Beall Melissa J
Full Text Available Abstract Background This study evaluated the exposure of dogs to three different Ehrlichia spp. in the south and central regions of the United States where vector-borne disease prevalence has been previously difficult to ascertain, particularly beyond the metropolitan areas. Methods Dog blood samples (n = 8,662 were submitted from 14 veterinary colleges, 6 private veterinary practices and 4 diagnostic laboratories across this region. Samples were tested for E. canis, E. chaffeensis and E. ewingii specific antibodies using peptide microtiter ELISAs. Results Overall, E. canis, E. chaffeensis and E. ewingii seroprevalence was 0.8%, 2.8%, and 5.1%, respectively. The highest E. canis seroprevalence (2.3% was found in a region encompassing Arkansas, Louisiana, Oklahoma, Tennessee and Texas. E. chaffeensis seroreactivity was 6.6% in the central region (Arkansas, Kansas, Missouri, and Oklahoma and 4.6% in the southeast region (Georgia, Maryland, North Carolina, South Carolina, Tennessee and Virginia. Seroreactivity to E. ewingii was also highest in the central region (14.6% followed by the southeast region (5.9%. The geospatial pattern derived from E. chaffeensis and E. ewingii seropositive samples was similar to previous reports based on E. chaffeensis seroreactivity in white-tailed deer and the distribution of human monocytic ehrlichiosis (HME cases reported by the CDC. Conclusions The results of this study provide the first large scale regional documentation of exposure to E. canis, E. chaffeensis and E. ewingii in pet dogs, highlighting regional differences in seroprevalence and providing the basis for heightened awareness of these emerging vector-borne pathogens by veterinarians and public health agencies.
Barlough, J E; Madigan, J E; Turoff, D R; Clover, J R; Shelly, S M; Dumler, J S
An ehrlichia was identified in the blood of a diseased llama (lama glama). Sequencing of its 16S rRNA gene showed the ehrlichia to be closely related to members of the Ehrlichia phagocytophila genogroup. The agent was also found in a pool of ticks (Ixodes pacificus) collected at the llama site. PMID:9157118
Taslima Taher Lina
Full Text Available Ehrlichia chaffeensis is a small, gram negative, obligately intracellular bacterium that preferentially infects mononuclear phagocytes. It is the etiologic agent of human monocytotropic ehrlichiosis (HME, an emerging life-threatening tick-borne zoonosis. Mechanisms by which E. chaffeensis establishes intracellular infection, and avoids host defenses are not well understood, but involve functionally relevant host-pathogen interactions associated with tandem and ankyrin repeat effector proteins. In this review, we discuss the recent advances in our understanding of the molecular and cellular mechanisms that underlie Ehrlichia host cellular reprogramming strategies that enable intracellular survival.
Heartwater is caused by Ehrlichia ruminantium and transmitted by ticks of the genus Amblyomma. It occurs in sub-Saharan Africa and in the Caribbean and affects domestic ruminants. There is general lack of information on the epidemiology of the disease in The Gambia. Results of a countrywide
Pennisi, Maria Grazia; Hofmann-Lehmann, Regina; Radford, Alan D; Tasker, Séverine; Belák, Sándor; Addie, Diane D; Boucraut-Baralon, Corine; Egberink, Herman; Frymus, Tadeusz; Gruffydd-Jones, Tim; Hartmann, Katrin; Horzinek, Marian C; Hosie, Margaret J; Lloret, Albert; Lutz, Hans; Marsilio, Fulvio; Thiry, Etienne; Truyen, Uwe; Möstl, Karin
OVERVIEW: Anaplasma species, Ehrlichia species and Rickettsia species are vector-borne pathogens infecting a wide variety of mammals, but causing disease in very few of them. Infection in cats: Anaplasma phagocytophilum is the most important feline pathogen among these rickettsial organisms, and
Inokuma, H; Brouqui, P; Drancourt, M; Raoult, D
The sequence of the citrate synthase gene (gltA) of 13 ehrlichial species (Ehrlichia chaffeensis, Ehrlichia canis, Ehrlichia muris, an Ehrlichia species recently detected from Ixodes ovatus, Cowdria ruminantium, Ehrlichia phagocytophila, Ehrlichia equi, the human granulocytic ehrlichiosis [HGE] agent, Anaplasma marginale, Anaplasma centrale, Ehrlichia sennetsu, Ehrlichia risticii, and Neorickettsia helminthoeca) have been determined by degenerate PCR and the Genome Walker method. The ehrlichial gltA genes are 1,197 bp (E. sennetsu and E. risticii) to 1,254 bp (A. marginale and A. centrale) long, and GC contents of the gene vary from 30.5% (Ehrlichia sp. detected from I. ovatus) to 51.0% (A. centrale). The percent identities of the gltA nucleotide sequences among ehrlichial species were 49.7% (E. risticii versus A. centrale) to 99.8% (HGE agent versus E. equi). The percent identities of deduced amino acid sequences were 44.4% (E. sennetsu versus E. muris) to 99.5% (HGE agent versus E. equi), whereas the homology range of 16S rRNA genes was 83.5% (E. risticii versus the Ehrlichia sp. detected from I. ovatus) to 99.9% (HGE agent, E. equi, and E. phagocytophila). The architecture of the phylogenetic trees constructed by gltA nucleotide sequences or amino acid sequences was similar to that derived from the 16S rRNA gene sequences but showed more-significant bootstrap values. Based upon the alignment analysis of the ehrlichial gltA sequences, two sets of primers were designed to amplify tick-borne Ehrlichia and Neorickettsia genogroup Ehrlichia (N. helminthoeca, E. sennetsu, and E. risticii), respectively. Tick-borne Ehrlichia species were specifically identified by restriction fragment length polymorphism (RFLP) patterns of AcsI and XhoI with the exception of E. muris and the very closely related ehrlichia derived from I. ovatus for which sequence analysis of the PCR product is needed. Similarly, Neorickettsia genogroup Ehrlichia species were specifically identified by
Full Text Available Canine Monocytic Ehrlichiosis is a zoonotic bacterial disease with worldwide distribution. With regards to the population of stray dogs, the disease is facilitated due to their lifestyle and the lack of anti-parasitic protection. The aim of this study was to provide serological data on the presence of a specific Ehrlichia canis IgG antibodies in stray dogs, originating from 7 municipalities in Serbia. During the period from April 2013 to June 2014, 217 canine sera were submitted to the laboratory of the Department of Infectious Diseases of Animals and Bees, Faculty of Veterinary Medicine in Belgrade. An immunofluorescent antibody test (IFAT was performed to detect antibodies to Ehrlichia canis (cut off, 1:50. Seropositive dogs were found in 5 out of 7 counties with a seroprevalence varying from 3.57% to 20% and an overall seroprevalence of 11.06% (24/217. There was no statistically significant difference between the prevalence of infection and the host age or gender. Results showed that stray dogs contribute to maintaining and spreading of Ehrlichia canis in Serbia. Due to the close relationship between people and dogs, it is of great importance to constantly monitor and improve prevention of this disease.
