%)和黑线姬鼠(59.57%)。562份肝脾标本中检出立克次体48份,阳性检出率为8.54%;其中无形体属占3.38%,斑疹伤寒群占1.78%,恙虫病东方体属占1.78%,埃立克体属占1.07%,斑点热群占0.53%。金东区和安吉县无形体属检出率较高为4.76%和4.27%,斑点热群仅在天台县检出。社鼠的立克次体阳性检出率最高为14.97%。同一鼠形动物可存在多种立克次体混合感染。结论浙江省不同类型地理环境鼠形动物中广泛存在立克次体感染,不同地理位?和不同鼠形动物的蜱源立克次体检出率不同。%Objective Toinvestigatethestatusoftick-borneRickettsiaeinfectionsamongmurine-likeanimalsin differentareasofZhejiangprovince.Methods Liverandspleensamplesofmurine-likeanimalscapturedthroughnight trapping method were collected from Anji,Jinhua and Tiantai County according to their geographic locations and historical detection of Rickettsiae .Nest-PCR tests were used to determine the presence of the 16S rRNA genes of Anaplasma and Ehrlichia ,and the heat shock protein genes (groEL)of Rickettsiae (including typhus and spotted fever group)and Orientiainthesesamples.Results Atotalof851murine-likeanimalsbelongingto14specieswerecaptured.The predominant species were Rattus confucianus (30

  8. Prevalence of haemoparasites in pet, working and stray dogs of Assam and North-East India: A hospital based study

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    K. Bhattacharjee

    2013-10-01

    Full Text Available Aim:This research work is aimed to find out the prevalence of haemoparasitic infections in different categories of dogs.Materials and Methods: Out of 2104 dogs registered in the Teaching Veterinary Clinical Complex of the College ofVeterinary Science, Khanapara, Guwahati during January 2009 to December 2010, blood of 424 cases suspected forhaemoparasites on the basis of clinical history were microscopically examined in wet blood film and giemsa stained bloodsmears.Results: The prevalence was 57.31% in the hospital population comprising pet (58.03% and working (54.54% dogs and63.64% in stray dog population. A total of 7 species viz. Babesia gibsoni (47.16%, Ehrlichia (Anaplasma platys (8.49%,Dirofilaria immitis (2.83%, Ehrlichia canis (2.12%, Babesia canis (1.41%, Hepatozoon canis (1.41% and Ehrlichiaewingii (0.47% in single or mixed infections were recorded. B. gibsoni was found to be most predominant haemoprotozoanspecies. However, B. canis, the large form of Babesia was detected in very few dogs. Similarly, inclusion of E. platys insideblood platelets, although rare in occurrence, was more easily detectable than that of E. canis inside the monocytes andlymphocytes. Infection with D. immitis in pet dogs (2.38% was comparatively lower than in working dogs (4.54% and foundin single and mixed infections with B. gibsoni, B. canis and E. platys. Hepatozoon canis was recorded in 6 hospital dogs eitherin single or mixed infection with B. gibsoni and E. platys. Six species of haemoparasites namely B. gibsoni (47.72%, D.immitis (27.27%, E. platys (4.54%, E. canis (2.27%, E. ewingii (2.27% and H. canis (2.27% were detected in blood ofstray dogs. Trypanosoma evansi as recorded in dogs from other parts of the country was not detected in this study.Conclusion: Present findings led to a significant conclusion that Assam and adjoining states of North East region of India arehighly enzootic for the vector borne haemoparasites of dog.

  9. Imported and travelling dogs as carriers of canine vector-borne pathogens in Germany

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    Lorentz Susanne

    2010-04-01

    Full Text Available Abstract Background With the import of pets and pets taken abroad, arthropod-borne diseases have increased in frequency in German veterinary practices. This is reflected by 4,681 dogs that have been either travelled to or relocated from endemic areas to Germany. The case history of these dogs and the laboratory findings have been compared with samples collected from 331 dogs living in an endemic area in Portugal. The various pathogens and the seroprevalences were examined to determine the occurrence of, and thus infection risk, for vector-borne pathogens in popular travel destinations. Results 4,681 dogs were examined serological for Leishmania infantum, Babesia canis and Ehrlichia canis. Buffy coats were detected for Hepatozoon canis and blood samples were examined for microfilariae via the Knott's test. The samples were sent in from animal welfare organizations or private persons via veterinary clinics. Upon individual requests, dogs were additionally examined serological for Anaplasma phagocytophilum, Borrelia burgdorferi and Rickettsia conorii. Overall B. canis was the most prevalent pathogen detected by antibody titers (23.4%, followed by L. infantum (12.2% and E. canis (10.1%. Microfilariae were detected in 7.7% and H. canis in 2.7% of the examined dogs. In 332/1862 dogs A. phagocytophilum, in 64/212 B. burgdorferi and in 20/58 R. conorii was detected. Of the 4,681 dogs, in total 4,226 were imported to Germany from endemic areas. Eighty seven dogs joined their owners for a vacation abroad. In comparison to the laboratory data from Germany, we examined 331 dogs from Portugal. The prevalence of antibodies/pathogens we detected was: 62.8% to R. conorii, 58% to B. canis, 30.5% to A. phagocytophilum, 24.8% to E. canis, 21.1% to H. canis (via PCR, 9.1% to L. infantum and 5.3% to microfilariae. Conclusions The examination of 4,681 dogs living in Germany showed pathogens like L. infantum that are non-endemic in Germany. Furthermore, the German

  10. Vector-borne pathogens in dogs and red foxes from the federal state of Brandenburg, Germany.

    Science.gov (United States)

    Liesner, Jana M; Krücken, Jürgen; Schaper, Roland; Pachnicke, Stefan; Kohn, Barbara; Müller, Elisabeth; Schulze, Christoph; von Samson-Himmelstjerna, Georg

