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Sample records for cholesterol-dependent anaplasma phagocytophilum

  1. Anaplasma phagocytophilum in White-tailed Deer

    OpenAIRE

    Massung, Robert F.; Courtney, Joshua W.; Hiratzka, Shannon L.; Virginia E. Pitzer; Smith, Gary; Dryden, Richard L.

    2005-01-01

    We examined the reservoir potential of white-tailed deer for Anaplasma phagocytophilum. Results suggest that white-tailed deer harbor a variant strain not associated with human infection, but contrary to published reports, white-tailed deer are not a reservoir for strains that cause human disease. These results will affect surveillance studies of vector and reservoir populations.

  2. 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.

  3. Lessons from Anaplasma phagocytophilum: Chromatin Remodeling by Bacterial Effectors

    OpenAIRE

    Rennoll-Bankert, Kristen E.; Dumler, J. Stephen

    2012-01-01

    Bacterial pathogens can alter global host gene expression via histone modifications and chromatin remodeling in order to subvert host responses, including those involved with innate immunity, allowing for bacterial survival. Shigella flexneri, Listeria monocytogenes, Chlamydia trachomatis, and Anaplasma phagocytophilum express effector proteins that modify host histones and chromatin structure. A. phagocytophilum modulates granulocyte respiratory burst in part by dampening transcription of se...

  4. 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

  5. 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

  6. The natural history of Anaplasma phagocytophilum.

    Science.gov (United States)

    Woldehiwet, Zerai

    2010-02-10

    Anaplasma phagocytophilum is the recently designated name replacing three species of granulocytic bacteria, Ehrlichia phagocytophila, Ehrlichia equi and the agent of human granulocytic ehrlichiosis, after the recent reorganization of the families Rickettsiaceae and Anaplasmataceae in the order Rickettsiales. Tick-borne fever (TBF), which is caused by the prototype of A. phagocytophilum, was first described in 1932 in Scotland. A similar disease caused by a related granulocytic agent was first described in horses in the USA in 1969; this was followed by the description of two distinct granulocytic agents causing similar diseases in dogs in the USA in 1971 and 1982. Until the discovery of human granulocytic anaplasmosis (HGA) in the USA in 1994, these organisms were thought to be distinct species of bacteria infecting specific domestic animals and free-living reservoirs. It is now widely accepted that the agents affecting different animal hosts are variants of the same Gram-negative obligatory intracellular bacterium, which is transmitted by hard ticks belonging to the Ixodes persulcatus complex. One of its fascinating features is that it infects and actively grows in neutrophils by employing an array of mechanisms to subvert their bactericidal activity. It is also able to survive within an apparently immune host by employing a complex mechanism of antigenic variation. Ruminants with TBF and humans with HGA develop severe febrile reaction, bacteraemia and leukopenia due to neutropenia, lymphocytopenia and thrombocytopenia within a week of exposure to a tick bite. Because of the severe haematological disorders lasting for several days and other adverse effects on the host's immune functions, infected animals and humans are more susceptible to other infections. PMID:19811878

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

    OpenAIRE

    G Javkhlan; B Enkhtaivan; B Baigal; Myagmarsuren, P.; Battur, B.; Battsetseg, B

    2014-01-01

    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 pers...

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

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    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.

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

    Directory of Open Access Journals (Sweden)

    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.

  10. Molecular characterization reveals distinct genospecies of Anaplasma phagocytophilum from diverse North American hosts

    OpenAIRE

    Rejmanek, Daniel; Bradburd, Gideon; Foley, Janet

    2012-01-01

    Anaplasma phagocytophilum is an emerging tick-borne pathogen that infects humans, domestic animals and wildlife throughout the Holarctic. In the far-western United States, multiple rodent species have been implicated as natural reservoirs for A. phagocytophilum. However, the presence of multiple A. phagocytophilum strains has made it difficult to determine which reservoir hosts pose the greatest risk to humans and domestic animals. Here we characterized three genetic markers (23S–5S rRNA inte...

  11. The pathogen-occupied vacuoles of Anaplasma phagocytophilum and Anaplasma marginale interact with the endoplasmic reticulum

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    Hilary Kay Truchan

    2016-03-01

    Full Text Available The genus Anaplasma consists of tick-transmitted obligate intracellular bacteria that invade white or red blood cells to cause debilitating and potentially fatal infections. A. phagocytophilum, a human and veterinary pathogen, infects neutrophils to cause granulocytic anaplasmosis. A. marginale invades bovine erythrocytes. Evidence suggests that both species may also infect endothelial cells in vivo. In mammalian and arthropod host cells, A. phagocytophilum and A. marginale reside in host cell derived pathogen-occupied vacuoles (POVs. While it was recently demonstrated that the A. phagocytophilum-occupied vacuole (ApV intercepts membrane traffic from the trans-Golgi network, it is unclear if it or the A. marginale-occupied vacuole (AmV interacts with other secretory organelles. Here, we demonstrate that the ApV and AmV extensively interact with the host endoplasmic reticulum (ER in endothelial, myeloid, and/or tick cells. ER lumen markers, calreticulin and protein disulfide isomerase, and the ER membrane marker, derlin-1, were pronouncedly recruited to the peripheries of both POVs. ApV association with the ER initiated early and continued throughout the infection cycle. Both the ApV and AmV interacted with the rough ER and smooth ER. However, only derlin-1-positive rough ER derived vesicles were delivered into the ApV lumen where they localized with intravacuolar bacteria. Transmission electron microscopy identified multiple ER-POV membrane contact sites on the cytosolic faces of both species’ vacuoles that corresponded to areas on the vacuoles’ lumenal faces where intravacuolar Anaplasma organisms closely associated. A. phagocytophilum is known to hijack Rab10, a GTPase that regulates ER dynamics and morphology. Yet, ApV-ER interactions were unhindered in cells in which Rab10 had been knocked down, demonstrating that the GTPase is dispensable for the bacterium to parasitize the ER. These data establish the ApV and AmV as pathogen

  12. SERUM ANTIBODIES TO BORRELIA BURGDORFERI, ANAPLASMA PHAGOCYTOPHILUM, AND BABESIA MICROTI IN RECAPTURED WHITE-FOOTED MICE

    OpenAIRE

    Magnarelli, Louis A.; Williams, Scott C.; Norris, Steven J; Fikrig, Erol

    2013-01-01

    A mark-release-recapture study was conducted during 2007 through 2010 in six, tick-infested sites in Connecticut, United States to measure changes in antibody titers for Borrelia burgdorferi sensu stricto, Anaplasma phagocytophilum, and Babesia microti in Peromyscus leucopus (white-footed mice). There was an overall recapture rate of 40%, but only four tagged mice were caught in ≥2 yr. Sera from 561 mice were analyzed for total antibodies to B. burgdorferi and A. phagocytophilum by using whol...

  13. Expression and Immunogenicity of Recombinant Immunoreactive Surface Protein 2 of Anaplasma phagocytophilum

    OpenAIRE

    Yu, Qiang; Chen, Chuang-Fu; Chen, Qiang; Zhang, Li-Juan

    2012-01-01

    Human granulocytic anaplasmosis (HGA), caused by Anaplasma phagocytophilum, is an emerging tick-borne zoonotic disease throughout the world. The first HGA cases in China were documented in 2008, and the greatest challenge posed by the disease is rapid and accurate diagnosis during the acute phage of illness. In this study, we successfully cloned and expressed an A. phagocytophilum immunoreactive surface protein (major surface protein 2 [MSP2]) and demonstrated that this recombinant protein ha...

  14. Nidicolous ticks of small mammals in Anaplasma phagocytophilum-enzootic sites in northern California

    OpenAIRE

    Foley, Janet; Rejmanek, Daniel; Fleer, Katryna; Nieto, Nathan

    2011-01-01

    Ixodes spp. tick-borne zoonotic diseases are present across the Holarctic in humans, domestic animals, and wildlife. Small mammals are reservoirs for the rickettsial pathogen Anaplasma phagocytophilum and tick vectors may include catholic-feeding bridge vectors as well as host-specialist or nidicolous ticks. Far western North American communities in which A. phagocytophilum is maintained are complex ecologically, with multiple reservoir host and tick species, multiple strains of the bacterial...

  15. Seroprevalence of Borrelia burgdorferi, Anaplasma phagocytophilum, Ehrlichia canis, and Dirofilaria immitis among dogs in Canada

    OpenAIRE

    Villeneuve, Alain; Goring, Jonas; Marcotte, Lynne; Overvelde, Sébastien

    2011-01-01

    The seropositivity of dogs to Borrelia burgdorferi, Anaplasma phagocytophilum, and Ehrlichia canis antibodies, and Dirofilaria immitis antigen was assessed in Canada. Borrelia burgdorferi had the highest seroprevalence, while that of Dirofilaria immitis has not changed significantly in the past 20 y. The risk for these vector-borne infectious agents in Canadian dogs is low but widespread with foci of higher prevalence.

  16. 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

    % 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...

  17. Anaplasma phagocytophilum Infection in Small Mammal Hosts of Ixodes Ticks, Western United States

    OpenAIRE

    Janet E. Foley; Nieto, Nathan C.; Adjemian, Jennifer; Dabritz, Haydee; Brown, Richard N.

    2008-01-01

    A total of 2,121 small mammals in California were assessed for Anaplasma phagocytophilum from 2006 through 2008. Odds ratios were >1 for 4 sciurids species and dusky-footed woodrats. High seroprevalence was observed in northern sites. Ten tick species were identified. Heavily infested rodent species included meadow voles, woodrats, deer mice, and redwood chipmunks.

  18. 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.

  19. Seroprevalence of Anaplasma phagocytophilum among healthy dogs and horses in Israel.

    Science.gov (United States)

    Levi, O; Waner, T; Baneth, G; Keysary, A; Bruchim, Y; Silverman, J; Harrus, S

    2006-03-01

    The presence of reacting antibodies to Anaplasma phagocytophilum has previously been demonstrated in Israel, both in humans and the golden jackal (Canis aureus syriacus). This study was undertaken to determine the seroprevalence of A. phagocytophilum antibodies in two additional potential hosts, domestic dogs and horses in order to investigate the possibility of exposure to the organism in Israel. Of 195 dogs tested, 9% were seroreactive with A. phagocytophilum antigen and 30% were seroreactive to Ehrlichia canis. Twenty-nine percent of the dogs seropositive for E. canis were also reactive to A. phagocytophilum. Two dogs had immunofluorescence antibody (IFA) antibody titres for A. phagocytophilum greater than E. canis. The equine serological survey (n = 300) revealed no seroreactive horses. The results presented in this study suggest that dogs in Israel could have been accidentally exposed to A. phagocytophilum, for example by ticks carried on migrating birds, however, the possibility of cross-reaction with E. canis should also be considered. In spite of the high prevalence of ticks on horses in Israel during the summer months, no evidence for exposure to A. phagocytophilum was apparent. PMID:16626404

  20. 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.

  1. Understanding the human ehrlichiosisis and standing out a causal agent: anaplasma phagocytophilum

    OpenAIRE

    Hernández-Ayazo Helí; Marien-Clarete Said

    2013-01-01

    Introduction: Anaplasma phagocytophilum is an obligatory intracytoplasmatic bacteriathat belong to Rickettsiales order and that has been recognized as a cause of humandisease. Most of symptomatic patients report exposure to ticks (vector) one or twoweeks before the beginning of the disease. They present a febrile disease, however theclinical status extends since asymptomatic infection until fatal disease.Clinical case: Male patient with prolonged fever and with negative proofs in the searchof...

  2. Reservoir Competence of the Redwood Chipmunk (Tamias Ochrogenys) for Anaplasma Phagocytophilum

    OpenAIRE

    Nieto, Nathan C.; Janet E. Foley

    2009-01-01

    Granulocytic anaplasmosis (GA) is an emerging tick-transmitted disease that persists in rodent- Ixodes ricinus-complex tick cycles across the Holarctic. Although the putative reservoir for anaplasmosis in the western United States is the dusky-footed woodrat (Neotoma fuscipes), this rodent was not shown reservoir-competent because of failure of infection from woodrats to other animals via ticks. Redwood chipmunks are common in habitats where Anaplasma phagocytophilum is common, have high PCR-...

  3. 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.

  4. 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

  5. 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.

  6. 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.

  7. Prevalence of Human-Active and Variant 1 Strains of the Tick-Borne Pathogen Anaplasma phagocytophilum in Hosts and Forests of Eastern North America

    OpenAIRE

    Keesing, Felicia; McHenry, Diana J.; Hersh, Michelle; Tibbetts, Michael; Jesse L Brunner; Killilea, Mary; LoGiudice, Kathleen; Kenneth A Schmidt; Ostfeld, Richard S.

    2014-01-01

    Anaplasmosis is an emerging infectious disease caused by infection with the bacterium Anaplasma phagocytophilum. In the eastern United States, A. phagocytophilum is transmitted to hosts through the bite of the blacklegged tick, Ixodes scapularis. We determined the realized reservoir competence of 14 species of common vertebrate hosts for ticks by establishing the probability that each species transmits two important strains of A. phagocytophilum (A. phagocytophilum human-active, which causes ...

  8. 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.

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

    Directory of Open Access Journals (Sweden)

    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.

  10. 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.

  11. First molecular detection and phylogenetic analysis of Anaplasma phagocytophilum in shelter dogs in Seoul, Korea.

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    Lee, Sukyee; Lee, Seung-Hun; VanBik, Dorene; Kim, Neung-Hee; Kim, Kyoo-Tae; Goo, Youn-Kyoung; Rhee, Man Hee; Kwon, Oh-Deog; Kwak, Dongmi

    2016-07-01

    In this study, the status of Anaplasma phagocytophilum infection was assessed in shelter dogs in Seoul, Korea, with PCR and phylogenetic analyses. Nested PCR on 1058 collected blood samples revealed only one A. phagocytophilum positive sample (female, age genetic variability of A. phagocytophilum was evaluated by genotyping, using the 16S rRNA, groEL, and msp2 gene sequences of the positive sample. BLASTn analysis revealed that the 16S rRNA, groEL, and msp2 genes had 99.6%, 99.9%, and 100% identity with the following sequences deposited in GenBank: a cat 16S rRNA sequence from Korea (KR021166), a rat groEL sequence from Korea (KT220194), and a water deer msp2 sequence from Korea (HM752099), respectively. Phylogenetic analyses classified the groEL gene into two distinct groups (serine and alanine), whereas the msp2 gene showed a general classification into two groups (USA and Europe) that were further subgrouped according to region. To the best of our knowledge, this study is the first to describe the molecular diagnosis of A. phagocytophilum in dogs reared in Korea. In addition, the high genetic identity of the 16S rRNA and groEL sequences between humans and dogs from the same region suggests a possible epidemiological relation. Given the conditions of climate change, tick ecology, and recent incidence of human granulocytic anaplasmosis in Korea, the findings of this study underscore the need to establish appropriate control programs for tick-borne diseases in Korea. PMID:27130537

  12. 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.

  13. 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

  14. Understanding the human ehrlichiosisis and standing out a causal agent: anaplasma phagocytophilum

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    Hernández-Ayazo Helí

    2013-06-01

    Full Text Available Introduction: Anaplasma phagocytophilum is an obligatory intracytoplasmatic bacteriathat belong to Rickettsiales order and that has been recognized as a cause of humandisease. Most of symptomatic patients report exposure to ticks (vector one or twoweeks before the beginning of the disease. They present a febrile disease, however theclinical status extends since asymptomatic infection until fatal disease.Clinical case: Male patient with prolonged fever and with negative proofs in the searchof hepatitis, malaria, dengue, leptospirosis and typhoid disease. In the peripheralblood smear with Wright’s stain, there were found granulocytes with intracytoplasmatic vacuoles called morulae (microcolonies of Anaplasma phagocytophilum. Diagnosticimpression was Human Granulocytic Anaplasmosis (HGA. Treatment with tetracyclinewas done and good evolution was reported.Conclusion: HGA should be a disease to have present in patients with prolonged febrileprofile. A detailed clinical history with information about exposure to ticks should begotten. It is important an early diagnosis to establish the differences with other similarinfections and to apply promptly indicated antibiotic therapy. Rev.cienc.biomed.2013;4(1:165-169RESUMEN:Introducción: Anaplasma phagocytophilum es una bacteria intracitoplasmática obligadaque pertenece al orden Rickettsiales, reconocido como causante de enfermedaden humanos. Pacientes sintomáticos reportan exposición a las garrapatas (vector unao dos semanas antes del inicio de la enfermedad. Presentan una enfermedad febril, noobstante el rango clínico se extiende desde infección asintomática hasta enfermedadfatal.Caso clínico: paciente masculino con cuadro febril prolongado. Pruebas negativas enla búsqueda de hepatitis, paludismo, dengue, leptospirosis y enfermedad tifoidea. Enel extendido de sangre periférica, con tinción de Wright se encontraron granulocitoscon vacuolas intracitoplasmáticas llamadas mórulas (microcolonias

  15. The first report of Anaplasma phagocytophilum and a novel Theileria spp. co-infection in a South African giraffe.

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    Zhang, Yan; Li, Tongyi; Cui, Yanyan; Wang, Jinhong; Lv, Yali; Wang, Rongjun; Jian, Fuchun; Zhang, Longxian; Wang, Jiantang; Yang, Guangcheng; Ning, Changshen

    2016-08-01

    Organisms of the genera Anaplasma and Theileria are important intracellular bacteria and parasites that cause various tick-borne diseases, threatening the health of numerous animals as well as human beings. In the present study, a 12-month-old male wild South African giraffe (Giraffa camelopardalis giraffa) originating from South Africa, and living in Zhengzhou Zoo (located in the urban district of Zhengzhou in the provincial capital of Henan), suddenly developed an unknown fatal disease and died 1day after the onset of the clinical signs. By microscopic examination of Giemsa-stained blood smears combined with nested PCR and DNA sequence analysis, Anaplasma phagocytophilum, Anaplasma bovis and a novel Theileria spp. were found in the blood of this giraffe. The six other Cervidae animals in the zoo and three ruminants living in the same colony house with them were found to be negative for both Anaplasma and Theileria in their blood specimens. We report on the first case of an A. phagocytophilum infection and the occurrence of a novel Theileria spp. in the blood of a giraffe. This is the first reported case of a multi-infection of A. bovis, A. phagocytophilum and Theileria spp. in a giraffe, as revealed by microscopic examination of blood smears and the results of nested PCR and DNA sequencing. PMID:27109775

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

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    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.

  17. The Prostaglandin E2-EP3 Receptor Axis Regulates Anaplasma phagocytophilum-Mediated NLRC4 Inflammasome Activation.

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    Wang, Xiaowei; Shaw, Dana K; Hammond, Holly L; Sutterwala, Fayyaz S; Rayamajhi, Manira; Shirey, Kari Ann; Perkins, Darren J; Bonventre, Joseph V; Velayutham, Thangam S; Evans, Sean M; Rodino, Kyle G; VieBrock, Lauren; Scanlon, Karen M; Carbonetti, Nicholas H; Carlyon, Jason A; Miao, Edward A; McBride, Jere W; Kotsyfakis, Michail; Pedra, Joao H F

    2016-08-01

    Rickettsial agents are sensed by pattern recognition receptors but lack pathogen-associated molecular patterns commonly observed in facultative intracellular bacteria. Due to these molecular features, the order Rickettsiales can be used to uncover broader principles of bacterial immunity. Here, we used the bacterium Anaplasma phagocytophilum, the agent of human granulocytic anaplasmosis, to reveal a novel microbial surveillance system. Mechanistically, we discovered that upon A. phagocytophilum infection, cytosolic phospholipase A2 cleaves arachidonic acid from phospholipids, which is converted to the eicosanoid prostaglandin E2 (PGE2) via cyclooxygenase 2 (COX2) and the membrane associated prostaglandin E synthase-1 (mPGES-1). PGE2-EP3 receptor signaling leads to activation of the NLRC4 inflammasome and secretion of interleukin (IL)-1β and IL-18. Importantly, the receptor-interacting serine/threonine-protein kinase 2 (RIPK2) was identified as a major regulator of the immune response against A. phagocytophilum. Accordingly, mice lacking COX2 were more susceptible to A. phagocytophilum, had a defect in IL-18 secretion and exhibited splenomegaly and damage to the splenic architecture. Remarkably, Salmonella-induced NLRC4 inflammasome activation was not affected by either chemical inhibition or genetic ablation of genes associated with PGE2 biosynthesis and signaling. This divergence in immune circuitry was due to reduced levels of the PGE2-EP3 receptor during Salmonella infection when compared to A. phagocytophilum. Collectively, we reveal the existence of a functionally distinct NLRC4 inflammasome illustrated by the rickettsial agent A. phagocytophilum. PMID:27482714

  18. The Prostaglandin E2-EP3 Receptor Axis Regulates Anaplasma phagocytophilum-Mediated NLRC4 Inflammasome Activation

    Science.gov (United States)

    Wang, Xiaowei; Shaw, Dana K.; Hammond, Holly L.; Sutterwala, Fayyaz S.; Rayamajhi, Manira; Shirey, Kari Ann; Perkins, Darren J.; Bonventre, Joseph V.; Velayutham, Thangam S.; Evans, Sean M.; Rodino, Kyle G.; VieBrock, Lauren; Scanlon, Karen M.; Carbonetti, Nicholas H.; Carlyon, Jason A.; Miao, Edward A.; McBride, Jere W.; Kotsyfakis, Michail

    2016-01-01

    Rickettsial agents are sensed by pattern recognition receptors but lack pathogen-associated molecular patterns commonly observed in facultative intracellular bacteria. Due to these molecular features, the order Rickettsiales can be used to uncover broader principles of bacterial immunity. Here, we used the bacterium Anaplasma phagocytophilum, the agent of human granulocytic anaplasmosis, to reveal a novel microbial surveillance system. Mechanistically, we discovered that upon A. phagocytophilum infection, cytosolic phospholipase A2 cleaves arachidonic acid from phospholipids, which is converted to the eicosanoid prostaglandin E2 (PGE2) via cyclooxygenase 2 (COX2) and the membrane associated prostaglandin E synthase-1 (mPGES-1). PGE2-EP3 receptor signaling leads to activation of the NLRC4 inflammasome and secretion of interleukin (IL)-1β and IL-18. Importantly, the receptor-interacting serine/threonine-protein kinase 2 (RIPK2) was identified as a major regulator of the immune response against A. phagocytophilum. Accordingly, mice lacking COX2 were more susceptible to A. phagocytophilum, had a defect in IL-18 secretion and exhibited splenomegaly and damage to the splenic architecture. Remarkably, Salmonella-induced NLRC4 inflammasome activation was not affected by either chemical inhibition or genetic ablation of genes associated with PGE2 biosynthesis and signaling. This divergence in immune circuitry was due to reduced levels of the PGE2-EP3 receptor during Salmonella infection when compared to A. phagocytophilum. Collectively, we reveal the existence of a functionally distinct NLRC4 inflammasome illustrated by the rickettsial agent A. phagocytophilum. PMID:27482714

  19. 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.

  20. 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

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

    Science.gov (United States)

    Oliva Chávez, Adela S; Fairman, James W; Felsheim, Roderick F; Nelson, Curtis M; Herron, Michael J; Higgins, LeeAnn; Burkhardt, Nicole Y; Oliver, Jonathan D; Markowski, Todd W; Kurtti, Timothy J; Edwards, Thomas E; Munderloh, Ulrike G

    2015-01-01

    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 mutations

  2. 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

  3. 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.

  4. 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

  5. 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.

  6. 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%.

  7. Molecular Survey of Anaplasma Species in Small Ruminants Reveals the Presence of Novel Strains Closely Related to A. phagocytophilum in Tunisia.

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    Ben Said, Mourad; Belkahia, Hanène; Alberti, Alberto; Zobba, Rosanna; Bousrih, Maha; Yahiaoui, Mouna; Daaloul-Jedidi, Monia; Mamlouk, Aymen; Gharbi, Mohamed; Messadi, Lilia

    2015-10-01

    A survey of Anaplasma species in small ruminants is still lacking in North African countries. In this study, the presence of A. phagocytophilum, A. phagocytophilum-related species, and A. ovis was investigated in a total of 563 healthy small ruminants (303 goats and 260 sheep), from 25 randomly selected flocks sampled in Tunisia. Anaplasma spp. and A. ovis overall infection rates were 95.0% and 93.8% in sheep and 69.6% and 65.3% in goats, respectively. A. phagocytophilum was not detected in any of tested animals. A total of 20 sheep (7.7%) and 144 goats (47.5%) were infected by Anaplasma strains genetically related to A. phagocytophilum. Both in sheep and goats A. ovis prevalence was higher in adults (≥2 years) than in young (rams (p = 0.010). The A. ovis infection rate was significantly lower in goats of the local breed (p = 0.049) and it was higher in goats infested by ticks than in not infested animals (p = 0.005). Genetic analysis of the msp4 gene of A. ovis indicated the presence of strains shared by Tunisian sheep and goats. Sequence analysis and phylogenetic studies on the basis of the 16S rRNA gene provided evidence for the circulation of at least two different potentially novel species genetically related to A. phagocytophilum in Tunisian small ruminants. These findings cause concern about specificity of serological tests used for detection of A. phagocytophilum in ruminants and provide additional information for elucidating pathogenesis and molecular epidemiology of A. phagocytophilum and related species. PMID:26394065

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

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    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

  9. Efficacy of sarolaner in the prevention of Borrelia burgdorferi and Anaplasma phagocytophilum transmission from infected Ixodes scapularis to dogs.

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    Honsberger, Nicole A; Six, Robert H; Heinz, Thomas J; Weber, Angela; Mahabir, Sean P; Berg, Thomas C

    2016-05-30

    The efficacy of sarolaner (Simparica™, Zoetis) to prevent transmission primarily of Borrelia burgdorferi and secondarily of Anaplasma phagocytophilum from infected wild-caught Ixodes scapularis to dogs was evaluated in a placebo-controlled laboratory study. Twenty-four purpose-bred laboratory Beagles seronegative for B. burgdorferi and A. phagocytophilum antibodies were allocated randomly to one of three treatment groups: placebo administered orally on Days 0 and 7, or sarolaner at 2mg/kg administered orally on Day 0 (28 days prior to tick infestation) or on Day 7 (21 days prior to tick infestation). On Day 28, each dog was infested with approximately 25 female and 25 male wild caught adult I. scapularis that were determined to have prevalence of 57% for B. burgdorferi and 6.7% for A. phagocytophilum by PCR. In situ tick counts were conducted on Days 29 and 30. On Day 33, all ticks were counted and removed. Acaricidal efficacy was calculated based on the reduction of geometric mean live tick counts in the sarolaner-treated groups compared to the placebo-treated group for each tick count. Blood samples collected from each dog on Days 27, 49, 63, 77, 91 and 104 were tested for the presence of B. burgdorferi and A. phagocytophilum antibodies using the SNAP(®) 4Dx(®) Plus Test, and quantitatively assayed for B. burgdorferi antibodies using an ELISA test. Skin biopsies collected on Day 104 were tested for the presence of B. burgdorferi by bacterial culture and PCR. Geometric mean live tick counts for placebo-treated dogs were 14.8, 12.8, and 19.1 on Days 29, 30, and 33, respectively. The percent reductions in mean live tick counts at 1, 2, and 5 days after infestation were 86.3%, 100%, and 100% for the group treated with sarolaner 21 days prior to infestation, and 90.9%, 97.1%, and 100% for the group treated with sarolaner 28 days prior to infestation. Geometric mean live tick counts for both sarolaner-treated groups were significantly lower than those for the

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

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    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

  11. 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.

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

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    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

  13. 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

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

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    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.

  15. 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.

  16. 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.

  17. 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.

  18. 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

  19. Are Apodemus spp. mice and Myodes glareolus reservoirs for Borrelia miyamotoi, Candidatus Neoehrlichia mikurensis, Rickettsia helvetica, R. monacensis and Anaplasma phagocytophilum?

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    Burri, C; Schumann, O; Schumann, C; Gern, L

    2014-04-01

    In Europe, in addition to Borrelia burgdorferi sensu lato and tick-borne encephalitis (TBE) virus, other zoonotic pathogens, like B. miyamotoi, a species related to the relapsing fever spirochaetes, Candidatus Neoehrlichia mikurensis (N. mikurensis), Rickettsia helvetica, Rickettsia monacensis, and Anaplasma phagocytophilum have been reported in the ixodid tick Ixodes ricinus. No study was conducted to identify reservoir hosts for these pathogens. Here, we investigated the role played by wild rodents in the natural transmission cycle of B. miyamotoi, N. mikurensis, R. helvetica, R. monacensis, and A. phagocytophilum in Switzerland. In 2011 and 2012, small mammals were captured in an area where these pathogens occur in questing ticks. Ixodes ricinus ticks infesting captured small mammals were analysed after their moult by PCR followed by reverse line blot to detect the different pathogens. Xenodiagnostic larvae were used to evaluate the role of rodents as reservoirs and analysed after their moult. Most of the 108 captured rodents (95.4%) were infested by I. ricinus ticks; 4.9%, 3.9%, 24.0%, and 0% of the rodents were infested by Borrelia, N. mikurensis, Rickettsia spp., and A. phagocytophilum-infected larvae, respectively. Borrelia afzelii, B. miyamotoi, N. mikurensis, Rickettsia spp., and A. phagocytophilum were detected in 2.8%, 0.17%, 2.6%, 6.8%, and 0% of the ticks attached to rodents, respectively. Borrelia afzelii was transmitted by 4 rodents to 41.2% of the xenodiagnostic ticks, B. miyamotoi by 3 rodents to 23.8%, and N. mikurensis was transmitted by 6 rodents to 41.0% of the xenodiagnostic ticks. None of the tested rodent transmitted Rickettsia spp. or A. phagocytophilum to I. ricinus xenodiagnostic larvae. This study showed that rodents are reservoir hosts for B. miyamotoi and N. mikurensis in Europe. PMID:24582511

  20. 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.

  1. Assessment of antibodies against surface and outer membrane proteins of Anaplasma phagocytophilum in Lyme borreliosis and tick-borne encephalitis paediatric patients.

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    Krbková, L; Homola, L; Hlaváčová, A; Mikolášek, P; Bednářová, J; Čermáková, Z

    2016-09-01

    To examine evidence of positive antibodies against immunogenic proteins of Anaplasma phagocytophilum in patients with other tick-borne infections and to diagnose possible co-infections, 412 serum specimens were tested by immunoblotting using three specific Anaplasma antigens: surface proteins p44 and Asp62 and outer membrane protein A (OmpA). In total, 284 serum samples from children with Lyme borreliosis and 12 serum samples from children with tick-borne encephalitis were tested. Sera from patients with viral aseptic meningitis (n = 47) and from blood donors (n = 69) were used as controls. Among all serum specimens from patients with tick-borne infections submitted for this study, six samples (2·0%) showed positive IgM reactions and seven samples (2·4%) were IgG positive for A. phagocytophilum by immunoblot. Borderline reactivity was found in 30 samples (10·14%) for IgM and 36 samples (12·2%) for IgG. The difference between patients and blood donors was statistically significant for IgM (P = 0·006) and for IgG (P = 0·0007) antibodies. A statistically significant result was obtained for IgG (P = 0·02) but not for IgM between patients and children with aseptic meningitis. Immunoblot using three specific antigens provides novel information about the positivity of antibodies to A. phagocytophilum in children with other tick-borne infections. Taking into account clinical and laboratory findings of children despite antibody positivity, no case of human granulocytic anaplasmosis was demonstrated. PMID:27180603

  2. A comparative study of clinical manifestations, haematological and serological responses after experimental infection with Anaplasma phagocytophilum in two Norwegian sheep breeds

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    Sandstedt Karin

    2011-02-01

    Full Text Available Abstract Background It has been questioned if the old native Norwegian sheep breed, Old Norse Sheep (also called Norwegian Feral Sheep, normally distributed on coastal areas where ticks are abundant, is more protected against tick-borne infections than other Norwegian breeds due to a continuously high selection pressure on pasture. The aim of the present study was to test this hypothesis in an experimental infection study. Methods Five-months-old lambs of two Norwegian sheep breeds, Norwegian White (NW sheep and Old Norse (ON sheep, were experimentally infected with a 16S rRNA genetic variant of Anaplasma phagocytophilum (similar to GenBank accession number M73220. The experiment was repeated for two subsequent years, 2008 and 2009, with the use of 16 lambs of each breed annually. Ten lambs of each breed were inoculated intravenously each year with 0.4 ml A. phagocytophilum-infected blood containing approximately 0.5 × 106 infected neutrophils/ml. Six lambs of each breed were used as uninfected controls. Half of the primary inoculated lambs in each breed were re-challenged with the same infectious dose at nine (2008 and twelve (2009 weeks after the first challenge. The clinical, haematological and serological responses to A. phagocytophilum infection were compared in the two sheep breeds. Results The present study indicates a difference in fever response and infection rate between breeds of Norwegian sheep after experimental infection with A. phagocytophilum. Conclusion Although clinical response seems to be less in ON-lambs compared to NW-lambs, further studies including more animals are needed to evaluate if the ON-breed is more protected against tick-borne infections than other Norwegian breeds.

  3. 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

  4. 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.

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

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    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.

  6. 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.

  7. 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

  8. 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.

  9. 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...

  10. 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.

  11. A serological survey of tick-borne pathogens in dogs in North America and the Caribbean as assessed by Anaplasma phagocytophilum, A. platys, Ehrlichia canis, E. chaffeensis, E. ewingii, and Borrelia burgdorferi species-specific peptides

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    Barbara A. Qurollo

    2014-10-01

    Full Text Available Introduction: Tick-borne pathogens cause a spectrum of disease manifestations in both dogs and humans. Recognizing regional and temporal shifts in exposure are important as tick distributions change. To better delineate regional exposure to canine tick-borne pathogens, an expanded set of species-specific peptides were used to detect Anaplasma phagocytophilum (Aph, Anaplasma platys (Apl, Ehrlichia canis (Ec, Ehrlichia chaffeensis (Ech, Ehrlichia ewingii (Eew, and Borrelia burgdorferi (Bb antibodies in canine serum. Methods: Archived canine serum samples (n=6,582 collected during 2008–2010 and in 2012 from the US, Canada, and the Caribbean were retrospectively screened for antibodies against Ehrlichia and Anaplasma species-specific peptides. Overall, regional and temporal seroprevalence rates were determined. Results: Overall Bb and Eew were the most seroprevalent pathogens. During 2008–2010, seroprevalence rates increased overall for Aph and Ech, and regionally, Bb and Aph seroprevalence rates increased in the South. Canada had unexpectedly high seroprevalence rates for Ec and Apl. The most common co-exposures were Eew+Ech, followed by Aph+Bb and Eew+Bb. Conclusions: This study demonstrated significant shifts in canine vector-borne disease seroprevalence rates. The use of specific peptides facilitated improved geographic delineation of tick-borne pathogen distributions among dogs, which may enhance epidemiological surveillance of vector-borne pathogens shared by dogs and humans.

  12. 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

  13. Reconstitution of Cholesterol-Dependent Vaginolysin into Tethered Phospholipid Bilayers

    DEFF Research Database (Denmark)

    Budvytyte, Rima; Pleckaityte, M.; Zvirbliene, A.;

    2013-01-01

    Functional reconstitution of the cholesterol-dependent cytolysin vaginolysin (VLY) from Gardnerella vaginalis into artificial tethered bilayer membranes (tBLMs) has been accomplished. The reconstitution of VLY was followed in real-time by electrochemical impedance spectroscopy (EIS). Changes of the...... EIS parameters of the tBLMs upon exposure to VLY solutions were consistent with the formation of water-filled pores in the membranes. It was found that reconstitution of VLY is a strictly cholesterol-dependent, irreversible process. At a constant cholesterol concentration reconstitution of VLY...... platform for the detection of the activity of VLY and possibly other cholesterol-dependent cytolysins....