U.S. Department of Health & Human Services — NNDSS - Table II. Ehrlichiosis and Anaplasmosis, Ehrlichia ewingii infection to Undetermined - 2018. In this Table, provisional cases of selected notifiable diseases...
U.S. Department of Health & Human Services — NNDSS - Table II. Ehrlichiosis and Anaplasmosis, Anaplasma phagocytophilum infection to Ehrlichia chaffeensis infection - 2018. In this Table, provisional cases of...
Romero, L E; Meneses, A I; Salazar, L; Jiménez, M; Romero, J J; Aguiar, D M; Labruna, M B; Dolz, G
The present study investigated Ehrlichia species in blood samples from dogs suspected of clinical ehrlichiosis, using molecular and isolation techniques in cell culture. From a total of 310 canine blood samples analyzed by 16S rRNA nested PCR, 148 (47.7%) were positive for Ehrlichia canis. DNA from Ehrlichia chaffeensis or Ehrlichia ewingii was not detected in any sample using species-specific primers in separated reactions. Leukocytes from five PCR-positive dogs were inoculated into DH82 cells; successful isolation of E. canis was obtained in four samples. Partial sequence of the dsb gene of eight canine blood samples (including the five samples for in vitro isolation) was obtained by PCR and their analyses through BLAST showed 100% of identity with the corresponding sequence of E. canis in GenBank. This study represents the first molecular diagnosis, isolation, and molecular characterization of E. canis in dogs from Costa Rica. Copyright © 2010 Elsevier Ltd. All rights reserved.
Skinner, Delaina; Mitcham, Jessica R; Starkey, Lindsay A; Noden, Bruce H; Fairbanks, W Sue; Little, Susan E
American black bears (Ursus americanus) are commonly infested with ticks throughout their range, but there are few surveys for tick-borne disease agents in bears. To characterize tick infestations and determine the prevalence of current infection with Babesia spp. and past or current infection with Ehrlichia spp. in newly re-established populations of black bears in east central and southeastern Oklahoma, US, we identified adult (n=1,048) and immature (n=107) ticks recovered from bears (n=62). We evaluated serum and whole blood samples from a subset (n=49) for antibodies reactive to, and characteristic DNA fragments of, Ehrlichia spp., as well as characteristic DNA fragments of Babesia spp. Amblyomma americanum, the most common tick identified, was found on a majority (56/62; 90%) of bears and accounted for 697/1,048 (66.5%) of all ticks recovered. Other ticks included Dermacentor variabilis (338/1,048; 32.3%) from 36 bears, Amblyomma maculatum (9/1,048; 0.9%) from three bears, and Ixodes scapularis (4/1,048; 0.4%) from three bears. Antibodies reactive to Ehrlichia spp. were detected in every bear tested (49/49; 100%); maximum inverse titers to Ehrlichia chaffeensis ranged from 64-4,096 (geometric mean titer 1,525). However, PCR failed to identify active infection with E. chaffeensis, Ehrlichia ewingii, or an Ehrlichia ruminantium-like agent. Infection with Babesia spp. was detected by PCR in 3/49 (6%) bears. Together these data confirm that tick infestations and infection with tick-borne disease agents are common in bears in the southern US. The significance of these infestations and infections to the health of bears, if any, and the identity of the Ehrlichia spp. responsible for the antibody reactivity seen, warrant further evaluation.
Ehrlichia are obligate intracellular pathogens, transmitted by ixodid ticks, of both animals and humans. Ehrlichiae are emerging diseases in the USA, and the discovery of new species proceeds more rapidly than the development of models to study these agents. Laboratory animals were evaluated as
Martínez-Vega, Pedro Pablo; Bolio-Gonzalez, Manuel Emilio; Rodríguez-Vivas, Roger Iván; Gutierrez-Blanco, Eduardo; Pérez-Osorio, Carlos; Villegas-Perez, Sandra Luz; Sauri-Arceo, Carlos Humberto
The objective of this study was to determine the seroprevalence to Ehrlichia spp. in dogs from Xcalak, Quintana Roo, Mexico, and the associated factors. Serum samples were obtained from 118 dogs and used in an indirect immunofluorescent assay test for the detection of antibodies against Ehrlichia spp. A questionnaire was used to obtain information about possible variables associated with seroprevalence. These variables were analyzed through Chi 2 test and logistic regression. Dog seroprevalence of antibodies against Ehrlichia spp. was 64% (75/118). Fifty-two percent (61/118) of dogs had tick infestation which was identified as Rhipicephalus sanguineus sensu lato . Anemia was observed in 36% of dogs. Leucopenia (2.5%), thrombocytopenia (70%), and hemorrhage (14%) were also observed. Thirty-one percent (23/75) of dogs with anemia, 4% (3/75) of dogs with leucopenia, 80% (60/75) of dogs with thrombocytopenia, 17% (13/75) of dogs with hemorrhages, and 59% (44/75) of dogs with ticks were positive for Ehrlichia spp. antibodies. The factors associated with seroprevalence were age (1-3 and >3 years old, OR = 7.77 and OR = 15.39, resp.), tick infestation (OR = 3.13), and thrombocytopenia (OR = 3.36). In conclusion, seroprevalence of Ehrlichia spp. was high in the community of Xcalak and its associated factors were age, tick infestation, and thrombocytopenia.
Pedro Pablo Martínez-Vega
Full Text Available The objective of this study was to determine the seroprevalence to Ehrlichia spp. in dogs from Xcalak, Quintana Roo, Mexico, and the associated factors. Serum samples were obtained from 118 dogs and used in an indirect immunofluorescent assay test for the detection of antibodies against Ehrlichia spp. A questionnaire was used to obtain information about possible variables associated with seroprevalence. These variables were analyzed through Chi2 test and logistic regression. Dog seroprevalence of antibodies against Ehrlichia spp. was 64% (75/118. Fifty-two percent (61/118 of dogs had tick infestation which was identified as Rhipicephalus sanguineus sensu lato. Anemia was observed in 36% of dogs. Leucopenia (2.5%, thrombocytopenia (70%, and hemorrhage (14% were also observed. Thirty-one percent (23/75 of dogs with anemia, 4% (3/75 of dogs with leucopenia, 80% (60/75 of dogs with thrombocytopenia, 17% (13/75 of dogs with hemorrhages, and 59% (44/75 of dogs with ticks were positive for Ehrlichia spp. antibodies. The factors associated with seroprevalence were age (1–3 and >3 years old, OR = 7.77 and OR = 15.39, resp., tick infestation (OR = 3.13, and thrombocytopenia (OR = 3.36. In conclusion, seroprevalence of Ehrlichia spp. was high in the community of Xcalak and its associated factors were age, tick infestation, and thrombocytopenia.