    2016-07-15

    Dirofilaria repens is endemic in eastern and southern European regions but was recently found in Germany in dogs, mosquitoes and one human patient. Since some of the positive dog and mosquito samples were collected in Brandenburg, it was aimed to systematically assess the prevalence of D. repens and other canine vector-borne pathogens in Brandenburg. Dog owners also received a questionnaire and were asked to provide more information about the dogs including travel history. In total, 1023 dog blood samples as well as 195 fox spleen and 179 fox blood samples were collected. DNA was analysed by PCR for the presence of filariae, piroplasms, anaplasmataceae and Rickettsia spp. Filariae were detected in six dogs (0.6%), two were positive for DNA from D. repens, two from Dirofilaria immitis and two from Acanthocheilonema reconditum. One of the D. repens positive dogs originated from an animal shelter in Brandenburg, but the origin of the other one remained unknown. Interestingly, both D. repens ITS-1 sequences showed 100% identity to a D. repens sample obtained from a Japanese woman that travelled in Europe and were 97% identical to a newly proposed species Dirofilaria sp. 'hongkongensis' described from Hong Kong. However, identity to other D. repens sequences from Thailand was considerably lower (81%). Identity of 12S rRNA and cytochrome oxidase I to D. repens samples from southern Europe was 99%. Due to the low number of Dirofilaria spp. positive dogs and since the origin of these was unknown, endemic occurrence of Dirofilaria in Brandenburg could not be confirmed. Anaplasma phagocytophilum was found in 15 dogs (1.5%), Candidatus Neoehrlichia mikurensis in three dogs (0.3%) and E. canis in one dog (0.1%), which was co-infected with D. repens. Rickettsia spp. were detected in 8 dogs (0.8%), seven were Rickettsia raoultii and one was Rickettsia felis. To the author's knowledge, R. raoultii DNA was detected for the first time in dogs in Germany in this study and Candidatus

  11. Diversifying selection and host adaptation in two endosymbiont genomes

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    Slatko Barton

    2007-04-01

    Full Text Available Abstract Background The endosymbiont Wolbachia pipientis infects a broad range of arthropod and filarial nematode hosts. These diverse associations form an attractive model for understanding host:symbiont coevolution. Wolbachia's ubiquity and ability to dramatically alter host reproductive biology also form the foundation of research strategies aimed at controlling insect pests and vector-borne disease. The Wolbachia strains that infect nematodes are phylogenetically distinct, strictly vertically transmitted, and required by their hosts for growth and reproduction. Insects in contrast form more fluid associations with Wolbachia. In these taxa, host populations are most often polymorphic for infection, horizontal transmission occurs between distantly related hosts, and direct fitness effects on hosts are mild. Despite extensive interest in the Wolbachia system for many years, relatively little is known about the molecular mechanisms that mediate its varied interactions with different hosts. We have compared the genomes of the Wolbachia that infect Drosophila melanogaster, wMel and the nematode Brugia malayi, wBm to that of an outgroup Anaplasma marginale to identify genes that have experienced diversifying selection in the Wolbachia lineages. The goal of the study was to identify likely molecular mechanisms of the symbiosis and to understand the nature of the diverse association across different hosts. Results The prevalence of selection was far greater in wMel than wBm. Genes contributing to DNA metabolism, cofactor biosynthesis, and secretion were positively selected in both lineages. In wMel there was a greater emphasis on DNA repair, cell division, protein stability, and cell envelope synthesis. Conclusion Secretion pathways and outer surface protein encoding genes are highly affected by selection in keeping with host:parasite theory. If evidence of selection on various cofactor molecules reflects possible provisioning, then both insect as

  12. The risk of vector-borne infections in sled dogs associated with existing and new endemic areas in Poland: Part 1: A population study on sled dogs during the racing season.

    Science.gov (United States)

    Bajer, Anna; Mierzejewska, Ewa J; Rodo, Anna; Bednarska, Malgorzata; Kowalec, Maciej; Welc-Falęciak, Renata

    2014-05-28

    The achievements of sled dogs in competitions depend both on their training and on their health. Vector-borne infections may lead to anaemia, affect joints or heart muscle or even cause death. Between December 2009 and October 2010, one hundred and twenty six individual blood samples were collected from 26 sled dog kennels situated in different regions of Poland. The majority of samples were taken during the racing season (winter 2009/10). The prevalences of 3 vector-borne infections- including 2 'old pathogens' Anaplasma phagocytophilum and Babesia canis, and 'new pathogen' Hepatozoon canis-were estimated in sled dogs using PCR and nested PCR. Additionally, 25 serum samples originating from a subset of 3 kennels situated in a tick-borne encephalitis (TBE) endemic area (Mazowiecki region), were tested for antibodies against the tick-borne encephalitis virus (TBEV). Because of the recently reported occurrence of Dirofilaria repens in Central Poland and that of fatal cases of unknown aetiology in two of the kennels, blood samples collected from dogs at these kennels in 2010 and in February-May 2013 and from two unaffected kennels were checked for evidence of presence of this parasite. Babesia canis DNA was detected in 11 sled dogs (4 with clinical babesiosis, 7 asymptomatic; 8.7%) inhabiting mainly endemic regions of Poland (9/11 cases). Three serum samples originating from one location tested positive for TBEV antibodies (total seroprevalence: 3/25=12%, local seroprevalence: 3/12=25%). The risk of TBEV infection was associated with previous B. canis infections. Dirofilaria repens DNA was detected in 15 dogs (44%). Prevalence was especially high in two sled dog kennels situated near Grodzisk Mazowiecki (50-57%). No blood samples tested positive for A. phagocytophilum or H. canis DNA. The present study has established that the prevalence of vector-borne pathogens in working sled dogs is significant in the endemic regions and has justified the important role of

  13. Zoonotic pathogens associated with Hyalomma aegyptium in endangered tortoises: evidence for host-switching behaviour in ticks?