  14. Cholesterol-dependent Conformational Plasticity in GPCR Dimers.

    Science.gov (United States)

    Prasanna, Xavier; Sengupta, Durba; Chattopadhyay, Amitabha

    2016-01-01

    The organization and function of the serotonin1A receptor, an important member of the GPCR family, have been shown to be cholesterol-dependent, although the molecular mechanism is not clear. We performed a comprehensive structural and dynamic analysis of dimerization of the serotonin1A receptor by coarse-grain molecular dynamics simulations totaling 3.6 ms to explore the molecular details of its cholesterol-dependent association. A major finding is that the plasticity and flexibility of the receptor dimers increase with increased cholesterol concentration. In particular, a dimer interface formed by transmembrane helices I-I was found to be sensitive to cholesterol. The modulation of dimer interface appears to arise from a combination of direct cholesterol occupancy and indirect membrane effects. Interestingly, the presence of cholesterol at the dimer interface is correlated with increased dimer plasticity and flexibility. These results represent an important step in characterizing the molecular interactions in GPCR organization with potential relevance to therapeutic interventions. PMID:27535203

  15. 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.

  16. 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. PMID:26951985

  17. Detection of Babesia and Anaplasma species in rabbits from Texas and Georgia, USA.

    Science.gov (United States)

    Yabsley, Michael J; Romines, Janean; Nettles, Victor F

    2006-01-01

    Rabbits have been shown to harbor a suite of zoonotic organisms, including a Babesia species, Borrelia burgdorferi, and Anaplasma phagocytophilum. In this study, we conducted a molecular survey for various tick-borne pathogens in three species of rabbits from Texas and Georgia. Of 18 black-tailed jackrabbits (Lepus californicus) tested from Texas, six (28%) were polymerase chain reaction (PCR) positive for Babesia, and nucleotide sequencing revealed two distinct species or strains. Two jackrabbits were infected with a Babesia species or strain (Babesia sp. A) that was nearly identical (99.9%) to a piroplasm previously detected in humans from Washington state, and the remaining four jackrabbits were infected with a Babesia species (Babesia sp. B) that was most similar (99.7%) to a Babesia species detected in cottontail rabbits from Massachusetts and humans from Kentucky and Missouri. Eleven (61%) black-tailed jackrabbits were positive for A. bovis, and one was positive for A. phagocytophilum. Two of four desert cottontails (Sylvilagus audubonii) from Texas were positive for the Babesia sp. B, and one desert cottontail each was positive for A. bovis and A. phagocytophilum. One of these desert cottontails was coinfected with the Babesia sp. B and A. phagocytophilum, and five jackrabbits were coinfected with Babesia species and A. bovis. Of 19 eastern cottontails (S. floridanus) from Georgia, only one (5.3%) was positive for A. phagocytophilum, and three (15.8%) were positive for A. bovis. No rabbits from Texas or Georgia were positive for Borrelia species. The only tick species detected on the Texas and Georgia rabbits was the rabbit tick, Haemaphysalis leporispalustris. These data extend the geographic and host range of these pathogens, and because both the Babesia species and A. phagocytophilum are potential zoonotic pathogens, it is important to be aware that these organisms are enzootic in parts of the southern United States. PMID:16584322

  18. Cholesterol-dependent hemolytic activity of Passiflora quadrangularis leaves

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    L.N. Yuldasheva

    2005-07-01

    Full Text Available Plants used in traditional medicine are rich sources of hemolysins and cytolysins, which are potential bactericidal and anticancer drugs. The present study demonstrates for the first time the presence of a hemolysin in the leaves of Passiflora quadrangularis L. This hemolysin is heat stable, resistant to trypsin treatment, has the capacity to froth, and acts very rapidly. The hemolysin activity is dose-dependent, with a slope greater than 1 in a double-logarithmic plot. Polyethylene glycols of high molecular weight were able to reduce the rate of hemolysis, while liposomes containing cholesterol completely inhibited it. In contrast, liposomes containing phosphatidylcholine were ineffective. The Passiflora hemolysin markedly increased the conductance of planar lipid bilayers containing cholesterol but was ineffective in cholesterol-free bilayers. Successive extraction of the crude hemolysin with n-hexane, chloroform, ethyl acetate, and n-butanol resulted in a 10-fold purification, with the hemolytic activity being recovered in the n-butanol fraction. The data suggest that membrane cholesterol is the primary target for this hemolysin and that several hemolysin molecules form a large transmembrane water pore. The properties of the Passiflora hemolysin, such as its frothing ability, positive color reaction with vanillin, selective extraction with n-butanol, HPLC profile, cholesterol-dependent membrane susceptibility, formation of a stable complex with cholesterol, and rapid erythrocyte lysis kinetics indicate that it is probably a saponin.

  19. Migratory and carnivorous birds in Brazil: reservoirs for Anaplasma and Ehrlichia species?

    Science.gov (United States)

    Machado, Rosangela Zacarias; André, Marcos Rogério; Werther, Karin; de Sousa, Eliane; Gavioli, Fernando Antônio; Alves Junior, José Roberto Ferreira

    2012-08-01

    In order to investigate new hosts for Anaplasmataceae agents in Brazil, we collected blood samples from 21 wild birds. Using molecular techniques, we detected the presence of Anaplasma phagocytophilum, Ehrlichia chaffeensis, and an Ehrlichia species closely related to Ehrlichia canis in carnivorous avian blood samples. In addition, an Ehrlichia species closely related to an Ehrlichia species found in wild felines in Brazil was also detected in a goose blood sample. Wild birds may play a role as carriers of Anaplasmataceae agents in Brazil. PMID:22607070

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

    Directory of Open Access Journals (Sweden)

    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. Sequence analysis of the msp4 gene of Anaplasma ovis strains

    Science.gov (United States)

    de la Fuente, J.; Atkinson, M.W.; Naranjo, V.; Fernandez de Mera, I. G.; Mangold, A.J.; Keating, K.A.; Kocan, K.M.

    2007-01-01

    Anaplasma ovis (Rickettsiales: Anaplasmataceae) is a tick-borne pathogen of sheep, goats and wild ruminants. The genetic diversity of A. ovis strains has not been well characterized due to the lack of sequence information. In this study, we evaluated bighorn sheep (Ovis canadensis) and mule deer (Odocoileus hemionus) from Montana for infection with A. ovis by serology and sequence analysis of the msp4 gene. Antibodies to Anaplasma spp. were detected in 37% and 39% of bighorn sheep and mule deer analyzed, respectively. Four new msp4 genotypes were identified. The A. ovis msp4 sequences identified herein were analyzed together with sequences reported previously for the characterization of the genetic diversity of A. ovis strains in comparison with other Anaplasma spp. The results of these studies demonstrated that although A. ovis msp4 genotypes may vary among geographic regions and between sheep and deer hosts, the variation observed was less than the variation observed between A. marginale and A. phagocytophilum strains. The results reported herein further confirm that A. ovis infection occurs in natural wild ruminant populations in Western United States and that bighorn sheep and mule deer may serve as wildlife reservoirs of A. ovis. ?? 2006.

  2. 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.

  3. Prevalence of Anaplasma, Bartonella and Borrelia Species in Haemaphysalis longicornis collected from goats in North Korea.

    Science.gov (United States)

    Kang, Jun-Gu; Ko, Sungjin; Smith, W Barney; Kim, Heung-Chul; Lee, In-Yong; Chae, Joon-Seok

    2016-06-30

    North Korea is located on the northern part of the Korean Peninsula in East Asia. While tick-borne pathogens of medical and veterinary importance have been reported from China and South Korea, they have not been reported from North Korea. To screen for zoonotic tick-borne pathogens in North Korea, ticks were collected from domestic goats. A total of 292 (27 nymph, 26 male, 239 female) Haemaphysalis (H.) longicornis were collected and assayed individually for selected tick-borne pathogens. A total of 77 (26.4%) were positive for Anaplasma bovis, followed by Bartonella (B.) grahamii (15, 5.1%), Anaplasma phagocytophilum (12, 4.1%), Bartonella henselae (10, 3.4%), and Borrelia spp. (3, 1.0%) based on 16S ribosomal RNA and ITS species-specific nested polymerase chain reaction. Using the groEL-based nested PCR, a total of 6 and 1 H. longicornis were positive for B. grahamii and B. henselae, respectively. All products were sequenced and demonstrated 100% identity and homology with previously reported sequences from other countries in GenBank. This is the first report of the detection of tick-borne pathogens in the North Korea and suggests that farm animals may act as reservoirs for zoonotic tick-borne pathogens. PMID:26645342

  4. Serological and molecular investigation of Ehrlichia spp. and Anaplasma spp. in ticks and blood of dogs, in the Thrace Region of Turkey.

    Science.gov (United States)

    Çetinkaya, Handan; Matur, Erdal; Akyazi, İbrahim; Ekiz, Elif Ergul; Aydin, Levent; Toparlak, Mufit

    2016-07-01

    In recent years, tick-borne diseases like ehrlichiosis and anaplasmosis became widespread worldwide threatening the health of both human and companion animals. Therefore, the aim of this study was to determine the presence of Anaplasma spp., and Ehrlichia spp. in dogs and ticks in the Thrace Region of Turkey. A total of 400 blood samples and 912 ticks were collected from dogs living in shelters that are located in four cities (Istanbul, Edirne, Tekirdag and Kirklareli) of the Thrace Region. Blood and buffy coat smears were prepared for microscopic examination. Hematologic and serologic analyses were performed using cell counter and commercial Snap3Dx test kit, respectively. Eight hundred fifty of collected ticks were classified as Rhipicephalus sanguineus, 33 as Rhipicephalus turanicus and 29 as Ixodes ricinus. After DNA extraction from blood samples and pooled ticks (127 tick pools, in total), nested PCR was performed to detect the DNA of Anaplasma spp., and Ehrlichia spp. The seroprevalence of Ehrlichia canis was 27.25% (109) by Snap3Dx test and the total molecular positivity was 11.75% (47) in dog blood samples and 21.25% (27) in tick pools by nested PCR. The frequencies of the infected blood samples with E. canis, Anaplasma phagocytophilum and Anaplasma platys were detected as 6%, 4% and 6%, respectively. E. canis and A. platys were detected in R. sanguineus pools with a ratio of 15.75% and 0.7%, respectively. In addition, A. platys was also detected in R. turanicus pools (0.7%). A. phagocytophilum was found only in I. ricinus pools (3.93%). Morulae of three species were detected in buffy coat and blood smears. While anemia was observed in dogs infected with E. canis and co-infected (with one or more species), thrombocytopenia was observed only in co-infected dogs. This is the first study providing evidence for the presence of Anaplasma spp. and Ehrlichia spp. in dogs and ticks in the Thrace Region of Turkey. Based on the results of the tests used in this study

  5. A new model for pore formation by cholesterol-dependent cytolysins.

    Directory of Open Access Journals (Sweden)

    Cyril F Reboul

    2014-08-01

    Full Text Available Cholesterol Dependent Cytolysins (CDCs are important bacterial virulence factors that form large (200-300 Å membrane embedded pores in target cells. Currently, insights from X-ray crystallography, biophysical and single particle cryo-Electron Microscopy (cryo-EM experiments suggest that soluble monomers first interact with the membrane surface via a C-terminal Immunoglobulin-like domain (Ig; Domain 4. Membrane bound oligomers then assemble into a prepore oligomeric form, following which the prepore assembly collapses towards the membrane surface, with concomitant release and insertion of the membrane spanning subunits. During this rearrangement it is proposed that Domain 2, a region comprising three β-strands that links the pore forming region (Domains 1 and 3 and the Ig domain, must undergo a significant yet currently undetermined, conformational change. Here we address this problem through a systematic molecular modeling and structural bioinformatics approach. Our work shows that simple rigid body rotations may account for the observed collapse of the prepore towards the membrane surface. Support for this idea comes from analysis of published cryo-EM maps of the pneumolysin pore, available crystal structures and molecular dynamics simulations. The latter data in particular reveal that Domains 1, 2 and 4 are able to undergo significant rotational movements with respect to each other. Together, our data provide new and testable insights into the mechanism of pore formation by CDCs.

  6. The Bacillus anthracis cholesterol-dependent cytolysin, Anthrolysin O, kills human neutrophils, monocytes and macrophages

    Directory of Open Access Journals (Sweden)

    Rest Richard F

    2006-06-01

    Full Text Available Abstract Background Bacillus anthracis is an animal and human pathogen whose virulence is characterized by lethal and edema toxin, as well as a poly-glutamic acid capsule. In addition to these well characterized toxins, B. anthracis secretes several proteases and phospholipases, and a newly described toxin of the cholesterol-dependent cytolysin (CDC family, Anthrolysin O (ALO. Results In the present studies we show that recombinant ALO (rALO or native ALO, secreted by viable B. anthracis, is lethal to human primary polymorphonuclear leukocytes (PMNs, monocytes, monocyte-derived macrophages (MDMs, lymphocytes, THP-1 monocytic human cell line and ME-180, Detroit 562, and A549 epithelial cells by trypan blue exclusion or lactate dehydrogenase (LDH release viability assays. ALO cytotoxicity is dose and time dependent and susceptibility to ALO-mediated lysis differs between cell types. In addition, the viability of monocytes and hMDMs was assayed in the presence of vegetative Sterne strains 7702 (ALO+, UT231 (ALO-, and a complemented strain expressing ALO, UT231 (pUTE544, and was dependent upon the expression of ALO. Cytotoxicity of rALO is seen as low as 0.070 nM in the absence of serum. All direct cytotoxic activity is inhibited by the addition of cholesterol or serum concentration as low as 10%. Conclusion The lethality of rALO and native ALO on human monocytes, neutrophils, macrophages and lymphocytes supports the idea that ALO may represent a previously unidentified virulence factor of B. anthracis. The study of other factors produced by B. anthracis, along with the major anthrax toxins, will lead to a better understanding of this bacterium's pathogenesis, as well as provide information for the development of antitoxin vaccines for treating and preventing anthrax.

  7. Arcanolysin is a cholesterol-dependent cytolysin of the human pathogen Arcanobacterium haemolyticum

    Directory of Open Access Journals (Sweden)

    Ratner Adam J

    2011-10-01

    Full Text Available Abstract Background Arcanobacterium haemolyticum is an emerging human pathogen that causes pharyngitis, wound infections, and a variety of occasional invasive diseases. Since its initial discovery in 1946, this Gram positive organism has been known to have hemolytic activity, yet no hemolysin has been previously reported. A. haemolyticum also displays variable hemolytic activity on laboratory blood agar that is dependent upon which species the blood is derived. Results Here we describe a cholesterol-dependent cytolysin (CDC secreted by A. haemolyticum, designated arcanolysin (aln, which is present in all strains (n = 52 tested by DNA dot hybridization. Among the known CDCs, ALN is most closely related to pyolysin (PLO from Trueperella (formerly Arcanobacterium pyogenes. The aln probe, however, did not hybridize to DNA from T. pyogenes. The aln open reading frame has a lower mol %G+C (46.7% than the rest of the A. haemolyticum genome (53.1% and is flanked by two tRNA genes, consistent with probable acquisition by horizontal transfer. The ALN protein (~ 64 kDa contains a predicted signal sequence, a putative PEST sequence, and a variant undecapeptide within domain 4, which is typically important for function of the toxins. The gene encoding ALN was cloned and expressed in Escherichia coli as a functional recombinant toxin. Recombinant ALN had hemolytic activity on erythrocytes and cytolytic activity on cultured cells from human, rabbit, pig and horse origins but was poorly active on ovine, bovine, murine, and canine cells. ALN was less sensitive to inhibition by free cholesterol than perfringolysin O, consistent with the presence of the variant undecapeptide. Conclusions ALN is a newly identified CDC with hemolytic activity and unique properties in the CDC family and may be a virulence determinant for A. haemolyticum.

  8. The cholesterol-dependent cytolysin family of gram-positive bacterial toxins.

    Science.gov (United States)

    Heuck, Alejandro P; Moe, Paul C; Johnson, Benjamin B

    2010-01-01

    The cholesterol-dependent cytolysins (CDCs) are a family of beta-barrel pore-forming toxins secreted by Gram-positive bacteria. These toxins are produced as water-soluble monomeric proteins that after binding to the target cell oligomerize on the membrane surface forming a ring-like pre-pore complex, and finally insert a large beta-barrel into the membrane (about 250 A in diameter). Formation of such a large transmembrane structure requires multiple and coordinated conformational changes. The presence of cholesterol in the target membrane is absolutely required for pore-formation, and therefore it was long thought that cholesterol was the cellular receptor for these toxins. However, not all the CDCs require cholesterol for binding. Intermedilysin, secreted by Streptoccocus intermedius only binds to membranes containing a protein receptor, but forms pores only if the membrane contains sufficient cholesterol. In contrast, perfringolysin O, secreted by Clostridium perfringens, only binds to membranes containing substantial amounts of cholesterol. The mechanisms by which cholesterol regulates the cytolytic activity of the CDCs are not understood at the molecular level. The C-terminus of perfringolysin O is involved in cholesterol recognition, and changes in the conformation of the loops located at the distal tip of this domain affect the toxin-membrane interactions. At the same time, the distribution of cholesterol in the membrane can modulate toxin binding. Recent studies support the concept that there is a dynamic interplay between the cholesterol-binding domain of the CDCs and the excess of cholesterol molecules in the target membrane. PMID:20213558

  9. 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.

  10. Molecular Characterization of Anaplasma and Ehrlichia Species in Different Cattle Breeds and Age Groups in Mbarara District (Western Uganda

    Directory of Open Access Journals (Sweden)

    Muhanguzi Dennis

    2010-07-01

    Full Text Available Anaplasma and Ehrlichia sp. (AEs cause significant economic losses to the livestock sector in Uganda. The aim of this study was to determine the prevalence of AEs in cattle from Kashaari county-Mbarara district (Uganda so as to compare the prevalence of AEs in different cattle breeds, age groups, sub county of origin and management systems and predictor(s of infection with AEs. Such information is deemed necessary to direct future tick-borne disease control programs. A single pair of primers w as used to am plify a 492-498bp fragment of the 16SRNA gene spanning the V1 region conserved for both AEs. PCR products were transferred onto the Reverse Line Blot (RLB membrane and AEs amplicons in the PCR products allowed to hybridize with AE species-specific oligonucleotides. The prevalence of Ehrlichia sp. was 5.1% (CI = 95%, 2.9-7.3% whereas that of Anaplasma species was 5.3 % (C I = 95%, 3-7.6%. Individual AEs detected include; A. bovis (5.1%, CI = 95%, 2.9-7.3%, E. ruminantium (4.5% , CI = 95%, 2.4-6.6%, A. marginale (3.7%, CI = 95%, 1.8-5.6%, A. (E. phagocytophilum (2.7%, CI = 95%, 1.1-4.3% , E. ovina/canis (2.7%, CI = 95%; 1.1-4.3%, E. sp. (omatjenne(1.9%, CI = 95 %, 0.5-3.3%. Cattle breed was found to be the best predictor of infection. To further understand bovine tick-borne parasites in Uganda, we recommend that studies covering a wider area and over longer periods, investigation of breed as a predictor of infection, molecular genetic characterization, transmission and pathogenicity studies on the different strains of AEs be carried out.

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

    Science.gov (United States)

    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. PMID:26611960

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

    Science.gov (United States)

    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.

  13. Cellular Functions and X-ray Structure of Anthrolysin O, a Cholesterol-dependent Cytolysin Secreted by Bacillus anthracis

    Energy Technology Data Exchange (ETDEWEB)

    Bourdeau, Raymond W.; Malito, Enrico; Chenal, Alexandre; Bishop, Brian L.; Musch, Mark W.; Villereal, Mitch L.; Chang, Eugene B.; Mosser, Elise M.; Rest, Richard F.; Tang, Wei-Jen; (CNRS-UMR); (Drexel-MED); (UC)

    2009-06-02

    Anthrolysin O (ALO) is a pore-forming, cholesterol-dependent cytolysin (CDC) secreted by Bacillus anthracis, the etiologic agent for anthrax. Growing evidence suggests the involvement of ALO in anthrax pathogenesis. Here, we show that the apical application of ALO decreases the barrier function of human polarized epithelial cells as well as increases intracellular calcium and the internalization of the tight junction protein occludin. Using pharmacological agents, we also found that barrier function disruption requires increased intracellular calcium and protein degradation. We also report a crystal structure of the soluble state of ALO. Based on our analytical ultracentrifugation and light scattering studies, ALO exists as a monomer. Our ALO structure provides the molecular basis as to how ALO is locked in a monomeric state, in contrast to other CDCs that undergo antiparallel dimerization or higher order oligomerization in solution. ALO has four domains and is globally similar to perfringolysin O (PFO) and intermedilysin (ILY), yet the highly conserved undecapeptide region in domain 4 (D4) adopts a completely different conformation in all three CDCs. Consistent with the differences within D4 and at the D2-D4 interface, we found that ALO D4 plays a key role in affecting the barrier function of C2BBE cells, whereas PFO domain 4 cannot substitute for this role. Novel structural elements and unique cellular functions of ALO revealed by our studies provide new insight into the molecular basis for the diverse nature of the CDC family.

  14. Differential endometrial cell sensitivity to a cholesterol-dependent cytolysin links Trueperella pyogenes to uterine disease in cattle.

    Science.gov (United States)

    Amos, Matthew R; Healey, Gareth D; Goldstone, Robert J; Mahan, Suman M; Düvel, Anna; Schuberth, Hans-Joachim; Sandra, Olivier; Zieger, Peter; Dieuzy-Labaye, Isabelle; Smith, David G E; Sheldon, Iain Martin

    2014-03-01

    Purulent disease of the uterus develops in 40% of dairy cows after parturition, when the epithelium of the endometrium is disrupted to expose the underlying stroma to bacteria. The severity of endometrial pathology is associated with isolation of Trueperella pyogenes. In the present study, T. pyogenes alone caused uterine disease when infused into the uterus of cattle where the endometrial epithelium was disrupted. The bacterium secretes a cholesterol-dependent cytolysin, pyolysin (PLO), and the plo gene was identical and the plo gene promoter was highly similar amongst 12 clinical isolates of T. pyogenes. Bacteria-free filtrates of the T. pyogenes cultures caused hemolysis and endometrial cytolysis, and PLO was the main cytolytic agent, because addition of anti-PLO antibody prevented cytolysis. Similarly, a plo-deletion T. pyogenes mutant did not cause hemolysis or endometrial cytolysis. Endometrial stromal cells were notably more sensitive to PLO-mediated cytolysis than epithelial or immune cells. Stromal cells also contained more cholesterol than epithelial cells, and reducing stromal cell cholesterol content using cyclodextrins protected against PLO. Although T. pyogenes or plo-deletion T. pyogenes stimulated accumulation of inflammatory mediators, such as IL-1beta, IL-6, and IL-8, from endometrium, PLO did not stimulate inflammatory responses by endometrial or hematopoietic cells, or in vitro organ cultures of endometrium. The marked sensitivity of stromal cells to PLO-mediated cytolysis provides an explanation for how T. pyogenes acts as an opportunistic pathogen to cause pathology of the endometrium once the protective epithelium is lost after parturition.

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

    Science.gov (United States)

    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.

  16. Complete Genome Sequence of Anaplasma marginale subsp. centrale

    Science.gov (United States)

    Anaplasma marginale subsp. centrale is a naturally attenuated subtype that has been used as a vaccine for a century. We sequenced the genome of this organism and compared it to those of virulent senso stricto A. marginale strains. The comparison markedly narrows the number of outer membrane protein ...

  17. Genetic diversity of anaplasma species major surface proteins and implications for anaplasmosis serodiagnosis and vaccine development

    OpenAIRE

    de la Fuente, J.; Lew, A.; Lutz, H.; Meli, M. L.; Hofmann-Lehmann, R.; Shkap, V; Molad, T; Mangold, A J; Almazán, C; Naranjo, V.; Gortázar, C.; Torina, A; Caracappa, S.; García-Pérez, A. L.; Barral, M.

    2005-01-01

    The genus Anaplasma (Rickettsiales: Anaplasmataceae) includes several pathogens of veterinary and human medical importance. An understanding of the diversity of Anaplasma major surface proteins (MSPs), including those MSPs that modulate infection, development of persistent infections, and transmission of pathogens by ticks, is derived in part, by characterization and phylogenetic analyses of geographic strains. Information concerning the genetic diversity of Anaplasma spp. MSPs will likely in...

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

    Science.gov (United States)

    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. PMID:27394441

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

    Science.gov (United States)

    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. PMID:27177475

  20. The cholesterol-dependent cytolysin signature motif: a critical element in the allosteric pathway that couples membrane binding to pore assembly.

    Directory of Open Access Journals (Sweden)

    Kelley J Dowd

    Full Text Available The cholesterol-dependent cytolysins (CDCs constitute a family of pore-forming toxins that contribute to the pathogenesis of a large number of Gram-positive bacterial pathogens.The most highly conserved region in the primary structure of the CDCs is the signature undecapeptide sequence (ECTGLAWEWWR. The CDC pore forming mechanism is highly sensitive to changes in its structure, yet its contribution to the molecular mechanism of the CDCs has remained enigmatic. Using a combination of fluorescence spectroscopic methods we provide evidence that shows the undecapeptide motif of the archetype CDC, perfringolysin O (PFO, is a key structural element in the allosteric coupling of the cholesterol-mediated membrane binding in domain 4 (D4 to distal structural changes in domain 3 (D3 that are required for the formation of the oligomeric pore complex. Loss of the undecapeptide function prevents all measurable D3 structural transitions, the intermolecular interaction of membrane bound monomers and the assembly of the oligomeric pore complex. We further show that this pathway does not exist in intermedilysin (ILY, a CDC that exhibits a divergent undecapeptide and that has evolved to use human CD59 rather than cholesterol as its receptor. These studies show for the first time that the undecapeptide of the cholesterol-binding CDCs forms a critical element of the allosteric pathway that controls the assembly of the pore complex.

  1. Down-regulation of lipid raft-associated onco-proteins via cholesterol-dependent lipid raft internalization in docosahexaenoic acid-induced apoptosis.

    Science.gov (United States)

    Lee, Eun Jeong; Yun, Un-Jung; Koo, Kyung Hee; Sung, Jee Young; Shim, Jaegal; Ye, Sang-Kyu; Hong, Kyeong-Man; Kim, Yong-Nyun

    2014-01-01

    Lipid rafts, plasma membrane microdomains, are important for cell survival signaling and cholesterol is a critical lipid component for lipid raft integrity and function. DHA is known to have poor affinity for cholesterol and it influences lipid rafts. Here, we investigated a mechanism underlying the anti-cancer effects of DHA using a human breast cancer cell line, MDA-MB-231. We found that DHA decreased cell surface levels of lipid rafts via their internalization, which was partially reversed by cholesterol addition. With DHA treatment, caveolin-1, a marker for rafts, and EGFR were colocalized with LAMP-1, a lysosomal marker, in a cholesterol-dependent manner, indicating that DHA induces raft fusion with lysosomes. DHA not only displaced several raft-associated onco-proteins, including EGFR, Hsp90, Akt, and Src, from the rafts but also decreased total levels of those proteins via multiple pathways, including the proteasomal and lysosomal pathways, thereby decreasing their activities. Hsp90 overexpression maintained its client proteins, EGFR and Akt, and attenuated DHA-induced cell death. In addition, overexpression of Akt or constitutively active Akt attenuated DHA-induced apoptosis. All these data indicate that the anti-proliferative effect of DHA is mediated by targeting of lipid rafts via decreasing cell surface lipid rafts by their internalization, thereby decreasing raft-associated onco-proteins via proteasomal and lysosomal pathways and decreasing Hsp90 chaperone function.

  2. Human cytomegalovirus entry into dendritic cells occurs via a macropinocytosis-like pathway in a pH-independent and cholesterol-dependent manner.

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    Fabienne Haspot

    Full Text Available Human cytomegalovirus (HCMV is a ubiquitous herpesvirus that is able to infect fibroblastic, epithelial, endothelial and hematopoietic cells. Over the past ten years, several groups have provided direct evidence that dendritic cells (DCs fully support the HCMV lytic cycle. We previously demonstrated that the C-type lectin dendritic cell-specific intercellular adhesion molecule-3-grabbing non-integrin (DC-SIGN has a prominent role in the docking of HCMV on monocyte-derived DCs (MDDCs. The DC-SIGN/HCMV interaction was demonstrated to be a crucial and early event that substantially enhanced infection in trans, i.e., from one CMV-bearing cell to another non-infected cell (or trans-infection, and rendered susceptible cells fully permissive to HCMV infection. Nevertheless, nothing is yet known about how HCMV enters MDDCs. In this study, we demonstrated that VHL/E HCMV virions (an endothelio/dendrotropic strain are first internalized into MDDCs by a macropinocytosis-like process in an actin- and cholesterol-dependent, but pH-independent, manner. We observed the accumulation of virions in large uncoated vesicles with endosomal features, and the virions remained as intact particles that retained infectious potential for several hours. This trans-infection property was specific to MDDCs because monocyte-derived macrophages or monocytes from the same donor were unable to allow the accumulation of and the subsequent transmission of the virus. Together, these data allowed us to delineate the early mechanisms of the internalization and entry of an endothelio/dendrotropic HCMV strain into human MDDCs and to propose that DCs can serve as a "Trojan horse" to convey CMV from entry sites to other locations that may favor the occurrence of either latency or acute infection.

  3. Molecular study on infection rates of Anaplasma ovis and Anaplasma marginale in sheep and cattle in West-Azerbaijan province, Iran.

    Science.gov (United States)

    Noaman, Vahid; Bastani, Davood

    2016-01-01

    This study was carried out to determine the presence and frequency of Anaplasma ovis and Anaplasma marginale in sheep and dairy cattle in West-Azerbaijan province, Iran. A total number of 200 blood samples were randomly collected via the jugular vein from apparently healthy cattle (100) and sheep (100). The extracted DNA from blood cells was screened using genus-specific (Anaplasma spp.) nested-polymerase chain reaction (PCR) based on 16S rRNA gene primer sets. Species-specific PCR was set up using major surface protein 4 (MSP4) gene primer set. None of cattle blood samples were positive for Anaplasma spp. by the first nested PCR. Five samples among the 100 sheep blood samples were both positive in the first nested PCR and A. ovis -specific PCR, based on MSP4 gene. In total, 5.00% of animals were A. ovis positive. This study identified a low prevalence of A. ovis in the blood of apparently healthy sheep in West Azerbaijan province. PMID:27482362

  4. 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.

  5. 新疆和硕地区荒漠景观中无形体感染的流行病学调查%Prevalence of Anaplasma spp. infection in a desert landscape region of Heshuo, Xinjiang

    Institute of Scientific and Technical Information of China (English)

    张桂林; 孙响; 赵焱; 刘晓明; 郑重; 孙毅; 刘然

    2013-01-01

    Objective To study the prevalence ot Anaptasmosts among human,domestic sheep and tick population in Heshuo area,southern Xinjiang and to investigate the diversity of Anaplasma species.Methods Ticks were captured from wild field and blood samples were collected from healthy residents and their domestic sheep.Indirect fluorescent assay was carried out to determine the presence of Anaplasma specific IgG antibodies in blood sample of human and goats,respectively.Nested PCR and sequence alignment of Anaplasma partial 16S rDNA were used to investigate the diversity of Anaplasma species.Results 43.31% (55/127) of human beings and 27.50% (55/200) of the goats were found positive forAnaplasma specific IgG.In total,367 ticks were captured,including 3 genus and 4 species,which mainly consisting of dominate Hyalomma (H.)asiaticum (47.41%) and Rhipicephalus (R.)pumilio (37.60%).5.00% (18/360) of the questing ticks and 4.49% (7/156) of the goat blood samples were found to have had 16S rDNA,representing Anaplasma sp.by nested PCR,but none was found from human beings.Results from sequencial alignment revealed that the positive amplicons were identified to be Anaplasma phagocytophilum (99.2% GU046565,99.5% GU064897 and 99.5% AB196721)and Anaplasma central (99.2%GU064903).Conclusion Human and zoonotic anaplasmosis,which were probably transmitted by H.asiaticum and R.pumilio,were co-circulating in the desert landscape of Heshuo area,Xinjiang.%目的 明确无形体病在新疆南部和硕地区人群、山羊与媒介蜱种群中流行状况,探明该地区无形体种属多样性.方法 采集当地健康人和山羊的血液样本,捕捉蜱样本;利用间接免疫荧光方法检测血清无形体特异性IgG抗体;通过巢式PCR方法检测无形体16S rDNA基因片段,通过序列比对分析确定无形体种属特征.结果 和硕地区农牧民、山羊血清中无形体特异性IgG抗体阳性率分别为43.31%(55/127)和27.50%(55/200)

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

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    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.

  7. 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

  8. 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 mature adults. No evidence was found within this herd to suggest increased risk for clinical bovine erythrocytic anaplasmosis. PMID:27656312

  9. 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.

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

    Science.gov (United States)

    Tucker, Thomas R; Aly, Sharif S; Maas, John; Davy, Josh S; 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 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

  11. Comparison of the Efficiency of Biological Transmission of Anaplasma marginale (Rickettsiales: Anaplasmataceae) by Dermacentor andersoni Stiles (Acari: Ixodidae) with Mechanical Transmission by the Horse Fly, Tabanus fuscicos

    Science.gov (United States)

    Mechanical transmission of Anaplasma marginale by horse flies (Tabanidae) is thought to be epidemiologically significant in some areas of the US. We compared the relative efficiencies of mechanical transmission of Anaplasma marginale by the horse fly, Tabanus fuscicostatus Hine during acute infectio...

  12. Evaluation de la participation d'Anaplasma phagocytophilum dans le syndrome "fièvre des montagnes" ou Belar Joa des ovins du Pays Basque français

    OpenAIRE

    Razimbaud, Fabrice

    2006-01-01

    Après une présentation des principales hémoparasitoses ovines, les méthodes de diagnostic épidémio-clinique et de laboratoire sont développées. L'impact des conditions climatiques et écologiques du milieu sur l'abondance des tiques vectrices d'hémoparasites est abordé. Le Belar Joa (en basque) ou "fièvre des montagnes" correspond à une maladie touchant les Ovins lors de leur première estive. Cette maladie est connue depuis longtemps par les bergers, mais son origine n'a jamais été élucidée....

  13. 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.

  14. First Molecular Characterization of Anaplasma marginale in Cattle and Rhipicephalus (Boophilus) microplus Ticks in Cebu, Philippines

    OpenAIRE

    YBAÑEZ, Adrian Patalinghug; Sivakumar, Thillaiampalam; YBAÑEZ, Rochelle Haidee Daclan; RATILLA, Jowarren Catingan; PEREZ, Zandro Obligado; GABOTERO, Shirleny Reyes; Hakimi, Hassan; KAWAZU, Shin-Ichiro; MATSUMOTO, Kotaro; Yokoyama, Naoaki; INOKUMA, Hisashi; 河津, 信一郎; 松本, 高太郎; 横山, 直明; 猪熊,壽

    2013-01-01

    Anaplasma marginale has been detected in the Philippines only by peripheral blood smear examination and serological methods. This study generally aimed to molecularly detect and characterize A. marginale in cattle and ticks in Cebu, Philippines. A total of 12 bovine blood samples and 60 Rhipicephalus (Boophilus) microplus ticks were collected on the Cebu Island in 2011. 16S rRNA-based screening-PCR and DNA sequencing revealed 8 cattle (66.7%) and 8 ticks (13.3%) to be positive for A. marginal...

  15. 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.

  16. Molecular characterization of two different strains of haemotropic mycoplasmas from a sheep flock with fatal haemolytic anaemia, concomitant Anaplasma ovis infection

    OpenAIRE

    Hornok, Sándor; Meli, Marina L; Erdős, András; Hajtós, István; Lutz, Hans; Hofmann-Lehmann, Regina

    2009-01-01

    Molecular characterization of two different strains of haemotropic mycoplasmas from a sheep flock with fatal haemolytic anaemia, concomitant Anaplasma ovis infection HUNGARY (Hornok, Sandor) HUNGARY Received: 2008-08-03 Revised: 2008-10-27 Accepted: 2008-10-29

  17. 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.

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

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    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.