Full Text Available Ehrlichia canis is a major pathogen in dogs throughout Africa, yet it has not been reported in Namibia. The aim of this study was to determine the seroprevalence of canine ehrlichiosis in central Namibia using the ImmunoComb assay (Biogal, Galed Laboratories. The study included 76 dogs that presented to the Rhino Park Veterinary Clinic in the north-western suburb of Khomasdal, Windhoek, Namibia, as well as 30 stray dogs from the Windhoek branch of the Society for the Prevention of Cruelty to Animals. Of the 106 dogs tested, 53.8% were seropositive at titres > 1:80. Dogs that presented with symptoms of E. canis infection had a significantly higher seroprevalence (86.6% compared with apparently healthy dogs (41.6% (P = 0.00. Location of habitation was significant (P < 0.017, with a high percentage of dogs exposed to E. canis living in the northern or north-western part of Windhoek. As the first study to serologically establish E. canis as a major pathogen in dogs in central Namibia, it is notable that the highest proportion of seropositive dogs came from low-income areas. Further investigation is necessary to describe the ecology of this important tick-borne pathogen of companion animals in Namibia.
KUBO, Shotaro; TATENO, Morihiro; ICHIKAWA, Yasuaki; ENDO, Yasuyuki
Tick-borne diseases are often encountered in canine clinical practice. In the present study, a molecular epidemiological survey of dogs in Japan was conducted to understand the prevalence and geographical distribution of Babesia spp., Hepatozoon spp., Ehrlichia spp. and Anaplasma spp. Pathogen-derived DNA in blood samples obtained from 722 dogs with a history of exposure to ticks and/or fleas was examined by PCR. The prevalence of Babesia gibsoni, Babesia odocoilei-like species, Hepatozoon canis and Ehrlichia spp./Anaplasma spp. was 2.4% (16/722), 0.1% (1/722), 2.5% (18/722) and 1.5% (11/722), respectively. While B. gibsoni and Ehrlichia spp./Anaplasma spp. were detected in the western part of Japan, H. canis was detected in Tohoku area in addition to western and central parts of Japan. PMID:25947226
Yu, X J; Crocquet-Valdes, P; Cullman, L C; Walker, D H
DNA encoding two repeat units of 120-kDa protein of Ehrlichia chaffeensis was cloned into the expression vector pGEX and expressed in Escherichia coli. The sensitivity and specificity of a dot blot assay for detection of human antibodies with the recombinant protein were 86 and 100%, respectively, compared with an indirect immunofluorescence assay.
Ísis Assis Braga
Full Text Available Ehrlichiosis is a worldwide distributed disease caused by different bacteria of the Ehrlichia genus that are transmitted by arthropod vectors. Its occurrence in dogs is considered endemic in several regions of Brazil. Regarding cats, however, few studies have been done and, consequently, there is not enough data available. In order to detect Ehrlichia spp. in cats from the central-western region of Brazil, blood and serum samples were collected from a regional population of 212 individuals originated from the cities of Cuiabá and Várzea Grande. These animals were tested by the Immunofluorescence Assay (IFA and the Polymerase Chain Reaction (PCR designed to amplify a 409 bp fragment of the dsb gene. The results obtained show that 88 (41.5% cats were seropositive by IFA and 20 (9.4% cats were positive by PCR. The partial DNA sequence obtained from PCR products yielded twenty samples that were found to match perfectly the Ehrlichia canis sequences deposited on GenBank. The natural transmission of Ehrlichia in cats has not been fully established. Furthermore, tick infestation was not observed in the evaluated cats and was not observed any association between age, gender and positivity of cats in both tests. The present study reports the first serological and molecular detection of E. canis in domestic cats located in the endemic area previously mentioned.
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
Cabezas-Cruz, A.; Valdés, James J.; de la Fuente, J.
Roč. 7, DEC 10 2014 (2014), s. 584 ISSN 1756-3305 R&D Projects: GA MŠk(CZ) EE2.3.30.0032 Institutional support: RVO:60077344 Keywords : Ehrlichia sp. UFMG-EV * Ehrlichia sp. UFMT-BV * E. mineirensis * Host-shift * Diversifying episodic selection Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 3.430, year: 2014
Gofton, Alexander W; Waudby, Helen P; Petit, Sophie; Greay, Telleasha L; Ryan, Una M; Irwin, Peter J
Anaplasma and Ehrlichia spp. are tick-borne pathogens that can cause severe disease in domestic animals, and several species are responsible for emerging zoonoses in the northern hemisphere. Until recently, the only members of these genera reported in Australia (A. marginale, A. centrale, and A. platys) were introduced from other continents, through the importation of domestic animals and their associated ticks. However, unique Anaplasma and Ehrlichia 16S rRNA gene sequences were recently detected for the first time in native Australian ticks, particularly in Amblyomma triguttatum subsp. ticks from southwest Western Australia (WA). We used molecular techniques to survey Am. triguttatum subsp. ticks from four allopatric populations in southern and western Australia for Anaplasma and Ehrlichia species, and described here the phylogeny of these novel organisms. An A. bovis variant (genotype Y11) was detected in ticks from two study sites; Yanchep National Park (12/280, 4.3%) and Barrow Island (1/69, 1.4%). Phylogenetic analysis of 16S rRNA and groEL gene sequences concluded that A. bovis genotype Y11 is a unique genetic variant, distinct from other A. bovis isolates worldwide. Additionally, a novel Ehrlichia species was detected in Am. triguttatum subsp. from three of the four study sites; Yanchep National Park (18/280, 6.4%), Bungendore Park (8/46, 17.4%), and Innes National Park (9/214, 4.2%), but not from Barrow Island. Phylogenetic analysis of 16S, groEL, gltA, and map1 gene sequences revealed that this Ehrlichia sp. is most closely related to, but clearly distinct from, E. ruminantium and Ehrlichia sp. Panola Mountain. We propose to designate this new species 'Candidatus Ehrlichia occidentalis'. Anaplasma bovis genotype Y11 and 'Candidatus E. occidentalis' are the first Anaplasma and Ehrlichia species to be recorded in native Australian ticks. Copyright © 2017 Elsevier GmbH. All rights reserved.
Zweygarth, E.; Schol, H.; Lis, K.; Cabezas Cruz, Alejandro; Thiel, C.; Silaghi, C.; Ribeiro, M.F.B.; Passos, L.M.F.
Roč. 60, NOV 2013 (2013), s. 86-92 ISSN 1865-1674 Grant - others:EU(XE) FP7-PEOPLE-ITN No.238511 Institutional support: RVO:60077344 Keywords : Ehrlichia * Rhipicephalus (Boophilus) microplus * in vitro culture * tick cell * DH82 * endothelial cell * cattle * 16S rRNA * Brazil Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 3.116, year: 2013
Watanabe, Malaika; Oikawa, T; Hiraoka, Hiroko; Kaneko, N; Itamoto, Kazuhito; Mizuno, Tohru; Okuda, Masaru; Inokuma, Hisashi
Three beagle dogs were inoculated with mice spleen/liver homogenate infected with Ehrlichia species detected from Ixodes ovatus (EIO) and one dog was used as a control. All three infected dogs did not show clinical signs of disease except for mild pyrexia throughout the 41-day study period. Splenomegaly was observed from Day 7 post-inoculation (p.i.) in two of the dogs. Hematological and biochemical abnormalities included mild thrombocytopenia, hypoproteinaemia, hypoalbuminaemia and increased...