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    Paștiu Anamaria I

    2012-12-01

    Full Text Available Abstract Background Hyalomma aegyptium is a hard-tick with a typical three-host life cycle. The main hosts are Palearctic tortoises of genus Testudo. However, other hosts can be used by immature ticks for feeding in natural conditions. Given this complex ecology and multiple host use, the circulation of pathogens by H. aegyptium between various hosts can be important from epidemiological point of view. The aim of this study was to evaluate the role of H. aegyptium as natural carrier of four important zoonotic pathogens. Methods From 2008 to 2011, 448 H. aegyptium ticks were collected from 45 Spur-thighed tortoises, Testudo graeca in Romania. DNA was extracted individually from each tick using a commercial kit. DNA was examined for the presence of specific sequences of Borrelia burgdorferi s.l., Anaplasma phagocytophilum, Ehrlichia canis and Coxiella burnetii by PCR, according to previously described protocols. Results PCR analysis of H. aegyptium revealed the presence of A. phagocytophilum (18.8%, E. canis (14.1% and C. burnetii (10%. 32.4% of the ticks were infected with at least one pathogen and 9.8% had co-infections. The stages most frequently infected were nymphs (50% followed by males (33.9% and females (27%. The number of tortoises which harboured infected ticks was 27/45 examined (60%. From all tested T. graeca, 40% harboured ticks infected with A. phagocytophilum, 46.7% had ticks infected with E. canis and 33.3% had ticks with C. burnetii. This study reports for the first time the presence of A. phagocytophilum and E. canis in H. aegyptium. Conclusions The presence and relatively high prevalence of three important zoonotic pathogens in H. aegyptium raises the question of their epidemiologic importance in disease ecology. As tortoises are unlikely to be reservoir hosts for A. phagocytophilum and E. canis and both these pathogens are common in H. aegyptium, this is an important indication for (1 a possible increased host

  14. Subset-directed antiviral treatment of 142 herpesvirus patients with chronic fatigue syndrome

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    A Martin Lerner

    2010-05-01

    Full Text Available A Martin Lerner1, Safedin Beqaj2, James T Fitzgerald3, Ken Gill4, Carol Gill4, James Edington41Department of Medicine, William Beaumont Hospital, Royal Oak; 2Wayne State University School of Medicine, Detroit; 3Department of Medical Education, University of Michigan Medical School, Ann Arbor, Michigan; 4The Dr A Martin Lerner Chronic Fatigue Syndrome Foundation, Beverly Hills, Michigan, USAPurpose: We hypothesized that chronic fatigue syndrome (CFS may be caused by single or multiple Epstein–Barr virus (EBV, cytomegalovirus (HCMV, or human herpesvirus 6 (HHV6 infection. To determine if CFS life-altering fatigue and associated findings including muscle aches, tachycardia at rest, chest aches, left ventricular dysfunction, syncope, and elevated herpesvirus serum antibody titers are reversed by long-term subset-directed valacyclovir and/or valganciclovir.Patients and methods: Data were collected at physician visits every 4–6 weeks from 142 CFS patients at one clinic from 2001 to 2007. To be included in this study, patients had to be followed for at least six months. The data captured included over 7000 patient visits and over 35,000 fields of information. Severity of fatigue was monitored by a validated Energy Index Point Score® (EIPS®. Baseline and follow-up serum antibody titers to EBV, HCMV, and HHV6, as well as coinfections with Borrelia burgdorferi, Anaplasma phagocytophila, Babesia microti, and antistreptolysin O, 24-hour ECG Holter monitors, 2D echocardiograms, cardiac dynamic studies, symptoms, and toxicity were captured and monitored. International criteria for CFS plus a specifically designed CFS diagnostic panel were used.Results and conclusions: The Group A herpesvirus CFS patients (no coinfections returned to a near-normal to normal life (P = 0.0001. The long-term EIPS value increased (primary endpoint, P < 0.0001 with subset-directed long-term valacyclovir and/or valganciclovir therapy. Secondary endpoints (cardiac, immunologic

  15. Abundance of questing ticks and molecular evidence for pathogens in ticks in three parks of Emilia-Romagna region of Northern Italy

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    Sara Aureli

    2015-09-01

    Full Text Available Introduction and objective. Infectious and parasitic diseases transmitted by ticks, such as Lyme diseases, granulocytic anaplasmosis and piroplasmosis, have been frequently reported in Europe, with increasing attention to them as an emerging zoonotic problem. The presented study was performed to assess the distribution and the density of questing ticks in three regional parks of Emilia-Romagna region of Northern Italy, and to seek molecular evidence of potential human pathogens in tick populations. Materials and Methods. In the period April-October 2010, 8,139 questing ticks were collected: 6,734 larvae, 1,344 nymphs and only a few adults – 28 females and 33 males. The abundance of[i] Ixodes ricinus[/i] questing ticks was compared among different sampling sites and related to microclimate parameters. 1,544 out of 8,139 ticks were examined for the presence of pathogens: PCR was used to detect piroplasms DNA and Real time Taqman PCR for [i]Anaplasma phagocytophilum[/i] and [i]Borrelia burgdorferi[/i] s.l. Results. The predominant species was [i]I. ricinus[/i] (overall abundance 1,075.9/100 m[sup]2[/sup] ; more rarely, [i]Dermacentor marginatus[/i] (n = 37 – 0.45%, [i]Scaphixodes frontalis[/i] (n = 13 – 0.16%, [i]Hyalomma[/i] spp. (n = 6 – 0.07% and [i]Ixodes acuminatus[/i] (n = 3 – 0.04% were also found. 28 out of 324 (8.6% samples of ticks were PCR-positive for piroplasm DNA. 11 amplicons of 18S rRNA gene were identical to each other and had 100% identity with[i] Babesia[/i] EU1 ([i]Babesia venatorum[/i] using BLAST analysis. Real time Taqman PCR gave positive results for [i]A. phagocytophilum[/i] in 23 out of 292 samples (7.9%, and for [i]B. burgdorferi[/i] s.l. in 78 out of 292 samples (26.7%. [i]I. ricinu[/i]s was the only species found positive for pathogens by molecular analysis; 16 tick samples were co-infected with at least 2 pathogens. Discussion. The peak of nymph presence was in May, and the higher prevalence of pathogens