  19. Molecular evidence of potential novel spotted fever group rickettsiae, Anaplasma and Ehrlichia species in Amblyomma ticks parasitizing wild snakes

    OpenAIRE

    Kho, Kai Ling; Koh, Fui Xian; Tay, Sun Tee

    2015-01-01

    Background Amblyomma ticks parasitize a wide range of animals in tropical regions. This study describes the identification of Amblyomma ticks from wild snakes in Malaysia and the detection of potential human pathogens such as Rickettsia, Anaplasma, Ehrlichia and bartonellae in the ticks. Findings Twenty one adult ticks (twelve A. varanense and nine Amblyomma helvolum ticks) identified from seven Python molurus snakes in Sepang and a pool of six A. helvolum ticks from a Naja sumatrana snake in...

  20. Evaluating the effectiveness of an inactivated vaccine from Anaplasma marginale derived from tick cell culture.

    Science.gov (United States)

    Lasmar, Pedro Veloso Facury; Carvalho, Antônio Último de; Facury Filho, Elias Jorge; Bastos, Camila Valgas; Ribeiro, Múcio Flávio Barbosa

    2012-01-01

    The protective efficacy of an inactivated vaccine from Anaplasma marginale that was cultured in tick cells (IDE8) for use against bovine anaplasmosis was evaluated. Five calves (Group 1) were inoculated subcutaneously, at 21-day intervals, with three doses of vaccine containing 3 × 10(9) A. marginale initial bodies. Five control calves received saline solution alone (Group 2). Thirty-two days after the final inoculation, all the calves were challenged with approximately 3 × 10(5) erythrocytes infected with A. marginale high-virulence isolate (UFMG2). The Group 1 calves seroconverted 14 days after the second dose of vaccine. After the challenge, all the animals showed patent rickettsemia. There was no significant difference (p > 0.05) between the Group 1 and 2 calves during the incubation period, patency period or convalescence period. All the animals required treatment to prevent death. The results suggest that the inactivated vaccine from A. marginale produced in IDE8 induced seroconversion in calves, but was not effective for preventing anaplasmosis induced by the UFMG2 isolate under the conditions of this experiment. PMID:22832750

  1. 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.

  2. 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. PMID:26797395

  3. Introduction to the alpha-proteobacteria: Wolbachia and Bartonella, Rickettsia, Brucella, Ehrlichia, and Anaplasma.

    Science.gov (United States)

    Bowman, Dwight D

    2011-11-01

    Wolbachia is an obligate intracellular endosymbiont and likely mutualist living within the heartworm Dirofilaria immitis and a number of other filarial nematodes in the family Onchocercidae. The bacterial infection is passed from worm to worm transovarially; the organisms are in ovarian cells, the developing microfilariae, and multiply and persist in all later developmental stages through the mosquito and into the next host. Besides being present in the ovaries of the adult worms, they also are present in large numbers within the hypodermal tissues of the nematode. It is now know that these bacteria that were first observed in heartworms more than 30 years ago are actually related to similar Wolbachia bacteria that are found in arthropods. Wolbachia is an alpha-proteobacteria, and this group includes a number of important arthropod-transmitted bacterial agents of dogs and cats: Rickettsia rickettsii, R. felis, Anaplasma platys, Ehrlichia canis, E. chaffeensis, and E. ewingii. Alpha-proteobacteria are also important as obligate intracellular mutualists in plants in which they are responsible for nitrogen fixation. Recent work on the treatment of heartworms in dogs with doxycycline stems from related work with the human filarial nematode Onchocerca volvulus that causes river blindness in people. PMID:22152604

  4. First molecular survey of Anaplasma bovis in small ruminants from Tunisia.

    Science.gov (United States)

    Ben Said, Mourad; Belkahia, Hanène; Karaoud, Maroua; Bousrih, Maha; Yahiaoui, Mouna; Daaloul-Jedidi, Monia; Messadi, Lilia

    2015-09-30

    To date, no information is available regarding the presence of Anaplasma bovis in the South Mediterranean area. In this study, prevalence, risk factors, and genetic diversity of A. bovis were assessed in small ruminants. A total of 563 healthy small ruminants (260 sheep and 303 goats), from 25 randomly selected flocks located in 5 localities from two bioclimatic areas in Tunisia, were investigated for the detection of A. bovis in blood by nested polymerase chain reaction (nPCR) assay. The overall infection rates of A. bovis were 42.7 and 23.8% in sheep and goats, respectively. Goats located in a sub-humid area were statistically more infected than those located in a humid area. A. bovis prevalence rate varied significantly according to sheep and goat flocks, and to the sheep breed. Infection with A. bovis was validated by sequencing. Sequence analysis based on the 16S rRNA gene showed that A. bovis from Tunisian goats and sheep clustered with other strain sequences detected from wild and domestic animals and published in GenBank. This study gives the first insight of presence of A. bovis DNA in small ruminants in Tunisia and suggests that these animal species may be playing an important role in the bovine anaplasmosis natural cycle caused by A. bovis in the South Mediterranean ecosystem. PMID:26088935

  5. Detección de Anaplasma spp. en Garrapatas de Myrmecophaga tridactyla de Vida Libre en la Sabana Inundable de Casanare‚ Colombia -resumen-

    OpenAIRE

    César Rojano-Bolaño; V Contreras-Cogollo; L Miranda-Cortés; R Ávila-Avilán; Jorge Miranda-Regino; Laura Hernández-Fernández; Salim Mattar-Velilla; Santiago Monsalve-Buriticá

    2014-01-01

    La anaplasmosis es una enfermedad infecciosa causada por rickettsias del género Anaplasma spp. Su relevancia esta principalmente determinada por el grado de afectación endémica en el ganado doméstico. Una amplia variedad de mamíferos silvestres también son susceptibles a la infección. Las bacterias pueden ser intracitoplasmáticas‚ dañando las estructuras de las células pudiendo causar anemia e ictericia. Algunas especies de Anaplasma spp han sido reportadas como microorganismos potencialmente...

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

    Science.gov (United States)

    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.

  7. 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.

  8. 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.

  9. Immunization-induced anaplasma marginale-specific T lymphocyte reponses impaired by A. marginale infection are restored after eliminating the infection with tetracycline

    Science.gov (United States)

    Infection of cattle with Anaplasma marginale fails to prime sustained effector/memory T-cell responses, and high bacterial load may induce antigen-specific CD4 T exhaustion and deletion. We tested the hypothesis that clearance of persistent infection restores the exhausted T-cell response. We show t...

  10. Independence of Anaplasma marginale Strains with High and Low Transmission Efficiencies in the Tick Vector following Simultaneous Acquisition by Feeding on a Superinfected Mammalian Reservoir Host

    Science.gov (United States)

    Strain superinfection occurs when a second pathogen strain infects a host already carrying a primary strain. Anaplasma marginale superinfection occurs when the second strain encodes a unique variant surface repertoire as compared to the primary strain and the epidemiologic consequences depend on th...

  11. Anaplasma marginale infection with persistent high-load bacteremia induces a dysfunctional memory CD4+ T lymphocyte response but sustained high IgG titers

    Science.gov (United States)

    Control of blood-borne infections is dependent on antigen-specific effector and memory T cells and high-affinity IgG responses. In chronic infections characterized by a high antigen load, it has been shown that antigen-specific T and B cells are vulnerable to downregulation and apoptosis. Anaplasma ...

  12. 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.

  13. 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

    OpenAIRE

    J.J.N. Ngeranwa; S.P. Shompole; E.H. Venter; Wambugu, A.; J.E. Crafford; Penzhorn, B.L.

    2008-01-01

    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 ...

  14. Cepas atenuadas de Babesia bovis e Babesia bigemina e de Anaplasma centrale como imunógenos no controle da tristeza parasitária bovina Attenuated strains of Babesia bovis and Babesia bigemina and of Anaplasma centrale as immunogens for tick borne disease control

    Directory of Open Access Journals (Sweden)

    Ana Maria Sastre Sacco

    2001-10-01

    Full Text Available Neste trabalho foram avaliadas cepas atenuadas de Babesia bovis e B. bigemina e Anaplasma centrale como imunógenos a serem utilizados no controle da Tristeza Parasitária Bovina. O processo de imunização demonstrou ser inócuo, imunogênico e eficiente, pelo menos no que diz respeito às babesias, pois protegeu os animais vacinados frente ao desafio com cepas heterólogas virulentas a campo, que provocou doença clínica e morte nos animais do grupo controle. O desafio a campo pelo Anaplasma marginale não se mostrou muito virulento ou patogênico, não sendo possível concluir que a imunização com A. centrale proteja os animais contra anaplasmose.Attenuated strains of Babesia bovis and B. bigemina and a strain of Anaplasma centrale were evaluated as immunogens against tick-borne disease. The immunization process was shown to be inocuous, immunogenic and efficient to protect cattle, at least against the babesias. Vaccinated animals proved to be protected when naturally challenged with heterologous and virulent strains, in contrast to control animals that showed clinical symptoms of the disease and had to be treated. The Anaplasma marginale challenge was not efficient since treatment was not necessary in the control animals, therefore it was not possible to conclude that immunization with A. centrale protected animals against anaplasmosis.

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

    Science.gov (United States)

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

    2016-03-01

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

  16. OCORRÊNCIA DE Anaplasma bovis (Donatien & Lestoquard, 1936, Dumler et al. 2001) NA REGIÃO DE BOM JESUS DO ITABAPOANA, RJ

    OpenAIRE

    Orlando Augusto Melo Junior; Antonio Peixoto Albernaz; José Tarcisio de Lima Thiebaut; Farlen Jose Bebber Miranda; Josias Alves Machado; Antonio Carlos Faber da Silva

    2010-01-01

    In the present study, 400 capillary blood smears collected from the ear and the tail of 200 bovines were microscopically examinedfor the presence of hemoparasites. The diagnosis was based on the direct research, semonstrating the etiologic agent inclusions. Eleven bovines (5.5%) were considered infected by Anaplasma bovis, confirming the presence of this hemoparasite in the city of Bom Jesus do Itabapoana, the first city of the northwesten region of the state of Rio de Janeiro to register suc...

  17. Vigilancia Epidemiológica de Rickettsia‚ Ehrlichia y Anaplasma en Fauna Silvestre ex situ e in situ de algunas Regiones de Colombia -resumen-

    OpenAIRE

    Santiago Monsalve-Buriticá; César Rojano-Bolaño; Jorge Miranda-Regino; Lizeth Quintana-Diosa; Laura Hernandez-Fernandez; M J Loaiza-Escobar; Luz Dary Acevedo-Cendales; Salim Mattar-Velilla; J Londoño-Londoño; J D Rodas-González

    2014-01-01

    Las enfermedades causadas por bacterias del orden de los Rickettsiales son trasmitidas por ectoparásitos originados en vida silvestre‚ sin embargo‚ en Colombia no existe una línea base del conocimiento en este sentido. Estos microorganismos intracelulares son causantes de enfermedades transmisibles en pacientes humanos; convirtiéndolas en zoonosis de tipo emergente. Este estudio ha permitido determinar la presencia de microorganismos del orden de los Rickettsiales (Ehrlichia spp.‚ Anaplasma s...

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

    Science.gov (United States)

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

    2016-03-01

    A survey was conducted to estimate the prevalence of Anaplasma, Babesia and Brucella spp. infections in cattle, goats and sheep in the Karamoja Region of Uganda and to identify possible risk factors existing in this semi-nomadic and pastoral area. Low cost laboratory tests were used to diagnose infections (Rose Bengal test for Brucella spp. antibodies and direct microscopic examination for Anaplasma and Babesia spp.). Multivariable logistic regression models were applied to identify possible risk factors linked to gender, animal species, age (only for cattle) and districts. A total of 3935 cattle, 729 goats and 306 sheep of five districts of the Karamoja Region were tested. Seroprevalence for Brucella was 9.2 % (CI, 95 %: 8.4-10), for Anaplasma 19.5 % (CI 95 %: 18.4-20.6) and for Babesia 16 % (CI 95 %: 15-17.1). Significant differences in infections prevalence were observed against risk factors associated with districts and species. Cattle were the species with higher risk of the infections. Female gender was identified as at risk only for Brucella spp. infection. Cattle more than one year old had greater likelihood to be Brucella seropositive. Co-infections of Anaplasma and Babesia spp. were statistically associated, especially in goats and sheep. Further studies to identify risk factors related to host species and geographical districts are needed. The influence on the semi-nomadic agro-pastoral system in Karamoja of animal raids and animal mixing should be further investigated. Findings were important to sensitize Karamojong undertaking measures on infection control, especially on cattle, which are their main source of food. PMID:26888206

  19. Detección de Anaplasma spp. en Garrapatas de Myrmecophaga tridactyla de Vida Libre en la Sabana Inundable de Casanare‚ Colombia -resumen-

    Directory of Open Access Journals (Sweden)

    César Rojano-Bolaño

    2014-12-01

    Full Text Available La anaplasmosis es una enfermedad infecciosa causada por rickettsias del género Anaplasma spp. Su relevancia esta principalmente determinada por el grado de afectación endémica en el ganado doméstico. Una amplia variedad de mamíferos silvestres también son susceptibles a la infección. Las bacterias pueden ser intracitoplasmáticas‚ dañando las estructuras de las células pudiendo causar anemia e ictericia. Algunas especies de Anaplasma spp han sido reportadas como microorganismos potencialmente zoonóticos‚ por lo tanto‚ la anaplasmosis‚ como otras enfermedades transmitidas por garrapatas‚ ha tomado una especial relevancia en la salud pública. El objetivo de este estudio fue detectar la presencia de Anaplasma spp.‚ en garrapatas colectadas en nueve osos palmeros (Myrmecophaga tridactyla de vida libre en el municipio de Pore‚ Casanare‚ entre los años 2013 y 2014‚ capturados con el método de búsqueda activa‚ y posteriormente fueron anestesiados utilizando un dardo con Ketamina (12 mg/kg y Xilacina (0‚5 mg/kg por vía intramuscular. Las garrapatas fueron colectadas directamente de los individuos‚ con ayuda de pinzas. En total se obtuvieron 204 ectoparásitos‚ que fueron almacenados en alcohol al 70%‚ y posteriormente fueron identificados como pertenecientes a la especie Amblyomma cajennense. Para este estudio se seleccionaron 55 pool de tres individuos y fueron analizadas por medio de la técnica de PCR en tiempo real. Se detectaron 2 pool de garrapatas positivos a este microorganismo (3‚6 % pertenecientes a dos osos palmeros. Los resultados preliminares de este estudio demuestran que Anaplasma spp. podría estar circulante entre garrapatas A. cajennense de animales silvestres en la sabana inundable del municipio de Pore‚ Casanare. Se hace necesario realizar nuevos muestreos para analizar un mayor número de garrapatas y determinar la especie de Anaplasma spp. presente.

  20. Seroprevalence of IgG antibodies against Anaplasma marginale in cattle from south Mozambique Soroprevalência de anticorpos de classe IgG contra Anaplasma marginale em bovinos da região Sul de Moçambique

    Directory of Open Access Journals (Sweden)

    António Amélia Mucalane Tembue

    2011-12-01

    Full Text Available The current study aimed to investigate the seroprevalence of IgG antibodies to Anaplasma marginale in cattle from Maputo, Gaza and Inhambane provinces, south Mozambique. A total of 809 serum samples from cattle were obtained and tested by indirect enzyme-linked immunosorbent assay (i-ELISA. The chi-square test at 5% significance was used to assess the association between seroprevalence and the variables gender, age and geographic origin of animals. The overall seropositivity was 76.5% (n = 619 and anti-A. marginale antibodies were detected in 89.1% (n = 156, 68.4% (n = 308 and 84.2% (n = 155 of the animals in the provinces of Maputo, Gaza and Inhambane, respectively. A significant association (p 12 and 24 and 36 months, respectively. These results indicate that in southern Mozambique there are areas of enzootic stability to A. marginale. Thus, epidemiological monitoring is required to monitor the immune status of animals in the region.O objetivo do presente estudo foi investigar a soroprevalência de anticorpos da classe IgG contra Anaplasma marginale em bovinos de corte da região Sul de Moçambique. Para esse efeito, 809 amostras de soro foram coletadas e avaliadas pelo ensaio imunoadsorção enzimático indireto (ELISA-i. O teste Qui-Quadrado, a 5% de significância, foi utilizado para avaliar a associação entre a soroprevalência e as variáveis sexo, idade e origem geográfica dos animais. A soropositividade geral foi de 76,5% (n = 619, e anticorpos anti-A. marginale foram detectados em 89,1% (n = 156, 68,4% (n = 308 e 84,2% (n = 155 dos animais nas províncias de Maputo, Gaza e Inhambane, respectivamente. Uma associação significativa (p 12 a 24 36 meses, respectivamente. Os resultados demonstram que, no Sul de Moçambique, existem áreas de estabilidade enzoótica para A. marginale, em animais maiores de 12 meses. Assim, monitoramento epidemiológico deve ser realizado para o acompanhamento do status imunológico dos animais na região.

  1. 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

    Directory of Open Access Journals (Sweden)

    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.

  2. Human Ehrlichiosis and Anaplasmosis in America

    Directory of Open Access Journals (Sweden)

    J Stephen Dumler

    2013-07-01

    Full Text Available A description of Ehrlichia and Anaplasma agents that have been linked to human disease is presented. A particular emphasis is given to Ehrlichia chaffeensis, Anaplasma phagocytophilum, and Ehrlichia canis. The clinical features associated with each agent, as well as the corresponding diagnosis and treatment are also described.

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

    Science.gov (United States)

    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

  4. Anaplasma marginale major surface protein 1a directs cell surface display of tick BM95 immunogenic peptides on Escherichia coli.

    Science.gov (United States)

    Canales, Mario; Almazán, Consuelo; Pérez de la Lastra, José M; de la Fuente, José

    2008-07-31

    The surface display of heterologous proteins on live Escherichia coli using anchoring motifs from outer membranes proteins has impacted on many areas of biochemistry, molecular biology and biotechnology. The Anaplasma marginale major surface protein 1a (MSP1a) contains N-terminal surface-exposed repeated peptides (28-289 amino acids) that are involved in pathogen interaction with host cell receptors and is surface-displayed when the recombinant protein is expressed in E. coli. Therefore, it was predicted that MSP1a would surface display on E. coli peptides inserted in the N-terminal repeats region of the protein. The Rhipicephalus (Boophilus) microplus BM86 and BM95 glycoproteins are homologous proteins that protect cattle against tick infestations. In this study, we demonstrated that a recombinant protein comprising tick BM95 immunogenic peptides fused to the A. marginale MSP1a N-terminal region is displayed on the E. coli surface and is recognized by anti-BM86 and anti-MSP1a antibodies. This system provides a novel approach to the surface display of heterologous antigenic proteins on live E. coli and suggests the possibility to use the recombinant bacteria for immunization studies against cattle tick infestations. PMID:18582976

  5. 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

    Directory of Open Access Journals (Sweden)

    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

  6. 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

  7. Prokaryotic expression and immunogenicity of recombinant AnkA protein from 3 Anaplasma phagocytophilum isolates%3株嗜吞噬细胞无形体AnkA蛋白原核表达及免疫原性差异性研究

    Institute of Scientific and Technical Information of China (English)

    姚娜; 陈创夫; 于强; 张丽娟

    2013-01-01

    In order to investigate the immunogenicity of recombinant AnkA proteins from 3 A. phagocytophilujn isolates, the partial ankA fragments of its strain Webster, strain Slovenia 1 567, and strain 96HK58 were amplified by PCR respectively and then inserted into the vector pET-30a and transformed into Escherichia coli BL21 to express with IPTG induction. The amino acid sequences of the expression products were identified by mass spectrum analysis. Polyclonal antibodies a-gainst the 3 recombinant AnkA proteins were produced by immuned mouse with the purified fusion proteins. The immunoreac-tivity of the 3 AnkA recombinant proteins was analyzed by Western blot using the serum from immunized mouse and HGA positive serum from human. The recombinant AnkA proteins were efficiently expressed. High titer antibodies against the 3 AnkA recombinant proteins were successfully produced and cross reacted among the 3 AnkA recombinant proteins and their corresponding antibodies. Apparent reaction was also observed among the 3 AnkA recombinant proteins and Chinese human HGA positive serum. Here we concluded that there was no immunogenicity difference among the 3 recombinant AnkA proteins of A. phagocylophilum strain Webster, strain Slovenia 1 567, and strain 96HE58.%目的 研究分析3株嗜吞噬细胞无形体AnkA蛋白的免疫原性.方法 利用原核表达PCR扩增了嗜吞噬细胞无形体Webster株,斯洛文尼亚1 567株,美国96HE58株ankA 基因部分序列,构建pET-30a-AnkA表达载体,转化至大肠杆菌BL21中,以IPTG诱导高效表达融合蛋白,经飞行质谱鉴定氨基酸序列,免疫BALB/c小鼠制备多克隆抗体.Western blot分析3株菌AnkA重组蛋白与免疫小鼠血清抗体相互间的免疫反应以及交叉反应性.同时检测3株菌AnkA重组蛋白与我国无形体病人血清抗体反应性.结果 表达的重组蛋白主要以可溶性形式存在,小鼠免疫获得高效价抗AnkA IgG抗体,3株菌重组蛋白免疫抗体间存在免疫交叉.3株菌重组蛋白与我国无形体病人血清抗体存在免疫反应.结论 成功原核表达并纯化了3株嗜吞噬细胞无形体AnkA蛋白,重组蛋白与我国无形体病人阳性血清存在免疫反应.3株嗜吞噬细胞无形体AnkA重组蛋白与免疫小鼠多克隆抗体间存在交叉反应.

  8. Silencing of genes involved in Anaplasma marginale-tick interactions affects the pathogen developmental cycle in Dermacentor variabilis

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    Almazán Consuelo

    2009-07-01

    Full Text Available Abstract Background The cattle pathogen, Anaplasma marginale, undergoes a developmental cycle in ticks that begins in gut cells. Transmission to cattle occurs from salivary glands during a second tick feeding. At each site of development two forms of A. marginale (reticulated and dense occur within a parasitophorous vacuole in the host cell cytoplasm. However, the role of tick genes in pathogen development is unknown. Four genes, found in previous studies to be differentially expressed in Dermacentor variabilis ticks in response to infection with A. marginale, were silenced by RNA interference (RNAi to determine the effect of silencing on the A. marginale developmental cycle. These four genes encoded for putative glutathione S-transferase (GST, salivary selenoprotein M (SelM, H+ transporting lysosomal vacuolar proton pump (vATPase and subolesin. Results The impact of gene knockdown on A. marginale tick infections, both after acquiring infection and after a second transmission feeding, was determined and studied by light microscopy. Silencing of these genes had a different impact on A. marginale development in different tick tissues by affecting infection levels, the densities of colonies containing reticulated or dense forms and tissue morphology. Salivary gland infections were not seen in any of the gene-silenced ticks, raising the question of whether these ticks were able to transmit the pathogen. Conclusion The results of this RNAi and light microscopic analyses of tick tissues infected with A. marginale after the silencing of genes functionally important for pathogen development suggest a role for these molecules during pathogen life cycle in ticks.

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

    Science.gov (United States)

    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.

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

    Science.gov (United States)

    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. PMID:25550150

  11. 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.

  12. 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.

  13. 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

  14. 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.

  15. 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.

    Science.gov (United States)

    Ngeranwa, J J N; Shompole, S P; Venter, E H; Wambugu, A; Crafford, J E; Penzhorn, B L

    2008-09-01

    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. PMID:19040134

  16. Vigilancia Epidemiológica de Rickettsia‚ Ehrlichia y Anaplasma en Fauna Silvestre ex situ e in situ de algunas Regiones de Colombia -resumen-

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    Santiago Monsalve-Buriticá

    2014-12-01

    Full Text Available Las enfermedades causadas por bacterias del orden de los Rickettsiales son trasmitidas por ectoparásitos originados en vida silvestre‚ sin embargo‚ en Colombia no existe una línea base del conocimiento en este sentido. Estos microorganismos intracelulares son causantes de enfermedades transmisibles en pacientes humanos; convirtiéndolas en zoonosis de tipo emergente. Este estudio ha permitido determinar la presencia de microorganismos del orden de los Rickettsiales (Ehrlichia spp.‚ Anaplasma spp.‚ y Rickettsia spp. en ejemplares silvestres de centros de conservación ex situ (Zoológicos‚ CAV y CAVR y en algunas regiones en condiciones in situ del país. De igual manera se identifica la infección de estos microorganismos en ectoparásitos vectores relacionados. Por medio de la técnica PCRrt‚ se detectó la presencia de Rickettsia spp.‚ Anaplasma spp.‚ y Ehrlichia spp.‚ en suero obtenido de mamíferos‚ y garrapatas colectadas de reptiles y mamíferos del Centro de Atención y Valoración de Fauna silvestre de la Corporación Autónoma Regional de los Valles del Sinú y San Jorge CAV-CVS en Montería (Córdoba‚ en osos hormigueros (Myrmecophaga tridactyla en Pore (Casanare‚ y en garrapatas obtenidas en ejemplares de titi gris (Saguinus leucopus y su hábitat en la vereda La Parroquia en Mariquita (Tolima. En Montería‚ los resultados obtenidos en mamíferos demostraron la ausencia de Rickettsia spp. en 72 muestras de suero‚ sin embargo se detectó un pool de garrapatas de la especie Amblyomma ovale positivo a este microorganismo‚ siendo el primer reporte en Colombia de ectoparásitos de Puma concolor positivos a Rickettsia spp.‚ mientras que en reptiles se detectaron 18 pools de garrapatas positivos a Rickettsia spp (90 %. En las garrapatas colectadas de nueve osos palmeros (Myrmecophaga tridactyla de vida libre se detectaron 12 pool de garrapatas positivos a este microorganismo (17‚6 %‚ y en ejemplares de

  17. 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.

  18. 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

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

    DEFF Research Database (Denmark)

    Michelet, Lorraine; Delannoy, Sabine; Devillers, Elodie;

    2014-01-01

    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...

  20. 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.

  1. Frequency of Anaplasma marginale (Theiler 1910 and Babesia sp in mestizo bovine Zebu, in the Municipality of Ixiamas county Abel Iturralde Department of The La Paz, Bolivia

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    Mercado Alvaro

    2011-08-01

    Full Text Available The present work one carries out in the Municipality of Ixiamas County Abel Iturralde of the department of Peace-Bolivia, between May and August of 2010. The objective was to determine the presence of Anaplama sp and Babesia sp, through sanguine smear, they took 160 samples of blood, 40 bovine of the four areas they have been evaluated. The results indicate the presence of Anaplasma sp 6,90%, 6,20% in females and 9,70% in males, this represents 2,50% for the area TO, 5,00% area B, 17,50% area C and 2,50% area D. According to the category 12,50% in calves, 0,00% in torillos, 4,80% in bulls, 6,20% in cows and 14,30% in heifers. The presence of Babesia sp 3,13%, with relationship to the sex 3,10% in females and 3,22% in males, being observed 2,50% in the area TO, 5,00% in the area B, 5,00% in the area C and 0,00% in the area D, according to the category 0,00% in calves, torillos 4,80% bulls, 2,73% vacates and 7,14% in heifers.

  2. Molecular Detection of Tick-Borne Rickettsiales in Goats and Sheep from Southeastern China.

    Science.gov (United States)

    Ge, Yan; Yin, Hongmei; Rikihisa, Yasuko; Pan, Weiqing; Yin, Hong

    2016-05-01

    Members from Rickettsiales such as Anaplasma phagocytophilum, Ehrlichia chaffeensis, and some spotted fever group (SFG) Rickettsiae are important tick-borne pathogens. One hundred goats and sheep from southeastern China were examined for the presence of Anaplasma, E. chaffeensis, and SFG Rickettsiae by PCR. A. phagocytophilum, Anaplasma bovis, and Anaplasma centrale were detected in 15, 49, and 16 samples, respectively. The A. phagocytophilum and A. centrale were highly homologous to strains from Japanese sika deer and Japanese cattle, respectively, whereas a diversity of A. bovis sequences were detected. New genetic variants of Anaplasma close to A. centrale have been revealed. No Ehrlichia was detected in this study. The presence of SFG Rickettsiae was determined in 26 samples. The coinfection with more than two pathogens tested in this study was as high as 29%. This study has molecularly characterized the circulation of Anaplasma and Rickettsiae in goats and sheep in southeastern China, which highlights the risk of contracting the pathogens upon tick exposure. PMID:26872274

  3. 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.

  4. Natural infection of calves by Anaplasma marginale in dairy herds of the Metalúrgica Region, Minas Gerais Infecção natural por Anaplasma marginale em bezerras de fazendas leiteiras da região Metalúrgica de Minas Gerais

    Directory of Open Access Journals (Sweden)

    Valeska S.P. Melo

    2001-12-01

    Full Text Available The dynamic of natural infections by Anaplasma marginale in calves was evaluated during a period of one year on two farms located in the Metalúrgica Region, State of Minas Gerais, Brazil. Blood samples were collected weekly for rickettsemia and packed cell volume (PCV determination. The animals born from March to July suffered the infection in October and November, independently of their age, whilst calves born from September to December acquired the infection during the first days of life. These animals presented patent rickettsemia from 30 days of life. During the patent period PCV decreased after one week of infection, ranging from 20 to 23%. It was concluded, that in the region studied, the transmission of A. marginale is influenced by climatic conditions, and that calves born during the dry season are more likely to acquire the infection when they are exposed to high transmission levels during the subsequent raining season.Realizou-se um estudo da dinâmica de infecções naturais por Anaplasma marginale, durante o período de um ano, em duas propriedades localizadas na Região Metalúrgica do Estado de Minas Gerais. Foram realizados esfregaços sangüíneos e determinação do volume globular (VG. Os animais que nasceram no período de março a julho sofreram a infecção em outubro e novembro, independentemente da idade, enquanto os que nasceram no período de setembro a dezembro adquiriram a infecção nos primeiros dias de vida, com parasitemia patente a partir de 30 dias de idade. Durante o período patente, o VG reduziu na primeira semana de infecção, variando entre 20 a 23%. Concluiu-se que a ocorrência da primo-infecção sofre influência das estações do ano e que bezerros nascidos na estação seca apresentam maior risco de infecção, quando expostos às condições de elevada transmissão durante a estação chuvosa subseqüente.

  5. Evaluating the effectiveness of an inactivated vaccine from Anaplasma marginale derived from tick cell culture Avaliação da eficácia de uma vacina inativada de Anaplasma marginale derivada de cultura de células de carrapato

    Directory of Open Access Journals (Sweden)

    Pedro Veloso Facury Lasmar

    2012-06-01

    Full Text Available The protective efficacy of an inactivated vaccine from Anaplasma marginale that was cultured in tick cells (IDE8 for use against bovine anaplasmosis was evaluated. Five calves (Group 1 were inoculated subcutaneously, at 21-day intervals, with three doses of vaccine containing 3 × 10(9 A. marginale initial bodies. Five control calves received saline solution alone (Group 2. Thirty-two days after the final inoculation, all the calves were challenged with approximately 3 × 10(5 erythrocytes infected with A. marginale high-virulence isolate (UFMG2. The Group 1 calves seroconverted 14 days after the second dose of vaccine. After the challenge, all the animals showed patent rickettsemia. There was no significant difference (p > 0.05 between the Group 1 and 2 calves during the incubation period, patency period or convalescence period. All the animals required treatment to prevent death. The results suggest that the inactivated vaccine from A. marginale produced in IDE8 induced seroconversion in calves, but was not effective for preventing anaplasmosis induced by the UFMG2 isolate under the conditions of this experiment.Foi avaliada a eficácia de uma vacina protetora para Anaplasma marginale cultivada em células de carrapato (IDE8 para uso contra a anaplasmose bovina. Cinco bezerros (Grupo 1 foram inoculados por via subcutânea com três doses, intervalados de 21 dias, de vacina contendo 3 × 10(9 corpúsculos iniciais de A. marginale inicial. Cinco bezerros do grupo controle receberam apenas solução salina (Grupo 2. Trinta e dois dias após a inoculação final, todos os bezerros foram desafiados com aproximadamente 3 × 10(5 eritrócitos infectados com isolado de A. marginale alta virulência (UFMG2. Os bezerros do Grupo 1 soroconverteram-se 14 dias após a segunda dose da vacina. Após o desafio, todos os animais mostraram riquestsemia patente. Não houve diferença significativa (p > 0,05 entre bezerros do Grupo 1 e 2 em período de incuba

  6. Seroprevalence of selected infectious agents in a free-ranging, low-density lion population in the Central Kalahari Game Reserves in Botswana

    OpenAIRE

    Ramsauer, S; Bay, G.; Meli, M; Hofmann-Lehmann, R.; Lutz, H.

    2007-01-01

    Twenty-one free-ranging Central Kalahari lions (Panthera leo) exhibited a high prevalence rate of feline herpesvirus (100%) and feline immunodeficiency virus (71.4%). Canine distemper virus and feline calicivirus occurred with a low prevalence. All individuals tested negative for feline coronavirus, feline parvovirus, feline leukemia virus, Ehrlichia canis, and Anaplasma phagocytophilum.

  7. Equine tick-borne infections in the Netherlands

    NARCIS (Netherlands)

    Butler, C.M.

    2012-01-01

    This thesis focuses on the emergence and establishment of equine tick-borne infections in the Netherlands, with particular attention to their diagnosis, clinical relevance and treatment. Four tick-borne agents (Borrelia burgdorferi, Theileria equi, Babesia caballi and Anaplasma phagocytophilum) appe

  8. Human anaplasmosis: the first Spanish case confirmed by PCR.

    Science.gov (United States)

    García, J C; Núñez, M J; Castro, B; Fraile, F J; López, A; Mella, M C; Blanco, A; Sieira, C; Loureiro, E; Portillo, A; Oteo, J A

    2006-10-01

    We report a case of human anaplasmosis (HA) fulfilling the confirmation criteria: epidemiologic data and clinical picture compatible with HA; presence of a morulae within polymorphonuclear leukocyte; and positive PCR assay for Anaplasma phagocytophilum: This case report shows the presence of HA in Spain. PMID:17114773

  9. 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

    Directory of Open Access Journals (Sweden)

    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

  10. 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

    Directory of Open Access Journals (Sweden)

    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

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

    OpenAIRE

    Elisabete S Correa; Giane R. Paludo; Scalon, Marcela C.; Josias A Machado; Ana Carolina Q Lima; Anderson Teixeira B Pinto; José Tarcísio L Thiebaut; Antônio P Albernaz

    2011-01-01

    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 ...

  12. A fluorescent glycolipid-binding peptide probe traces cholesterol dependent microdomain-derived trafficking pathways.

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    Steffen Steinert

    Full Text Available BACKGROUND: The uptake and intracellular trafficking of sphingolipids, which self-associate into plasma membrane microdomains, is associated with many pathological conditions, including viral and toxin infection, lipid storage disease, and neurodegenerative disease. However, the means available to label the trafficking pathways of sphingolipids in live cells are extremely limited. In order to address this problem, we have developed an exogenous, non-toxic probe consisting of a 25-amino acid sphingolipid binding domain, the SBD, derived from the amyloid peptide Abeta, and conjugated by a neutral linker with an organic fluorophore. The current work presents the characterization of the sphingolipid binding and live cell trafficking of this novel probe, the SBD peptide. SBD was the name given to a motif originally recognized by Fantini et al in a number of glycolipid-associated proteins, and was proposed to interact with sphingolipids in membrane microdomains. METHODOLOGY/PRINCIPAL FINDINGS: In accordance with Fantini's model, optimal SBD binding to membranes depends on the presence of sphingolipids and cholesterol. In synthetic membrane binding assays, SBD interacts preferentially with raft-like lipid mixtures containing sphingomyelin, cholesterol, and complex gangliosides in a pH-dependent manner, but is less glycolipid-specific than Cholera toxin B (CtxB. Using quantitative time-course colocalization in live cells, we show that the uptake and intracellular trafficking route of SBD is unlike that of either the non-raft marker Transferrin or the raft markers CtxB and Flotillin2-GFP. However, SBD traverses an endolysosomal route that partially intersects with raft-associated pathways, with a major portion being diverted at a late time point to rab11-positive recycling endosomes. Trafficking of SBD to acidified compartments is strongly disrupted by cholesterol perturbations, consistent with the regulation of sphingolipid trafficking by cholesterol. CONCLUSIONS/SIGNIFICANCE: The current work presents the characterization and trafficking behavior of a novel sphingolipid-binding fluorescent probe, the SBD peptide. We show that SBD binding to membranes is dependent on the presence of cholesterol, sphingomyelin, and complex glycolipids. In addition, SBD targeting through the endolysosomal pathway in neurons is highly sensitive to cholesterol perturbations, making it a potentially useful tool for the analysis of sphingolipid trafficking in disease models that involve changes in cholesterol metabolism and storage.