Seroprevalencia de Leptospira sp., Rickettsia sp. Ehrlichia sp. en trabajadores rurales del departamento de Sucre, Colombia Seroprevalence of Leptospira sp., Rickettsia sp. and Ehrlichia sp. in rural workers of Sucre, Colombia
Full Text Available Objective. Determinar la seroprevalencia de Leptospira sp., Rickettsia sp. y Ehrlichia sp. en trabajadores de áreas rurales del departamento de Sucre. Material y métodos. Se realizó un estudio escriptivo, prospectivo, de corte transversal, que pretendió determinar la seroprevalencia e Leptospira sp., Rickettsia sp. y Ehrlichia sp. en 90 trabajadores de áreas rurales del departamento de Sucre. Se estableció la presencia de anticuerpos séricos anti-IgM específicos anti-Leptospira por la técnica de ELISA indirecta. Para la determinación de Rickettsia sp. y Ehrlichia sp. se uso la técnica de inmunofluorescencia indirecta. Resultados. La población evaluada estaba compuesta por 27 (30% ordeñadores, 21 (23% jornaleros, 18 (20% profesionales del campo y 24 (27% que realizaban otras actividades. Ventidós (24% muestras resultaron positivas en alguna de las pruebas. De éstas, 12 (13,3% fueron positivas para Leptospira sp., 7 (7,8% para Rickettsia sp. y 3 (3,3% ara Ehrlichia sp. Conclusión. Este fue el primer estudio que se llevó a cabo en el departamento de Sucre y permitió demostrar que existe una prevalencia importante de Leptospira p.,Rickettsia sp. y Ehrlichia sp.. Los factores de riesgo ocupacional fueron factores determinantes en la seropositividad.Objective. To determine the seroprevalence of Leptospira sp., Rickettsia sp. and Ehrlichia sp. in agricultural workers of Sucre. Methods. A descriptive prospective cross-sectional study was conducted in ninety rural workers of Sucre. Presence of serum antibodies anti-IgM specific anti-Leptospira by indirect ELISA was established. For the determination of Rickettsia and Ehrlichia indirect inmunoflorescence was used. Results.The population was composed by 27 (30% milkers, 21 (23% day workers, 18 farm professionals (20% and 24 (26% workers in others activities. A total of 22 (24% samples were positive to some test. Twelve (13.3% were positive to Leptospira sp., seven (7.8% to Rickettsia sp
Almeida, Aliny P; Souza, Tayse D; Marcili, Arlei; Labruna, Marcelo B
This study evaluated infection by vector-borne agents in 58 crab-eating fox (Cerdocyon thous L.) that were road-killed in an Atlantic rainforest reserve in the state of Espírito Santo, southeastern Brazil. Spleen, lung, or blood samples collected from the foxes were tested in the laboratory by a battery of polymerase chain reaction (PCR) assays targeting bacteria of the genera Rickettsia, Borrelia, Coxiella, Anaplasma, and Ehrlichia; and protozoa of the genera Babesia, Hepatozoon, and Leishmania. Of the targeted organisms, evidence of infection in the foxes was detected for Ehrlichia and Hepatozoon organisms only. Overall, six (10.3%) foxes were infected by an ehrlichial agent closely related to an ehrlichial agent recently detected in free-ranging Jaguars [(Panthera onca (L.)] in central-western Brazil, and to Ehrlichia ruminantium. For Hepatozoon, 28 (48.3%) foxes were infected by an agent closely related to Hepatozoon sp. Curupira 2 and H. americanum; and one (1.7%) fox was infected by an organism closely related to reptile-associated Hepatozoon agents. Finally, 11 (19.0%) foxes were found infested by Amblyomma cajennense (F.) nymphs, which were all PCR negative for the range of vector-borne agents cited above. Because the haplotypes found in free-ranging foxes are genetically closely related to pathogens of great veterinary importance, namely E. ruminantium and H. americanum, it is highly desirable to know if these novel organisms have any important role as agents of diseases in domestic animals and wildlife in Brazil.
Full Text Available Rickettsial organisms resembling Ehrlichia ruminantium (the causative organism of heartwater were demonstrated in brain smears and formalin-fixed brain sections derived from a buffalo calf that died on a private game reserve in northern KwaZulu-Natal. The possibility that the tick-free environment of a quarantine boma may have affected the calf 's immunity, is discussed. These findings suggest that monitoring heartwater in wild ruminants and making brain smears as a routine during post mortem evaluations of wild ruminants, should be encouraged.
Faria, Joice Lara Maia; Dagnone, Ana Sílvia; Munhoz, Thiago Demarchi; João, Carolina Franchi; Pereira, Wanderson Adriano Biscola; Machado, Rosângela Zacarias; Tinucci-Costa, Mirela
The aim of this study was to compare the detection of Ehrlichia canis morulae and DNA by nPCR in whole blood and spleen aspiration. The sample included 40 dogs showing thrombocytopenia associated to clinical signs suggestive of canine ehrlichiosis. Morulae detection showed that in 35 of the dogs studied, 17 had morulae in spleen tissue, and two in buffy coat smears. E. canis DNA was detected in 29/40 blood samples. We verified that morulae detection is more efficient in cytological preparations from spleen aspiration. On the other hand, nPCR on spleen and blood samples were equally efficient for disease diagnosis.
Trout Fryxell, Rebecca T; Hendricks, Brain M; Pompo, Kimberly; Mays, Sarah E; Paulsen, Dave J; Operario, Darwin J; Houston, Allan E
Ehrlichiosis and rickettsiosis are two common bacterial tick-borne diseases in the southeastern United States. Ehrlichiosis is caused by ehrlichiae transmitted by Amblyomma americanum and rickettsiosis is caused by rickettsiae transmitted by Amblyomma maculatum and Dermacentor variabilis. These ticks are common and have overlapping distributions in the region. The objective of this study was to identify Anaplasma, Ehrlichia, and Rickettsia species associated with questing ticks in a Rocky Mountain spotted fever (RMSF) hotspot, and identify habitats, time periods, and collection methods for collecting questing-infected ticks. Using vegetation drags and CO 2 -baited traps, ticks were collected six times (May-September 2012) from 100 sites (upland deciduous, bottomland deciduous, grassland, and coniferous habitats) in western Tennessee. Adult collections were screened for Anaplasma and Ehrlichia (simultaneous polymerase chain reaction [PCR]) and Rickettsia using genus-specific PCRs, and resulting positive amplicons were sequenced. Anaplasma and Ehrlichia were only identified within A. americanum (Ehrlichia ewingii, Ehrlichia chaffeensis, Panola Mountain Ehrlichia, and Anaplasma odocoilei sp. nov.); more Ehrlichia-infected A. americanum were collected at the end of June regardless of habitat and collection method. Rickettsia was identified in three tick species; "Candidatus Rickettsia amblyommii" from A. americanum, R. parkeri and R. andeanae from A. maculatum, and R. montanensis ( = montana) from D. variabilis. Overall, significantly more Rickettsia-infected ticks were identified as A. americanum and A. maculatum compared to D. variabilis; more infected-ticks were collected from sites May-July and with dragging. In this study, we report in the Tennessee RMSF hotspot the following: (1) Anaplasma and Ehrlichia are only found in A. americanum, (2) each tick species has its own Rickettsia species, (3) a majority of questing-infected ticks are collected May-July, (4) A
Full Text Available Canine leishmaniasis (CanL due to Leishmania infantum and canine monocytic ehrilichiosis (CME due to Ehrlichia canis are common diseases with zoonotic potential in the Mediterranean area. Their prevalence in R. Macedonia as a neighboring Mediterranean county is expected. In both diseases similar clinical symptoms can be manifested in dogs such as: lethargy, anorexia, weight loss, epistaxis, fever, pale mucous membranes, enlarged lymph nodes, splenomegaly, ocular signs. This case report present an atypical case of 11 year old female Samoyed with starting single clinical symptom epistaxys. Initial diagnostic procedures revealed the presence only of CanL, which was diagnosed using indirect immunofluorescence method and ELISA. First laboratory findings showed normal hematological and renal profiles. Dog was put on a treatment with Allopurinol (20mg/kg, p/o for at least 9 months. Termination of the therapy after 6 months brought a numerous clinical symptoms involving weakness, dehydration, pale mucous membranes lost pupilar reflex, uremic breath and biochemical parameters revealed a renal failure. Using a commercial ELISA kit Ehrlichia canis as a co infection was diagnosed. Most probably the second infectious agent was induced in the past 6 months, causing more severe pathological effects than CanL infection alone.