  16. Borrelia Diversity and Co-infection with Other Tick Borne Pathogens in Ticks

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    Raileanu, Cristian; Moutailler, Sara; Pavel, Ionuţ; Porea, Daniela; Mihalca, Andrei D.; Savuta, Gheorghe; Vayssier-Taussat, Muriel

    2017-01-01

    Identifying Borrelia burgdorferi as the causative agent of Lyme disease in 1981 was a watershed moment in understanding the major impact that tick-borne zoonoses can have on public health worldwide, particularly in Europe and the USA. The medical importance of tick-borne diseases has long since been acknowledged, yet little is known regarding the occurrence of emerging tick-borne pathogens such as Borrelia spp., Anaplasma phagocytophilum, Rickettsia spp., Bartonella spp., “Candidatus Neoehrlichia mikurensis”, and tick-borne encephalitis virus in questing ticks in Romania, a gateway into Europe. The objective of our study was to identify the infection and co-infection rates of different Borrelia genospecies along with other tick-borne pathogens in questing ticks collected from three geographically distinct areas in eastern Romania. We collected 557 questing adult and nymph ticks of three different species (534 Ixodes ricinus, 19 Haemaphysalis punctata, and 4 Dermacentor reticulatus) from three areas in Romania. We analyzed ticks individually for the presence of eight different Borrelia genospecies with high-throughput real-time PCR. Ticks with Borrelia were then tested for possible co-infections with A. phagocytophilum, Rickettsia spp., Bartonella spp., “Candidatus Neoehrlichia mikurensis”, and tick-borne encephalitis virus. Borrelia spp. was detected in I. ricinus ticks from all sampling areas, with global prevalence rates of 25.8%. All eight Borrelia genospecies were detected in I. ricinus ticks: Borrelia garinii (14.8%), B. afzelii (8.8%), B. valaisiana (5.1%), B. lusitaniae (4.9%), B. miyamotoi (0.9%), B. burgdorferi s.s (0.4%), and B. bissettii (0.2%). Regarding pathogen co-infection 64.5% of infected I. ricinus were positive for more than one pathogen. Associations between different Borrelia genospecies were detected in 9.7% of ticks, and 6.9% of I. ricinus ticks tested positive for co-infection of Borrelia spp. with other tick-borne pathogens. The

  17. Seroprevalence and molecular biological investigation of commonly known Rickettsia species in rodents in Qujing Prefecture, Yunnan Province, China%云南省曲靖市啮齿动物的立克次体血清学和分子流行病学调查

    Institute of Scientific and Technical Information of China (English)

    章域震; 王勇; 杨卫红; 朱向晖; 皮自林; 冯云; 何丽芳; 张海林; 张丽娟

    2015-01-01

    目的 调查云南省曲靖市啮齿动物感染立克次体状况.方法 用鼠笼法捕鼠,采集鼠血清和脾脏.用间接免疫荧光试验(IFA)检测鼠血清中7种常见立克次体的IgG抗体;用巢氏PCR方法检测鼠脾中嗜单核细胞埃立克体和嗜吞噬细胞无形体16S rRNA基因片段部份序列.结果 2012年7-9月在曲靖市捕获啮齿动物3种592只,褐家鼠(Rattus norvegicus)和黄胸鼠(Rattus flavipectus)构成比分别为61.49%和35.47%.鼠血清中贝氏柯克斯体(Coxiella burentii)、莫氏立克次体(Rickettsia typhi)、恙虫病东方体(Orientia tsutsugamushi)、西伯利亚立克次体(Rickettsia sibirica)、嗜吞噬细胞无形体(Anaplasma phagocytophilum)和嗜单核细胞埃立克体(Ehrlichia chafeensis)的抗体阳性率依次为21.45%、7.60%、7.09%、3.38%、1.18%和0.51%;未检测到猫立克次体(Rickettsia felis)抗体.从5份褐家鼠脾脏标本中检测到16S rRNA基因序列,同源性和进化分析表明,1株为嗜吞噬细胞无形体,4株为莫氏立克次体.结论曲靖市啮齿动物中存在莫氏立克次体和人粒细胞无形体流行,褐家鼠可能是主要宿主;同时还存在恙虫病东方体、贝氏柯克斯体、斑点热立克次体和埃立克体的感染.当地疾控和医疗机构应加强立克次体病的监测.

  18. 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

  19. A review of Hyalomma scupense (Acari, Ixodidae in the Maghreb region: from biology to control

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    2014-01-01

    Full Text Available Hyalomma scupense (syn. Hyalomma detritum is a two-host domestic endophilic tick of cattle and secondarily other ungulates in the Maghreb region (Africa. This species transmits several pathogens, among which two are major livestock diseases: Theileria annulata and Theileria equi. Various other pathogens are also transmitted by this tick species, such as Anaplasma phagocytophilum and Ehrlichia bovis. Hyalomma scupense is common in sub-humid and semi-arid areas of several regions in the world, mainly in the Maghreb region. In this region, adults attach to animals during the summer season; larvae and nymphs attach to their hosts during autumn, but there is a regional difference in H. scupense phenology. There is an overlap between immature and adult ticks, leading in some contexts to a dramatic modification of the epidemiology of tick-borne diseases. This tick species attaches preferentially to the posterior udder quarters and thighs. Tick burdens can reach 130 ticks per animal, with a mean of 60 ticks. Calves are 70 times less infested than adult cattle. The control can be implemented through six options: (i rehabilitation of the farm buildings by roughcasting and smoothing the outer and inner surfaces of the enclosures and walls. This control option should be recommended to be combined with a thorough cleaning of the farm and its surrounding area. With regard to Theileria annulata infection, this control option is the most beneficial. (ii Acaricide application to animals during the summer season, targeting adults. (iii Acaricide application during the autumn period for the control of the immature stages. (iv Acaricide application to the walls: many field veterinarians have suggested this option but it is only partially efficient since nymphs enter deep into the cracks and crevices. It should be used if there is a very high tick burden or if there is a high risk of tick-borne diseases. (v Manual tick removal: this method is not efficient since the

  20. Survey of ticks collected in Mississippi for Rickettsia, Ehrlichia, and Borrelia species.

    Science.gov (United States)

    Goddard, Jerome; Sumner, John W; Nicholson, William L; Paddock, Christopher D; Shen, John; Piesman, Joseph

    2003-12-01

    % identity with an endosymbiont of Acanthoamoeba. Other sequences represented in the top 50 Basic Local Alignment Search (BLAST) scores were primarily from soil bacteria, although some similarity to several Anaplasma species and Ehrlichia risticii was indicated. The significance of this finding remains undetermined.