  13. The insertion and transport of anandamide in synthetic lipid membranes are both cholesterol-dependent.

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    Eric Di Pasquale

    Full Text Available BACKGROUND: Anandamide is a lipid neurotransmitter which belongs to a class of molecules termed the endocannabinoids involved in multiple physiological functions. Anandamide is readily taken up into cells, but there is considerable controversy as to the nature of this transport process (passive diffusion through the lipid bilayer vs. involvement of putative proteic transporters. This issue is of major importance since anandamide transport through the plasma membrane is crucial for its biological activity and intracellular degradation. The aim of the present study was to evaluate the involvement of cholesterol in membrane uptake and transport of anandamide. METHODOLOGY/PRINCIPAL FINDINGS: Molecular modeling simulations suggested that anandamide can adopt a shape that is remarkably complementary to cholesterol. Physicochemical studies showed that in the nanomolar concentration range, anandamide strongly interacted with cholesterol monolayers at the air-water interface. The specificity of this interaction was assessed by: i the lack of activity of structurally related unsaturated fatty acids (oleic acid and arachidonic acid at 50 nM on cholesterol monolayers, and ii the weak insertion of anandamide into phosphatidylcholine or sphingomyelin monolayers. In agreement with these data, the presence of cholesterol in reconstituted planar lipid bilayers triggered the stable insertion of anandamide detected as an increase in bilayer capacitance. Kinetics transport studies showed that pure phosphatidylcholine bilayers were weakly permeable to anandamide. The incorporation of cholesterol in phosphatidylcholine bilayers dose-dependently stimulated the translocation of anandamide. CONCLUSIONS/SIGNIFICANCE: Our results demonstrate that cholesterol stimulates both the insertion of anandamide into synthetic lipid monolayers and bilayers, and its transport across bilayer membranes. In this respect, we suggest that besides putative anandamide protein-transporters, cholesterol could be an important component of the anandamide transport machinery. Finally, this study provides a mechanistic explanation for the key regulatory activity played by membrane cholesterol in the responsiveness of cells to anandamide.

  14. Helicobacter pylori lipopolysaccharide modification, Lewis antigen expression, and gastric colonization are cholesterol-dependent

    OpenAIRE

    McGee David J; Hildebrandt Ellen

    2009-01-01

    Abstract Background Helicobacter pylori specifically takes up cholesterol and incorporates it into the bacterial membrane, yet little is currently known about cholesterol's physiological roles. We compared phenotypes and in vivo colonization ability of H. pylori grown in a defined, serum-free growth medium, F12 with 1 mg/ml albumin containing 0 to 50 μg/ml cholesterol. Results While doubling times were largely unaffected by cholesterol, other overt phenotypic changes were observed. H. pylori ...

  15. Cholesterol-dependent retention of GPI-anchored proteins in endosomes.

    OpenAIRE

    Mayor, S; Sabharanjak, S; Maxfield, F R

    1998-01-01

    Several cell surface eukaryotic proteins have a glycosylphosphatidylinositol (GPI) modification at the Cterminal end that serves as their sole means of membrane anchoring. Using fluorescently labeled ligands and digital fluorescence microscopy, we show that contrary to the potocytosis model, GPI-anchored proteins are internalized into endosomes that contain markers for both receptor-mediated uptake (e.g. transferrin) and fluid phase endocytosis (e.g. dextrans). This was confirmed by immunogol...

  16. 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

    Directory of Open Access Journals (Sweden)

    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

  17. Epidemiology and control of human granulocytic anaplasmosis: a systematic review.

    Science.gov (United States)

    Jin, Hongtao; Wei, Feng; Liu, Quan; Qian, Jun

    2012-04-01

    Granulocytic anaplasmosis caused by Anaplasma phagocytophilum is an emerging tick-borne zoonosis worldwide. The obligate intracellular pathogen is transmitted by Ixodes ticks and infects neutrophils in humans and animals, resulting in clinical symptoms ranging from asymptomatic seroconversion to mild, severe, or fatal disease. Since the initial description of human granulocytic anaplasmosis (HGA) in the United States in 1990, HGA has been increasingly recognized in America, Europe, and Asia. This review describes the epidemiology, diagnosis, and treatment of HGA and provides background information on the potential vectors and reservoirs of A. phagocytophilum. PMID:22217177

  18. 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.

  19. Outbreak of anaplasmosis associated with the presence of different Anaplasma marginale strains in dairy cattle in the states of São Paulo and Goiás, Brazil.

    Science.gov (United States)

    Machado, Rosangela Zacarias; da Silva, Jenevaldo Barbosa; André, Marcos Rogério; Gonçalves, Luiz Ricardo; Matos, Carlos Antonio; Obregón, Dasiel

    2015-01-01

    The present study reports the genetic diversity of Anaplasma marginale during anaplasmosis outbreaks in rural properties of the states of Goiás and São Paulo, Brazil. Mortality rates of 3.5% (37/1,050) in calves, 4.7% (45/954) in heifers and 1.1% (25/2,200) in lactating cows were observed in a cattle herd of the municipality of Mambaí, state of Goiás, central-western Brazil. In a cattle herd from the municipality of Lins, state of São Paulo, in southeastern Brazil, none of the animals died, despite presenting clinical signs suggestive of bovine anaplasmosis and exhibiting a drastic decrease in milk production. Thus, blood samples were collected from 100 animals with clinical signs suggestive of bovine anaplasmosis in the municipalities of Mambaí and Lins. Based on the microsatellite structure of the MSP1a of A. marginale, the genotypes E and H were observed in Lins, and the C, D and E genotypes were found in Mambaí. The analysis of the tandem repeat structures of the MSP1a showed nine different strains (τ-10 -15, α-β2, α-β3-13, α-β2 192, τ-β-100, α-β2-Γ, 193-β-100, 191-13-Γ and 191-13-18) in Lins and two (α-β3-Γ and E-F-φ2-F2) in Mambaí. Three new tandem repeats of MSP1a (191, 192 and 193) were described. The τ-10-15 and α-β3-Γ strains were predominantly associated with the occurrence of clinical anaplasmosis and mortality in calves, heifers and lactating cows. PMID:26648009

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

    Directory of Open Access Journals (Sweden)

    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

  1. Tickborne fever associated with abortion outbreak in dairy cows.

    Science.gov (United States)

    2016-08-20

    Anaplasma phagocytophilum detected in aborting cows on rough grazingLead poisoning in bullocksPersistent bovine viral diarrhoea virus infection and colisepticaemia in a 20-hour-old calfAbortion due to bovine herpesvirus 1 in a four-year-old cowTickborne fever in lambsInfectious sinusitis due to Mycoplasma gallisepticum in pheasants These are among matters discussed in the disease surveillance report for May 2016 from SAC Consulting: Veterinary Services (SAC C VS). PMID:27550334

  2. Detection of Borrelia burgdorferi sensu lato and spotted fever group rickettsiae in hard ticks (Acari, Ixodidae) parasitizing bats in Poland

    OpenAIRE

    Piksa, Krzysztof; Stańczak, Joanna; Biernat, Beata; Górz, Andrzej; Nowak-Chmura, Magdalena; Siuda, Krzysztof

    2016-01-01

    A total of 491 Ixodes vespertilionis and 8 Ixodes ricinus collected from bats and cave walls in southern Poland between 2010 and 2012 were examined by the polymerase chain reaction for tick-transmitted pathogens. PCR analysis for Borrelia burgdorferi s.l., Rickettsia spp., and Anaplasma phagocytophilum yielded negative results for all I. vespertilionis. DNA of Rickettsia helvetica was detected in three specimens of I. ricinus attached to Rhinolophus hipposideros or Myotis myotis, while Borrel...

  3. 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

    OpenAIRE

    Mahan, S. M.; B.H. Simbi; Burridge, M J

    2004-01-01

    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 a...

  4. Assessment of Polymicrobial Infections in Ticks in New York State

    OpenAIRE

    Tokarz, Rafal; Jain, Komal; Bennett, Ashlee; Briese, Thomas; Lipkin, W. Ian

    2010-01-01

    Ixodes scapularis ticks are clinically important hematophagous vectors. A single tick bite can lead to a polymicrobial infection. We determined the prevalence of polymicrobial infection with Borrelia burgdorferi, Anaplasma phagocytophilum, Babesia microti, Borrelia miyamotoi, and Powassan virus in 286 adult ticks from the two counties in New York State where Lyme disease is endemic, utilizing a MassTag multiplex polymerase chain reaction assay. Seventy-one percent of the ticks harbored at lea...

  5. Coexistence of Pathogens in Host-Seeking and Feeding Ticks within a Single Natural Habitat in Central Germany▿ †

    OpenAIRE

    Franke, Jan; Fritzsch, Julia; Tomaso, Herbert; Straube, Eberhard; Dorn, Wolfram; Hildebrandt, Anke

    2010-01-01

    The importance of established and emerging tick-borne pathogens in Central and Northern Europe is steadily increasing. In 2007, we collected Ixodes ricinus ticks feeding on birds (n = 211) and rodents (n = 273), as well as host-seeking stages (n = 196), in a habitat in central Germany. In order to find out more about their natural transmission cycles, the ticks were tested for the presence of Lyme disease borreliae, Anaplasma phagocytophilum, spotted fever group (SFG) rickettsiae, Francisella...

  6. First evidence of feline herpesvirus, calicivirus, parvovirus, and Ehrlichia exposure in Brazilian free-ranging felids.

    OpenAIRE

    Filoni, C.; Catão-Dias, J L; Bay, G.; E. L. Durigon; Jorge, R S P; Lutz, H.; Hofmann-Lehmann, R.

    2006-01-01

    Serum samples from 18 pumas (Puma concolor), one ocelot (Leopardus pardalis), and two little spotted cats (Leopardus tigrinus) collected from free-ranging animals in Brazil between 1998 and 2004 were tested by indirect immunofluorescence (IFA) for antibodies to feline herpesvirus 1 (FHV 1), calicivirus (FCV), coronavirus (FCoV), parvo-virus (FPV), Ehrlichia canis, Anaplasma pha-gocytophilum, and Bartonella henselae. Serum samples also were tested, by Western blot and ELISA, for feline leukemi...

  7. Agents of Human Anaplasmosis and Lyme Disease at Camp Ripley, Minnesota

    OpenAIRE

    Johnson, Russell C.; Kodner, Carrie; Jarnefeld, Janet; Eck, Deborah K.; Xu, Yaning

    2011-01-01

    The transmission dynamics of Anaplasma phagocytophilum (Ap) and Borrelia burgdorferi (Bb) among Ixodes scapularis (Is) and mammalian hosts was investigated at Camp Ripley, an area representative of central Minnesota. Prevalence of white-footed mouse infection with Ap and Bb were 20% and 42%, respectively, with a coinfection level of 14%. Peak levels of infection with both agents occurred in May. The average levels of seropositivity to Ap and Bb were 29.3% and 48%, respectively. Of the mice in...

  8. The ecology of tick-transmitted infections in the redwood chipmunk (Tamias ochrogenys)

    OpenAIRE

    Janet E. Foley; Nieto, Nathan C.

    2011-01-01

    The redwood chipmunk contributes to the maintenance of tick-borne diseases in northern California. The range of redwood chipmunks overlaps that of western black-legged ticks and tick-borne disease, including granulocytic anaplasmosis and Lyme borreliosis. Chipmunks have high Anaplasma phagocytophilum PCR- and seroprevalence, are infested with a diversity of Ixodes spp. ticks, and are reservoir competent for Borrelia burgdorferi. We hypothesized that chipmunks could maintain tick-borne disease...

  9. 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.

  10. Sphingolipid and cholesterol dependence of alphavirus membrane fusion - Lack of correlation with lipid raft formation in target liposomes

    NARCIS (Netherlands)

    Waarts, BL; Bittman, R; Wilschut, J

    2002-01-01

    Semliki Forest virus (SFV) and Sindbis virus (SIN) are enveloped viruses that infect their host cells by receptor-mediated endocytosis and subsequent fusion from within acidic endosomes. Fusion of the viral envelope requires the presence of both cholesterol and sphingolipids in the target membrane.

  11. Cholesterol-dependent conformational changes of P-glycoprotein are detected by the 15D3 monoclonal antibody.

    Science.gov (United States)

    Gutay-Tóth, Zsuzsanna; Fenyvesi, Ferenc; Bársony, Orsolya; Szente, Lajos; Goda, Katalin; Szabó, Gábor; Bacsó, Zsolt

    2016-03-01

    The 15D3 mouse monoclonal antibody (mAb) binds an uncharacterized extracellular epitope of the ATP Binding Cassette (ABC) transporter human P-glycoprotein (Pgp). Depletion of cell plasma membrane cholesterol by using methyl-β-cyclodextrin or other chemically modified β-cyclodextrins decreased the Pgp binding affinity of 15D3 mAb. UIC2 mAb, which is known to distinguish two conformers of this ABC transporter, binds only a fraction of cell surface Pgps. UIC2 mAb non-reactive pools of Pgp can be identified with other extracellular mAbs such as 15D3. Cyclosporin A (CsA) can shift non-reactive Pgps into their UIC2-reactive conformation: a phenomenon called the "UIC2 shift". Competition studies proposed these two mAbs share overlapping epitopes and can reveal conformational changes of Pgp that correlate (r=0.97) with the cholesterol content of cells. An apparent increase in competition of these mAbs suggested a conformational change similar to those found in the presence of CsA. However, the reason turned out not to be the UIC2-shift because cholesterol removal from the plasma membrane (PM) reduced the amount of detectable Pgps by 15D3 mAb. This study showed that 15D3 mAb bound to a conformation sensitive epitope of Pgp that was responsive to PM cholesterol levels. These conformational changes were gradual and not as great as the changes observed between the two conformers recognized by the UIC2 mAb.

  12. Lipoplex-mediated transfection of mammalian cells occurs through the cholesterol-dependent clathrin-mediated pathway of endocytosis

    NARCIS (Netherlands)

    Zuhorn, IS; Kalicharan, R; Hoekstra, D

    2002-01-01

    Synthetic amphiphiles are widely used as a carrier system. However, to match transfection efficiencies as obtained for viral vectors, further insight is required into the properties of lipoplexes that dictate transfection efficiency, including the mechanism of delivery. Although endocytosis is often

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

    Directory of Open Access Journals (Sweden)

    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.

  14. Coexistence of emerging bacterial pathogens in Ixodes ricinus ticks in Serbia*

    Directory of Open Access Journals (Sweden)

    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.

  15. 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.

  16. Theileria sp. OT3 and other tick-borne pathogens in sheep and ticks in Italy: molecular characterization and phylogeny.

    Science.gov (United States)

    Giangaspero, A; Marangi, M; Papini, R; Paoletti, B; Wijnveld, M; Jongejan, F

    2015-02-01

    PCR Reverse Line Blot (RLB) hybridization and sequencing were used to determine the dynamics of infection with tick-borne pathogens in one hundred apparently healthy sheep in Italy. Blood samples were tested once prior to the onset of the grazing season (June 2010) and once after the end of the grazing season (August 2010). Ticks collected from sheep and from the vegetation were also tested by PCR/RLB. Before grazing, 56% of the sheep harbored several tick-borne pathogens: Anaplasma ovis was the most prevalent (41%), followed by A. ovis co-infected with Theileria sp. OT3 (14%). After grazing, 87% of sheep were positive for A. ovis alone (41%), co-infected with Theileria sp. OT3 (8%) or co-infected with Babesia motasi (5%). Other sheep were infected with Anaplasma phagocytophilum alone (20%), co-infected with B. motasi (7%) or with Theileria sp. OT3 (5%) (p<0.001). After grazing, sheep were significantly more infected with tick-borne pathogens than before grazing. Ticks collected were all Haemaphysalis punctata (n-89) and 36% were positive for A. ovis, Ehrlichia ovina and A. ovis combined with A. phagocytophilum. Phylogenetic analysis including isolates from countries in the Mediterranean Basin show circulation of the same variants of Theileria sp. OT3, whereas two different geographical origins for the isolates of A. ovis and A. phagocytophilum were identified. This is the first report from Italy of Theileria sp. OT3 in sheep, whereas the detection of Ehrlichia ovina in ticks is worth noting, and the presence of A. phagocytophilum in sheep and in ticks poses a potential public health risk. PMID:25448422

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

    Directory of Open Access Journals (Sweden)

    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.

  18. Molecular screening for bacteria and protozoa in great cormorants (Phalacrocorax carbo sinensis) nesting in Slovakia, central Europe.

    Science.gov (United States)

    Víchová, Bronislava; Reiterová, Katarína; Špilovská, Silvia; Blaňarová, Lucia; Hurníková, Zuzana; Turčeková, Ĺudmila

    2016-09-01

    This study brings the data about the occurrence of bacterial and protozoan pathogens in 32 great cormorants (Phalacrocorax carbo sinensis), representing approximately 20% of the population nesting in the surroundings of water basin Liptovská Mara (northern part of Central Slovakia). A survey revealed the presence of tick-borne bacteria Anaplasma phagocytophilum (6.25%) and parasitic protozoa Toxoplasma gondii (3.1%). These data indicate an infectious status of the great cormorant population nesting in Slovakia; they might suggest a degree of environmental contamination by infectious agents and demonstrate the role of migratory seabirds in the circulation and dispersal of pathogens with zoonotic potential. PMID:27447224

  19. Detection of Borrelia burgdorferi sensu lato and spotted fever group rickettsiae in hard ticks (Acari, Ixodidae) parasitizing bats in Poland.

    Science.gov (United States)

    Piksa, Krzysztof; Stańczak, Joanna; Biernat, Beata; Górz, Andrzej; Nowak-Chmura, Magdalena; Siuda, Krzysztof

    2016-04-01

    A total of 491 Ixodes vespertilionis and 8 Ixodes ricinus collected from bats and cave walls in southern Poland between 2010 and 2012 were examined by the polymerase chain reaction for tick-transmitted pathogens. PCR analysis for Borrelia burgdorferi s.l., Rickettsia spp., and Anaplasma phagocytophilum yielded negative results for all I. vespertilionis. DNA of Rickettsia helvetica was detected in three specimens of I. ricinus attached to Rhinolophus hipposideros or Myotis myotis, while Borrelia garinii was found in one tick parasitizing Myotis daubentonii. These pathogens were recorded for the first time in hard ticks that parasitized bats. PMID:26833325

  20. 人粒细胞无形体病预防控制技术指南(试行)%Guideline of human granulocytic anaplasmosis prevention and control technique (draft)

    Institute of Scientific and Technical Information of China (English)

    中华人民共和国卫生部办公厅

    2010-01-01

    @@ 人粒细胞无形体病(human granulocytic anaplasmosis,HGA)是由嗜吞噬细胞无形体(anaplasma phagocytophilum,曾称为"人粒细胞埃立克体,human granulocytic ehrlichiae")侵染人末梢血中性粒细胞引起,以发热伴白细胞、血小板减少和多脏器功能损害为主要临床表现的蜱传疾病.

  1. Human Granulocytic Anaplasmosis: First Reported Case in Canada

    Directory of Open Access Journals (Sweden)

    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.

  2. 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.

  3. 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.

  4. Passive Surveillance of Ixodes scapularis (Say), Their Biting Activity, and Associated Pathogens in Massachusetts.

    Science.gov (United States)

    Xu, Guang; Mather, Thomas N; Hollingsworth, Craig S; Rich, Stephen M

    2016-08-01

    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

  5. 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. PMID:23473225

  6. Niemann Pick type C cells show cholesterol dependent decrease of APP expression at the cell surface its increased processing through the ?-secretase pathway

    OpenAIRE

    Malnar, Martina; KOŠIČEK, MARKO; Mitterreiter, Stefan; Omerbašić, Damir; Lichtenthaler, Stefan F.; Goate, Alison; Hećimović, Silva

    2010-01-01

    Abstract The link between cholesterol and Alzheimer's disease has recently been revealed in Niemann Pick type C disease. We found that NPC1-/- cells show decreased expression of APP at the cell surface and increased processing of APP through the ?-secretase pathway resulting in increased C99, sAPP? and intracellular A?40 levels. This effect is dependent on increased cholesterol levels, since cholesterol depletion reversed cell surface APP expression and lowered A?/C99 levels in NPC...

  7. Ehrlichiosis y anaplasmosis en Costa Rica

    Directory of Open Access Journals (Sweden)

    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.

  8. Molecular survey of tick-borne pathogens in Ixodid ticks collected from hunted wild animals in Tuscany, Italy

    Institute of Scientific and Technical Information of China (English)

    Valentina; Virginia; Ebani; Fabrizio; Bertelloni; Barbara; Turchi; Dario; Filogari; Domenico; Cerri

    2015-01-01

    Objective: To determine the prevalence of zoonotic tick-borne bacteria in feeding ticks removed from hunted wild animals. Methods: PCR was executed on DNA extracted from 77 tick pools to detect Anaplasma phagocytophilum, Bartonella spp., Borrelia burgdorferi sensu lato, Coxiella burnetii and Rickettsia spp. Results: A total of 432 ticks were collected: 30(6.94%) Haemaphysalis punctata, 72(16.7%) Dermacentor marginatus and 330(76.38%) Ixodes ricinus. For each animal one or two pools of 3 ticks of the same species was constituted. Seventy-seven tick pools were examined by PCR: 58(75.32%) resulted infected and among them 14(18.18%) showed co-infections. In particular, 29(37.66%) pools were positive for Bartonella spp., 23(29.87%) for Anaplasma phagocytophilum, 16(20.78%) for Rickettsia spp., and 5(6.49%) for Borrelia burgdorferi s.l. All samples were negative for Coxiella burnetii. Conclusions: The results demonstrate the presence of several zoonotic tick-borne pathogens in the studied area, and underline the risk of exposure to infections for hunters not only during the outdoor activity, but also when they manipulate hunted animals infested by infected ticks.

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

    Directory of Open Access Journals (Sweden)

    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.

  10. Prevalence of borreliosis, anaplasmosis, ehrlichiosis and Dirofilaria immitis in dogs and vectors in Voronezh Reserve (Russia).

    Science.gov (United States)

    Volgina, N S; Romashov, B V; Romashova, N B; Shtannikov, A V

    2013-12-01

    Most of the dogs studied for the prevalence of CVBD have previously received acaricidal and insecticidal treatments. In the present work, a very specific population of dogs (Group 1) that had never been treated against ticks and mosquitoes was studied. Moreover, the territory occupied by this population has also never been treated, because it is a protected area--Voronezh Natural Reserve. Canine patients from veterinary clinics (Group 2) that had been treated against VBD vectors were studied for comparison. Eighty-two dogs (Group 1) were enrolled in June, 2008. Blood samples were tested using the IDEXX SNAP(®) 4Dx(®) test. A specific heartworm antigen was detected in 12.2% samples. The seroprevalence for Anaplasma phagocytophilum was found to be 34.1%. The antibodies to Borrelia C6 peptide and to Ehrlichia canis were detected in 2.4% of the samples. Almost all dogs with infections had no clinical signs. Only 3 mixed-infected dogs showed non-specific clinical signs. During the tick season, 358 Ixodes ricinus were collected; the prevalence of Borrelia burgdorferi s.l. and Anaplasma phagocytophilum was 21.9% and 0.6%, respectively. Four hundred and forty dogs (Group 2) were studied for comparison. Antibodies to B. burgdorferi s.l. were detected only in one dog, seroprevalence for A. phagocytophilum represented 1.1%, no E. canis seropositive dogs were identified, and 8.2% dogs were found infected with Dirofilaria immitis. Fifty-six percent of dogs with dirofilariosis had clinical signs. All dogs with anaplasmosis showed specific clinical signs--fever, anemia, splenitis. Three dogs died within a few days. PMID:24054985

  11. 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.

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

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    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.

  13. 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.

  14. 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.

  15. First evidence of feline herpesvirus, calicivirus, parvovirus, and Ehrlichia exposure in Brazilian free-ranging felids.

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    Filoni, Claudia; Catão-Dias, José Luiz; Bay, Gert; Durigon, Edison Luiz; Jorge, Rodrigo Silva Pinto; Lutz, Hans; Hofmann-Lehmann, Regina

    2006-04-01

    Serum samples from 18 pumas (Puma concolor), one ocelot (Leopardus pardalis), and two little spotted cats (Leopardus tigrinus) collected from free-ranging animals in Brazil between 1998 and 2004 were tested by indirect immunofluorescence (IFA) for antibodies to feline herpesvirus 1 (FHV 1), calicivirus (FCV), coronavirus (FCoV), parvo-virus (FPV), Ehrlichia canis, Anaplasma pha-gocytophilum, and Bartonella henselae. Serum samples also were tested, by Western blot and ELISA, for feline leukemia virus (FeLV) specific antibodies and antigen, respectively, by Western blot for antibodies to feline immunodeficiency virus (FIV), and by indirect ELISA for antibodies to puma lentivirus (PLV). Antibodies to FHV 1, FCV, FCoV, FPV, FeLV, FIV, PLV or related viruses, and to B. henselae were detected. Furthermore, high-titered antibodies to E. canis or a closely related agent were detected in a puma for the first time. PMID:16870878

  16. 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

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

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    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.

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

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    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.

  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. 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.

  1. 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

  2. Multiple myeloma in a dog with multiple concurrent infectious diseases and persistent polyclonal gammopathy.

    Science.gov (United States)

    Geigy, Caroline; Riond, Barbara; Bley, Carla Rohrer; Grest, Paula; Kircher, Patrick; Lutz, Hans

    2013-03-01

    A 12-year-old, spayed female, mixed-breed dog was presented for acute hematuria, stranguria, polyuria, and polydipsia, as well as lameness for 8 days. Previous medical history included treatment for infection with Ehrlichia canis, Anaplasma phagocytophilum, Leishmania infantum, and Dirofilaria immitis 6.5 years prior to presentation. Besides persistently increased antibody titers to E canis and A phagocytophilum, polyclonal gammopathy with a monoclonal spike and moderate hypercalcemia were observed. There was marked hematuria, and Staphylococcus aureus was cultured from urine. Two weeks after successful treatment of the urinary tract infection, radiographs showed an extensive destructive monostotic lesion of the right humerus. Cytologic examination of fine-needle aspirates of this lesion revealed a neoplastic round cell population suggestive of multiple myeloma. The dog was treated with melphalan and prednisolone for suspected multiple myeloma and doxycycline for suspected ehrlichiosis and anaplasmosis. Treatments lead to resolution of the clinical signs, hypercalcemia, and monoclonal gammopathy, and there was radiographic improvement of bone lesions; polyclonal gammopathy persisted. About one year after presentation the dog was still in clinical remission. This is a rare report of a dog with suspected multiple myeloma and a history of multiple chronic infectious diseases, suggesting that chronic infection and uncontrolled long-term stimulation of the immune system could contribute to the pathogenesis of multiple myeloma. PMID:23278475

  3. Rodents as carriers of tick-borne zoonotic diseases and their ecological impact

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    Paulauskas, A.

    2011-10-01

    Full Text Available Rodents as pests are known to be reservoirs of tick-borne zoonotic infection of viral, bacterial and protozoan origin and are important hosts of the immature stages of Ixodes ticks. A total 493 rodents were live-trapped in different habitat in two biogeographical regions Lithuania and Norway. The ecological impact of different species of rodents in particular habitats was evaluated according to their infestation with ticks and presence of Borrelia burgdorferi s.l. and Anaplasma phagocytophilum infection. Results of the present study demonstrate that although the infestation rate varied between rodent species it was dependent on sampling location and not correlated with infection prevalence in rodents. We observed that mice in Lithuania were more frequently infested with ticks than voles, but, the higher prevalence of B. burgdorferi s.l. infection was detected in voles, than in mice. The overall prevalence of infection in Lithuania was higher than in Norway. In Norway, prevalence of infection in A. flavicollis and M. glareolus was not significantly different. The present study confirmed evidence that the rodent impact on maintenance of Lyme borreliosis depends on biogeographical region and habitat type, and that the zoonotic reservoirs of B. afzelii are M. arvalis, M. glareolus, and A. flavicollis in Lithuania, and A. flavicollis, A. sylvaticus and M. glareolus in Norway. In our present study A. phagocytophilum was not detected in any of the rodents.

  4. 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

    .) burneti and Nss-rRNA gene from Babesia were amplified with nested polymerase chain reaction (nested PCR),respectively.Results Among 204 lxodes persulcatus,104 were positive for tick-borne pathogens with the positive rate of 50.98%,and among them the positive rates of B.burgdorferi,spotted fever group Rickettsia and Anaplasma phagocytophilum were 34.31% (n =70),28.92% (n =59),9.31% (n =19),respectively.And no C.burnetii and Babesia were detected.The overall co-infection rate was 19.12% (39/204),the co-infection rate was 16.18%(33/204) for B.garinii and spotted fever group Rickettsia,4.90% (10/204) for B.burgdorferi and Anaplasma phagocytophilum,2.94%(6/204) for spotted fever group Rickettsia and Anaplasma phagocytophilum and 2.45% (5/204) for B.burgdorferi,Anaplasma phagocytophilum and spotted fever group Rickettsia.Conclusion The results indicated that the natural co-infections of B.garinii,B.afzelii,Anaplasma phagocytophilum and spotted fever group Rickettsia existed in Charles Hilary Ixodes persulcatus collected in Xinjiang.

  5. 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.

  6. Fusion of raft-like lipid bilayers operated by a membranotropic domain of the HSV-type I glycoprotein gH occurs through a cholesterol-dependent mechanism.

    Science.gov (United States)

    Vitiello, Giuseppe; Falanga, Annarita; Petruk, Ariel Alcides; Merlino, Antonello; Fragneto, Giovanna; Paduano, Luigi; Galdiero, Stefania; D'Errico, Gerardino

    2015-04-21

    A wealth of evidence indicates that lipid rafts are involved in the fusion of the viral lipid envelope with the target cell membrane. However, the interplay between these sterol- and sphingolipid-enriched ordered domains and viral fusion glycoproteins has not yet been clarified. In this work we investigate the molecular mechanism by which a membranotropic fragment of the glycoprotein gH of the Herpes Simplex Virus (HSV) type I (gH625) drives fusion of lipid bilayers formed by palmitoyl oleoyl phosphatidylcholine (POPC)-sphingomyelin (SM)-cholesterol (CHOL) (1 : 1 : 1 wt/wt/wt), focusing on the role played by each component. The comparative analysis of the liposome fusion assays, Dynamic Light Scattering (DLS), spectrofluorimetry, Neutron Reflectivity (NR) and Electron Spin Resonance (ESR) experiments, and Molecular Dynamics (MD) simulations shows that CHOL is fundamental for liposome fusion to occur. In detail, CHOL stabilizes the gH625-bilayer association by specific interactions with the peptide Trp residue. The interaction with gH625 causes an increased order of the lipid acyl chains, whose local rotational motion is significantly hampered. SM plays only a minor role in the process, favoring the propagation of lipid perturbation to the bilayer inner core. The stiffening of the peptide-interacting bilayer leaflet results in an asymmetric perturbation of the membrane, which is locally destabilized thus favoring fusion events. Our results show that viral fusion glycoproteins are optimally suited to exert a high fusogenic activity on lipid rafts and support the relevance of cholesterol as a key player of membrane-related processes.

  7. Cholesterol-Dependent Energy Transfer between Fluorescent Proteins—Insights into Protein Proximity of APP and BACE1 in Different Membranes in Niemann-Pick Type C Disease Cells

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    Bjoern von Einem

    2012-11-01

    Full Text Available Förster resonance energy transfer (FRET -based techniques have recently been applied to study the interactions between β-site APP-cleaving enzyme-GFP (BACE1-GFP and amyloid precursor protein-mRFP (APP-mRFP in U373 glioblastoma cells. In this context, the role of APP-BACE1 proximity in Alzheimer’s disease (AD pathogenesis has been discussed. FRET was found to depend on intracellular cholesterol levels and associated alterations in membrane stiffness. Here, NPC1 null cells (CHO-NPC1−/−, exhibiting increased cholesterol levels and disturbed cholesterol transport similar to that observed in Niemann-Pick type C disease (NPC, were used to analyze the influence of altered cholesterol levels on APP-BACE1 proximity. Fluorescence lifetime measurements of whole CHO-wild type (WT and CHO-NPC1−/− cells (EPI-illumination microscopy, as well as their plasma membranes (total internal reflection fluorescence microscopy, TIRFM, were performed. Additionally, generalized polarization (GP measurements of CHO-WT and CHO-NPC1−/− cells incubated with the fluorescence marker laurdan were performed to determine membrane stiffness of plasma- and intracellular-membranes. CHO-NPC1−/− cells showed higher membrane stiffness at intracellular- but not plasma-membranes, equivalent to cholesterol accumulation in late endosomes/lysosomes. Along with higher membrane stiffness, the FRET efficiency between BACE1-GFP and APP-mRFP was reduced at intracellular membranes, but not within the plasma membrane of CHO-NPC1−/−. Our data show that FRET combined with TIRF is a powerful technique to determine protein proximity and membrane fluidity in cellular models of neurodegenerative diseases.