Oliveira, Ana Cristina; Luz, Maria Francisca; Granada, Sara; Vilhena, Hugo; Nachum-Biala, Yaarit; Lopes, Ana Patrícia; Cardoso, Luís; Baneth, Gad
Molecular identification of tick-borne pathogen infection in cats from Africa is scarce. The presence of bacterial (Anaplasma and Ehrlichia) and protozoal (Babesia and Hepatozoon) agents was investigated in blood samples from 102 domestic cats from Luanda, Angola, by polymerase chain reaction and DNA sequencing. Three cats (2.9%) were found infected with Ehrlichia canis, three (2.9%) with Hepatozoon felis and one (1.0%) with Anaplasma bovis. The prevalence of infections with one single agent was 4.9%, and that of infection with two agents (i.e. E. canis and H. felis) was 1.0%. In total, six cats (5.9%) were found infected with at least one of the detected tick-borne agents. This is the first report of A. bovis, E. canis and H. felis in cats from Angola. To the best of our knowledge, A. bovis is also being reported for the first time in domestic cats outside of Japan. Cats are at a low to moderate risk of being infected with tick-borne agents in Luanda.
Bekloo, Ahmad Jafar; Bakhshi, Hasan; Soufizadeh, Ayoub; Sedaghat, Mohammad Mehdi; Bekloo, Romina Jafar; Ramzgouyan, Maryam Roya; Chegeni, Asadollah Hosseini; Faghihi, Faezeh; Telmadarraiy, Zakkyeh
Ticks serve as important vectors of some pathogens of medical importance all over the world and identification of their rate of infection plays an important role for further control of diseases. In the current study, we investigated on ticks collected from north of Iran where raising and caring livestock are the main task of the people in order to find evidences of infection of Babesia, Theileria, Anaplasma and Ehrlichia microbial agents. Totally, 609 hard tick species from two genera Hyalomma and Rhipicephalus including; Hy. scupense, Hy. dromedarii, Hy. rufipes, Hy. marginatum, Hy. asiaticum, Hy. anatolicum, R. bursa, R. sanguineus and R. turanicus were identified. Molecular analysis revealed the presence of Anaplasma, Ehrlichia, Babesia and Theileria microorganism agents in all collected tick species except Hy. asiaticum and R. turanicus. To the best of our knowledge, this is the first report on identification of B. occultans in Hyalomma anatolicum and B. ovis in Hyalomma sp in Iran. Copyright © 2017 Elsevier B.V. All rights reserved.
Dredla, Brynn; Freeman, William D
Thunderclap headache is a sudden and severe headache that can occur after an aneurysmal subarachnoid hemorrhage (SAH). Subarachnoid hemorrhage is a medical emergency that requires prompt attention and hospitalization. Patients with thunderclap headache often undergo a noncontrast head computed tomography (CT) scan to ascertain SAH bleeding and, if the scan is negative, then undergo a lumbar puncture to look for cerebrospinal fluid (CSF) red blood cells (RBCs), which would be consistent with an aneurysmal leak. If the initial CT is negative and CSF is positive for RBCs, patients are usually admitted to the hospital for evaluation of intracranial aneurysm. We encountered a patient with thunderclap headache whose initial head CT was negative for SAH and whose CSF tested positive for RBCs. The patient was referred to our center for evaluation and management of aneurysmal SAH. However, on careful review of the patient's medical history, serum laboratory values, and spinal fluid values, the patient was diagnosed with Ehrlichia chaffeensis meningitis. While Ehrlichia meningitis is rare, it is important to recognize the clinical clues that could help avoid formal cerebral angiography, a costly and potentially unnecessary procedure. We present how this case represented a cognitive framing bias and anchoring heuristic as well as steps that medical providers can use to prevent such cognitive errors in diagnosis.
Bouza-Mora, Laura; Dolz, Gaby; Solórzano-Morales, Antony; Romero-Zuñiga, Juan José; Salazar-Sánchez, Lizbeth; Labruna, Marcelo B; Aguiar, Daniel M
This study focuses on the detection and identification of DNA and antibodies to Ehrlichia spp. in samples of blood bank donors in Costa Rica using molecular and serological techniques. Presence of Ehrlichia canis was determined in 10 (3.6%) out of 280 blood samples using polymerase chain reaction (PCR) targeting the ehrlichial dsb conserved gene. Analysis of the ehrlichial trp36 polymorphic gene in these 10 samples revealed substantial polymorphism among the E. canis genotypes, including divergent tandem repeat sequences. Nucleotide sequences of dsb and trp36 amplicons revealed a novel genotype of E. canis in blood bank donors from Costa Rica. Indirect immunofluorescence assay (IFA) detected antibodies in 35 (35%) of 100 serum samples evaluated. Thirty samples showed low endpoint titers (64-256) to E. canis, whereas five sera yielded high endpoint titers (1024-8192); these five samples were also E. canis-PCR positive. These findings represent the first report of the presence of E. canis in humans in Central America. Copyright Â© 2016 Elsevier GmbH. All rights reserved.