  1. Prevalence of Neoehrlichia mikurensis in ticks and rodents from North-west Europe

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    Jahfari Setareh

    2012-07-01

    Full Text Available Abstract Background Neoehrlichia mikurensis s an emerging and vector-borne zoonosis: The first human disease cases were reported in 2010. Limited information is available about the prevalence and distribution of Neoehrlichia mikurensis in Europe, its natural life cycle and reservoir hosts. An Ehrlichia-like schotti variant has been described in questing Ixodes ricinus ticks, which could be identical to Neoehrlichia mikurensis. Methods Three genetic markers, 16S rDNA, gltA and GroEL, of Ehrlichia schotti-positive tick lysates were amplified, sequenced and compared to sequences from Neoehrlichia mikurensis. Based on these DNA sequences, a multiplex real-time PCR was developed to specifically detect Neoehrlichia mikurensis in combination with Anaplasma phagocytophilum in tick lysates. Various tick species from different life-stages, particularly Ixodes ricinus nymphs, were collected from the vegetation or wildlife. Tick lysates and DNA derived from organs of wild rodents were tested by PCR-based methods for the presence of Neoehrlichia mikurensis. Prevalence of Neoehrlichia mikurensis was calculated together with confidence intervals using Fisher's exact test. Results The three genetic markers of Ehrlichia schotti-positive field isolates were similar or identical to Neoehrlichia mikurensis. Neoehrlichia mikurensis was found to be ubiquitously spread in the Netherlands and Belgium, but was not detected in the 401 tick samples from the UK. Neoehrlichia mikurensis was found in nymphs and adult Ixodes ricinus ticks, but neither in their larvae, nor in any other tick species tested. Neoehrlichia mikurensis was detected in diverse organs of some rodent species. Engorging ticks from red deer, European mouflon, wild boar and sheep were found positive for Neoehrlichia mikurensis. Conclusions Ehrlichia schotti is similar, if not identical, to Neoehrlichia mikurensis. Neoehrlichia mikurensis is present in questing Ixodes ricinus ticks throughout the

  2. Detection of Rickettsia and Ehrlichia spp. in Ticks Associated with Exotic Reptiles and Amphibians Imported into Japan.

    Science.gov (United States)

    Andoh, Masako; Sakata, Akiko; Takano, Ai; Kawabata, Hiroki; Fujita, Hiromi; Une, Yumi; Goka, Koichi; Kishimoto, Toshio; Ando, Shuji

    2015-01-01

    One of the major routes of transmission of rickettsial and ehrlichial diseases is via ticks that infest numerous host species, including humans. Besides mammals, reptiles and amphibians also carry ticks that may harbor Rickettsia and Ehrlichia strains that are pathogenic to humans. Furthermore, reptiles and amphibians are exempt from quarantine in Japan, thus facilitating the entry of parasites and pathogens to the country through import. Accordingly, in the current study, we examined the presence of Rickettsia and Ehrlichia spp. genes in ticks associated with reptiles and amphibians originating from outside Japan. Ninety-three ticks representing nine tick species (genera Amblyomma and Hyalomma) were isolated from at least 28 animals spanning 10 species and originating from 12 countries (Ghana, Jordan, Madagascar, Panama, Russia, Sri Lanka, Sudan, Suriname, Tanzania, Togo, Uzbekistan, and Zambia). None of the nine tick species are indigenous in Japan. The genes encoding the common rickettsial 17-kDa antigen, citrate synthase (gltA), and outer membrane protein A (ompA) were positively detected in 45.2% (42/93), 40.9% (38/93), and 23.7% (22/93) of the ticks, respectively, by polymerase chain reaction (PCR). The genes encoding ehrlichial heat shock protein (groEL) and major outer membrane protein (omp-1) were PCR-positive in 7.5% (7/93) and 2.2% (2/93) of the ticks, respectively. The p44 gene, which encodes the Anaplasma outer membrane protein, was not detected. Phylogenetic analysis showed that several of the rickettsial and ehrlichial sequences isolated in this study were highly similar to human pathogen genes, including agents not previously detected in Japan. These data demonstrate the global transportation of pathogenic Rickettsia and Ehrlichia through reptile- and amphibian-associated ticks. These imported animals have potential to transfer pathogens into human life. These results highlight the need to control the international transportation of known and

  3. Detection of Rickettsia and Ehrlichia spp. in Ticks Associated with Exotic Reptiles and Amphibians Imported into Japan.

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    Masako Andoh

    Full Text Available One of the major routes of transmission of rickettsial and ehrlichial diseases is via ticks that infest numerous host species, including humans. Besides mammals, reptiles and amphibians also carry ticks that may harbor Rickettsia and Ehrlichia strains that are pathogenic to humans. Furthermore, reptiles and amphibians are exempt from quarantine in Japan, thus facilitating the entry of parasites and pathogens to the country through import. Accordingly, in the current study, we examined the presence of Rickettsia and Ehrlichia spp. genes in ticks associated with reptiles and amphibians originating from outside Japan. Ninety-three ticks representing nine tick species (genera Amblyomma and Hyalomma were isolated from at least 28 animals spanning 10 species and originating from 12 countries (Ghana, Jordan, Madagascar, Panama, Russia, Sri Lanka, Sudan, Suriname, Tanzania, Togo, Uzbekistan, and Zambia. None of the nine tick species are indigenous in Japan. The genes encoding the common rickettsial 17-kDa antigen, citrate synthase (gltA, and outer membrane protein A (ompA were positively detected in 45.2% (42/93, 40.9% (38/93, and 23.7% (22/93 of the ticks, respectively, by polymerase chain reaction (PCR. The genes encoding ehrlichial heat shock protein (groEL and major outer membrane protein (omp-1 were PCR-positive in 7.5% (7/93 and 2.2% (2/93 of the ticks, respectively. The p44 gene, which encodes the Anaplasma outer membrane protein, was not detected. Phylogenetic analysis showed that several of the rickettsial and ehrlichial sequences isolated in this study were highly similar to human pathogen genes, including agents not previously detected in Japan. These data demonstrate the global transportation of pathogenic Rickettsia and Ehrlichia through reptile- and amphibian-associated ticks. These imported animals have potential to transfer pathogens into human life. These results highlight the need to control the international transportation of known