  8. Prevalence of vector-borne pathogens in dogs from Haiti.

    Science.gov (United States)

    Starkey, Lindsay A; Newton, Kassie; Brunker, Jill; Crowdis, Kelly; Edourad, Emile Jean Pierre; Meneus, Pedro; Little, Susan E

    2016-07-15

    Canine vector-borne pathogens are common on some Caribbean islands, but survey data in Haiti are lacking. To determine the prevalence of selected vector-borne pathogens in dogs from Haiti, we tested blood samples collected from 210 owned dogs, 28 (13.3%) of which were infested with Rhipicephalus sanguineus ticks at the time of blood collection. No other tick species were identified on these dogs. A commercially available ELISA identified antibodies to Ehrlichia spp. in 69 (32.9%), antibodies to Anaplasma spp. in 37 (17.6%), and antigen of Dirofilaria immitis in 55 (26.2%); antibodies to Borrelia burgdorferi were not detected in any sample. Molecular assays of whole blood from 207 of the dogs confirmed infection with Ehrlichia canis (15; 7.2%), Anaplasma platys (13; 6.3%), D. immitis (46; 22.2%), Wolbachia spp. (45; 21.7%), Babesia vogeli (16; 7.7%), and Hepatozoon canis (40; 19.3%), but Anaplasma phagocytophilum, Babesia canis, Babesia rossi, Babesia gibsoni, Ehrlichia chaffeensis, Ehrlichia ewingii, or Hepatozoon americanum were not detected. Co-infection with two or more vector-borne pathogens was detected by serology in 42 (20.0%) dogs and by molecular assays in 22 (10.6%) dogs; one dog was co-infected with B. vogeli and E. canis as detected by PCR with D. immitis detected by serology (antigen). Overall, evidence of past or current infection with at least one vector-borne pathogen was identified in 142/210 (67.6%) dogs in this study, underscoring the common nature of these pathogens, some of which are zoonotic, in Haiti. PMID:27270383

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

    Science.gov (United States)

    Quiroz-Castañeda, Rosa Estela; 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

  10. 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

  11. Molecular study on selected vector-borne infections in urban stray colony cats in northern Italy.

    Science.gov (United States)

    Spada, Eva; Proverbio, Daniela; Galluzzo, Paola; Della Pepa, Alessandra; Perego, Roberta; Bagnagatti De Giorgi, Giada; Ferro, Elisabetta

    2014-08-01

    Feline vector-borne diseases can be caused by a range of pathogens transmitted by arthropods. Many of these infections have zoonotic implications, and stray cats are potential sentinels for human and pet health. This study investigated the prevalence of selected vector-borne infections in stray colony cats in Milan. Blood samples from 260 stray cats were evaluated, using conventional polymerase chain reaction tests (cPCRs), for the presence of DNA associated with Rickettsia species, Anaplasma phagocytophilum and Ehrlichia species. Positive cPCR results occurred in 127/260 subjects (48.9%; 95% confidence interval [CI] = 40.7-58.1), with a prevalence of 31.9% (83/260, 95% CI = 25.4-39.6) for Rickettsia species, 17.7% (46/260, 95% CI= 13.0-23.6) for A phagocytophilum, and 5.4% (14/260, 95% CI = 2.9-9.0) for Ehrlichia species. There was no statistical association between a positive PCR test for vector-borne infections surveyed and colony location, age, gender, body condition score or complete blood count abnormalities, nor feline immunodeficiency virus, feline leukaemia virus or Toxoplasma gondii status. The only variable linked to positive PCR results was detection of signs of ocular infection and PCR positivity for Rickettsia species (P = 0.04, odds ratio [OR] = 2.2, 95% CI = 1.1-4.4, P = 0.02). There is a significant prevalence of vector-borne infections with zoonotic potential in urban stray cats in Milan. Thus, dogs and pet cats with outdoor access should be monitored and treated for ectoparasites on a regular basis to minimise risks of disease and the potential transmission of zoonotic agents to people. PMID:24319060

  12. Molecular evidence for bacterial pathogens in Ixodes ricinus ticks infesting Shetland ponies.

    Science.gov (United States)

    Skotarczak, Bogumiła; Wodecka, Beata; Rymaszewska, Anna; Adamska, Małgorzata

    2016-06-01

    Ixodes ricinus has the potential to transmit zoonotic pathogens to humans and domestic animals. The feeding I. ricinus (n = 1737) collected from 49 Shetland ponies and questing ones from vegetation (n = 371) were tested for the presence and differentiation of the bacterial species. DNA of I. ricinus ticks was examined with PCR and sequencing analysis to identify species of Borrelia burgdorferi sensu lato (Bbsl), Anaplasma phagocytophilum and Rickettsia spp. Altogether, 24.3 % I. ricinus of the infested horses and 12.4 % ticks from vegetation carried at least one pathogen species. Horse-feeding ticks (19.2 %) were significantly more frequently infected with Borrelia spp. than questing ticks (4.8 %). Among Bbsl species, in I. ricinus infesting ponies, B. garinii, B. afzelii, B. burgdorferi sensu stricto, B. valaisiana and B. lusitanie and one species, B. miyamotoi related to relapsing fever group, were detected. The 73 flaB gene sequences of Borrelia obtained from feeding I. ricinus have been deposited in GenBank. Among Rickettsia species, two were identified: R. helvetica which was dominant and R. monacensis. Infections with more than one pathogenic species, involving mostly Bbsl and R. helvetica were detected in 6.3 % of infected ticks collected from horses. Shetland ponies may play an important role in the epidemiological cycle of Bbsl and probably could contribute to the natural cycle of A. phagocytophilum and R. helvetica as host for infected ticks. The awareness about these infectious agents in ticks from ponies might be an important criterion for the risk assessment of human diseases, especially as these animals are maintained for recreational purposes. PMID:26920921

  13. 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

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

    Science.gov (United States)

    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

  15. 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.

  16. 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.

  17. 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. PMID:27638113

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

    Science.gov (United States)

    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. PMID:24889035

  19. 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.

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

    Science.gov (United States)

    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.

  1. 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

  2. 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

  3. 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.

  4. 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.

  5. 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. PMID:26726019

  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. Vector-borne pathogens in ticks and EDTA-blood samples collected from client-owned dogs, Kiev, Ukraine.

    Science.gov (United States)

    Hamel, Dietmar; Silaghi, Cornelia; Zapadynska, Svitlana; Kudrin, Anton; Pfister, Kurt

    2013-02-01

    Due to the availability of adequate habitats in urban environments, e.g. city parks and recreational green areas, ticks from such settings may also carry pathogens of veterinary and public health concern. Thus, tick-borne infections may readily be identified in companion animals residing in urbanised areas. To investigate the presence of vector-borne pathogens in Kiev, Ukraine, 52 engorged adult ticks, 33 Dermacentor reticulatus and 19 Ixodes ricinus, were collected from 15 dogs in the spring of 2010, and further 23 canine EDTA-blood samples were obtained in the spring of 2011 from client-owned patients presented in a veterinary clinic in Kiev. DNA of 9 pathogens was detected by PCR in ticks and canine EDTA-blood samples: Babesia canis canis, Anaplasma phagocytophilum, Rickettsia helvetica, Ri. monacensis, Ri. raoultii, and Dirofilaria repens (by proxy) were identified in engorged ticks and B. c. canis, Hepatozoon canis, Di. immitis, Di. repens, and Mycoplasma haemocanis in canine EDTA-blood samples. This is the first description of Ri. raoultii in the Ukraine. This study adds information on the occurrence of vector-borne pathogens of veterinary and public health importance in Kiev, Ukraine. PMID:23069260

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

    Directory of Open Access Journals (Sweden)

    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.

  9. 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.

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

    Science.gov (United States)

    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

  11. Development of a multiplex PCR assay for simultaneous detection of Theileria annulata, Babesia bovis and Anaplasma marginale in cattle

    OpenAIRE

    Bilgiç, Huseyin B.; KARAGENÇ, Tülin; Simuunza, Martin; Shiels, Brian; Tait, Andy; Eren, Hasan; Weir, William

    2013-01-01

    Tropical theileriosis, bovine babesiosis and anaplasmosis are tick-borne protozoan diseases that impose serious constraints on the health and productivity of domestic cattle in tropical and sub-tropical regions of the world. A common feature of these diseases is that, following recovery from primary infection, animals become persistent carriers of the pathogen and continue to play a critical role in disease epidemiology, acting as reservoirs of infection. This study describes development and ...

  12. Vector-borne disease surveillance in puerto rico: pathogen prevalence rates in canines ? Implications for public health and the u.s. Military ? Applying the one health concept.

    Science.gov (United States)

    McCown, Michael E; Opel, Taylor; Grzeszak, Benjamin

    2013-01-01

    Vector-borne diseases (VBDs) make up a large number of emerging infectious and zoonotic diseases. Vectors such as ticks, fleas, and mosquitoes parasitize dogs, thus making canine populations adequate reservoirs for infectious disease and zoonoses. The U.S. military deploys its personnel and Military Working Dogs (MWDs) throughout the world with possible risk of exposure to VBDs. Canine VBDs continue to have veterinary and public health significance for the host nations as well as for deployed U.S. personnel and MWDs. Thus, ongoing and consistent disease surveillance is an essential component to preserve health. The purpose of this study was to survey dogs from multiple cities and varying regions throughout Puerto Rico to determine the prevalence of ehrlichiosis (Ehrlichia canis), anaplasmosis (Anaplasma phagocytophilum), Lyme disease (Borrelia burgdorferi), and heartworm disease (Dirofilaria immitis) from May to July 2012. Canine blood samples (1?3 ml) from the cities of San Juan (n = 629), Guaynabo (n = 50), Ponce (n = 20) and Vieques Island (n = 53) were obtained and tested on-site using an IDEXX SNAP? 4Dx? (enzyme-linked immunosorbent assay) test kit. Prevalence for single or multiple disease status was calculated for each site. The overall period prevalence of VBD in Puerto Rico in the shelter population was 57.7% (71/123). In Guaynabo, the VBD prevalence was 30% (15/50); 2 (13%) of these positive dogs had VBD co-infection. In the coastal port city of Ponce, it was 60% (12/20); 6 (50%) dogs were infected by two or more VBDs. On Vieques Island, it was 83% (44/53); 27 (61%) dogs were coinfected. Conversely, samples collected at the Fort Buchanan Veterinary Clinic in the capitol city of San Juan resulted in a VBD prevalence of 8.9% (56/629). Lyme disease was not detected in any sample. This study showed the presence of D. immitis, E. canis, and A. phagocytophilum in all four sites of Puerto Rico, emphasizing the value of surveillance for VBDs to determine disease

  13. 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.

  14. 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

  15. 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.

  16. 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

  17. 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.

  18. 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. PMID:22835153

  19. Zoonotic Infections Among Employees from Great Smoky Mountains and Rocky Mountain National Parks, 2008–2009

    Science.gov (United States)

    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

    2012-01-01

    Abstract 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. PMID:22835153

  20. 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.

  1. 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.

  2. 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.

  3. 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.

  4. 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

  5. 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

  6. 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

  7. 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

  8. 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.

  9. 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

  10. 9 CFR 130.18 - User fees for veterinary diagnostic reagents produced at NVSL or other authorized site (excluding...

    Science.gov (United States)

    2010-01-01

    ...-Sept. 30, 2011 Beginning Oct. 1, 2011 Anaplasma card test antigen 2 mL $95.00 $97.00 $99.00 $101.00 $103.00 Anaplasma card test kit without antigen Kit 127.00 130.00 133.00 136.00 139.00 Anaplasma CF....00 32.00 Bacterial ELISA antigens 1 mL 29.00 30.00 31.00 31.00 32.00 Bacterial or...

  11. 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% ...

  12. 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% ...

  13. 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% ...

  14. 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% ...

  15. 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% ...

  16. 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% ...

  17. Effect of milk production system on the enzootic stability to Anaplasma marginale and Babesia bovis in calves in the Campo das Vertentes region of Minas Gerais state, BrazilEfeito do sistema de produção de leite sobre a estabilidade enzoótica para Anaplasma marginale e Babesia bovis em bezerras na região do Campo das Vertentes de Minas Gerais, Brasil

    Directory of Open Access Journals (Sweden)

    Christiane Maria Barcellos Magalhães Rocha

    2012-04-01

    Full Text Available We conducted a cross-sectional observational study, in order to determine the frequency of anti-A. marginale and anti-B. bovis antibodies in calves from four to 12 months of age from ten farms that producing B type milk and an equal number that produce raw milk refrigerated, located in the Campo das Vertentes region of Minas Gerais state, in the period September 2008 to August 2009. Blood smears were performed, serologic testing by indirect immunofluorescence technique (IFAT, given the packed cell volume, rickettsemia, and the clinical scores of animals infected by A. marginale. In the farms that produce B type milk, the overall average frequency of seropositive calves was 94.47% (166/176 and 89.20% (157/176 for A. marginale and B. bovis, respectively. Already on the farms that produce raw milk refrigerated, the overall average frequency of A. marginale was 92.59% (149/161 and for B. bovis from 86.30% (139/161, and there was no significant difference (p > 0.05 in the frequency of calves infected for both hemoparasitic between the two systems of milk production. Statistically significant (p 0.05 among calves from properties that produce B type milk and raw milk refrigerated. The results of this study indicate that, in the Campos das Vertentes region of Minas Gerais, the production system does not interfere with the enzootic stability for A. marginale and B. bovis in calves from dairy farms B milk or raw milk refrigerated, with low probability of anaplasmosis and/or babesiosis in adults animals.Foi realizado um estudo observacional do tipo transversal, com o objetivo de determinar a frequência de anticorpos anti-A. marginale e B. bovis em 337 bezerras com idade entre quatro a 12 meses, oriundas de dez propriedades produtoras de leite B e igual número de fazendas de leite cru refrigerado (leite C, na região do Campo das Vertentes de Minas Gerais, no período de setembro de 2008 a agosto de 2009. Foram realizados esfregaços sanguíneos, sorologia por meio da reação de imunofluorescência indireta (RIFI, determinado o volume globular e riquetsemia, e os escores clínicos das bezerras infectadas por A. marginale. Nas propriedades de leite B (LB, a frequência média global de bezerras soropositivas para A. marginale e B. bovis foi de 94,47% (166/176 e 89,20% (157/176, respectivamente. Nas fazendas de leite cru refrigerado (LCR, a frequência média global de bezerras soropositivas para A. marginale foi de 92,59% (149/161 e para B. bovis de 86,30% (139/161, não havendo diferença (p > 0,05 na frequência de animais infectados para ambos hemoparasitos entre os dois sistemas de produção de leite. Entretanto, houve diferença (p 0,05 entre os animais de propriedades de LB e LCR. Os resultados deste estudo indicam que, na região do Campo das Vertentes de Minas Gerais, o sistema de produção não interfere na estabilidade enzoótica para A. marginale e B. bovis em bezerras provenientes de fazendas produtoras de Leite B ou leite cru refrigerado, com baixo risco de ocorrência de anaplasmose e/ou babesiose em animais adultos.

  18. 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

  19. 边缘无浆体MSP1α与MSP1β基因的克隆表达及其对牛红细胞的黏附特性%Cloning and expression of MSP1α and MSP1β proteins of Anaplasma marginale and their adherence to bovine red cells

    Institute of Scientific and Technical Information of China (English)

    倪宏波; 钱爱东; 姜海芳

    2013-01-01

    目的 原核表达边缘无浆体(Anaplasmamarginale,A.marginale)MSP1α和MSP1β蛋白,并检测其对牛红细胞的黏附作用.方法 通过PCR法扩增MSP1α和MSP1β基因,插入表达载体pGEX-6p-1中,转化感受态大肠埃希菌(E.coli)BL21(DE3),IPTG诱导表达,表达的重组MSP1α和MSP1β蛋白经透析法纯化后,分别免疫新西兰白兔和商品蛋鸡,制备免疫兔血清及抗MSP1α和MSP1p IgY抗体,Western blot法分析其免疫原性,间接免疫荧光法(immunofluorescence assay,IFA)检测其在大肠埃希菌外膜上的表达,血凝试验(hemagglutination test,HA)和血凝抑制试验(hemagglutination inhibition test,HI)检测其对牛红细胞的黏附作用.结果 重组表达质粒pGEX-6p-1-MSP1α和pGEX-6p-1-MSP1p经PCR、双酶切及测序证实构建正确;表达的重组MSP1α和MSP1β蛋白相对分子质量分别约为58 000和107 000,主要以包涵体形式存在,表达量分别占菌体总蛋白的31%和14%;纯化的重组MSP1α和MSP1β蛋白纯度分别可达93%和91%,可与免疫兔血清发生反应;抗MSP1α和MSP1p IgY抗体可分别与表达MSP1α和MSP1β蛋白的重组菌发生反应;重组MSP1α和MSP1p蛋白对牛红细胞具有明显的黏附作用,吸附率可达56.4%和52.7%.结论 MSP1α和MSP1β基因可在大肠埃希菌外膜上大量表达,并对牛红细胞具有明显的吸附作用,为进一步揭示牛边缘无浆体病的作用机理奠定了基础.

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

    Directory of Open Access Journals (Sweden)

    Shiwen Wang

    2012-02-01

    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.

  1. Surveillance of zoonotic and infectious diseases in Ecuador: implications for special operations forces medical operations, personnel, and canines.

    Science.gov (United States)

    McCown, Michael; Monterroso, Victor H; Grzeszak, Benjamin

    2011-01-01

    Vector-borne diseases (VBD) make up a large number of emerging infectious and zoonotic diseases. Ticks, fleas, and mosquitoes are effective vectors parasitizing canines, making dogs adequate reservoirs for zoonoses. The U.S. military deploys personnel and government-owned animals around the world with possible risk of exposure to VBD. Canine VBD have veterinary and public health significance for the host nations as well as for the U.S. troops and its working animals deployed in the theater of operations. These factors make disease surveillance a great importance. The objective of this work was to survey canines from the cities of Manta and Guayaquil in Ecuador to determine prevalence of heartworm disease (D. immitis), ehrlichi os is (E. canis), Lyme disease (B. burgdorf eri), and anapl asmosis (A. phagocytophilum). Canine blood samples (1-3ml) collected from the cities of Manta (n=50) and Guayaquil (n=50) were tested on site using a SNAPR 4DxR Test Kit. Prevalence for single or multiple disease status was calculated for each city. In the city of Manta the overall prevalence of diseases was 78%; 52% for E. canis alone, and 26% for co-infection with E. canis and A. phagocytophilum. The overall prevalence for the city of Guayaquil was 88%; 40% for E. canis alone, 22% for A. phagocytophilum alone, and 26% for co-infection with E. canis and A. phagocytophilum. Neither heartworm disease nor Lyme disease was detected in any samp le. In conclusion, this study showed the extensive presence of E. canis and A. phagocytophilum in both cities in Ecuador, emphasizing the value of surveillance for zoonotic diseases to determine disease prevalence and risk assessments, as well as to implement control measures. PMID:22173599

  2. 75 FR 18836 - Findings of Research Misconduct

    Science.gov (United States)

    2010-04-13

    ..., 16A, 17A, 18, 19C, and 20A, which are included in her thesis, ``Cholesterol-dependent mechanism(s) of... institution sends a copy of the certification to ORI; and (4) That she will write letters, approved by ORI,...

  3. 21 CFR 522.1660a - Oxytetracycline solution, 200 milligrams/milliliter.

    Science.gov (United States)

    2010-04-01

    ... cattle, dairy cattle, and calves including prerumenative (veal) calves—(i) Amounts and indications for... intravenously for treatment of anaplasmosis caused by Anaplasma marginale, severe foot-rot, and advanced cases.... Not for use in lactating dairy animals. (2) Swine—(i) Amounts and indications for use—(A) Sows: 3...

  4. Prevalence of Endoglobular Hemotropic Parasites in Pure Gyr Cattle in Córdoba, Colombia

    Directory of Open Access Journals (Sweden)

    Rafael Blanco Martínez

    2015-12-01

    Full Text Available Bovine parasitic sadness produces significant losses in Colombia and it is associated with the presence of ticks. It is caused by microscopic endoglobular hemotropic parasites such as Anaplasma spp. and Babesia spp. In this study, 131 pure Gyr cows were studied from four cattle farms in Córdoba, Colombia. A blood sample of 5 ml was collected from the coccygeal vein for hematocrit determination and for blood smears stained with Wright’s stain, in order to assess intracellular parasitic forms morphologically compatible with Anaplasma spp. and Babesia spp. Chi-square test was used to determine whether the variables of body condition, mucous color, sex and production system (grazing, semi-confinement, and confinement were independent from the frequency of endoglobular hemotropic parasites. The study found that 24.43% of the sampled animals were positive for endoglobular hemotropic parasites; 20.61% (27/131 of them were positive for Anaplasma spp.; 3.05% (4/131 for Babesia spp., and 0.76% (1/131 for both Anaplasma spp. and Babesia spp. No significant differences (p > 0.05 were found for variables of mucous color, sex and production system (grazing, semi-confinement, and confinement. This allowed to register for the first time the prevalence of infection by endoglobular hemotropic parasites in Bos indicus cattle, of the Gyr breed specifically.

  5. Immunisation of smallholder dairy cattle against anaplasmosis and babesiosis in Malawi

    DEFF Research Database (Denmark)

    Tjørnehøj, Kirsten; Lawrence, J. A.; Kafuwa, P. T.;

    1997-01-01

    A field study was conducted in the Southern Region of Malawi to evaluate the possible benefits of immunisation of improved dairy cattle against Anaplasma marginale, Babesia bigemina and Babesia bovis. Friesian crossbred heifers were immunised when they were being reared on Government farms...

  6. The characterization and manipulation of the bacterial microbiome of the Rocky Mountain wood tick, Dermacentor andersoni

    Science.gov (United States)

    Background: In North America, ticks are the most economically impactful vectors of human and animal pathogens. The Rocky Mountain wood tick, Dermacentor andersoni (Acari: Ixodidae), transmits Rickettsia rickettsii and Anaplasma marginale to humans and cattle, respectively. In recent years, studies h...

  7. 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 tic...

  8. Molecular survey on the presence of zoonotic arthropod-borne pathogens in wild red deer (Cervus elaphus).

    Science.gov (United States)

    Ebani, Valentina Virginia; Rocchigiani, Guido; Bertelloni, Fabrizio; Nardoni, Simona; Leoni, Alessandro; Nicoloso, Sandro; Mancianti, Francesca

    2016-08-01

    To estimate the prevalence of some zoonotic tick-borne pathogens in red deer (Cervus elaphus) living in Italian areas with high risk of arthropod exposure, blood samples from 60 red deer were tested by PCR for A. phagocytophilum, Borrelia burgdorferi s.l., Coxiella burnetii, Francisella tularensis, and piroplasms. Thirty-four (56.67%) animals resulted positive for one or more pathogens. In particular, 24 (40%) red deer were positive for A. phagocytophilum, 16 (26.67%) for Babesia divergens, 6 (10%) for C. burnetii, 2 (3.33%) for B. burgdorferi s.l. No positive reaction was observed for F. tularensis. Thirteen (21.67%) animals resulted co-infected by two or three pathogens. Red deer is confirmed as competent reservoir of A. phagocytophilum and B. divergens, but not of B. burgdorferi. This is the first report of C. burnetii-positive red deer in central Italy. Hunters may be at risk of infection both through infected ticks and during the infected cervids carcasses dressing. PMID:27477510

  9. Regulation of the membrane insertion and conductance activity of the metamorphic chloride intracellular channel protein CLIC1 by cholesterol.

    Directory of Open Access Journals (Sweden)

    Stella M Valenzuela

    Full Text Available The Chloride Intracellular ion channel protein CLIC1 has the ability to spontaneously insert into lipid membranes from a soluble, globular state. The precise mechanism of how this occurs and what regulates this insertion is still largely unknown, although factors such as pH and redox environment are known contributors. In the current study, we demonstrate that the presence and concentration of cholesterol in the membrane regulates the spontaneous insertion of CLIC1 into the membrane as well as its ion channel activity. The study employed pressure versus area change measurements of Langmuir lipid monolayer films; and impedance spectroscopy measurements using tethered bilayer membranes to monitor membrane conductance during and following the addition of CLIC1 protein. The observed cholesterol dependent behaviour of CLIC1 is highly reminiscent of the cholesterol-dependent-cytolysin family of bacterial pore-forming proteins, suggesting common regulatory mechanisms for spontaneous protein insertion into the membrane bilayer.

  10. Atorvastatin induces thrombomodulin expression in the aorta of cholesterol-fed rabbits and in TNFa-treated human aortic endothelial cells

    OpenAIRE

    Lin, Shing-Jong; Hsieh, Fang-Yu; Chen, Yung-Hsiang; Lin, Chia-Chi; Kuan, I-I; Wang, Shu-Huei; Wu, Chau-Chung; Chien, Hsiung-Fei; Lin, Fen-Yen; Chen, Yuh-Lien

    2009-01-01

    Expression of functionally active thrombomodulin (TM) on endothelial cells is critical for vascular thromboresistance. 3-Hydroxyl-3-methyl coenzyme A reductase inhibitors (statins) can protect the vasculature from inflammation and atherosclerosis caused by cholesterol-dependent and cholesterolindependent mechanisms. In the present study, the effects of atorvastatin on TM expression in the aorta of cholesterol-fed rabbits and in TNFa-treated human aortic endothelial ...

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

    Directory of Open Access Journals (Sweden)

    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.

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

    Directory of Open Access Journals (Sweden)

    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.

  13. 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.

  14. Use of serum C-reactive protein as an early marker of inflammatory activity in canine type II immune-mediated polyarthritis: case report

    OpenAIRE

    Kristensen Annemarie T; Jessen Lisbeth; Houser Geoffrey A; Jensen Asger; Kjelgaard-Hansen Mads

    2006-01-01

    Abstract Background Monitoring systemic inflammatory activity during steroid therapy of canine immune-mediated polyarthritis (IMPA) is difficult and mainly relies on clinical signs. Case presentation Canine serum C-reactive protein (CRP) was measured serially and blinded during a 27-week follow-up period of a case of Anaplasma phagocytophilia induced type II immune-mediated polyarthritis. Conclusion WBC was, as expected, observed not to reflect the inflammatory activity during steroid treatme...

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

    OpenAIRE

    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...

  16. Risks of suffering tick-borne diseases in sheep translocated to a tick infested area: a laboratory approach for the investigation of an outbreak.

    Science.gov (United States)

    Hurtado, Ana; Barandika, Jesús F; Oporto, Beatriz; Minguijón, Esmeralda; Povedano, Inés; García-Pérez, Ana L

    2015-02-01

    This study was designed to investigate an outbreak of high mortality that occurred in naïve Assaf sheep introduced into a Latxa sheep flock in the Basque Country, a region where piroplasmosis is endemic. To identify the causes of this outbreak, a panel of different methods, including traditional pathological, biopathological and parasitological analyses combined with recently developed molecular methods, was used. These novel molecular methods included a multiplex real-time PCR assay to screen for the presence of the most important tick-borne pathogens (piroplasms and anaplasmas), followed by a second species-specific multiplex real-time PCR assay for the identification of Anaplasma-positive samples. The identification of piroplasm-positive samples was carried out by a multiplexed microsphere-based suspension array using a Luminex(®) xMAP technology-based procedure. Anaplasmas and/or piroplasms were detected in 7/10 lambs and 11/13 ewes, with Babesia ovis being detected in 12 of the 23 animals, Theileria ovis in 6 and Anaplasma ovis in 4, both as single and mixed infections. Most of the animals infected with B. ovis had a marked decrease in the values of the red blood cell parameters. Ticks collected from the animals were identified as Riphicephalus bursa, recognised vector of B. ovis. Other haemolytic pathologies (clostridial disease, copper poisoning and leptospirosis) were ruled out and, considering all clinical, laboratory and epidemiological data, babesiosis by B. ovis was diagnosed. A detailed description of the clinical outcome, with ca. 60% of mortality, laboratory results and epidemiological findings are provided. The implications of the introduction of naïve animals into a piroplasmosis endemic area are discussed. PMID:25257849

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

    Directory of Open Access Journals (Sweden)

    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.

  18. Comparison of a Real-Time PCR Method with Serology and Blood Smear Analysis for Diagnosis of Human Anaplasmosis: Importance of Infection Time Course for Optimal Test Utilization

    OpenAIRE

    Schotthoefer, A. M.; Meece, J. K.; Ivacic, L. C.; Bertz, P. D.; Zhang, K.; Weiler, T.; Uphoff, T S; Fritsche, T R

    2013-01-01

    Anaplasmosis and ehrlichiosis are emerging tick-borne diseases with clinically similar presentations caused by closely related pathogens. Currently, laboratories rely predominantly on blood smear analysis (for the detection of intracellular morulae) and on serologic tests, both of which have recognized limitations, for diagnostic purposes. We compared the performance of a published real-time PCR assay that incorporates melt curve analysis to differentiate Anaplasma and Ehrlichia species with ...

  19. Comparative genomics reveals multiple pathways to mutualism for tick-borne pathogens

    OpenAIRE

    Lockwood, Svetlana; Brayton, Kelly A.; Broschat, Shira L.

    2016-01-01

    Background Multiple important human and livestock pathogens employ ticks as their primary host vectors. It is not currently known whether this means of infecting a host arose once or many times during evolution. Results In order to address this question, we conducted a comparative genomics analysis on a set of bacterial pathogens from seven genera – Borrelia, Rickettsia, Anaplasma, Ehrlichia, Francisella, Coxiella, and Bartonella, including species from three different host vectors – ticks, l...

  20. 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.

  1. 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

  2. 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

  3. Spatial and temporal control of signaling through lipid rafts.

    Science.gov (United States)

    Golub, Tamara; Wacha, Stefan; Caroni, Pico

    2004-10-01

    Sphingolipid- and cholesterol-dependent microdomains (rafts) order proteins at biological membranes and have been implicated in most signaling processes at the cell surface, but the principles and mechanisms through which lipid rafts influence signaling are not well understood. Recent studies have revealed how lipid rafts are rapidly redistributed and assembled locally in response to extracellular signals, and how components of raft-based signaling domains undergo rapid and regulated rearrangements influencing signal quality, duration, and strength. These findings highlight the exquisitely dynamic properties of signaling domains based on lipid rafts, and suggest that processes of raft trafficking and assembly take central roles in mediating spatial and temporal control of signaling.

  4. 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.

  5. 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...

  6. Use of serum C-reactive protein as an early marker of inflammatory activity in canine type II immune-mediated polyarthritis: case report

    Directory of Open Access Journals (Sweden)

    Kristensen Annemarie T

    2006-06-01

    Full Text Available Abstract Background Monitoring systemic inflammatory activity during steroid therapy of canine immune-mediated polyarthritis (IMPA is difficult and mainly relies on clinical signs. Case presentation Canine serum C-reactive protein (CRP was measured serially and blinded during a 27-week follow-up period of a case of Anaplasma phagocytophilia induced type II immune-mediated polyarthritis. Conclusion WBC was, as expected, observed not to reflect the inflammatory activity during steroid treatment in a clinical useful manner, whereas, CRP is suggested a valuable unbiased marker of inflammatory activity during steroid treatment in this case.

  7. Use of serum C-reactive protein as an early marker of inflammatory activity in canine type II immune-mediated polyarthritis: case report

    Science.gov (United States)

    Kjelgaard-Hansen, Mads; Jensen, Asger Lundorff; Houser, Geoffrey A; Jessen, Lisbeth Rem; Kristensen, Annemarie T

    2006-01-01

    Background Monitoring systemic inflammatory activity during steroid therapy of canine immune-mediated polyarthritis (IMPA) is difficult and mainly relies on clinical signs. Case presentation Canine serum C-reactive protein (CRP) was measured serially and blinded during a 27-week follow-up period of a case of Anaplasma phagocytophilia induced type II immune-mediated polyarthritis. Conclusion WBC was, as expected, observed not to reflect the inflammatory activity during steroid treatment in a clinical useful manner, whereas, CRP is suggested a valuable unbiased marker of inflammatory activity during steroid treatment in this case. PMID:16987405

  8. Choque séptico por ehrliquiosis

    OpenAIRE

    Viviana Hernández-de Mezerville; Juan I Padilla-Cuadra

    2007-01-01

    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 visualiz...

  9. Control de Rhipicephalus microplus (Acari: Ixodidae) mediante el uso del hongo entomopatógeno Metarhizium anisopliae (Hypocreales: Clavicipitaceae). Revisión

    OpenAIRE

    Melina Maribel Ojeda-Chi; Roger Ivan Rodríguez-Vivas; Edelmira Galindo-Velasco; Roberto Lezama-Gutiérrez; Carlos Cruz-Vázquez

    2011-01-01

    Las infestaciones de la garrapata del ganado, Rhipicephalus microplus, producen el mayor problema global de ectoparásitos en ganado de regiones tropicales y subtropicales, provocan importantes pérdidas económicas en la producción de carne, leche y pieles, además incrementan los gastos derivados de los programas de control, y son capaces de transmitir Babesia bovis, B. bigemina y Anaplasma marginale. El control de R. microplus se basa principalmente en el uso de ixodicidas, sin embargo, su uso...

  10. Dicty_cDB: Contig-U11805-1 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available deria ambifaria AMMD chr... 330 6e-89 CP000614_2179( CP000614 |pid:none) Burkholderia vietnamiensis G4 ...172 2e-41 CP000615_628( CP000615 |pid:none) Burkholderia vietnamiensis G4 ch... 1...15 |pid:none) Yersinia enterocolitica subsp. ... 331 4e-89 (B5XNJ7) RecName: Full=Cysteine desulfurase; EC=2...) Anaplasma marginale str. St. Mar... 358 2e-97 CR382129_705( CR382129 |pid:none) Yarrowia lipolytica strain... EA301845 ) Sequence 23 from patent US 7285399. 58 1e-14 4 ( BD312725 ) Composition and method utili

  11. Bacterial cytolysin during meningitis disrupts the regulation of glutamate in the brain, leading to synaptic damage.

    Directory of Open Access Journals (Sweden)

    Carolin Wippel

    Full Text Available Streptococcus pneumoniae (pneumococcal meningitis is a common bacterial infection of the brain. The cholesterol-dependent cytolysin pneumolysin represents a key factor, determining the neuropathogenic potential of the pneumococci. Here, we demonstrate selective synaptic loss within the superficial layers of the frontal neocortex of post-mortem brain samples from individuals with pneumococcal meningitis. A similar effect was observed in mice with pneumococcal meningitis only when the bacteria expressed the pore-forming cholesterol-dependent cytolysin pneumolysin. Exposure of acute mouse brain slices to only pore-competent pneumolysin at disease-relevant, non-lytic concentrations caused permanent dendritic swelling, dendritic spine elimination and synaptic loss. The NMDA glutamate receptor antagonists MK801 and D-AP5 reduced this pathology. Pneumolysin increased glutamate levels within the mouse brain slices. In mouse astrocytes, pneumolysin initiated the release of glutamate in a calcium-dependent manner. We propose that pneumolysin plays a significant synapto- and dendritotoxic role in pneumococcal meningitis by initiating glutamate release from astrocytes, leading to subsequent glutamate-dependent synaptic damage. We outline for the first time the occurrence of synaptic pathology in pneumococcal meningitis and demonstrate that a bacterial cytolysin can dysregulate the control of glutamate in the brain, inducing excitotoxic damage.

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

    Directory of Open Access Journals (Sweden)

    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.

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

    Directory of Open Access Journals (Sweden)

    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.

  14. Lipid Rafts Identified on Synaptic Vesicles from Rat Brain

    Institute of Scientific and Technical Information of China (English)

    HE Li; L(U) Jihua; ZHOU Qinghua; SUI Senfang

    2006-01-01

    For a long time, lipid rafts have been thought to participate in regulating neurotransmitter release. However,the existence of lipid rafts on synaptic vesicles (SVs) and the mechanism by which exocytosis-relative proteins distribute on this structure have not been fully investigated. There is also much controversial data concerning rafts on SVs and synaptic vesicle proteins which makes the results difficult to interpret. This study systematically analyzed the existence and properties of lipid rafts on purified SVs by sucrose density gradient centrifugation, cholesterol depletion, and temperature variation. The data reveals that typical lipid rafts on SVs are both cholesterol dependent and temperature sensitive. Previous confusing results may have been caused by improper treatment or side effects of particular reagent. We also screened the lateral distribution of major exocytosis-related SV proteins and found that only the synaptobrevin (syb) and synaptotagmin (syt) produce detectable association with lipid rafts in 1% Triton X-100.

  15. Cytotoxic action of triterpene glycosides from sea cucumbers from the genus Cucumaria on mouse spleen lymphocytes. Inhibition of nonspecific esterase.

    Science.gov (United States)

    Aminin, Dmitry L; Silchenko, Alexandra S; Avilov, Sergey A; Stepanov, Vadim G; Kalinin, Vladimir I

    2009-06-01

    Four triterpene glycosides from sea cucumbers belonging to the genus Cucumaria, okhotoside A(1)-1 (1), cucumarioside A(0)-1 (2), frondoside A (3) and cucumarioside A(2)-2 (4) inhibit the activity of nonspecific esterase of mouse spleen lymphocytes. The dependence of the inhibitory activity of the glycosides on their structure is similar to that for hemolytic activity. The absence of inhibitory activity for the preparation Cumaside, which is a complex of cucumarioside A(2)-2 and related compounds with cholesterol, shows a cholesterol-dependent character of the inhibitory action of the glycosides. The effective inhibitory concentrations of frondoside A and cucumarioside A(2)-2 are significantly higher than the immunomodulatory doses of these glycosides. PMID:19634320

  16. A Common Fold Mediates Vertebrate Defense and Bacterial Attack

    Energy Technology Data Exchange (ETDEWEB)

    Rosado, Carlos J.; Buckle, Ashley M.; Law, Ruby H.P.; Butcher, Rebecca E.; Kan, Wan-Ting; Bird, Catherina H.; Ung, Kheng; Browne, Kylie A.; Baran, Katherine; Bashtannyk-Puhalovich, Tanya A.; Faux, Noel G.; Wong, Wilson; Porter, Corrine J.; Pike, Robert N.; Ellisdon, Andrew M.; Pearce, Mary C.; Bottomley, Stephen P.; Emsley, Jonas; Smith, A. Ian; Rossjohn, Jamie; Hartland, Elizabeth L.; Voskoboinik, Ilia; Trapani, Joseph A.; Bird, Phillip I.; Dunstone, Michelle A.; Whisstock, James C. (PMCI-A); (Monash); (Nottingham)

    2008-10-02

    Proteins containing membrane attack complex/perforin (MACPF) domains play important roles in vertebrate immunity, embryonic development, and neural-cell migration. In vertebrates, the ninth component of complement and perforin form oligomeric pores that lyse bacteria and kill virus-infected cells, respectively. However, the mechanism of MACPF function is unknown. We determined the crystal structure of a bacterial MACPF protein, Plu-MACPF from Photorhabdus luminescens, to 2.0 angstrom resolution. The MACPF domain reveals structural similarity with poreforming cholesterol-dependent cytolysins (CDCs) from Gram-positive bacteria. This suggests that lytic MACPF proteins may use a CDC-like mechanism to form pores and disrupt cell membranes. Sequence similarity between bacterial and vertebrate MACPF domains suggests that the fold of the CDCs, a family of proteins important for bacterial pathogenesis, is probably used by vertebrates for defense against infection.