Full Text Available Objective: To ascertain the prevalence of the Anaplasma/Ehrlichia infections in tick population within four provinces of Iran. Methods: A total of 384 tick specimens were collected from domestic animals inhabiting in four provinces (East Azerbaijan, Gilan, South Khorasan and Yazd. Specimens were identified based on morphological analysis. The detection of Anaplasma spp./Ehrlichia spp. within tick samples was carried out by nested PCR amplification of the 16S ribosomal RNA gene accompanied by DNA sequencing and analysis for verification. Results: A total of 10 tick species were identified as follows: Ornithodoros lahorensis (O. lahorensis (44.8%, Hyalomma dromedarii (15.6%, Dermacentor marginatus (13.5%, Hyalomma anatolicum (11.2%, Hyalomma asiaticum (5.7%, Hyalomma marginatum (4.9%, Rhipicephalus sanguineus (2.3%, Hyalomma detritum (1.0%, Dermacentor niveus (0.5% and Argas persicus (0.3%. The percentage distribution of Anaplasma/Ehrlichia was 55.5% (213 across 384 studied ticks. Conclusions: To the best of our knowledge, this is the first report of Anaplasma ovis infection in O. lahorensis in Iran. We also conjecture the prevalence of Ehrlichia spp. in Yazd Province based on sequencing results; also, it is suggested that O. lahorensis is a potential vector in the studied area. This survey highlights the importance of Argasidae family to verify and correlate their threat in causing anaplasmosis and other diseases in animals.
Zweygarth, E.; Cabezas Cruz, Alejandro; Josemans, A.I.; Oosthuizen, M.C.; Matjila, P.T.; Lis, K.; Broniszewska, M.; Schöl, H.; Ferrolho, J.; Grubhoffer, Libor; Passos, L.M.F.
Roč. 5, č. 4 (2014), s. 423-431 ISSN 1877-959X Institutional support: RVO:60077344 Keywords : Ehrlichia canis * In vitro culture * IDE8 tick cells * DH82 * 16S rRNA * gp36 Subject RIV: FN - Epidemiology, Contagious Diseases ; Clinical Immunology Impact factor: 2.718, year: 2014
Felipe da Silva Krawczak
Full Text Available INTRODUCTION: The present study was designed to assess the occurrence of co-infection or cross-reaction in the serological techniques used for detecting the anti-Leishmania spp., -Babesia canis vogeli and -Ehrlichia canis antibodies in urban dogs from an area endemic to these parasites. METHODS: The serum samples from dogs were tested for the Babesia canis vogeli strain Belo Horizonte antigen and Ehrlichia canis strain São Paulo by immunofluorescence antibody test (IFAT and by anti-Leishmania immunoglobulin G (IgG antibody detection to assess Leishmania infection. We used the following four commercial kits for canine visceral leishmaniasis: ELISA, IFAT, Dual Path Platform (DPP (Bio Manguinhos(r/FIOCRUZ/MS and a rK39 RDT (Kalazar Detect Canine Rapid Test; Inbios. RESULTS : Of 96 serum samples submitted to serological assays, 4 (4.2% were positive for Leishmania as determined by ELISA; 12 (12.5%, by IFAT; 14 (14.6% by rK39 RDT; and 20 (20.8%, by DPP. Antibodies against Ehrlichia and Babesia were detected in 23/96 (23.9% and 30/96 (31.2% samples, respectively. No significant association was identified between the results of tests for detecting Babesia or Ehrlichia and those for detecting Leishmania (p-value>0.05. CONCLUSIONS: In the present study, we demonstrated co-infection with Ehrlichia or Babesia and Leishmania in dogs from Minas Gerais (Brazil; we also found that the serological tests that were used did not cross-react.
Kawahara, Makoto; Ito, Tadahiko; Suto, Chiharu; Shibata, Shinichiro; Rikihisa, Yasuko; Hata, Kazuhisa; Hirai, Katsuya
In metropolitan Tokyo, the Ehrlichia muris seropositivity rate of 24 wild mice was 63% in Hinohara Village, but in the surrounding areas, it was 0 to 5%. This finding suggests that the reservoir of E. muris is focal. Among the 15 seropositive mice, ehrlichiae were isolated from 9 Apodemus speciosus mice and 1 A. argenteus mouse, respectively. Five ehrlichial isolates were obtained from 10 ticks (Haemaphysalis flava) collected in Asuke Town, Aichi Prefecture, where the E. muris type strain had...
Full Text Available El objetivo del estudio fue determinar la frecuencia y factores asociados a la infección por Ehrlichia canis en perros. Se recolectaron muestras de sangre de 150 perros infestados con garrapatas en 10 consultorios veterinarios de la ciudad de Huánuco en Perú, los perros fueron seleccionados al azar sin distinción de raza, edad ni sexo. Se detectó anticuerpos contra Ehrlichia canis mediante inmunoensayo cromatográfico. El 51,3% de perros estuvieron infectados por Ehrlichia canis. En el análisis multivariado se encontraron asociados a la presencia Ehrlichia canis, el mal estado de salud del perro (p=0,049, un promedio mayor de infestación por garrapatas (p=0,018, perros de edad adulta (p=0,038. La frecuencia de Ehrlichia canis en perros de esta ciudad es alta. Se recomienda el control de la garrapata marrón del perro (Rhipicephalus sanguineus vector de la Ehrlichia canis
Al Izzi, Salah; Martin, Donald S; Chan, Roxanne Y Y; Leutenegger, Christian M
A 12-month-old male neutered mixed breed dog was presented with a history of diarrhea, lethargy, emaciation, polydypsia, and sniffling. Physical examination findings included pale mucous membranes, increased heart and respiratory rates, and normal rectal temperature (38°C). Hematologic abnormalities included anemia and thrombocytopenia. Biochemical abnormalities included hypoalbuminemia, hyperbilirubinemia, and elevated ALP and ALT activities. A SNAP 4Dx test result was positive for Ehrlichia canis. Babesia canis vogeli organisms were found in the peripheral blood films, while morulae of E canis were not seen. Real-time polymerase chain reaction testing confirmed the presence of both B c vogeli and E canis organisms, and also was positive for Anaplasma platys infection. The dog recovered following treatment with doxycycline and imidocarb dipropionate, with normal hematology and biochemical profiles. © 2013 American Society for Veterinary Clinical Pathology.
Magnarelli, L A; Stafford, K C; Ijdo, J W; Fikrig, E; Oliver, J H; Hutcheson, H J; Boone, J L
Serum samples, collected from Peromyscus leucopus (white-footed mouse) or Peromyscus gossypinus (cotton mouse) during 1987 through 1990 in Florida, Georgia, Maryland, Mississippi, and North Carolina (USA), and in 1997 in southern Connecticut were analyzed by indirect fluorescent antibody (IFA) staining methods or Western blot procedures for antibodies to granulocytic ehrlichiae. Of the 82 sera from white-footed mice in Connecticut tested by IFA methods with either the BDS or NCH-1 strain of the human granulocytic ehrlichiosis (HGE) agent, 45 (55%) and 42 (51%) of the samples contained antibodies to these strains, respectively, at concentrations ranging from 1:80 to 1:2560. One (2%) of 43 sera from P. leucopus captured in Assateague Island (Maryland) had a titer of 1:80, while three (20%) of 15 sera from P. gossypinus, captured in Sapelo Island (Georgia) and four (40%) of 10 sera from cotton mice caught in Amelia Island (Florida) had antibodies to the NCH-1 strain at titers of 1:160 to 1:1,280. Fifty-five sera from P. leucopus in Cape Hatteras (North Carolina) and 30 sera from P. gossypinus in Mississippi were negative. Western blot analyses confirmed seropositivity for 19 (95%) of 20 mouse sera positive by IFA staining methods, including samples from both mouse species captured in Connecticut, Maryland, or Florida. There were key banding patterns to proteins having molecular masses of about 44, 80, 105, 110, or 120 kDa. Both serologic assays can be used to determine if mice have been exposed to granulocytic ehrlichiae. These rodents also may be useful in surveillance programs to identify endemic sites for HGE and in performing laboratory studies on immune responses to the etiologic agent.