  4. Sero-epidemiologic investigation on tick-borne diseases of humans and domestic animals in Zhejiang province%浙江省人和家畜蜱媒传染病血清流行病学调查

    Institute of Scientific and Technical Information of China (English)

    柴程良; 陆群英; 孙继民; 姜理平; 凌锋; 张丽娟; 郑寿贵; 张宏; 葛君华

    2010-01-01

    heilongjiangii, Orientia tsutsugamushi, R. typhi, Anaplasma phagocytos, Ehrlichia chaffeensis, Bartonella, R. hainan and Coxiella burnetii in these samples.Results Six hundred and eighty-three blood samples including 579 from humans and 104 from domestic animals(53 from cattles and 51 from sheep)were collected from the three sites. Antibody positive rates of Orientia tsutsugamushi, R. typhi, Ehrlichia chaffeensis and Coxiella burnetii were significantly different between these sites. IgG from all the 8 pathogens were detected in samples from humans. It was found that the sero-prevalence rates of R. typhi, Bartonella and C. burnetii(20.7%,10.9%, 5.5%)of adults were higher than those of other Rickettsiae under investigation. The seroprevalence of R. typhi increased along with age. IgG from the 7 pathogens were detected in samples from domestic animals except for Anaplasma phagocytos. The sero-prevalence rates of R. typhi, Bartonella and R. hainan(69.2%, 51.0%, 22.1%)of adults were higher than those of other Rickettsiae investigated. Conclusion Tick-borne diseases did spread widely in humans and domestic animals from different rural areas of Zhejiang province. The sero-prevalence rates of R. typhi,B. henselae, R. hainan and C. burnetii were higher than that from other pathogens.

  5. 云南省保山市一起地方性斑疹伤寒暴发的调查%An outbreak of endemic typhus in Baoshan city, Yunnan province

    Institute of Scientific and Technical Information of China (English)

    亚红祥; 张海林; 周济华; 段恒先; 赵孝泽; 孙桂娟; 郑维斌; 黄文丽

    2011-01-01

    Objective To understand the epidemiologic characteristics of endemic typhus in Baoshan city. Methods Epidemiological data were collected and characteristics were analyzed. IgG antibody(Ab) of Rickettsia mooseri and Orientia tsutsuganushi in serum of patients were tested using both Weil-Felix and IFA method. The Rickettsia mooseri gltA gene, Rickettsia prowazekii gltA gene,Orientia tsutsugamushi 56 kDa protein gene, SFGR ompA gene, Ehrlichia sp. 16S rRNA gene and Anaplasma sp. 16S rRNA gene in spleen of mice were examined by PCR. Results Fifty- eight endemic typhus cases were found in Longyang district of Baoshan city, during July to August, 2009.Among them, 48 cases were confirmed by clinical diagnosis and 10 cases by laboratory tests. The Ab of Orientia tsutsugamushi Karp serotype was detected in 3 cases from laboratory diagnosis. The spleen samples from 85 Rattns flavipectus were tested using PCR. Of them, 3 samples for Rickettsia mooseri gltA gene showed positive (positive rate was 3.5% ), and the homology of 3 Rickettsia mooseri and Rickettsia mooseri Wilmington strain (GenBank U59714.1) was 100% through comparing gene sequence. The results of PCR for detecting Rickettsia prowazekii, Orientia tsutsugamushi, SFGR,Anaplasma sp. and Ehrlichia. sp were all negative. Conclusion The outbreak of endemic typhus was confirmed in Longyang district of Baoshan city through epidemiological data, clinical diagnosis and laboratory tests. Rickettsia mooseri DNA was detected in the dominant Raw flavipectus, suggesting that endemic typhus did exist in the local areas.%目的 了解云南省保山市一起地方性斑疹伤寒暴发的流行病学特征.方法 收集患者的流行病学资料.应用外斐反应和间接免疫荧光方法同时检测患者血清中莫氏立克次体、恙虫病东方体IgG抗体.用PCR检测鼠类脾脏标本中莫氏立克次体、普氏立克次体gltA基因,恙虫病东方体56kDa蛋白基因,斑点热群立克次体ompA基因,埃立克体16S

  6. Tristeza parasitária bovina na região sul do Rio Grande do Sul: estudo retrospectivo de 1978-2005 Tick fever in southern Brazil: a retrospective study of 1978-2005

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    Milton Begeres de Almeida