  17. Aptamer-based single-molecule imaging of insulin receptors in living cells

    Science.gov (United States)

    Chang, Minhyeok; Kwon, Mijin; Kim, Sooran; Yunn, Na-Oh; Kim, Daehyung; Ryu, Sung Ho; Lee, Jong-Bong

    2014-05-01

    We present a single-molecule imaging platform that quantitatively explores the spatiotemporal dynamics of individual insulin receptors in living cells. Modified DNA aptamers that specifically recognize insulin receptors (IRs) with a high affinity were selected through the SELEX process. Using quantum dot-labeled aptamers, we successfully imaged and analyzed the diffusive motions of individual IRs in the plasma membranes of a variety of cell lines (HIR, HEK293, HepG2). We further explored the cholesterol-dependent movement of IRs to address whether cholesterol depletion interferes with IRs and found that cholesterol depletion of the plasma membrane by methyl-β-cyclodextrin reduces the mobility of IRs. The aptamer-based single-molecule imaging of IRs will provide better understanding of insulin signal transduction through the dynamics study of IRs in the plasma membrane.

  18. Internalization of Trichosanthin via Different Endocytic Mechanisms

    Institute of Scientific and Technical Information of China (English)

    ZHANG Fan; SUI Senfang

    2005-01-01

    Trichosanthin (TCS) is a plant toxin with ribosome-inactivating activity. TCS can be internalized by the host cells and then attacks the ribosomes resulting in cell death. However, the manner for endocytic uptake of TCS is not well understood. The present work investigates the endocytosis pathway of TCS in human choriocarcinoma cells. The different endocytic mechanisms are interfered by potassium depletion, cholesterol-extraction/addition, or treatments of various drugs. The experiments detect their effects on the TCS-uptake. The results show that a large portion of the TCS can be internalized by clathrin-dependent, as well as by clathrin-independent but cholesterol-dependent endocytosis in human choriocarcinoma cells.

  19. Crystal structure of listeriolysin O reveals molecular details of oligomerization and pore formation

    Science.gov (United States)

    Köster, Stefan; van Pee, Katharina; Hudel, Martina; Leustik, Martin; Rhinow, Daniel; Kühlbrandt, Werner; Chakraborty, Trinad; Yildiz, Özkan

    2014-04-01

    Listeriolysin O (LLO) is an essential virulence factor of Listeria monocytogenes that causes listeriosis. Listeria monocytogenes owes its ability to live within cells to the pH- and temperature-dependent pore-forming activity of LLO, which is unique among cholesterol-dependent cytolysins. LLO enables the bacteria to cross the phagosomal membrane and is also involved in activation of cellular processes, including the modulation of gene expression or intracellular Ca2+ oscillations. Neither the pore-forming mechanism nor the mechanisms triggering the signalling processes in the host cell are known in detail. Here, we report the crystal structure of LLO, in which we identified regions important for oligomerization and pore formation. Mutants were characterized by determining their haemolytic and Ca2+ uptake activity. We analysed the pore formation of LLO and its variants on erythrocyte ghosts by electron microscopy and show that pore formation requires precise interface interactions during toxin oligomerization on the membrane.

  20. Tethered bilayer membranes as a complementary tool for functional and structural studies: The pyolysin case.

    Science.gov (United States)

    Preta, Giulio; Jankunec, Marija; Heinrich, Frank; Griffin, Sholeem; Sheldon, Iain Martin; Valincius, Gintaras

    2016-09-01

    We demonstrate the use of tethered bilayer lipid membranes (tBLMs) as an experimental platform for functional and structural studies of membrane associated proteins by electrochemical techniques. The reconstitution of the cholesterol-dependent cytolysin (CDC) pyolysin (PLO) from Trueperella pyogenes into tBLMs was followed in real-time by electrochemical impedance spectroscopy (EIS). Changes of the EIS parameters of the tBLMs upon exposure to PLO solutions were consistent with the dielectric barrier damage occurring through the formation of water-filled pores in membranes. Parallel experiments involving a mutant version of PLO, which is able to bind to the membranes but does not form oligomer pores, strengthen the reliability of this methodology, since no change in the electrochemical impedance was observed. Complementary atomic force microscopy (AFM) and neutron reflectometry (NR) measurements revealed structural details of the membrane bound PLO, consistent with the structural transformations of the membrane bound toxins found for other cholesterol dependent cytolysins. In this work, using the tBLMs platform we also observed a protective effect of the dynamin inhibitor Dynasore against pyolysin as well as pneumolysin. An effect of Dynasore in tBLMs, which was earlier observed in experiments with live cells, confirms the biological relevance of the tBLMs models, as well as demonstrates the potential of the electrochemical impedance spectroscopy to quantify membrane damage by the pore forming toxins. In conclusion, tBLMs are a reliable and complementary method to explore the activity of CDCs in eukaryotic cells and to develop strategies to limit the toxic effects of CDCs. PMID:27211243

  1. Canine vector-borne diseases in Brazil

    Directory of Open Access Journals (Sweden)

    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.

  2. 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

    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......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......, 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...

  3. 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.

  4. 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.

  5. 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.

  6. Sir Arnold Theiler and the discovery of anaplasmosis : a centennial perspective : tick-borne diseases

    Directory of Open Access Journals (Sweden)

    G.H. Palmer

    2009-09-01

    Full Text Available Sir Arnold Theiler's research in 1908/09 led to the discovery of the first rickettsial pathogen, Anaplasma marginale, and set the stage for his development and implementation of an effective live vaccine based on a less virulent strain, A. marginale ss. centrale. His 1910 report, describing A. marginale, is among the classic monographs in infectious disease research, presenting not only observations in exacting detail but also highlighting the deductive reasoning leading to association of a new pathogen with a specific disease. With a centennial perspective and both conceptual frameworks and molecular tools unimaginable in Theiler's time, the significance of several observations in the original report - cyclic bacteremia, strain superinfection, and taxonomic position - is now clear and highlight the broad applicability of key principles of pathogen biology.

  7. New Insight into Immunity and Immunopathology of Rickettsial Diseases

    Directory of Open Access Journals (Sweden)

    Pasquale Mansueto

    2012-01-01

    Full Text Available Human rickettsial diseases comprise a variety of clinical entities caused by microorganisms belonging to the genera Rickettsia, Orientia, Ehrlichia, and Anaplasma. These microorganisms are characterized by a strictly intracellular location which has, for long, impaired their detailed study. In this paper, the critical steps taken by these microorganisms to play their pathogenic roles are discussed in detail on the basis of recent advances in our understanding of molecular Rickettsia-host interactions, preferential target cells, virulence mechanisms, three-dimensional structures of bacteria effector proteins, upstream signalling pathways and signal transduction systems, and modulation of gene expression. The roles of innate and adaptive immune responses are discussed, and potential new targets for therapies to block host-pathogen interactions and pathogen virulence mechanisms are considered.

  8. 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

    Directory of Open Access Journals (Sweden)

    K.B. Stevens

    2007-09-01

    Full Text Available 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.

  9. 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

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

    Science.gov (United States)

    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

  11. 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

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

    Science.gov (United States)

    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

  13. 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.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.

  14. [Vector-borne parasites of dogs on the Islands of Cabo Verde].

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    Kirchner, Marlene; Brunner, Adrian; Edelhofer, Renate; Joachim, Anja

    2008-01-01

    During an animal welfare campaign on the Islands of Cape Verde (in the capital Praia on the island of Santiago) a total of 57 dogs were examined for ectoparasites and blood parasites. 84% of the animals were infested with arthropods, mostly ticks. Haemotropic parasites were examined via blood smear, polymerase chain reaction (PCR) and/or serology. 23 dogs had single infections, five multiple infections with haemoparasites. In eight dogs Ehrlichia canis and in one Amaplasma phagocytophilum were detected by PCR, while 43.1% and 36.2% respectively were serologically positive. In 23 blood smears Babesia gibsoni, but no Babesia canis could be detected. In four cases Hepatozoon canis was found in the smears. All animals were negative for Dirofilaria larvae or antibodies against Leishmania. Arthropod-infested animals more frequently harboured babesiae (44%), ehrlichiae (19%) or H. canis (8%) compared to non-infested animals (20%, 0%, 0%). In bitches and dogs aged one year or less babesiae were more frequently detected (48% of the bitches and 13% of the male dogs; 45% of animals less than one year and 26% of the older ones). Due to the high infection rates the import of animals from Cabo Verde to Central Europe must be seen as critical, since an endemisation of (sub-)tropical infectious agents cannot be excluded. PMID:19066773

  15. External quality assessment of the detection of rickettsioses inChina

    Institute of Scientific and Technical Information of China (English)

    Lijuan Zhang; Daxin Ni; Zijian Feng

    2010-01-01

    Objective:To report a training course on the laboratory diagnoses of rickettsioses that10 provincial/cityCDCs participated in laboratory external quality assurance(EQA)based on the serological specific antibodies detection and rapidPCR amplifying targeted genes of rickettsiae. Methods: An EQA program to evaluate the following laboratory procedures was developed to detect rickettsiae: (1)immunofluorescent assay (IFA) to detect specific antibodies ofA. phagocytophilum, R. heilongjiangensis andO. tsutsugamshi respectively.(2) Two sets of nested PCRwere used amplifying groEL genes for most members of the familyRickettsiaceae and amplifying16SrRNA genes for the most members of familyanaplasmae, respectively. A scoring scheme based on the distribution of the median antibody titer values of the serologic assays was designed and a ranking list of the scores of the PCRresults based on the detected minimal copy numbers of referenceDNA was created.Results: Among nine laboratories who reported the results on time, eight laboratories gave acceptable serologic results, the other one provided an unacceptable antibody titer (1:2 vs median1:64) results forO. tsutsugamshi. The limits of detection(LOD)for thePCR amplifying for five referencesDNA ranged from 1copy/μL to106 copy/μL.Conclusions:We successfully trained and popularized modern diagnostic methods of rickettsiae in10 provincialCDCs in China and first conducted the EQA projects and evaluated the results.

  16. Seroprevalences of vector-transmitted infections of small-holder dairy cattle in coastal Kenya.

    Science.gov (United States)

    Maloo, S H; Thorpe, W; Kioo, G; Ngumi, P; Rowlands, G J; Perry, B D

    2001-11-01

    A cross-sectional study was carried out from July to September 1989 in Kaloleni Division, Coast Province, Kenya to estimate the prevalence of vector-transmitted diseases in small-holder dairy cattle and to identify the risk factors associated with different management systems. One hundred and thirty of the 157 herds with dairy cattle in Kaloleni Division were surveyed. These were from three agro-ecological zones (coconut-cassava, cashew nut-cassava and livestock-millet), comprised two management systems (stall-feeding and herded grazing) and were herds with either dairy cattle only or with Zebu and dairy cattle. A formal questionnaire sought answers to questions on cattle health and management practices. A total of 734 dairy and 205 Zebu cattle in 78 dairy and 52 mixed (dairy and Zebu) herds were sampled and screened for haemoparasites (Trypanosoma, Anaplasma, Babesia, and Theileria infections). Sera were tested for antibodies to Theileria parva, using the schizonts-antigen indirect fluorescent-antibody (IFA) test and to antibodies for Babesia bigemina and antigens to Anaplasma marginale by enzyme-linked immunosorbent assay (ELISA). Packed-cell volume (PCV) also was measured. Tick-control measures were practised by all except three of the farmers. Despite this, overall seroprevalence to T. parva was >70%--suggesting either that control practices were not strictly implemented or they were ineffective. The seroprevalence of T. parva in adult cattle kept in stall-feeding systems in the coconut-cassava zone was significantly lower (57+/-8% (S.E.)) than in herded-grazing systems (79+/-3%) and there was no association between antibody prevalence and age of cattle in this zone. Antibody prevalences in cattle in the cashew nut-cassava and the drier livestock-millet zone increased with age. Cattle in herded-grazing systems had an overall lower seroprevalence of T. parva infection in the livestock-millet zone (45+/-6%) than in the other two zones. Analysis was confined to

  17. Identification of vector-borne pathogens in dogs and cats from Southern Brazil.

    Science.gov (United States)

    Malheiros, J; Costa, M M; do Amaral, R B; de Sousa, K C M; André, M R; Machado, R Z; Vieira, M I B

    2016-07-01

    Dogs and cats are often infected with vector-borne pathogens and play a crucial role as reservoirs and hosts in their life cycles. The aim of the present study was to investigate the occurrence of vector-borne pathogens among dogs and cats in the northwestern region of Rio Grande do Sul (RS) State, Brazil. One hundred and ten blood samples were collected from dogs (n=80) and cats (n=30). Laboratory analysis were carried out through stained blood smears, indirect enzyme-linked immunosorbent assay (ELISA) for Babesia vogeli and Ehrlichia canis (only for dogs) and polymerase chain reaction (PCR) aiming the detection of pathogens. The following pathogens were screened by PCR among dogs and cats: Babesia spp. and Hepatozoon spp. (18S rRNA gene), Anaplasma spp. (16S rRNA gene), and Ehrlichia spp. (dsb gene for dogs and 16S rRNA gene for cats) and Bartonella spp. (nuoG gene only for cats). Using blood smears structures morphologically compatible with piroplasms were found in 5.45% (6/110) of the samples. Anti-B. vogeli and anti-E. canis antibodies were detected in 91% (73/80) and 9% (7/80) of the dogs, respectively. All the seropositive dogs to E. canis were also to B. vogeli. Nineteen (17.3%) animals were positive to hemoparasites by PCR. After sequencing Rangelia vitalii 6/80 (7.5%), B. vogeli 3/80 (4%), Hepatozoon spp. 1/80 (1%), and Anaplasma spp. 1/80 (1%) were found in the dogs, and B. vogeli 2/30 (7%) and Bartonella spp. 6/30 (20%) were detected in the screened cats. No sample was positive for genes dsb and 16S rRNA of Ehrlichia spp. Only those animals which were positive for R. vitalii showed findings compatible with rangeliosis, such as anemia (100%), thrombocytopenia (67%), jaundice (50%), external bleeding (50%), and anorexia (50%). This is the first time that B. vogeli detected among cats in Southern Brazil. PMID:27266811

  18. 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.

  19. 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.

  20. 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.

  1. Cholesterol binding by the bacterial type III translocon is essential for virulence effector delivery into mammalian cells.

    Science.gov (United States)

    Hayward, Richard D; Cain, Robert J; McGhie, Emma J; Phillips, Neil; Garner, Matthew J; Koronakis, Vassilis

    2005-05-01

    A ubiquitous early step in infection of man and animals by enteric bacterial pathogens like Salmonella, Shigella and enteropathogenic Escherichia coli (EPEC) is the translocation of virulence effector proteins into mammalian cells via specialized type III secretion systems (TTSSs). Translocated effectors subvert the host cytoskeleton and stimulate signalling to promote bacterial internalization or survival. Target cell plasma membrane cholesterol is central to pathogen-host cross-talk, but the precise nature of its critical contribution remains unknown. Using in vitro cholesterol-binding assays, we demonstrate that Salmonella (SipB) and Shigella (IpaB) TTSS translocon components bind cholesterol with high affinity. Direct visualization of cell-associated fluorescently labelled SipB and parallel immunogold transmission electron microscopy revealed that cholesterol levels limit both the amount and distribution of plasma membrane-integrated translocon. Correspondingly, cholesterol depletion blocked effector translocation into cultured mammalian cells by not only the related Salmonella and Shigella TTSSs, but also the more divergent EPEC system. The data reveal that cholesterol-dependent association of the bacterial TTSS translocon with the target cell plasma membrane is essential for translocon activation and effector delivery into mammalian cells.

  2. Listeria monocytogenes listeriolysin O and phosphatidylinositol-specific phospholipase C affect adherence to epithelial cells.

    Science.gov (United States)

    Krawczyk-Balska, Agata; Bielecki, Jacek

    2005-09-01

    Listeria monocytogenes, a foodborn intracellular animal and human pathogen, produces several exotoxins contributing to virulence. Among these are listeriolysin O (LLO), a pore-forming cholesterol-dependent hemolysin, and a phosphatidylinositol-specific phospholipase C (PI-PLC). LLO is known to play an important role in the escape of bacteria from the primary phagocytic vacuole of macrophages, and PI-PLC supports this process. Evidence is accumulating that LLO and PI-PLC are multifunctional virulence factors with many important roles in the host-parasite interaction other than phagosomal membrane disruption. LLO and PI-PLC may induce a number of host cell responses by modulating signal transduction of infected cells via intracellular Ca2+ levels and the metabolism of phospholipids. This would result in the activation of host phospholipase C and protein kinase C. In the present study, using Bacillus sub tilis strains expressing LLO, PI-PLC, and simultaneously LLO and PI-PLC, we show that LLO and PI-PLC enhance bacterial binding to epithelial cells Int407, with LLO being necessary and PI-PLC playing an accessory role. The results of this work suggest that these two listerial proteins act on epithelial cells prior to internalization. PMID:16391652

  3. Binding and Endocytosis of Bovine Hololactoferrin by the Parasite Entamoeba histolytica

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    Guillermo Ortíz-Estrada

    2015-01-01

    Full Text Available Entamoeba histolytica is a human parasite that requires iron (Fe for its metabolic function and virulence. Bovine lactoferrin (B-Lf and its peptides can be found in the digestive tract after dairy products are ingested. The aim of this study was to compare virulent trophozoites recently isolated from hamster liver abscesses with nonvirulent trophozoites maintained for more than 30 years in cultures in vitro regarding their interaction with iron-charged B-Lf (B-holo-Lf. We performed growth kinetics analyses of trophozoites in B-holo-Lf and throughout several consecutive transfers. The virulent parasites showed higher growth and tolerance to iron than nonvirulent parasites. Both amoeba variants specifically bound B-holo-Lf with a similar Kd. However, averages of 9.45 × 105 and 6.65 × 106 binding sites/cell were found for B-holo-Lf in nonvirulent and virulent amoebae, respectively. Virulent amoebae bound more efficiently to human and bovine holo-Lf, human holo-transferrin, and human and bovine hemoglobin than nonvirulent amoebae. Virulent amoebae showed two types of B-holo-Lf binding proteins. Although both amoebae endocytosed this glycoprotein through clathrin-coated vesicles, the virulent amoebae also endocytosed B-holo-Lf through a cholesterol-dependent mechanism. Both amoeba variants secreted cysteine proteases cleaving B-holo-Lf. These data demonstrate that the B-Lf endocytosis is more efficient in virulent amoebae.

  4. Modeling Signal Transduction and Lipid Rafts in Immune Cells

    Science.gov (United States)

    Prasad, Ashok

    2011-03-01

    Experimental evidence increasingly suggests that lipid rafts are nanometer sized cholesterol dependent dynamic assemblies enriched in sphingolipids and associated proteins. Lipid rafts are dynamic structures that break-up and reform on a relatively short time-scale, and are believed to facilitate the interactions of raft-associated proteins. The role of these rafts in signaling has been controversial, partly due to controversies regarding the existence and nature of the rafts themselves. Experimental evidence has indicated that in several cell types, especially T cells, rafts do influence signal transduction and T cell activation. Given the emerging consensus on the biophysical character of lipid rafts, the question can be asked as to what roles they possibly play in signal transduction. Here we carry out simulations of minimal models of the signal transduction network that regulates Src-family kinase dynamics in T cells and other cell types. By separately treating raft-based biochemical interactions, we find that rafts can indeed putatively play an important role in signal transduction, and in particular may affect the sensitivity of signal transduction. This illuminates possible functional consequences of membrane heterogeneities on signal transduction and points towards mechanisms for spatial control of signaling by cells.

  5. Effects of Suilysin on Streptococcus suis-Induced Platelet Aggregation

    Science.gov (United States)

    Zhang, Shengwei; Wang, Junping; Chen, Shaolong; Yin, Jiye; Pan, Zhiyuan; Liu, Keke; Li, Lin; Zheng, Yuling; Yuan, Yuan; Jiang, Yongqiang

    2016-01-01

    Blood platelets play important roles during pathological thrombocytopenia in streptococcal toxic shock syndrome (STSS). Streptococcus suis (S. suis) an emerging human pathogen, can cause STSS similarly to S. pyogenes. However, S. suis interactions with platelets are poorly understood. Here, we found that suilysin (SLY), different from other bacterial cholesterol-dependent cytolysins (CDCs), was the sole stimulus that induced platelet aggregation. Furthermore, the inside-out activation of GPIIb/IIIa of platelets mediated SLY-induced platelet aggregation. This process was triggered by Ca2+ influx that depend on the pore forming on platelets by SLY. Additionally, although SLY induced α-granule release occurred via the MLCK-dependent pathway, PLC-β-IP3/DAG-MLCK and Rho-ROCK-MLCK signaling were not involved in SLY-induced platelet aggregation. Interestingly, the pore dependent Ca2+ influx was also found to participate in the induction of platelet aggregation with pneumolysin (PLY) and streptolysin O (SLO), two other CDCs. It is possible that the CDC-mediated platelet aggregation we observed in S. suis is a similar response mechanism to that used by a wide range of bacteria. These findings might lead to the discovery of potential therapeutic targets for S. suis-associated STSS. PMID:27800304

  6. The Pore-Forming Haemolysins of Bacillus Cereus: A Review

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    Vincent Sanchis

    2013-06-01

    Full Text Available The Bacillus cereus sensu lato group contains diverse Gram-positive spore-forming bacteria that can cause gastrointestinal diseases and severe eye infections in humans. They have also been incriminated in a multitude of other severe, and frequently fatal, clinical infections, such as osteomyelitis, septicaemia, pneumonia, liver abscess and meningitis, particularly in immuno-compromised patients and preterm neonates. The pathogenic properties of this organism are mediated by the synergistic effects of a number of virulence products that promote intestinal cell destruction and/or resistance to the host immune system. This review focuses on the pore-forming haemolysins produced by B. cereus: haemolysin I (cereolysin O, haemolysin II, haemolysin III and haemolysin IV (CytK. Haemolysin I belongs to the cholesterol-dependent cytolysin (CDC family whose best known members are listeriolysin O and perfringolysin O, produced by L. monocytogenes and C. perfringens respectively. HlyII and CytK are oligomeric ß-barrel pore-forming toxins related to the α-toxin of S. aureus or the ß-toxin of C. perfringens. The structure of haemolysin III, the least characterized haemolytic toxin from the B. cereus, group has not yet been determined.

  7. Dispatched and Scube Mediate the Efficient Secretion of the Cholesterol-Modified Hedgehog Ligand

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    Hanna Tukachinsky

    2012-08-01

    Full Text Available The Hedgehog (Hh signaling pathway plays critical roles in metazoan development and in cancer. How the Hh ligand is secreted and spreads to distant cells is unclear, given its covalent modification with a hydrophobic cholesterol molecule, which makes it stick to membranes. We demonstrate that Hh ligand secretion from vertebrate cells is accomplished via two distinct and synergistic cholesterol-dependent binding events, mediated by two proteins that are essential for vertebrate Hh signaling: the membrane protein Dispatched (Disp and a member of the Scube family of secreted proteins. Cholesterol modification is sufficient for a heterologous protein to interact with Scube and to be secreted in a Scube-dependent manner. Disp and Scube recognize different structural aspects of cholesterol similarly to how Niemann-Pick disease proteins 1 and 2 interact with cholesterol, suggesting a hand-off mechanism for transferring Hh from Disp to Scube. Thus, Disp and Scube cooperate to dramatically enhance the secretion and solubility of the cholesterol-modified Hh ligand.

  8. An isoprenylation and palmitoylation motif promotes intraluminal vesicle delivery of proteins in cells from distant species.

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    Clara L Oeste

    Full Text Available The C-terminal ends of small GTPases contain hypervariable sequences which may be posttranslationally modified by defined lipid moieties. The diverse structural motifs generated direct proteins towards specific cellular membranes or organelles. However, knowledge on the factors that determine these selective associations is limited. Here we show, using advanced microscopy, that the isoprenylation and palmitoylation motif of human RhoB (-CINCCKVL targets chimeric proteins to intraluminal vesicles of endolysosomes in human cells, displaying preferential co-localization with components of the late endocytic pathway. Moreover, this distribution is conserved in distant species, including cells from amphibians, insects and fungi. Blocking lipidic modifications results in accumulation of CINCCKVL chimeras in the cytosol, from where they can reach endolysosomes upon release of this block. Remarkably, CINCCKVL constructs are sorted to intraluminal vesicles in a cholesterol-dependent process. In the lower species, neither the C-terminal sequence of RhoB, nor the endosomal distribution of its homologs are conserved; in spite of this, CINCCKVL constructs also reach endolysosomes in Xenopus laevis and insect cells. Strikingly, this behavior is prominent in the filamentous ascomycete fungus Aspergillus nidulans, in which GFP-CINCCKVL is sorted into endosomes and vacuoles in a lipidation-dependent manner and allows monitoring endosomal movement in live fungi. In summary, the isoprenylated and palmitoylated CINCCKVL sequence constitutes a specific structure which delineates an endolysosomal sorting strategy operative in phylogenetically diverse organisms.

  9. Arcanobacterium pyogenes: Virulence factors, importance in mastitis etiology and therapeutic (impossibilities

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    Milanov Dubravka

    2011-01-01

    Full Text Available Arcanobacterium pyogenes is an opportunistic pathogen, a causative agent of suppurative infections of organs and tissues in economically important livestock species. Most frequently this bacteria is isolated from inflamed lung lesions in pigs and cattle, in samples of uterine mucus of cows with endometritis and milk from cows with clinical mastitis. A. pyogenes possesses a number of virulence factors: cholesterol-dependent cytolysin (pyolysin, two neuraminidases, several proteases, extracellular matrix-binding proteins, DNases, fimbriae. The virulence factors are well studied in laboratory conditions, but the role of these factors in the pathogenesis of A. pyogenes infections remains to be elucidated. Lately, the ability of A. pyogenes to form biofilm in vivo has also been implicated as a virulence factor and a possible cause of therapeutic failure. Despite the fact that A. pyogenes milk isolates in cows with mastitis in vitro are very sensitive to β-lactam drugs and tetracycline, experience has shown that therapy is usually ineffective, prognosis is poor and the affected quarter is lost for milk production.

  10. Helical crystallization on nickel-lipid nanotubes: perfringolysin O as a model protein.

    Science.gov (United States)

    Dang, Thanh X; Milligan, Ronald A; Tweten, Rodney K; Wilson-Kubalek, Elizabeth M

    2005-11-01

    To facilitate purification and subsequent structural studies of recombinant proteins the most widely used genetically encoded tag is the histidine tag (His-tag) which specifically binds to N-nitrilotriacetic-acid-chelated nickel ions. Lipids derivatized with a nickel-chelating head group can be mixed with galactosylceramide glycolipids to prepare lipid nanotubes that bind His-tagged proteins. In this study, we use His-tagged perfringolysin O (PFO), a soluble toxin secreted by the bacterial pathogen Clostridium perfringens, as a model protein to test the utility of nickel-lipid nanotubes as a tool for structural studies of His-tagged proteins. PFO is a member of the cholesterol dependent cytolysin family (CDC) of oligomerizing, pore-forming toxins found in a variety of Gram-positive bacterial pathogens. CDC pores have been difficult to study by X-ray crystallography because they are membrane associated and vary in size. We demonstrate that both a wild-type and a mutant form of PFO form helical arrays on nickel-lipid containing nanotubes. Cryo-electron microscopy and image analysis of the helical arrays were used to reconstruct a 3D density map of wild-type PFO. This study suggests that the use of nickel-lipid nanotubes may offer a general approach for structural studies of recombinant proteins and may provide insights into the molecular interactions of proteins that have a natural affinity for a membrane surface.

  11. Characterization of endocytosis and exocytosis of cationic nanoparticles in airway epithelium cells

    Energy Technology Data Exchange (ETDEWEB)

    Dombu, Christophe Youta; Kroubi, Maya; Zibouche, Rima; Matran, Regis; Betbeder, Didier, E-mail: dbetbeder@aol.com [EA 4483, IFR 114, Laboratoire de Physiologie, Faculte de Medecine Pole Recherche, Universite de Lille 2, 1 place de Verdun, 59045 Lille Cedex (France)

    2010-09-03

    A major challenge of drug delivery using colloids via the airway is to understand the mechanism implied in their interactions with epithelial cells. The purpose of this work was to characterize the process of endocytosis and exocytosis of cationic nanoparticles (NPs) made of maltodextrin which were developed as a delivery system for antigens in vaccine applications. Confocal microscopy demonstrated that these NP are rapidly endocytosed after as little as 3 min incubation, and that the endocytosis was also faster than NP binding since most of the NPs were found in the middle of the cells around the nuclei. A saturation limit was observed after a 40 min incubation, probably due to an equilibrium becoming established between endocytosis and exocytosis. Endocytosis was dramatically reduced at 4 deg. C compared with 37 deg. C, or by NaN{sub 3} treatment, both results suggesting an energy dependent process. Protamine pretreatment of the cells inhibited NPs uptake and we found that clathrin pathway is implied in their endocytosis. Cholesterol depletion increased NP uptake by 300% and this phenomenon was explained by the fact that cholesterol depletion totally blocked NP exocytosis. These results suggest that these cationic NPs interact with anionic sites, are quickly endocytosed via the clathrin pathway and that their exocytosis is cholesterol dependent, and are similar to those obtained in other studies with viruses such as influenza.

  12. Structure of the poly-C9 component of the complement membrane attack complex

    Science.gov (United States)

    Dudkina, Natalya V.; Spicer, Bradley A.; Reboul, Cyril F.; Conroy, Paul J.; Lukoyanova, Natalya; Elmlund, Hans; Law, Ruby H. P.; Ekkel, Susan M.; Kondos, Stephanie C.; Goode, Robert J. A.; Ramm, Georg; Whisstock, James C.; Saibil, Helen R.; Dunstone, Michelle A.

    2016-02-01

    The membrane attack complex (MAC)/perforin-like protein complement component 9 (C9) is the major component of the MAC, a multi-protein complex that forms pores in the membrane of target pathogens. In contrast to homologous proteins such as perforin and the cholesterol-dependent cytolysins (CDCs), all of which require the membrane for oligomerisation, C9 assembles directly onto the nascent MAC from solution. However, the molecular mechanism of MAC assembly remains to be understood. Here we present the 8 Å cryo-EM structure of a soluble form of the poly-C9 component of the MAC. These data reveal a 22-fold symmetrical arrangement of C9 molecules that yield an 88-strand pore-forming β-barrel. The N-terminal thrombospondin-1 (TSP1) domain forms an unexpectedly extensive part of the oligomerisation interface, thus likely facilitating solution-based assembly. These TSP1 interactions may also explain how additional C9 subunits can be recruited to the growing MAC subsequent to membrane insertion.

  13. Structural basis of complement membrane attack complex formation

    Science.gov (United States)

    Serna, Marina; Giles, Joanna L.; Morgan, B. Paul; Bubeck, Doryen

    2016-02-01

    In response to complement activation, the membrane attack complex (MAC) assembles from fluid-phase proteins to form pores in lipid bilayers. MAC directly lyses pathogens by a `multi-hit' mechanism; however, sublytic MAC pores on host cells activate signalling pathways. Previous studies have described the structures of individual MAC components and subcomplexes; however, the molecular details of its assembly and mechanism of action remain unresolved. Here we report the electron cryo-microscopy structure of human MAC at subnanometre resolution. Structural analyses define the stoichiometry of the complete pore and identify a network of interaction interfaces that determine its assembly mechanism. MAC adopts a `split-washer' configuration, in contrast to the predicted closed ring observed for perforin and cholesterol-dependent cytolysins. Assembly precursors partially penetrate the lipid bilayer, resulting in an irregular β-barrel pore. Our results demonstrate how differences in symmetric and asymmetric components of the MAC underpin a molecular basis for pore formation and suggest a mechanism of action that extends beyond membrane penetration.

  14. Mechanisms linking bacterial infections of the bovine endometrium to disease and infertility.

    Science.gov (United States)

    Carneiro, Luísa Cunha; Cronin, James Graham; Sheldon, Iain Martin

    2016-03-01

    Bacterial infections of the endometrium after parturition commonly cause metritis and endometritis in dairy cattle, and these diseases are important because they compromise animal welfare and incur economic costs, as well as delaying or preventing conception. Here we highlight that uterine infections cause infertility, discuss which bacteria cause uterine disease, and review the evidence for mechanisms of inflammation and tissue damage in the endometrium. Bacteria cultured from the uterus of diseased animals include Escherichia coli, Trueperella pyogenes, and several anaerobic species, but their causative role in disease is challenged by the discovery of many other bacteria in the uterine disease microbiome. Irrespective of the species of bacteria, endometrial cell inflammatory responses to infection initially depend on innate immunity, with Toll-like receptors binding pathogen-associated molecular patterns, such as lipopolysaccharide and bacterial lipopeptides. In addition to tissue damage associated with parturition and inflammation, endometrial cell death is caused by a cholesterol-dependent cytolysin secreted by T. pyogenes, called pyolysin, which forms pores in plasma membranes of endometrial cells. However, endometrial cells surprisingly do not sense damage-associated molecular patterns, but a combination of infections followed by cell damage leads to release of the intracellular cytokine interleukin (IL)-1 alpha from endometrial cells, which then acts to scale inflammatory responses. To develop strategies to limit the impact of uterine disease on fertility, future work should focus on determining which bacteria and virulence factors cause endometritis, and understanding how the host response to infection is regulated in the endometrium. PMID:26952747

  15. Perfringolysin O: The Underrated Clostridium perfringens Toxin?

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    Stefanie Verherstraeten

    2015-05-01

    Full Text Available The anaerobic bacterium Clostridium perfringens expresses multiple toxins that promote disease development in both humans and animals. One such toxin is perfringolysin O (PFO, classically referred to as θ toxin, a pore-forming cholesterol-dependent cytolysin (CDC. PFO is secreted as a water-soluble monomer that recognizes and binds membranes via cholesterol. Membrane-bound monomers undergo structural changes that culminate in the formation of an oligomerized prepore complex on the membrane surface. The prepore then undergoes conversion into the bilayer-spanning pore measuring approximately 250–300 Å in diameter. PFO is expressed in nearly all identified C. perfringens strains and harbors interesting traits that suggest a potential undefined role for PFO in disease development. Research has demonstrated a role for PFO in gas gangrene progression and bovine necrohemorrhagic enteritis, but there is limited data available to determine if PFO also functions in additional disease presentations caused by C. perfringens. This review summarizes the known structural and functional characteristics of PFO, while highlighting recent insights into the potential contributions of PFO to disease pathogenesis.