Lina, Taslima T.; Dunphy, Paige S.; Luo, Tian; McBride, Jere W.
ABSTRACT Ehrlichia chaffeensis preferentially targets mononuclear phagocytes and survives through a strategy of subverting innate immune defenses, but the mechanisms are unknown. We have shown E.?chaffeensis type 1 secreted tandem repeat protein (TRP) effectors are involved in diverse molecular pathogen-host interactions, such as the TRP120 interaction with the Notch receptor-cleaving metalloprotease ADAM17. In the present study, we demonstrate E.?chaffeensis, via the TRP120 effector, activat...
Full Text Available Abstract Pathogens transmitted by ticks are an emerging problem worldwide, this study aimed to diagnose the causal agents of infection in dogs presenting suspected hemoparasitoses. Fifty-eight dogs with clinical signs such as depression, hemorrhagic diathesis and fever were evaluated regarding clinical presentation, hemogram, blood smears and serological tests, using the indirect immunofluorescence method for the agents Babesia vogeli and Ehrlichia canis and conventional PCR for Babesia spp. (gene 18S rRNA, Rangelia vitalii (gene 18S rRNA and Ehrlichia spp. (gene dsb. Five (8.6% of the 58 dogs were serologically positive for Babesia spp. and three (5.1% for E. canis. Four dogs (6.8% were positive for R. vitalii through the molecular diagnosis. The PCR products were sequenced and the DNA from R. vitalii was found to be 99% genetically identical to samples of R. vitalii that had been isolated in Brazil. No presence of Babesia spp. or E. canis was observed through PCR on the dogs evaluated here. The results indicate the presence of R. vitalii and exposure to Babesia spp. and Ehrlichia spp. among the dogs analyzed.
Sebastian, Patrick S; Tarragona, Evelina L; Bottero, María N Saracho; Mangold, Atilio J; Mackenstedt, Ute; Nava, Santiago
The aim of this study was to get an overview about the occurrence of bacteria from the genus Ehrlichia and Rickettsia in ixodid ticks with medical importance in Argentina. Therefore, in 2013 and 2014, free-living ticks were collected in different provinces of northern Argentina. These ticks were determined as Amblyomma sculptum, Amblyomma neumanni, Amblyomma parvum, Amblyomma triste, Amblyomma ovale, Amblyomma tonelliae and Haemaphysalis juxtakochi. All samples were tested to determine the infection with Ehrlichia spp. and Rickettsia spp. by PCR assays. Rickettsial DNA was detected in all tested tick species, with the exception of A. tonelliae. 'Candidatus Rickettsia amblyommii', 'Candidatus Rickettsia andeanae', and Rickettsia parkeri were found in A. neumanni, A. parvum, and A. triste, respectively. Another rickettsial species, Rickettsia bellii, was found in A. sculptum, A. ovale and H. juxtakochi. None of the tested ticks showed infection with Ehrlichia. The results of the study demonstrate that Rickettsia species belonging to the spotted fever group are associated with various species of Amblyomma throughout a wide area of northern Argentina, where cases of Amblyomma ticks biting humans are common.
Parola, Philippe; Cornet, Jean-Paul; Sanogo, Yibayiri Osée; Miller, R Scott; Thien, Huynh Van; Gonzalez, Jean-Paul; Raoult, Didier; Telford III, Sam R; Wongsrichanalai, Chansuda
A total of 650 ticks, including 13 species from five genera, were collected from animals, from people, or by flagging of the vegetation at sites on the Thai-Myanmar border and in Vietnam. They were tested by PCR to detect DNA of bacteria of the order RICKETTSIALES: Three Anaplasma spp. were detected in ticks collected in Thailand, including (i) Anaplasma sp. strain AnDa465, which was considered a genotype of Anaplasma platys (formerly Ehrlichia platys) and which was obtained from Dermacentor auratus ticks collected from dogs; (ii) Anaplasma sp. strain AnAj360, which was obtained from Amblyomma javanense ticks collected on a pangolin; and (iii) Anaplasma sp. strain AnHl446, which was closely related to Anaplasma bovis and which was detected in Haemaphysalis lagrangei ticks collected from a bear. Three Ehrlichia spp. were identified, including (i) Ehrlichia sp. strain EBm52, which was obtained from Boophilus microplus ticks collected from cattle from Thailand; (ii) Ehrlichia sp. strain EHh324, which was closely related to Ehrlichia chaffeensis and which was detected in Haemaphysalis hystricis ticks collected from wild pigs in Vietnam; and (iii) Ehrlichia sp. strain EHh317, which was closely related to Ehrlichia sp. strain EBm52 and which was also detected in H. hystricis ticks collected from wild pigs in Vietnam. Two Rickettsia spp. were detected in Thailand, including (i) Rickettsia sp. strain RDla420, which was detected in Dermacentor auratus ticks collected from a bear, and (ii) Rickettsia sp. strain RDla440, which was identified from two pools of Dermacentor larvae collected from a wild pig nest. Finally, two bacteria named Eubacterium sp. strain Hw124 and Eubacterium sp. strain Hw191 were identified in Haemaphysalis wellingtoni ticks collected from chicken in Thailand; these strains could belong to a new group of bacteria.