    2006-12-01

    Full Text Available Foi realizado um estudo retrospectivo dos casos de tristeza parasitária bovina (TPB ocorridos no sul do Rio Grande do Sul, área de influência do Laboratório Regional de Diagnóstico (LRD da Faculdade de Veterinária da Universidade Federal de Pelotas entre 1978 e 2005. De um total de 4.884 materiais de bovinos provenientes de necropsias realizadas e órgãos ou sangue enviados ao laboratório 231 (4,7% tiveram o diagnóstico de TPB. Desses 231 surtos foram resgatados os dados de 221 diagnósticos dos quais 91 (41,1% foram causados por Babesia bovis, 11 (4,9% por Babesia bigemina, e 65 (29,41% por Anaplasma marginale. Em outros 33 (14,93% surtos de babesiose não foi informada a espécie de Babesia e em 21 (9,5% surtos foi detectada infecção mista por Babesia sp e A. marginale. Os índices gerais médios de morbidade, mortalidade e letalidade, resgatados em 149 dos 221 surtos da doença, foram de 11,17%, 6,81% e 70,04%, respectivamente. Verificou-se que, na região estudada, a maioria dos surtos ocorre durante os meses de verão e outono, e que os animais com um a três anos de idade são os mais afetados. Os sinais clínicos nos surtos caracterizaram-se por apatia, orelhas caídas, debilidade, febre, anorexia e emagrecimento. Os valores de hematócrito eram baixos. Hemoglobinúria foi frequentemente observada nos casos de babesiose. Sinais neurológicos estavam presentes nos casos de babesiose por B. bovis e se caracterizaram por transtornos da locomoção, tremores musculares, agressividade e quedas com movimentos de pedalagem. As lesões macroscópicas principais relatadas nos casos de babesiose foram esplenomegalia, hepatomegalia, fígado amarelo, hemoglobinúria, icterícia, hemorragias cardíacas e bile espessa. Congestão do córtex cerebral foi relatada nos casos de babesiose por B. bovis. Nesta região, com população de bovinos de aproximadamente 2.630.000 cabeças as perdas anuais por morte de bovinos pela enfermidade podem ser

  7. [Transfer of exotic ticks (Acari: ixodida) on reptiles (Reptilia) imported to Poland].

    Science.gov (United States)

    2009-01-01

    (V. salvator and V. exanthematicus) and 92 specimens pythons (P. regius) were examined, with detailed descriptions of where the parasite was feeding on the body of the host. Among the 434 specimens of ticks collected from the monitor lizards, the majority were attached on the host's legs (40.5%), on the trunk (29.3%), on the head (20.3%), with fewest on the tail (9.9%). Also, 430 specimens of ticks were collected from the bodies of pythons. They mostly parasitized along the whole length of the back (54.4%) and on the stomach side of the trunk (29.8%), less frequently in the area of the cloaca (5.6%), around the eyes (3.7%), in the nostril openings (0.9%) and on the remainder of the head (5.6%). On the hosts, ticks were found at different development stages, but adult development stages dominated. The most frequent were males (999 specimens), then adult females (552 specimens), nymphs (508 specimens) and larvae (45 specimens). During the research, 13 cases of anomalies of morphological structure were confirmed for ticks Amblyomma flavomaculatum, Amblyomma latum and Hyalomma aegyptium. Asymmetries and deformations of the general body shape were observed, as were anomalies concerning structures on the surface of the body and anomalies of the legs. For the first time in Poland, epidemiological tests were carried out in the direction of the infection of exotic ticks gathered from reptiles with micro-organisms which pose a threat for the health of people and animals. For this purpose, molecular techniques - polymerase chain reaction (PCR) and DNA sequencing were used. The isolates from 345 ticks, were examined for the presence of DNA of Anaplasma phagocytophilum, which is the etiological factor in human granulocytic anaplasmosis, and Rickettsia spp. from the spotted fever group, causing human rickettsiosis. This study confirmed the presence of Anaplasma phagocytophilum in two ticks of Amblyomma flavomaculatum (constituting 0.6% of all the ticks investigated) feeding on

  8. Surveillance and study of tick-borne diseases in Zhejiang province, China%浙江省蜱媒传染病监测研究

    Institute of Scientific and Technical Information of China (English)

    凌锋; 龚震宇; 柴程良; 林君芬; 孙继民; 侯娟; 吴海霞; 刘起勇

    2013-01-01

    Objective To investigate the current situation of surveillance and study on the tick - borne diseases in Zhejiang province, China. Methods By literature research and surveillance data analysis, brief descriptive statistic analysis was performed on the literature and annual surveillance data on the tick-borne diseases in Zhejiang that have been published since 2000. Results Through ten years surveillance and study involving serological and molecular biological methods, more than 10 tick-borne diseases or pathogens, including Lyme disease, Anaplasma phagocytophilum, Ehrlichia chaffeensis, spotted fever, Bartonella, Rickettsia mooseri, and Boehner kirk austenite, were confirmed to exist in Zhejiang. Besides, the various tick species and tick-borne diseases were found in as many as 23 counties (county-level cities, districts) in Hangzhou, Jinhua, Lishui, Taizhou, Huzhou, Wenzhou, Quzhou, and Shaoxing in Zhejiang. Conclusion The ticks and tick-borne diseases diversified and widespread in Zhejiang province. Although some achievements have been made in the surveillance and study on tick-borne diseases, the surveillance is neither sufficient nor systematic since lack of proper planning. The related information cannot be fully integrated and utilized for risk assessment and early-waming. It is necessary to establish a platform for tick-borne disease surveillance.%目的 了解浙江省蜱媒传染病监测研究现状.方法 采用文献研究法和监测资料分析相结合,对2000年以来发表的有关浙江省蜱媒传染病的文献及其历年监测资料做简要描述性统计分析.结果 通过近10年来的监测研究,浙江省已通过血清学或分子生物学方法证实存在莱姆病、嗜吞噬细胞无形体、查菲埃立克体、斑点热、巴尔通体等10多种蜱媒传染病,且蜱种群和蜱媒传染病分布较广,涉及到浙江省杭州、金华、丽水、台州、湖州、温州、衢州和绍兴8个市的23个县(市、区).结论 浙江

  9. Research on the pathogenic infection of Haemaphysalis longicornis ticks parasitizing goats in Yiyuan County, Shandong Province%山东沂源羊体寄生长角血蜱携带病原调查研究

    Institute of Scientific and Technical Information of China (English)