  16. An integrative approach to understanding pyrethroid resistance in Rhipicephalus microplus and R. decoloratus ticks.

    Science.gov (United States)

    Wyk, Roelof Dj van; Baron, Samantha; Maritz-Olivier, Christine

    2016-06-01

    Rhipicephalus microplus and Rhipicephalus decoloratus species occur in regions with savannah and temperate climates, typically in grassland and wooded areas used as cattle pasture. Both species are associated with the transmission of Anaplasma and Babesia spp., impacting livestock health and quality of livestock-associated products. In Africa, tick control is predominantly mediated with the use of acaricides, such as synthetic pyrethroids. After several years on the market, reports of resistance to synthetic pyrethroids escalated but limited field data and validation studies have been conducted to determine the extent of acaricide resistance in Africa. Without this data, knowledge-based tick control will remain problematic and selection pressure will remain high increasing the rate of resistance acquisition. To date, several pyrethroid resistance associated single nucleotide polymorphisms (SNPs) have been reported for arthropods within the voltage-gated sodium channel. Three SNPs have been identified within this channel in pyrethroid resistant R. microplus ticks, but none has been reported for R. decoloratus. This study is the first to report the presence of a shared SNP within the voltage-gated sodium channel in both R. microplus and R. decoloratus, which is directly linked to pyrethroid resistance in R. microplus. As the mode of action by which these SNPs mediate pyrethroid resistance remains unknown, this study aims to set hypotheses by means of predictive structural modelling. This not only paves the way forward to elucidating the underlying biological mechanisms involved in pyrethroid resistance, but also improvement of existing acaricides and ultimately sustainable tick control management. PMID:26851840

  17. Pharmacological characterization of a tyramine receptor from the southern cattle tick, Rhipicephalus (Boophilus) microplus.

    Science.gov (United States)

    Gross, Aaron D; Temeyer, Kevin B; Day, Tim A; Pérez de León, Adalberto A; Kimber, Michael J; Coats, Joel R

    2015-08-01

    The southern cattle tick (Rhipicephalus (Boophilus) microplus) is a hematophagous external parasite that vectors the causative agents of bovine babesiosis or cattle tick fever, Babesia bovis and B. bigemina, and anaplasmosis, Anaplasma marginale. The southern cattle tick is a threat to the livestock industry in many locations throughout the world. Control methods include the use of chemical acaricides including amitraz, a formamidine insecticide, which is proposed to activate octopamine receptors. Previous studies have identified a putative octopamine receptor from the southern cattle tick in Australia and the Americas. Furthermore, this putative octopamine receptor could play a role in acaricide resistance to amitraz. Recently, sequence data indicated that this putative octopamine receptor is probably a type-1 tyramine receptor (TAR1). In this study, the putative TAR1 was heterologously expressed in Chinese hamster ovary (CHO-K1) cells, and the expressed receptor resulted in a 39-fold higher potency for tyramine compared to octopamine. Furthermore, the expressed receptor was strongly antagonized by yohimbine and cyproheptadine, and mildly antagonized by mianserin and phentolamine. Tolazoline and naphazoline had agonistic or modulatory activity against the expressed receptor, as did the amitraz metabolite, BTS-27271; however, this was only observed in the presence of tyramine. The southern cattle tick's tyramine receptor may serve as a target for the development of anti-parasitic compounds, in addition to being a likely target of formamidine insecticides. PMID:25958152

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

    Science.gov (United States)

    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

  19. 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.

  20. 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

  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. Molecular evidence of Ehrlichia canis and Rickettsia massiliae in ixodid ticks of carnivores from South Hungary.

    Science.gov (United States)

    Hornok, Sándor; Fuente, José; Horváth, Gábor; Fernández de Mera, Isabel G; Wijnveld, Michiel; Tánczos, Balázs; Farkas, Róbert; Jongejan, Frans

    2013-03-01

    To monitor the emergence of thermophilic, Mediterranean ixodid tick species and tick-borne pathogens in southern Hungary, 348 ticks were collected from shepherd dogs, red foxes and golden jackals during the summer of 2011. Golden jackals shared tick species with both the dog and the red fox in the region. Dermacentor nymphs were collected exclusively from dogs, and the sequence identification of these ticks indicated that dogs are preferred hosts of both D. reticulatus and D. marginatus nymphs, unlike previously reported. Subadults of three ixodid species were selected for reverse line blot hybridisation (RLB) analysis to screen their vector potential for 40 pathogens/groups. Results were negative for Anaplasma, Babesia and Theileria spp. Investigation of D. marginatus nymphs revealed the presence of Ehrlichia canis, Rickettsia massiliae and Borrelia afzelii for the first time in this tick species. These findings broaden the range of those tick-borne agents, which are typically transmitted by Rhipicephalus sanguineus, but may also have Dermacentor spp. as potential or alternative vectors. Ehrlichiacanis was also newly detected in Ixodes canisuga larvae from red foxes. In absence of transovarial transmission in ticks this implies that Eurasian red foxes may play a reservoir role in the epidemiology of canine ehrlichiosis. PMID:23439290

  3. 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.

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

    Science.gov (United States)

    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.

  5. 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.

  6. 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).

  7. 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.

  8. 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.

  9. 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.

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

    Directory of Open Access Journals (Sweden)

    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.

  11. Transport of Ixodid ticks and tick-borne pathogens by migratory birds.

    Directory of Open Access Journals (Sweden)

    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.

  12. Prevalence of parasitic infections in cattle of Ludhiana district, Punjab.

    Science.gov (United States)

    Singh, N K; Singh, Harkirat; Jyoti; Haque, M; Rath, S S

    2012-10-01

    A total of 862 cattle were sampled for both haematological (703) and coprological (159) investigations at the Large Animal Clinics, GADVASU, Ludhiana, Punjab, India. Examination of Giemsa-stained peripheral blood smears exhibited that 22.9 % (161/703) of cattle were infected with haematozoa comprising Theileria annulata (14.65 %), Trypanosoma evansi (0.28 %), Babesia bigemina (1.56 %) and Anaplasma marginale (8.53 %) while mixed infection appeared in 2.13 % (15/703) animals. The prevalence of total haemoparasites and A. marginale infections were significantly higher (p Balantidium coli, strongyles and Trichuris spp. were detected from 3.77, 1.88, 3.77, 2.52, 10.69 and 1.26 % of examined animals. Except coccidiosis, there was no significant variation of GI parasitic infections in relation to sex as Eimeria spp. were found higher (p < 0.01) in males. The prevalence of Trichuris spp. was significantly higher (p < 0.05) in younger animals <6 months of age. The present work emphasized that strongyles and T. annulata were the most prevalent GI and haemoparasites, respectively. PMID:24082538

  13. Nucleic acid probes as a diagnostic method for tick-borne hemoparasites of veterinary importance.

    Science.gov (United States)

    Figueroa, J V; Buening, G M

    1995-03-01

    An increased number of articles on the use of nucleic acid-based hybridization techniques for diagnostic purposes have been recently published. This article reviews nucleic acid-based hybridization as an assay to detect hemoparasite infections of economic relevance in veterinary medicine. By using recombinant DNA techniques, selected clones containing inserts of Anaplasma, Babesia, Cowdria or Theileria genomic DNA sequences have been obtained, and they are now available to be utilized as specific, highly sensitive DNA or RNA probes to detect the presence of the hemoparasite DNA in an infected animal. Either in an isotopic or non-isotopic detection system, probes have allowed scientists to test for--originally in samples collected from experimentally infected animals and later in samples collected in the field--the presence of hemoparasites during the prepatent, patent, convalescent, and chronic periods of the infection in the host. Nucleic acid probes have given researchers the opportunity to carry out genomic analysis of parasite DNA to differentiate hemoparasite species and to identify genetically distinct populations among and within isolates, strains and clonal populations. Prevalence of parasite infection in the tick vector can now be accomplished more specifically with the nucleic acid probes. Lately, with the advent of the polymerase chain reaction technique, small numbers of hemoparasites can be positively identified in the vertebrate host and tick vector. These techniques can be used to assess the veterinary epidemiological situation in a particular geographical region for the planning of control measures. PMID:7597795

  14. Enzyme-Linked Immunosorbent Assay (Elisa Based Detection of Antibodies to Mycoplasma bovis in Cattle Naturally Infected with Haemoparasites in Institutional farms in Sokoto State, Nigeria

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    F.M. Tambuwal

    2011-01-01

    Full Text Available This was a cross-sectional study involving cattle from four (4 institutional farms (Prison farm, Livestock Investigation and Breeding Centre (LIBC, Usmanu Danfodiyo University Teaching and Research (UDUTRF and Kebbe Cattle Ranch (KCR in Sokoto state, Nigeria. A total of 62 cattle comprising 49 females and 13 males were randomly selected and bled from a total population of 205. The cattle sampled were local breeds comprising Gudali, Rahaji, White-Fulani and their crosses. They were aged 1-10 years and are managed semi-intensively. The enzyme-linked immunosorbent assay was used for the detection M. bovis antibody. Of the 62 cattle screened, M. bovis antibody was detected in 41(66%. Also, 24 out of the 41 M. bovis positive cattle were found infected with haemoparasites. Similarly, 11 out of the 21 serologically negative cattle were infected with one or more haemoparasites. Seven (17%, 3 (7.3% and 7 (17% of the M. bovis positive cattle were infected with Babesia bigemina, Anaplasma marginale, and or B. bigemina and A. marginale, respectively. In the overall, 27 of the 62 screened cattle were infected with one of blood parasites or a combination of both. However, there is no significant statistical relationship (p> 0.05 between the number of cattle positive for M. bovis and the presence of haemoparasites among the examined cattle.

  15. New records and human parasitism by Ornithodoros mimon (Acari: Argasidae) in Brazil.

    Science.gov (United States)

    Labruna, Marcelo B; Marcili, Arlei; Ogrzewalska, Maria; Barros-Battesti, Darci M; Dantas-Torres, Filipe; Fernandes, André A; Leite, Romario C; Venzal, Jose M

    2014-01-01

    The bat tick Ornithodoros mimon Kohls, Clifford & Jones is currently known by only few reports in Bolivia, Uruguay, Argentina, and the state of São Paulo in southeastern Brazil. Here, we expand the distribution of O. mimon in Brazil to the states of Minas Gerais (southeastern region), Goiás (central-western), Pernambuco, and Rio Grande do Norte (northeastern). Ticks were collected on human dwellings, where there had been repeated complains of tick bites on persons during the night. Tick bites were generally followed by intense inflammatory reactions that lasted for several weeks at the bite site. Bats and opossums were reported to inhabit the attic of the infested houses. In addition, a free-ranging opossum (Didelphis albiventris Lund) trapped in Rio Grande do Norte was found infested by argasid larvae. Based on morphological and/or molecular analysis, all ticks were identified as O. mimon. From one of the sites (Tiradentes, state of Minas Gerais), 20 field-collected nymphs were tested by a battery of polymerase chain reaction protocols targeting tick-borne microorganisms of the genera Babesia, Hepatozoon, Rickettsia, Borrelia, Anaplasma, Ehrlichia, and Coxiella; no tick specimen was found infected by any of these microorganism genera. The current study expands northwards the distribution of O. mimon, which has been shown to be very harmful to humans because of the intense inflammatory response that usually occurs after tick bites. PMID:24605480

  16. 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.

  17. 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.

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

    Science.gov (United States)

    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.

  19. Prevalence of Haemoprotozoan infections in bovines of Shimoga region of Karnataka state.

    Science.gov (United States)

    Krishna Murthy, C M; Ananda, K J; Adeppa, J

    2016-09-01

    A study was conducted to record the prevalence of Haemoprotozoan infections in bovines of Shimoga region for a period of 1 year from April 2012 to March 2013. A total of 300 blood samples were examined for the presence of haemoprotozoan parasites, of which 215 from cattle and 85 from buffaloes were examined by Giemsa staining technique. Out of 300 blood samples examined, 130 (43.3 %) were found positive for Haemoprotozoan infections. Out of 215 cattle blood samples examined, 62 (28.8 %) were positive for Theileria annulata, 27 (12.5 %) were harbored Babesia bigemina, 15 (6.9 %) were found positive for Trypanosoma evansi and 06 (2.7 %) samples showed Anaplasma marginale. Among 85 buffalo samples examined, 11 (12.9 %) were showed Theileria spp, 04 (4.7 %) found positive for B. bigemina, 03 (3.5 %) were found positive for T. evansi and 02 (2.3 %) were positive for A. marginale. Among haemoprotozoan parasites, the highest prevalence was observed with T. annulata followed by B. bigemina and T. evansi infection. The lowest prevalence was observed with A. marginale infection. PMID:27605804

  20. The bacterial microbiome of Dermacentor andersoni ticks influences pathogen susceptibility.

    Science.gov (United States)

    Gall, Cory A; Reif, Kathryn E; Scoles, Glen A; Mason, Kathleen L; Mousel, Michelle; Noh, Susan M; Brayton, Kelly A

    2016-08-01

    Ticks are of medical importance owing 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. Although ticks host pathogenic bacteria, they also harbor bacterial endosymbionts that have a role in tick physiology, survival, as well as pathogen acquisition and transmission. The goal of this study was to characterize the bacterial microbiome and examine the impact of microbiome disruption on pathogen susceptibility. The bacterial microbiome of two populations of D. andersoni with historically different susceptibilities to A. marginale was characterized. In this study, the microbiome was disrupted and then ticks were exposed to A. marginale or Francisella novicida to determine whether the microbiome correlated with pathogen susceptibility. Our study showed that an increase in proportion and quantity of Rickettsia bellii in the microbiome was negatively correlated to A. marginale levels in ticks. Furthermore, a decrease in Francisella endosymbionts was associated with lower F. novicida infection levels, demonstrating a positive pathogen-endosymbiont relationship. We demonstrate that endosymbionts and pathogens have varying interactions, and suggest that microbiome manipulation may provide a possible method for biocontrol by decreasing pathogen susceptibility of ticks. PMID:26882265

  1. 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.

  2. 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.

  3. Evaluation of an indirect ELISA for the diagnosis of Babesia bovis in Uruguay

    International Nuclear Information System (INIS)

    In initially establishing the FAO/IAEA indirect ELISA for the detection of antibodies to Babesia bovis, the optical density (OD) values of sera from known positive or negative local cattle were compared to the OD values obtained from the negative and positive reference sera provided with the ELISA kit. The percentage of false positive and negative sera were 2.53% and 2.97% respectively. The cut-off values for the negative reference serum in the kit were compared with those of a local negative population. These values were found to be similar. The specificity of the test was evaluated by testing 30 sera from animals experimentally infected with Anaplasma marginale and 30 sera from animals infected with Babesia bigemina. These were no cross-reaction either between A. marginale and B. bovis or between B. bigemina and B. bovis. A serological survey using this ELISA kit was carried out on animals from an enzootic area and an area free from the vecot Boophilus microplus. 53 out of 282 animals (18.8%) in the enzootic area were positive whilst all the animals (113) from the free area were negative. This study would indicate that the FAO/IAEA ELISA kit has a sensitivity of around 98% and specificity of 97%. (author). 8 refs, 2 figs, 2 tabs

  4. Tick and tick borne protozoan diseases of livestock in the selected hilly areas of Bangladesh

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    U.K. Mohanta

    2011-12-01

    Full Text Available To study the tick and tick borne protozoan diseases of livestock in the hilly areas of Bangladesh, an attempt was made to collect tick and blood samples from cattle, goat and gayal (Bos frontalis from different areas of the three hill districts. In this study, two species of ticks namely, Boophilus microplus (92% and Amblyomma testudinarium (21.6% and two species of blood protozoa like Babesia bigemina (16.63% and Anaplasma marginale (14.94% were recorded. Seasonal prevalence of ticks was highest in summer (97% in comparison to rainy (95% and winter (86% season. On the other hand, the seasonal prevalence of blood protozoa was highest in rainy season (45.45% in comparison to summer (27.87% and winter (16.55%. Again, animals aged more than 2 (two years of age (52% found to be more susceptible to blood protozoan diseases than animals aged between 1-2 years of age (33.97%. But none of the animals under one year of age were found to be infected with blood protozoan diseases.

  5. 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...

  6. Control of tick infestations in cattle vaccinated with bacterial membranes containing surface-exposed tick protective antigens.

    Science.gov (United States)

    Almazán, Consuelo; Moreno-Cantú, Orlando; Moreno-Cid, Juan A; Galindo, Ruth C; Canales, Mario; Villar, Margarita; de la Fuente, José

    2012-01-01

    Vaccines containing the Rhipicephalus (Boophilus) microplus BM86 and BM95 antigens protect cattle against tick infestations. Tick subolesin (SUB), elongation factor 1a (EF1a) and ubiquitin (UBQ) are new candidate protective antigens for the control of cattle tick infestations. Previous studies showed that R. microplus BM95 immunogenic peptides fused to the Anaplasma marginale major surface protein (MSP) 1a N-terminal region (BM95-MSP1a) for presentation on the Escherichia coli membrane were protective against R. microplus infestations in rabbits. In this study, we extended these results by expressing SUB-MSP1a, EF1a-MSP1a and UBQ-MSP1a fusion proteins on the E. coli membrane using this system and demonstrating that bacterial membranes containing the chimeric proteins BM95-MSP1a and SUB-MSP1a were protective (>60% vaccine efficacy) against experimental R. microplus and Rhipicephalus annulatus infestations in cattle. This system provides a novel, simple and cost-effective approach for the production of tick protective antigens by surface display of antigenic protein chimera on the E. coli membrane and demonstrates the possibility of using recombinant bacterial membrane fractions in vaccine preparations to protect cattle against tick infestations. PMID:22085549

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

    Directory of Open Access Journals (Sweden)

    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.

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

    Science.gov (United States)

    Alvåsen, Karin; Johansson, Sandra M; Höglund, Johan; Ssuna, Richard; Emanuelson, Ulf

    2016-01-01

    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. PMID:27543039

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

    Directory of Open Access Journals (Sweden)

    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.

  10. 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.

  11. Helicobacter pylori's cholesterol uptake impacts resistance to docosahexaenoic acid.

    Science.gov (United States)

    Correia, Marta; Casal, Susana; Vinagre, João; Seruca, Raquel; Figueiredo, Ceu; Touati, Eliette; Machado, José C

    2014-05-01

    Helicobacter pylori colonizes half of the world population and is associated with gastric cancer. We have previously demonstrated that docosahexaenoic acid (DHA), an n-3 polyunsaturated fatty acid known for its anti-inflammatory and antitumor effects, directly inhibits H. pylori growth in vitro and in mice. Nevertheless, the concentration of DHA shown to reduce H. pylori mice gastric colonization was ineffective in vitro. Related to the auxotrophy of H. pylori for cholesterol, we hypothesize that other mechanisms, in addition to DHA direct antibacterial effect, must be responsible for the reduction of the infection burden. In the present study we investigated if DHA affects also H. pylori growth, by reducing the availability of membrane cholesterol in the epithelial cell for H. pylori uptake. Levels of cholesterol in gastric epithelial cells and of cholesteryl glucosides in H. pylori were determined by thin layer chromatography and gas chromatography. The consequences of epithelial cells' cholesterol depletion on H. pylori growth were assessed in liquid cultures. We show that H. pylori uptakes cholesterol from epithelial cells. In addition, DHA lowers cholesterol levels in epithelial cells, decreases its de novo synthesis, leading to a lower synthesis of cholesteryl glucosides by H. pylori. A previous exposition of H. pylori to cholesterol influences the bacterium response to the direct inhibitory effect of DHA. Overall, our results suggest that a direct effect of DHA on H. pylori survival is modulated by its access to epithelial cell cholesterol, supporting the notion that cholesterol enhances the resistance of H. pylori. The cholesterol-dependent resistance of H. pylori to antimicrobial compounds raises new important aspects for the development of new anti-bacterial strategies. PMID:24447914

  12. Listeriolysin O is necessary and sufficient to induce autophagy during Listeria monocytogenes infection.

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    Nicole Meyer-Morse

    Full Text Available BACKGROUND: Recent studies have suggested that autophagy is utilized by cells as a protective mechanism against Listeria monocytogenes infection. METHODOLOGY/PRINCIPAL FINDINGS: However we find autophagy has no measurable role in vacuolar escape and intracellular growth in primary cultured bone marrow derived macrophages (BMDMs deficient for autophagy (atg5-/-. Nevertheless, we provide evidence that the pore forming activity of the cholesterol-dependent cytolysin listeriolysin O (LLO can induce autophagy subsequent to infection by L. monocytogenes. Infection of BMDMs with L. monocytogenes induced microtubule-associated protein light chain 3 (LC3 lipidation, consistent with autophagy activation, whereas a mutant lacking LLO did not. Infection of BMDMs that express LC3-GFP demonstrated that wild-type L. monocytogenes was encapsulated by LC3-GFP, consistent with autophagy activation, whereas a mutant lacking LLO was not. Bacillus subtilis expressing either LLO or a related cytolysin, perfringolysin O (PFO, induced LC3 colocalization and LC3 lipidation. Further, LLO-containing liposomes also recruited LC3-GFP, indicating that LLO was sufficient to induce targeted autophagy in the absence of infection. The role of autophagy had variable effects depending on the cell type assayed. In atg5-/- mouse embryonic fibroblasts, L. monocytogenes had a primary vacuole escape defect. However, the bacteria escaped and grew normally in atg5-/- BMDMs. CONCLUSIONS/SIGNIFICANCE: We propose that membrane damage, such as that caused by LLO, triggers bacterial-targeted autophagy, although autophagy does not affect the fate of wild-type intracellular L. monocytogenes in primary BMDMs.

  13. Intermedilysin induces EGR-1 expression through calcineurin/NFAT pathway in human cholangiocellular carcinoma cells

    Energy Technology Data Exchange (ETDEWEB)

    Susilowati, Heni [Department of Oral Microbiology, Institute of Health Biosciences, The University of Tokushima Graduate School, Tokushima 770-8504 (Japan); Okamura, Hirohiko [Department of Histology and Oral Histology, Institute of Health Biosciences, The University of Tokushima Graduate School, Tokushima 770-8504 (Japan); Hirota, Katsuhiko, E-mail: hirota@dent.tokushima-u.ac.jp [Department of Oral Microbiology, Institute of Health Biosciences, The University of Tokushima Graduate School, Tokushima 770-8504 (Japan); Shono, Masayuki [Support Center for Advanced Medical Sciences, The University of Tokushima, Tokushima 770-8504 (Japan); Yoshida, Kaya [Department of Fundamental Oral Health Science, School of Oral Health and Welfare, Institute of Health Biosciences, The University of Tokushima Graduate School, Tokushima 770-8504 (Japan); Murakami, Keiji [Department of Oral Microbiology, Institute of Health Biosciences, The University of Tokushima Graduate School, Tokushima 770-8504 (Japan); Tabata, Atsushi; Nagamune, Hideaki [Department of Biological Science and Technology, Life System, Institute of Technology and Science, The University of Tokushima Graduate School, Tokushima 770-8506 (Japan); Haneji, Tatsuji [Department of Histology and Oral Histology, Institute of Health Biosciences, The University of Tokushima Graduate School, Tokushima 770-8504 (Japan); Miyake, Yoichiro [Department of Oral Microbiology, Institute of Health Biosciences, The University of Tokushima Graduate School, Tokushima 770-8504 (Japan)

    2011-01-07

    Research highlights: {yields} ILY leads to the accumulation of [Ca{sup 2+}]i in the nucleus in HuCCT1 cells. {yields} ILY induced activation of NFAT1 through a calcineurin-dependent pathway. {yields} Calcineuri/NFAT pathway is involved in EGR-1 expression in response to ILY treatment. -- Abstract: Intermedilysin (ILY) is a cholesterol-dependent cytolysin produced by Streptococcus intermedius, which is associated with human brain and liver abscesses. Although intrahepatic bile duct cells play a valuable role in the pathogenesis of liver abscess, the molecular mechanism of ILY-treated intrahepatic bile duct cells remains unknown. In this study, we report that ILY induced a nuclear accumulation of intracellular calcium ([Ca{sup 2+}]i) in human cholangiocellular cells HuCCT1. We also demonstrate that 10 ng/ml ILY induced NFAT1 dephosphorylation and its nuclear translocation in HuCCT1 cells. In contrast to the result that ILY induced NF-{kappa}B translocation in human hepatic HepG2 cells, ILY did not affect NF-{kappa}B localization in HuCCT1 cells. Dephosphorylation and nuclear translocation of NFAT1 caused by ILY were prevented by [Ca{sup 2+}]i calcium chelator, BAPTA/AM, and calcineurin inhibitors, cyclosporine A and tacrolimus. ILY induced early growth response-1 (EGR-1) expression and it was inhibited by the pre-treatment with cyclosporine A, indicating that the calcineurin/NFAT pathway was involved in EGR-1 expression in response to ILY. ILY-induced calcineurin/NFAT1 activation and sequential EGR-1 expression might be related to the pathogenesis of S. intermedius in human bile duct cells.

  14. The impact of pneumolysin on the macrophage response to Streptococcus pneumoniae is strain-dependent.

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    Richard M Harvey

    Full Text Available Streptococcus pneumoniae is the world's leading cause of pneumonia, bacteremia, meningitis and otitis media. A major pneumococcal virulence factor is the cholesterol-dependent cytolysin, which has the defining property of forming pores in cholesterol-containing membranes. In recent times a clinically significant and internationally successful serotype 1 ST306 clone has been found to express a non-cytolytic variant of Ply (Ply306. However, while the pneumococcus is a naturally transformable organism, strains of the ST306 clonal group have to date been virtually impossible to transform, severely restricting efforts to understand the role of non-cytolytic Ply in the success of this clone. In this study isogenic Ply mutants were constructed in the D39 background and for the first time in the ST306 background (A0229467 to enable direct comparisons between Ply variants for their impact on the immune response in a macrophage-like cell line. Strains that expressed cytolytic Ply were found to induce a significant increase in IL-1β release from macrophage-like cells compared to the non-cytolytic and Ply-deficient strains in a background-independent manner, confirming the requirement for pore formation in the Ply-dependent activation of the NLRP3 inflammasome. However, cytolytic activity in the D39 background was found to induce increased expression of the genes encoding GM-CSF (CSF2, p19 subunit of IL-23 (IL23A and IFNβ (IFNB1 compared to non-cytolytic and Ply-deficient D39 mutants, but had no effect in the A0229467 background. The impact of Ply on the immune response to the pneumococcus is highly dependent on the strain background, thus emphasising the importance of the interaction between specific virulence factors and other components of the genetic background of this organism.

  15. Roles for lipid heterogeneity in immunoreceptor signaling.

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    Holowka, David; Baird, Barbara

    2016-08-01

    Immune receptors that specifically recognize foreign antigens to activate leukocytes in adaptive immune responses belong to a family of multichain cell surface proteins. All of these contain immunoreceptor tyrosine-based activation motifs in one or more subunits that initiate signaling cascades following stimulated tyrosine phosphorylation by Src-family kinases. As highlighted in this review, lipids participate in this initial activation step, as well as in more downstream signaling steps. We summarize evidence for cholesterol-dependent ordered lipids serving to regulate the store-operated Ca(2+) channel, Orai1, and we describe the sensitivity of Orai1 coupling to the ER Ca(2+) sensor, STIM1, to inhibition by polyunsaturated fatty acids. Phosphoinositides play key roles in regulating STIM1-Orai1 coupling, as well as in the stimulated Ca(2+) oscillations that are a consequence of IgE receptor signaling in mast cells. They also participate in the coupling between the plasma membrane and the actin cytoskeleton, which regulates immune receptor responses in T cells, B cells, and mast cells, both positively and negatively, depending on the cellular context. Recent studies show that other phospholipids with mostly saturated acylation also participate in coupling between receptors and the actin cytoskeleton. Lipid heterogeneity is a central feature of the intimate relationship between the plasma membrane and the actin cytoskeleton. The detailed nature of these interactions and how they are dynamically regulated to initiate and propagate receptor-mediated cell signaling are challenging questions for further investigation. This article is part of a Special Issue entitled: The cellular lipid landscape edited by Tim P. Levine and Anant K. Menon. PMID:26995463

  16. Defensive effect of lansoprazole in dementia of AD type in mice exposed to streptozotocin and cholesterol enriched diet.

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    Rupinder K Sodhi

    Full Text Available The present study investigates the potential of lansoprazole (a proton pump inhibitor and agonist of liver x receptors in experimental dementia of AD type. Streptozotocin [STZ, 3 mg/kg, injected intracerebroventricular (i.c.v, and high fat diet (HFD, administered for 90 days] were used to induce dementia in separate groups of Swiss mice. Morris water maze (MWM test was performed to assess learning and memory of the animals. A battery of biochemical and histopathological studies were also performed. Extent of oxidative stress was measured by estimating the levels of brain reduced glutathione (GSH and thiobarbituric acid reactive species (TBARS. Brain acetylcholinestrase (AChE activity and serum cholesterol levels were also estimated. The brain level of myeloperoxidase (MPO was measured as a marker of inflammation. STZ and HFD produced a marked decline in MWM performance of the animals, reflecting impairment of learning and memory. STZ/HFD treated mice exhibited a marked accentuation of AChE activity, TBARS and MPO levels along with a fall in GSH levels. Further, the stained micrographs of STZ/HFD treated mice indicated pathological changes, severe neutrophilic infiltration and amyloid deposition. Lansoprazole treatment significantly attenuated STZ and HFD -induced memory deficits, biochemical and histopathological alterations. It also prevented HFD-induced rise in the cholesterol level. Therefore, the findings demonstrate potential of lansoprazole in memory dysfunctions which may probably be attributed to its anti-cholinesterase, anti-oxidative and anti-inflammatory effects. Moreover, both cholesterol-dependent as well as cholesterol-independent effects of lansoprazole appear to play a role. In addition study indicates the role of liver x receptors in dementia.

  17. Role of spike protein conformational changes in fusion of Semliki Forest virus.

    Science.gov (United States)

    Justman, J; Klimjack, M R; Kielian, M

    1993-12-01

    The alphavirus Semliki Forest virus (SFV) and a number of other enveloped animal viruses infect cells via a membrane fusion reaction triggered by the low pH within endocytic vesicles. In addition to having a low pH requirement, SFV fusion and infection are also strictly dependent on the presence of cholesterol in the host cell membrane. A number of conformational changes in the SFV spike protein occur following low-pH treatment, including dissociation of the E1-E2 dimer, conformational changes in the E1 and E2 subunits, and oligomerization of E1 to a homotrimer. To allow the ordering of these events, we have compared the kinetics of these conformational changes with those of fusion, using pH treatment near the fusion threshold and low-temperature incubation to slow the fusion reaction. Dimer dissociation, the E1 conformational change, and E1 trimerization all occur prior to the mixing of virus and cell membranes. Studies of cells incubated at 20 degrees C showed that as with virus fusion, E1 trimerization occurred in the endosome before transport to lysosomes. However, unlike the strictly cholesterol-dependent membrane fusion reaction, the E1 homotrimer was produced in vivo during virus uptake by cholesterol-depleted cells or in vitro by low-pH treatment of virus in the presence of artificial liposomes with or without cholesterol. Purified, lipid-free spike protein rosettes were assayed to determine the requirement for virus membrane cholesterol in E1 homotrimer formation. Spike protein rosettes were found to undergo E1 oligomerization upon exposure to low pH and target liposomes and showed an enhancement of oligomerization with cholesterol-containing membranes. The E1 homotrimer may represent a perfusion complex that requires cholesterol to carry out the final coalescence of the viral and target membranes. PMID:8230478

  18. Na+ Influx Induced by New Antimalarials Causes Rapid Alterations in the Cholesterol Content and Morphology of Plasmodium falciparum.

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    Sudipta Das

    2016-05-01

    Full Text Available Among the several new antimalarials discovered over the past decade are at least three clinical candidate drugs, each with a distinct chemical structure, that disrupt Na+ homeostasis resulting in a rapid increase in intracellular Na+ concentration ([Na+]i within the erythrocytic stages of Plasmodium falciparum. At present, events triggered by Na+ influx that result in parasite demise are not well-understood. Here we report effects of two such drugs, a pyrazoleamide and a spiroindolone, on intraerythrocytic P. falciparum. Within minutes following the exposure to these drugs, the trophozoite stage parasite, which normally contains little cholesterol, was made permeant by cholesterol-dependent detergents, suggesting it acquired a substantial amount of the lipid. Consistently, the merozoite surface protein 1 and 2 (MSP1 and MSP2, glycosylphosphotidylinositol (GPI-anchored proteins normally uniformly distributed in the parasite plasma membrane, coalesced into clusters. These alterations were not observed following drug treatment of P. falciparum parasites adapted to grow in a low [Na+] growth medium. Both cholesterol acquisition and MSP1 coalescence were reversible upon the removal of the drugs, implicating an active process of cholesterol exclusion from trophozoites that we hypothesize is inhibited by high [Na+]i. Electron microscopy of drug-treated trophozoites revealed substantial morphological changes normally seen at the later schizont stage including the appearance of partial inner membrane complexes, dense organelles that resemble "rhoptries" and apparent nuclear division. Together these results suggest that [Na+]i disruptor drugs by altering levels of cholesterol in the parasite, dysregulate trophozoite to schizont development and cause parasite demise.

  19. Lipid nanocapsules containing the non-ionic surfactant Solutol HS15 inhibit the transport of calcium through hyperforin-activated channels in neuronal cells.

    Science.gov (United States)

    Chauvet, Sylvain; Barras, Alexandre; Boukherroub, Rabah; Bouron, Alexandre

    2015-12-01

    Hyperforin is described as a natural antidepressant inhibiting the reuptake of neurotransmitters and also activating cation channels. However the blood-brain barrier limits the access to the brain of this biomolecule. To circumvent this problem it was envisaged to encapsulate hyperforin into biomimetic lipid nano-carriers like lipid nanocapsules (LNCs). When testing the safety of 25 nm LNCs it appeared that they strongly blocked hyperforin-activated Ca2+ channels of cultured cortical neurons. This inhibition was due to one of their main component: solutol HS15 (polyoxyethylene-660-12-hydroxy stearate), a non-ionic soluble surfactant. Solutol HS15 rapidly depresses in a concentration-dependent manner the entry of Ca2+ through hyperforin-activated channels without influencing store-operated channels. This effect is mimicked by Brij58 but not by PEG600, indicating that the lipid chain of Solutol HS15 is important in determining its effects on the channels. The inhibition of the Ca2+ fluxes depends on the cellular cholesterol content; it is stronger after depleting cholesterol with methyl-β-cyclodextrin and is nearly absent on cells cultured in a cholesterol-rich medium. When chronically applied for 24 h, Solutol HS15 slightly up-regulates the entry of Ca2+ through hyperforin-activated channels. Similar observations were made when testing 25 nm lipid nanocapsules containing the surfactant Solutol HS15. Altogether, this study shows that Solutol HS15 perturbs in a cholesterol-dependent manner the activity of some neuronal channels. This is the first demonstration that LNCs containing this surfactant can influence cellular calcium signaling in the brain, a finding that can have important clinical implications.

  20. Lipid rafts in epithelial brush borders: atypical membrane microdomains with specialized functions.

    Science.gov (United States)

    Danielsen, E Michael; Hansen, Gert H

    2003-10-31

    Epithelial cells that fulfil high-throughput digestive/absorptive functions, such as small intestinal enterocytes and kidney proximal tubule cells, are endowed with a dense apical brush border. It has long been recognized that the microvillar surface of the brush border is organized in cholesterol/sphingolipid-enriched membrane microdomains commonly known as lipid rafts. More recent studies indicate that microvillar rafts, in particular those of enterocytes, have some unusual properties in comparison with rafts present on the surface of other cell types. Thus, microvillar rafts are stable rather than transient/dynamic, and their core components include glycolipids and the divalent lectin galectin-4, which together can be isolated as "superrafts", i.e., membrane microdomains resisting solubilization with Triton X-100 at physiological temperature. These glycolipid/lectin-based rafts serve as platforms for recruitment of GPI-linked and transmembrane digestive enzymes, most likely as an economizing effort to secure and prolong their digestive capability at the microvillar surface. However, in addition to microvilli, the brush border surface also consists of membrane invaginations between adjacent microvilli, which are the only part of the apical surface sterically accessible for membrane fusion/budding events. Many of these invaginations appear as pleiomorphic, deep apical tubules that extend up to 0.5-1 microm into the underlying terminal web region. Their sensitivity to methyl-beta-cyclodextrin suggests them to contain cholesterol-dependent lipid rafts of a different type from the glycolipid-based rafts at the microvillar surface. The brush border is thus an example of a complex membrane system that harbours at least two different types of lipid raft microdomains, each suited to fulfil specialized functions. This conclusion is in line with an emerging, more varied view of lipid rafts being pluripotent microdomains capable of adapting in size, shape, and content to

  1. 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

  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. 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

  4. Effectiveness of enrofloxacin for the treatment of experimentally-induced bovine anaplasmosis Eficácia da enrofloxacina no tratamento da anaplasmose bovina experimental

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    Elias Jorge Facury-Filho

    2012-03-01

    Full Text Available Four groups of six Holstein calves were inoculated with 3.6 × 10(7 erythrocytes parasitized with Anaplasma marginale. The criteria for treatment of calves were increasing A. marginale rickettsemia and 30% reduction of baseline packed cell volume (PCV of each animal. Group 1 (G1 received 7.5 mg.kg-1 of enrofloxacin in a single dose; Group 2 (G2 7.5 mg.kg-1 of enrofloxacin twice every three days; Group 3 (G3 20 mg.kg-1 of long-acting oxytetracycline in a single dose; and Group 4 (G4 a single dose of PBS. Physical examinations, blood smears and PCV were performed daily. On day treatment, G1, G2 and G3 animals had a mean rickettsemia of 17, 23 and 12%, respectively. At 2 days after treatment (DAT G1 and G2 animals showed a significant reduction of rickettsemia (p Quatro grupos de seis bezerros da raça Holandesa foram inoculados com 3,6 × 10(7 eritrócitos parasitados com Anaplasma marginale. Os critérios para o tratamento dos bezerros foram aumento da rickettsemia do A. marginale e redução de 30% do valor basal de volume globular (VG de cada animal. O Grupo 1 (G1 recebeu 7,5 mg.kg-1 de enrofloxacina em dose única; o Grupo 2 (G2, 7,5 mg.kg-1 de enrofloxacina duas vezes a cada três dias; o Grupo 3 (G3, 20 mg.kg-1 de oxitetraciclina de longa ação em dose única; e o Grupo 4 (G4 uma única dose de PBS. Exames físicos, esfregaço sanguíneo e VG foram realizadas diariamente. No dia do tratamento, os animais G1, G2 e G3 apresentaram rickettsemia média de 17, 23 e 12%, respectivamente. Nos primeiros 2 dias após o tratamento (DAT os animais do G1 e G2 mostraram uma redução significativa de rickettsemia (p < 0,05 em relação ao G3. Animais do G3 tiveram altas taxas de rickettsemia nos 2 DAT e uma diminuição lenta até à estabilização em 9 de DAT. O VG médio no G1 e G2 aumentou e estabilizou após 7 e 8 DAT, respectivamente. A estabilização do VG do G3 foi aos 13 DAT. A enrofloxacina e a oxitetraciclina foram efetivas no tratamento da

  5. Parasites of cattle in the trans-boundary areas of Ogun State, Nigeria

    International Nuclear Information System (INIS)

    Full text: An assessment of the parasites of cattle along a major trans-boundary route was carried out in Yewa division of Ogun State, Nigeria between February 2006 and July 2007. The location of study is bound to the west by the Republic of Benin, with which it shares 155 kilometres of International boundary, within latitude 6 deg. 151 N and 7 deg. 581 N in a deciduous/derived Savannah zone of Nigeria. A total of 62 herds of between 56 and 98 heads of cattle entering Nigeria from neighbouring West African Countries by hoof along the Iwoye - Imeko - Olodo - Abeokuta route were observed for various haemo- and ecto-parasites. About 5ml of blood was collected from the jugular vein of each of 245 randomly selected animals into sterile bijou bottles containing 250il of 200 mM disodium ethylene diamine tetra acetic acid (Na2EDTA) as anticoagulant. Animals infested with ectoparasites were noted and the parasites on each of the animals were collected into universal bottles containing 1% formaldehyde. The pus expressed from skin nodules on 18 animals and skin scabs from 15 animals were also collected for Parasitology and Microbiology respectively. All the samples were analysed at the College of veterinary medicine, University of Agriculture, Abeokuta. The blood samples were screened for the presence of trypanosomes using the buffy coat centrifugation method and for other blood parasites by examination of Giemsa-stained thin blood smear. Of the 245 blood samples analysed, 123 (50.20%) were positive for blood protozoans. Of these, Babesia spp. infection accounted for 50 (40.65%); Trypanosoma spp. accounted for 29 (23.58%); Anaplasma spp. 14 (11.38%); mixed Anaplasma spp. and Babesia spp. 9 (7.32%) and mixed Babesia spp. and Trypanosoma spp. 21 (17.07%). All the animals sampled had varying degrees of tick infestation ranging from a total count of 26 to 280 ticks per cattle. Of the 3756 ticks collected, 1080 (28.75%) were Rhipicephalus appendiculatus, 917 (24.41%) were

  6. Seroprevalence and risk factors for cattle anaplasmosis, babesiosis, and trypanosomiasis in a Brazilian semiarid region Soroprevalência e fatores de risco para anaplasmose, babesiose e tripanosomíase bovina em uma região semiárida do Brasil

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    Valéria Medeiros de Mendonça Costa

    Full Text Available The seroprevalence of Anaplasma marginale, Babesia bigemina, Babesia bovis and Trypanosoma vivax and the risk factors for these infections were investigated in 509 cows on 37 farms in the semiarid region of Paraíba, northeastern Brazil. Cow serum samples were tested by means of immunofluorescence assay (IFA against each specific antigen. The mean seroprevalence values per farm were 15.0% (range: 0-75% for A. marginale, 9.5% (range: 0-40% for B. bigemina and 26.9% (range: 0-73.7% for B. bovis. All cows tested negative for T. vivax. Higher prevalence for A. marginale was significantly associated with less frequent acaricide spraying per year and with higher use of injectable antihelminthics. Presence of cows positive for B. bigemina was significantly associated with acaricide use and with presence of horse flies on the farm. Both occurrence and higher prevalence of B. bovis were significantly associated with recent observations of ticks on cattle. Overall, the present results indicate that the region investigated is an enzootically unstable area for A. marginale, B. bigemina and B. bovis, since most animals were seronegative to at least one agent.A soroprevalência de Anaplasma marginale, Babesia bigemina, Babesia bovis e Trypanosoma vivax, assim como os fatores de risco para estas infecções, foram investigadas em 37 fazendas (total de 509 vacas da região semiárida da Paraíba, nordeste do Brasil. A presença de anticorpos nos soros dos animais foi detectada pela técnica de imunofluorescência indireta, utilizando antígenos específicos. Os valores médios de soroprevalência por fazenda foram 15,0% (0-75% para A. marginale, 9,5% (0-40% para B. bigemina, e 26,9% (0-73,7% para B. bovis. Todas as vacas foram soronegativas para T. vivax. As maiores prevalências de A. marginale foram significativamente associadas com menor uso de carrapaticidas por ano e com uso mais frequente de antihelmínticos injetáveis. A soroprevalência de B. bigemina

  7. 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

  8. Bmcystatin, a cysteine proteinase inhibitor characterized from the tick Boophilus microplus

    International Nuclear Information System (INIS)

    The bovine tick Rhipicephalus (Boophilus) microplus is a blood-sucking animal, which is responsible for Babesia spp and Anaplasma marginale transmission for cattle. From a B. microplus fat body cDNA library, 465 selected clones were sequenced randomly and resulted in 60 Contigs. An open reading frame (ORF) contains 98 amino acids named Bmcystatin, due to 70% amino acid identity to a classical type 1 cystatin from Ixodes scapularis tick (GenBank Accession No. DQ066227). The Bmcystatin amino acid sequence analysis showed two cysteine residues, theoretical pI of 5.92 and Mr of 11kDa. Bmcystatin gene was cloned in pET 26b vector and the protein expressed using bacteria Escherichia coli BL21 SI. Recombinant Bmcystatin (rBmcystatin) purified by affinity chromatography on Ni-NTA-agarose column and ionic exchange chromatography on HiTrap Q column presented molecular mass of 11kDa, by SDS-PAGE and the N-terminal amino acid sequenced revealed unprocessed N-terminal containing part of pelB signal sequence. Purified rBmcystatin showed to be a C1 cysteine peptidase inhibitor with Ki value of 0.1 and 0.6nM for human cathepsin L and VTDCE (vitellin degrading cysteine endopeptidase), respectively. The rBmcystatin expression analyzed by semi-quantitative RT-PCR confirmed the amplification of a specific DNA sequence (294bp) in the fat body and ovary cDNA preparation. On the other hand, a protein band was detected in the fat body, ovary, and the salivary gland extracts using anti-Bmcystatin antibody by Western blot. The present results suggest a possible role of Bmcystatin in the ovary, even though the gene was cloned from the fat body, which could be another site of this protein synthesis

  9. Cloning and sequence analysis of partial genomic DNA coding for HtrA-type serine protease of Wolbachia from human lymphatic filarial parasite, Wuchereria bancrofti

    Science.gov (United States)

    Dhamodharan, R; Hoti, SL; Sivapragasam, G; Das, MK

    2011-01-01

    Background: Periplasmic serine proteases of HtrA type of Wolbachia have been shown to play a role in the pathogenesis of filarial disease. Aims: This study was aimed to sequence Wb-HtrA serine protease and analyze its phylogenetic position by comparing with other filarial and non-filarial nematode homologs. Materials and Methods: Partial HtrA gene fragment was amplified from DNA isolated from periodic and sub-periodic Wuchereria bancrofti parasites collected from Pondicherry and Nicobar islands, respectively. The amplicons were sequenced, and sequence homology and phylogenetic relationship with other filarial and non-filarial nematodes were analyzed. Results: Partial orthologue of HtrA-type serine protease from Wolbachia of W. bancrofti was amplified, cloned and sequenced. The deduced amino acid sequence exhibited 87%, 81% and 74% identity with the homologous Wolbachia proteases identified from Brugia malayi, Onchocerca volvulus and Drosophila melanogaster, respectively. The Wb-HtrA has arthologues in several proteobacteria with very high homology and hence is highly conserved not only among Wolbachia of filarial parasites but also across proteobacteria. The phylogenetic tree constructed using Neighbor-Joining method showed two main clusters: cluster-I containing bacteria that dwell in diverse habitats such as soil, fresh and marine waters and plants and cluster-II comprising Anaplasma sp. and Erlichia, and Wolbachia endosymbionts of insects and nematodes, in distinct groups. Conclusions: HtrA-type serine protease from Wolbachia of W. bancrofti is highly conserved among filarial parasites. It will be of interest to know whether filarial Wolbachia HtrA type of serine protease might influence apoptosis and lymphatic epithelium, thereby playing a role in the filarial pathogenesis. Such information will be useful for identifying targets for the development of newer drugs for filariasis treatment, especially for preventing lymphatic pathology. PMID:23508470

  10. 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.

  11. 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.

  12. 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

  13. 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.

  14. 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

  15. 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.

  16. 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

  17. 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.

  18. Dog overpopulation and burden of exposure to canine distemper virus and other pathogens on Santa Cruz Island, Galapagos.

    Science.gov (United States)

    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. PMID

  19. 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.

  20. Incidence of Cercopithifilaria bainae in dogs and probability of co-infection with other tick-borne pathogens.

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    Rafael Antonio Nascimento Ramos

    Full Text Available BACKGROUND: Cercopithifilaria bainae is a filarioid parasite that infects dogs, being transmitted by Rhipicephalus sanguineus group ticks in many countries of the Mediterranean basin. This study assessed the incidence density rate (IDR of infection by C. bainae in dogs and the probability of co-infection with other tick-borne pathogens (i.e., Anaplasma platys, Babesia vogeli and Hepatozoon canis, in an area of high endemicity in southern Italy. METHODOLOGY/PRINCIPAL FINDINGS: From March 2011 to October 2012, a field study involving 58 young dogs naturally exposed to tick infestation was conducted. Skin and blood samples obtained from each dog six times during an 18-month period were tested for C. bainae by parasite detection within skin snip sediments, with subsequent confirmation through PCR and DNA sequencing. Dogs examined monthly for ticks and A. platys, B. vogeli and H. canis were microscopically and/or molecularly diagnosed and after the first and the second summer seasons, the IDR for positive animal-month at risk was 3.8% and 1.7% in November 2011 and October 2012, respectively. All 58 C. bainae-infected dogs were simultaneously infected with at least one other tick-borne pathogen. After the first summer season (assessment in November 2011, a C. bainae-infected dog had a 33% probability of being infected with H. canis or A. platys, whereas after the second tick season (assessment in October 2012 the probability of co-infection was 78%, 22% and 11% for H. canis, A. platys and B. vogeli, respectively. CONCLUSIONS: Our data indicate that tick-infested dogs are at risk of acquiring infection by C. bainae. In addition, the detection of C. bainae microfilariae indicates a prior tick exposure and, should stimulate testing for other tick-borne disease causing pathogens.

  1. 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.

  2. 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.

  3. Bacterial community in Haemaphysalis ticks of domesticated animals from the Orang Asli communities in Malaysia.

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    Khoo, Jing-Jing; Chen, Fezshin; Kho, Kai Ling; Ahmad Shanizza, Azzy Iyzati; Lim, Fang-Shiang; Tan, Kim-Kee; Chang, Li-Yen; AbuBakar, Sazaly

    2016-07-01

    Ticks are vectors in the transmission of many important infectious diseases in human and animals. Ticks can be readily found in the semi-forested areas such as the settlements of the indigenous people in Malaysia, the Orang Asli. There is still minimal information available on the bacterial agents associated with ticks found in Malaysia. We performed a survey of the bacterial communities associated with ticks collected from domestic animals found in two Orang Asli villages in Malaysia. We collected 62 ticks, microscopically and molecularly identified as related to Haemaphysalis wellingtoni, Haemaphysalis hystricis and Haemaphysalis bispinosa. Bacterial 16s rRNA hypervariable region (V6) amplicon libraries prepared from the tick samples were sequenced on the Ion Torrent PGM platform. We detected a total of 392 possible bacterial genera after pooling and sequencing 20 samples, indicating a diverse bacterial community profile. Dominant taxa include the potential tick endosymbiont, Coxiella. Other dominant taxa include the tick-associated pathogen, Rickettsia, and environmental bacteria such as Bacillus, Mycobacterium, Sphingomonas and Pseudomonas. Other known tick-associated bacteria were also detected, including Anaplasma, Ehrlichia, Rickettsiella and Wolbachia, albeit at very low abundance. Specific PCR was performed on selected samples to identify Rickettsia and Coxiella. Sequence of Rickettsia felis, which causes spotted fever in human and cats, was identified in one sample. Coxiella endosymbionts were detected in three samples. This study provides the baseline knowledge of the microbiome of ticks in Malaysia, focusing on tick-associated bacteria affecting the Orang Asli communities. The role of the herein found Coxiella and Rickettsia in tick physiology or disease transmission merits further investigation. PMID:27132518

  4. 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

  5. Comparative Metagenomic Profiling of Symbiotic Bacterial Communities Associated with Ixodes persulcatus, Ixodes pavlovskyi and Dermacentor reticulatus Ticks.

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    Alexander Kurilshikov

    Full Text Available Ixodes persulcatus, Ixodes pavlovskyi, and Dermacentor reticulatus ticks inhabiting Western Siberia are responsible for the transmission of a number of etiological agents that cause human and animal tick-borne diseases. Because these ticks are abundant in the suburbs of large cities, agricultural areas, and popular tourist sites and frequently attack people and livestock, data regarding the microbiomes of these organisms are required. Using metagenomic 16S profiling, we evaluate bacterial communities associated with I. persulcatus, I. pavlovskyi, and D. reticulatus ticks collected from the Novosibirsk region of Russia. A total of 1214 ticks were used for this study. DNA extracted from the ticks was pooled according to tick species and sex. Sequencing of the V3-V5 domains of 16S rRNA genes was performed using the Illumina Miseq platform. The following bacterial genera were prevalent in the examined communities: Acinetobacter (all three tick species, Rickettsia (I. persulcatus and D. reticulatus and Francisella (D. reticulatus. B. burgdorferi sensu lato and B. miyamotoi sequences were detected in I. persulcatus and I. pavlovskyi but not in D. reticulatus ticks. The pooled samples of all tick species studied contained bacteria from the Anaplasmataceae family, although their occurrence was low. DNA from A. phagocytophilum and Candidatus Neoehrlichia mikurensis was first observed in I. pavlovskyi ticks. Significant inter-species differences in the number of bacterial taxa as well as intra-species diversity related to tick sex were observed. The bacterial communities associated with the I. pavlovskyi ticks displayed a higher biodiversity compared with those of the I. persulcatus and D. reticulatus ticks. Bacterial community structure was also diverse across the studied tick species, as shown by permutational analysis of variance using the Bray-Curtis dissimilarity metric (p = 0.002. Between-sex variation was confirmed by PERMANOVA testing in I

  6. Cardiovascular Disease Death Before Age 65 in 168 Countries Correlated Statistically with Biometrics, Socioeconomic Status, Tobacco, Gender, Exercise, Macronutrients, and Vitamin K

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    Agutter, Paul S

    2016-01-01

    Background Nutrition researchers recently recognized that deficiency of vitamin K2 (menaquinone: MK-4–MK-13) is widespread and contributes to cardiovascular disease (CVD). The deficiency of vitamin K2 or vitamin K inhibition with warfarin leads to calcium deposition in the arterial blood vessels. Methods Using publicly available sources, we collected food commodity availability data and derived nutrient profiles including vitamin K2 for people from 168 countries. We also collected female and male cohort data on early death from CVD (ages 15–64 years), insufficient physical activity, tobacco, biometric CVD risk markers, socioeconomic risk factors for CVD, and gender. The outcome measures included (1) univariate correlations of early death from CVD with each risk factor, (2) a multiple regression-derived formula relating early death from CVD (dependent variable) to macronutrient profile, vitamin K1 and K2 and other risk factors (independent variables), (3) for each risk factor appearing in the multiple regression formula, the portion of CVD risk attributable to that factor, and (4) similar univariate and multivariate analyses of body mass index (BMI), fasting blood sugar (FBS) (simulated from diabetes prevalence), systolic blood pressure (SBP), and cholesterol/ HDL-C ratio (simulated from serum cholesterol) (dependent variables) and dietary and other risk factors (independent variables). Results Female and male cohorts in countries that have vitamin K2 24 µg/day of vitamin K2 per 2000 kcal/day (n = 72). A multiple regression-derived formula relating early death from CVD to dietary nutrients and other risk factors accounted for about 50% of the variance between cohorts in early CVD death. The attributable risks of the variables in the CVD early death formula were: too much alcohol (0.38%), too little vitamin K2 (6.95%), tobacco (6.87%), high blood pressure (9.01%), air pollution (9.15%), early childhood death (3.64%), poverty (7.66%), and male gender (6

  7. Effect of Probiotics on Blood Lipid Concentrations: A Meta-Analysis of Randomized Controlled Trials.

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    Cho, Young Ae; Kim, Jeongseon

    2015-10-01

    Previous clinical studies have reported mixed results regarding the effect of probiotics on lipid metabolism. Therefore, we conducted a meta-analysis of randomized controlled trials to quantify the direction and magnitude of the potential effect of probiotics on blood lipid concentrations.Eligible studies were randomized, placebo-controlled trials whose interventions were probiotic products containing live bacteria. The studies reported net changes in lipid profiles (total cholesterol, LDL cholesterol, HDL cholesterol, and triglycerides) and their associated standard deviations (or the data to calculate them). The probiotic products did not contain prebiotics or other active ingredients, and the full article was accessible in English.The pooled mean net change in lipid profiles and 95% confidence intervals (95% CIs) were calculated. Q statistics and I were calculated to examine heterogeneity. Potential sources of heterogeneity were investigated via subgroup and sensitivity analyses, and publication biases were estimated.A total of 30 randomized controlled trials with 1624 participants (828 in intervention groups and 796 in placebo groups) were included in this analysis. Subjects treated with probiotics demonstrated reduced total cholesterol and LDL cholesterol compared to control subjects by 7.8 mg/dL (95% CI: -10.4, -5.2) and 7.3 mg/dL (95% CI: -10.1, -4.4), respectively. There was no significant effect of probiotics on HDL cholesterol or triglycerides. The effect of probiotics on total cholesterol and LDL cholesterol depended on a variety of factors. The significant effects were greater for higher baseline total cholesterol levels, longer treatment durations, and certain probiotic strains. In addition, these associations seem stronger in studies supported by probiotics companies.The studies included in this meta-analysis showed significant heterogeneity as indicated by the Q statistics and I. In addition, industry sponsorship may affect study findings

  8. Uptake of long chain fatty acids is regulated by dynamic interaction of FAT/CD36 with cholesterol/sphingolipid enriched microdomains (lipid rafts

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    Herrmann Thomas

    2008-08-01

    Full Text Available Abstract Background Mechanisms of long chain fatty acid uptake across the plasma membrane are important targets in treatment of many human diseases like obesity or hepatic steatosis. Long chain fatty acid translocation is achieved by a concert of co-existing mechanisms. These lipids can passively diffuse, but certain membrane proteins can also accelerate the transport. However, we now can provide further evidence that not only proteins but also lipid microdomains play an important part in the regulation of the facilitated uptake process. Methods Dynamic association of FAT/CD36 a candidate fatty acid transporter with lipid rafts was analysed by isolation of detergent resistant membranes (DRMs and by clustering of lipid rafts with antibodies on living cells. Lipid raft integrity was modulated by cholesterol depletion using methyl-β-cyclodextrin and sphingolipid depletion using myriocin and sphingomyelinase. Functional analyses were performed using an [3H]-oleate uptake assay. Results Overexpression of FAT/CD36 and FATP4 increased long chain fatty acid uptake. The uptake of long chain fatty acids was cholesterol and sphingolipid dependent. Floating experiments showed that there are two pools of FAT/CD36, one found in DRMs and another outside of these domains. FAT/CD36 co-localized with the lipid raft marker PLAP in antibody-clustered domains at the plasma membrane and segregated away from the non-raft marker GFP-TMD. Antibody cross-linking increased DRM association of FAT/CD36 and accelerated the overall fatty acid uptake in a cholesterol dependent manner. Another candidate transporter, FATP4, was neither present in DRMs nor co-localized with FAT/CD36 at the plasma membrane. Conclusion Our observations suggest the existence of two pools of FAT/CD36 within cellular membranes. As increased raft association of FAT/CD36 leads to an increased fatty acid uptake, dynamic association of FAT/CD36 with lipid rafts might regulate the process. There is no

  9. Survey of selected pathogens and blood parameters of northern yellowstone elk: Wolf sanitation effect implications

    Science.gov (United States)

    Barber-Meyer, S. M.; White, P.J.; Mech, L.D.

    2007-01-01

    The restoration or conservation of predators could reduce seroprevalences of certain diseases in prey if predation selectively removes animals exhibiting clinical signs. We assessed disease seroprevalences and blood parameters of 115 adult female elk (Cervus elaphus) wintering on the northern range of Yellowstone National Park [YNP] during 2000-2005 and compared them to data collected prior to wolf (Canis lupus) restoration (WR) in 1995 and to two other herds in Montana to assess this prediction. Blood parameters were generally within two standard deviations of the means observed in other Montana herds (Gravelly-Snowcrest [GS] and Garnet Mountain [GM]), but Yellowstone elk had higher seroprevalences of parainfluenza-3 virus (95% CI YNP = 61.1-78.6, GS = 30.3-46.5) and bovine-virus-diarrhea virus type 1 (95% CI YNP = 15.9-31.9, GM = 0). In comparisons between pre-wolf restoration [pre-WR] (i.e., prior to 1995) seroprevalences with those post-wolf restoration [post-WR] in Yellowstone, we found lower seroprevalences for some disease-causing agents post-wolf restoration (e.g., bovine-virus-diarrhea virus type-1 [95% CI pre-WR = 73.1-86.3, post-WR = 15.9-31.9] and bovine-respiratory syncytial virus [95% CI pre-WR = 70.0-83.8, post-WR = 0]), but similar (e.g., Brucella abortus [95% CI pre-WR = 0-4.45, post-WR = 0-4.74] and epizootic hemorrhagic disease virus [95% CI pre-WR = 0, post-WR = 0]) or higher for others (e.g., Anaplasma marginale [95% CI pre-WR = 0, post-WR = 18.5-38.7] and Leptospira spp. [95% CI pre-WR = 0.5-6.5, post-WR = 9.5-23.5]). Though we did not detect an overall strong predation effect through reduced disease seroprevalence using retrospective comparisons with sparse data, our reference values will facilitate future assessments of this issue.

  10. Fifty-six Cases with Mediterranean Spotted Fever: Evaluation of Tick-Borne Spotted Diseases in Turkey

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    Özlem Altuntaş Aydın

    2009-12-01

    Full Text Available Background and Design: There are very few clinical series about Mediterranean Spotted Fever (MSF reported from Turkey. Fifty-six patients admitted with fever and rash were diagnosed as MSF in our hospital by clinical, epidemiological signs and Weil-Felix test. We aimed to take attention to the similarities and differencies with another tick-borne infection Crimean-Congo Haemorrhagic Fever (CCHF reported lately in Turkey. Material and Method: Between 1989-2006 years 56 cases were diagnosed as rickettsioses based on epidemiological, clinical and laboratory data. We evaluated the patients retrospectively using by “Diagnosis guide for tick transmitted infections in Europe” prepared by ESCMID Study group for Coxiella, Anaplasma, Rickettsia and Barthonella (ESCAR. We compared two diseases according to the Fisher’s exact test in clinical and laboratory characteristics. Results: Twenty-nine female and 27 male cases reached to 25 point according to ESCAR diagnostic scoring were included into the present study. Age distribution was 20-80 years (mean 43, duration of hospitalization was 1-38 days (mean 8 days. Tache noire was established in most of the cases (64%. All of the cases were recovered by 7-10 days of doxycyclin treatment except one case stayed in hospital for 38 days and died. When it is compared statistically haemorrhagic manifestations, history of tick exposure, nausea and vomiting, thrombocytopenia and leukopenia rates were found higher in CCHF (p<0.05. Conclusion: Whereas MSF and CCHF have similar seasonal and clinical features and transmitted by the tick, the treatment regimen and mortality rates differ. In Istanbul we realised that some of the cases reported as possible CCHF were found to be MSF later, for this reason in patients who has fever and maculopapular rash with a history of tick exposure, MSF should always be kept in mind. The differential diagnosis of CCHF and MSF which can cause epidemics in our country should be done by

  11. 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.

  12. Cloning and sequence analysis of the 16S rRNA gene of Eperythrozoon wenyonii in cow%牛附红细胞体16SrRNA基因的克隆测序及分析

    Institute of Scientific and Technical Information of China (English)

    周作勇; 聂奎; 胡世君; 周荣琼; 唐成

    2011-01-01

    无菌采集自然感染附红细胞体的牛血液,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1 500 bp的扩增片段,将其克隆到pMD18-T载体后进行测序和分析.结果,所克隆的牛附红细胞体基因片段大小为1 454 bp,GenBank登录号为FJ375309(丰都株).序列比对结果显示,牛附红细胞体丰都株与武汉株(AY946266)最高,达99.7%,与支原体科代表种同源性为60.7%~76.2%,而与立克次氏体科的立克次氏体和无形体科的无形体同源性仅为51.4%~56.4%,表明牛附红细胞体应归为支原体科,附红细胞体属,而不应属于立克次氏体目,无浆体科.对国内外牛温氏附红细胞体的亲缘关系分析表明,牛温氏附红细胞体无明显的地域性差别趋势.%we cloned and sequenced 16S rRNA gene of Eperythrozoon wenyonii in cow from Fengdu with universal primer of haemotrophic bacteria. The result showed that the sequence length of Eperythrozoon wenyonii was 1 454 bp,GenBank accession number was FJ375309 (Fengdu strain). The determinated sequence was most similar to that of AY946266 ( Wuhan strain ) (with the homology of 99. 7%). Phylogenetic analysis indicated that it was more similar to those of Mycoplasma spp. in Mycoplasmataceae (homology level between 60. 7% and 76.2 % ), but was farther from Rickettsia spp. in Riekettsiaceae and Anaplasma spp. in Anaplasmataceae ( homology level between 51.4% .and 56.4%). These data indicated that E. wenyoni seems to belong to Mycoplasmataceae. Eperythrozoon, but not Rickettsiales,Anaplasmataceae. Genetic relationship analysis of Eperythrozoon wenyonii isolates indicated that there was any apparente tendency of geographical grouping of Eperythrozoon wenyonii in the world.

  13. 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

  14. Detection of Rickettsia and Ehrlichia spp. in Ticks Associated with Exotic Reptiles and Amphibians Imported into Japan.

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    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

  15. 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

  16. Vector-borne pathogens in dogs from Costa Rica: first molecular description of Babesia vogeli and Hepatozoon canis infections with a high prevalence of monocytic ehrlichiosis and the manifestations of co-infection.

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    Rojas, Alicia; Rojas, Diana; Montenegro, Víctor; Gutiérrez, Ricardo; Yasur-Landau, Daniel; Baneth, Gad

    2014-01-31

    Infection with canine vector-borne pathogens was evaluated in dogs from four different regions of Costa Rica by PCR. Demographic data, clinical signs, packed cell volume values, and the presence of tick infestation were recorded for each dog. Forty seven percent (69/146) of the dogs were infected with at least one pathogen and 12% were co-infected with two pathogens. Ehrlichia canis was detected in 34%, Anaplasma platys in 10%, Babesia vogeli in 8%, and Hepatozoon canis in 7.5% of the blood samples. No infection was detected with Leishmania spp. in blood, skin scrapings or conjunctival swabs. Thirty percent of the dogs presented at least one clinical sign compatible with vector-borne disease, and of those, 66% were infected with a pathogen. Subclinical infections were determined in 58% of the infected dogs including 82% (9/11), 58% (29/50), 42% (5/12) and 36% (5/14) of the dogs with H. canis, E. canis, B. vogeli and A. platys infections, respectively. A distinct relationship was found between infection and anemia. The mean PCV values were 34.4% in dogs with no infection, 31.5% in those who had a single infection and 23% in those with co-infection. Co-infected dogs had significantly lower PCV values compared to non-infected and single-infected dogs (pticks, 82% of them were infested with Rhipicephalus sanguineus sensu lato and 18% with Amblyomma ovale. Dogs infected with A. platys, B. vogeli, or E. canis were significantly associated with R. sanguineus s.l. infestation (pdescription of infections with B. vogeli and H. canis in Costa Rica as well as in Central America. The results of this study indicate that multiple vector-borne pathogens responsible for severe diseases infect dogs in Costa Rica and therefore, increased owner and veterinarian awareness are needed. Moreover, prevention of tick infestation is recommended to decrease the threat of these diseases to the canine population. PMID:24315693

  17. The emergence and maintenance of vector-borne diseases in the khyber pakhtunkhwa province, and the federally administered tribal areas of pakistan.

    Science.gov (United States)

    Nieto, Nathan C; Khan, Khalid; Uhllah, Ghufran; Teglas, Mike B

    2012-01-01

    , outbreaks of Dengue hemorrhagic fever have been reported in the region, and high prevalence of cattle infected and co-infected with multiple species of hemoparasites (Theileria, Babesia, Anaplasma). The emergence of which has followed the increased density of the rural population due to an influx of refugees from violent conflicts in Afghanistan and is exacerbated by an already impoverished society and wide diversity of potential arthropod vectors. These human outbreaks may be exacerbated by episodes of social upheaval but are also tied to the historically close association of people in the region with their livestock and subsequent zoonosis that result from spillover from co-habitation with infected domestic animals. PMID:22934007

  18. 中国10个省级疾病预防控制中心无形体检测实验室质量控制分析%Evaluation on external qualitv control assessment for laboratory diagnosis of Anaplasmosis in China

    Institute of Scientific and Technical Information of China (English)

    张丽娟; 吴建英; 岳永杰; 张铮; 李文涓; 邓红; 夏依旦; 禹惠兰; 王誓闻; 汪鹏; 姜霞; 钱振宇; 刘桂艳; 雷露; 刘学升; 雷霞; 南晓伟

    2011-01-01

    Objective To evaluate the results of the external quality assessment (EQA) on the laboratory diagnosis of Anaplasmosis and differential diagnosis between Anaplasma and other rickettsiae in 10 provincial/municipal CDCs laboratories in China. Methods The micro-indirect immunofluoresce assay (mIFA) recommended by WHO Rickettsial Diseases Collaborating Center was adopted as a standard serological diagnosis technique. The performances of two nested PCR assays, one testing for most members of family Rickettsia to amplify groEL gene and other targeting members of family Anaplasma to amplify 16S rRNA gene were evaluated. The laboratories involved received a package containing primers and reference templates ( P1,P2, P3, P4, P5 ) for PCR tests and SI and S2 reference sera for serological assays.Each laboratory was required to perform these procedures based on its own laboratory condition including equipments and some reagents except the reference materials provided. Results Seven laboratories from the 10 CDCs reported the results for testing S1 and S2 references, and the results in 5 of them (71.4%) were qualified according to the assessment criteria. Eight laboratories reported PCR results, but the significant differences on specificity and sensitivity of PCR between them were found, 62. 5% (5/8) of laboratories provided acceptable results for P2 and P3 references and 50%(4/8) of laboratories provided acceptable results for P4, P5, P6 references. For repeatability evaluation, all the laboratories reported acceptable results according to the statistical procedures described. Conclusion This EQA demonstrated all the laboratory involved had systematic errors in their performances. It is necessary for these laboratories to improve their test condition, strengthen the internal quality control and take part in the external quality control activity regularly to improve the diseases diagnosis and differential diagnosis capacity.%目的 对10个省、市级疾病预防控制

  19. 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

    Directory of Open Access Journals (Sweden)

    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

  20. 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

  1. 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.

  2. 恙虫病东方体Karp株56-kDa外膜抗原基因片段的原核表达及免疫原性研究%Prokaryotic expression and immunogenicity of major outer membrane protein 56-kDa antigen of Orientia tsutsugamushi strain Karp

    Institute of Scientific and Technical Information of China (English)

    王园园; 陈强; 于强; 张丽娟

    2012-01-01

    Objective To express the 56-kDa protein of Orientia tsutsugamushi strain Karp and evaluate the application of the recombi-nant protein in immunological diagnosis. Methods The fragment of 56-kDa gene was cloned into pET30a( + ) and then transformed to host bacteria Escherichia coli Rossetta. Specificity of the recombinant protein was assessed by ELISA with rabbit antisera against common member of order Rickettsiae and ten kinds of other clinical pathogens. Immunogenicity of the recombinant protein was studied in BALB/c mice. Results After induction with IPTG and analysis by SDS-PAGE, an approximately 46-kD band was observed and the test via western blot. Mass-spectrum proved that the recombinant protein was expressed successfully. Specificity analysis demonstrated that all sera except antisera against 0. Tsutsugamushi strain TA763, TH817 and Ehrlichia, B. Quintana, A. Phagocytophilum were negative. The purified protein was used to immunize BALB/c male mice and then the antisera (polyclonal antibody) were harvested. According to IFA and ELISA test, the titer of the polyclonal antibody reaches to 1:1 600. Conclusion The recombinant 56-kDa protein in this study exhibited excellent antigenicity and specificity, and should prove valuable for developing a simple and rapid diagnostic test and preparing the vaccine for 0. Tsutsugamushi.%目的 探讨恙虫病东方体56-kDa蛋白在大肠埃希菌中的表达及作为诊断性抗原的可行性.方法 用PCR扩增恙虫病东方体Karp菌株编码56 -kDa蛋白长约1 100bp的DNA,克隆至表达载体pET30a(+).表达产物经SDS-PAGE、质谱分析及western blot鉴定分析.重组蛋白质纯化后作为包被抗原,用方阵实验确定最佳浓度包被,ELISA法检测其与立克次体目16种成员及10种致病菌阳性血清的反应性.用此重组蛋白质免疫小鼠后制备抗血清,分别用ELISA及间接免疫荧光法(IFA)检测抗血清的效价.结果 重组质粒经PCR及测序分析证明,56-kDa外膜抗