    刘相叶; 郑辰; 李向阳; 颜超; 华慧; 汤仁仙; 郑葵阳

    2015-01-01

    目的:了解山东省沂源县境内寄生于山羊体表的长角血蜱携带病原体的情况。方法2014年9月—2015年7月从该地区山羊体表采集不同生活史时期的长角血蜱。利用PCR扩增特异性基因的方法,分别检测长角血蜱感染嗜吞噬细胞无形体、巴贝斯虫、伯氏疏螺旋体以及巴尔通体的情况。结果山东沂源地区长角血蜱中嗜吞噬细胞无形体的感染率为48.1%,巴贝斯虫的感染率为40.7%,其中24.1%的长角血蜱存在嗜吞噬细胞无形体和巴贝斯虫的双重感染,但所有长角血蜱体内未检测到伯氏疏螺旋体和巴尔通体。结论山东沂源羊体寄生的长角血蜱存在较高的嗜吞噬细胞无形体和巴贝斯虫感染率。因此,该地区应加强对人粒细胞无形体病和巴贝斯虫病的防控。%Objective To investigate the tick-borne pathogens transmitted by Haemaphysalis longicornis ticks par-asitizing goats in Yiyuan County, Shandong province.Methods H.longicornis ticks were collected from goats from Sep-tember 2014 to July 2015.Through amplification of specific genes by PCR, tick-borne pathogens including Anaplasma phagocytophilum, Babesia species, Borellia burgdorferi and Bartonella species were detected.Results The infection rates of A.phagocytophilum and Babesia sp.in H.longicornis ticks were 48.1%and 40.7%, respectively.Meanwhile, 24.1% of H.longicornis ticks were infected with both A.phagocytophilum and Babesia sp.However, neither Borellia burgdorferi nor Bartonella sp.was detected in the ticks.Conclusion High infection rates of A.phagocytophilum and Babesia sp.were found in H.longicornis ticks parasitizing in goats in Yiyuan County.Therefore, urgent measures should be promptly taken to enhance the prevention and control of human granulocytic anaplasmosis and babesiosis.

  10. Occurrence and identification of risk areas of Ixodes ricinus-borne pathogens: a cost-effectiveness analysis in north-eastern Italy

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    Capelli Gioia

    2012-03-01

    Full Text Available Abstract Background Ixodes ricinus, a competent vector of several pathogens, is the tick species most frequently reported to bite humans in Europe. The majority of human cases of Lyme borreliosis (LB and tick-borne encephalitis (TBE occur in the north-eastern region of Italy. The aims of this study were to detect the occurrence of endemic and emergent pathogens in north-eastern Italy using adult tick screening, and to identify areas at risk of pathogen transmission. Based on our results, different strategies for tick collection and pathogen screening and their relative costs were evaluated and discussed. Methods From 2006 to 2008 adult ticks were collected in 31 sites and molecularly screened for the detection of pathogens previously reported in the same area (i.e., LB agents, TBE virus, Anaplasma phagocytophilum, Rickettsia spp., Babesia spp., "Candidatus Neoehrlichia mikurensis". Based on the results of this survey, three sampling strategies were evaluated a-posteriori, and the impact of each strategy on the final results and the overall cost reductions were analyzed. The strategies were as follows: tick collection throughout the year and testing of female ticks only (strategy A; collection from April to June and testing of all adult ticks (strategy B; collection from April to June and testing of female ticks only (strategy C. Results Eleven pathogens were detected in 77 out of 193 ticks collected in 14 sites. The most common microorganisms detected were Borrelia burgdorferi sensu lato (17.6%, Rickettsia helvetica (13.1%, and "Ca. N. mikurensis" (10.5%. Within the B. burgdorferi complex, four genotypes (i.e., B. valaisiana, B. garinii, B. afzelii, and B. burgdorferi sensu stricto were found. Less prevalent pathogens included R. monacensis (3.7%, TBE virus (2.1%, A. phagocytophilum (1.5%, Bartonella spp. (1%, and Babesia EU1 (0.5%. Co-infections by more than one pathogen were diagnosed in 22% of infected ticks. The prevalences of infection

  11. Molecular detection of rickettsias DNA in ticks around northeast region of China%东北部分地区蜱携带几种病原立克次体的分子检测

    Institute of Scientific and Technical Information of China (English)

    吴琼; 冯立; 王洪军; 吴益民; 付学奇; 王卓; 张志强

    2012-01-01

    Objective To understand the prevalence of tick-borne rickettsias around northeast region of China. Methods Wild ticks were captured and polymerase chain reaction (PCR) was used to amplify the rickettsias DNA of spotted fever group, human granulocytic ehrlichia and Ehrlichia chaff eensis carried by ticks. Results Ehrlichia chaff eensis DNA was found in I. Persulcatus and D. silvarum with the positive rates of 3.29% and 1.88% respectively. Meanwhile, human granulocytic ehrlichia DNA was amplified from I. Persulcatus with the positive rate of 2. 95%. However spotted fever group Rickettsiae was mainly found in H. concinna, with as high as 6.67% ticks carrying this rickettsiae, higher than that in I. Persulcatus and D. silvarum which were only 1. 69% and 2. 36%. Conclusions 7. Persulcatus and D. silvarum are possibly the main hosts of human granulocytic ehrlichia and Ehrlichia chaff eensis , while H. concinna mainly carries spotted fever group Rickettsiae in northeast region of China, indicating that it might be the natural foci of spotted feve, human granulocytic anaplasma and Ehrlichiosis.%目的 为了解东北地区蜱中携带立克次体的情况.方法 应用人工小时布旗法采集东北部分地区不同生境的游离蜱,采用PCR作斑点热立克次体、查菲埃立克体、人粒细胞无形体DNA检测.结果 从东北地区全沟硬蜱和森林革蜱检出查菲埃立克体DNA,全沟硬蜱平均阳性率为3.29%,森林革蜱为1.88%;从全沟硬蜱检出人粒细胞无形体DNA,平均阳性率为2.98%;从嗜群血蜱检出斑点热立克次体DNA,其平均阳性率(6.67%)明显高于全沟硬蜱(1.69%)和森林革蜱(2.36%).结论 东北地区全沟硬蜱和森林革蜱可能是查菲埃立克体、人粒细胞无形体的主要携带媒介,嗜群血蜱主要携带斑点热立克次体,提示东北地区存在三种蜱媒立克次体病的自然疫源地.