Attipa, Charalampos; Solano-Gallego, Laia; Papasouliotis, Kostas; Soutter, Francesca; Morris, David; Helps, Chris; Carver, Scott; Tasker, Séverine
In the Mediterranean basin, Leishmania infantum is a major cause of disease in dogs, which are frequently co-infected with other vector-borne pathogens (VBP). However, the associations between dogs with clinical leishmaniosis (ClinL) and VBP co-infections have not been studied. We assessed the risk of VBP infections in dogs with ClinL and healthy controls. We conducted a prospective case-control study of dogs with ClinL (positive qPCR and ELISA antibody for L. infantum on peripheral blood) and clinically healthy, ideally breed-, sex- and age-matched, control dogs (negative qPCR and ELISA antibody for L. infantum on peripheral blood) from Paphos, Cyprus. We obtained demographic data and all dogs underwent PCR on EDTA-blood extracted DNA for haemoplasma species, Ehrlichia/Anaplasma spp., Babesia spp., and Hepatozoon spp., with DNA sequencing to identify infecting species. We used logistic regression analysis and structural equation modelling (SEM) to evaluate the risk of VBP infections between ClinL cases and controls. From the 50 enrolled dogs with ClinL, DNA was detected in 24 (48%) for Hepatozoon spp., 14 (28%) for Mycoplasma haemocanis, 6 (12%) for Ehrlichia canis and 2 (4%) for Anaplasma platys. In the 92 enrolled control dogs, DNA was detected in 41 (45%) for Hepatozoon spp., 18 (20%) for M. haemocanis, 1 (1%) for E. canis and 3 (3%) for A. platys. No Babesia spp. or "Candidatus Mycoplasma haematoparvum" DNA was detected in any dog. No statistical differences were found between the ClinL and controls regarding age, sex, breed, lifestyle and use of ectoparasitic prevention. A significant association between ClinL and E. canis infection (OR = 12.4, 95% CI: 1.5-106.0, P = 0.022) was found compared to controls by multivariate logistic regression. This association was confirmed using SEM, which further identified that younger dogs were more likely to be infected with each of Hepatozoon spp. and M. haemocanis, and dogs with Hepatozoon spp. were more likely to
Full Text Available Abstract Background Ehrlichia ruminantium is the causal agent of heartwater, a fatal tropical disease affecting ruminants with important economic impacts. This bacterium is transmitted by Amblyomma ticks and is present in sub-Saharan Africa, islands in the Indian Ocean and the Caribbean, where it represents a threat to the American mainland. Methods An automated DNA extraction method was adapted for Amblyomma ticks and a new qPCR targeting the pCS20 region was developed to improve E. ruminantium screening capacity and diagnosis. The first step in the preparation of tick samples, before extraction, was not automated but was considerably improved by using a Tissue Lyser. The new pCS20 Sol1 qPCR and a previously published pCS20 Cow qPCR were evaluated with the OIE standard pCS20 nested PCR. Results pCS20 Sol1 qPCR was found to be more specific than the nested PCR, with a 5-fold increase in sensitivity (3 copies/reaction vs 15 copies/reaction, was less prone to contamination and less time-consuming. As pCS20 Sol1 qPCR did not detect Rickettsia, Anasplasma and Babesia species or closely related species such as Panola Mountain Ehrlichia, E. chaffeensis and E. canis, its specificity was also better than Cow qPCR. In parallel, a tick 16S qPCR was developed for the quality control of DNA extraction that confirmed the good reproducibility of the automated extraction. The whole method, including the automated DNA extraction and pCS20 Sol1 qPCR, was shown to be sensitive, specific and highly reproducible with the same limit of detection as the combined manual DNA extraction and nested PCR, i.e. 6 copies/reaction. Finally, 96 samples can be tested in one day compared to the four days required for manual DNA extraction and nested PCR. Conclusions The adaptation of an automated DNA extraction using a DNA/RNA viral extraction kit for tick samples and the development of a new qPCR increased the accuracy of E. ruminantium epidemiological studies, as well as the
Mylonakis, Mathio E; Koutinas, Alex F; Baneth, Gad; Polizopoulou, Zoe; Fytianou, Anna
A 5-month-old, female, mongrel dog was admitted to the Clinic of Companion Animal Medicine, Aristotle University of Thessaloniki, Greece, with depression, anorexia, fever, peripheral lymphadenopathy, splenomegaly, oculonasal discharge, nonregenerative anemia, and mild thrombocytopenia. Cytology of Giemsa-stained buffy coat, bone marrow, and lymph node aspiration smears revealed numerous morulae in mononuclear leukocytes and in neutrophils, and Hepatozoon canis gamonts in neutrophils. The dog was seropositive to Ehrlichia canis (immunofluorescence assay [IFA]) and Hepatozoon canis (ELISA) but not to Anaplasma phagocytophilum (IFA). A nested polymerase chain reaction performed on bone marrow aspirates was positive for E canis. This method was not applied for the detection of A phagocytophilum. Treatment with doxycycline and imidocarb dipropionate resulted in both clinical and parasitologic cure. This is the first reported case of a mixed infection with E canis, H canis, and presumptive A phagocytophilum. The findings emphasize the value of cytology in offering a quick and inexpensive diagnosis in mixed tick-borne infections of dogs.
Criado-Fornelio, Angel; Martín-Pérez, T; Verdú-Expósito, C; Reinoso-Ortiz, S A; Pérez-Serrano, J
Wildlife species are involved in the transmission of diverse pathogens. This study aimed to monitor raccoons (Procyon lotor), American minks (Neovison vison), and red foxes (Vulpes vulpes) as potential reservoirs in central Spain. Specifically, 200 spleen and fecal samples (from 194 raccoons, 3 minks, and 3 foxes) were analyzed molecularly by PCR/qPCR and sequencing for the presence of piroplasmids, Hepatozoon spp., Toxoplasma gondii, and Ehrlichia canis infections in the Community of Madrid (Spain). Biological samples were obtained in the years 2014, 2015, and 2016. No pathogen DNA was found in fecal samples. In contrast, analysis of raccoon spleen samples revealed that Toxoplasma was the most prevalent pathogen (prevalence 3.6 ± 2.6%), followed by Hepatozoon canis and E. canis (each with a prevalence of 2.57 ± 2.2%). Hepatozoon canis was also diagnosed in all three of the analyzed foxes. Analysis of yearly prevalence showed that tick-borne pathogens were less frequent in raccoon in 2015, a dry and warm year compared both to 2014 and 2016. These data suggest that fecal PCR assays are unsuitable for detection of DNA of non-erythrocytic pathogens. Furthermore, they demonstrate that the raccoon (an invasive species often living in proximity to domestic areas) and the red fox are putative reservoirs for pathogenic organisms in the Community of Madrid.
Muangchuen, Ajima; Chaumpluk, Piyasak; Suriyasomboon, Annop; Ekgasit, Sanong
Canine monocytic ehrlichiosis (CME) is a major thick-bone disease of dog caused by Ehrlichia canis. Detection of this causal agent outside the laboratory using conventional methods is not effective enough. Thus an assay for E. canis detection based on the p30 outer membrane protein gene was developed. It was based on the p30 gene amplification using loop-mediated isothermal DNA amplification (LAMP). The primer set specific to six areas within the target gene were designed and tested for their sensitivity and specificity. Detection of DNA signals was based on modulation of gold nanoparticles' surface properties and performing DNA/DNA hybridization using an oligonucleotide probe. Presence of target DNA affected the gold colloid nanoparticles in terms of particle aggregation with a plasmonic color change of the gold colloids from ruby red to purple, visible by the naked eye. All the assay steps were completed within 90 min including DNA extraction without relying on standard laboratory facilities. This method was very specific to target bacteria. Its sensitivity with probe hybridization was sufficient to detect 50 copies of target DNA. This method should provide an alternative choice for point of care control and management of the disease.
Carolina G. Sosa-Gutiérrez
Full Text Available Objective. Phylogenetic characterization of Ehrlichia canis in dogs naturally infected and ticks, diagnosed by PCR and sequencing of 16SrRNA gene; compare different isolates found in American countries. Materials and methods. Were collected Blood samples from 139 dogs with suggestive clinical manifestations of this disease and they were infested with ticks; part of 16SrRNA gene was sequenced and aligned, with 17 sequences reported in American countries. Two phylogenetic trees were constructed using the Maximum likelihood method, and Maximum parsimony. Results. They were positive to E. canis 25/139 (18.0% dogs and 29/139 (20.9% ticks. The clinical manifestations presented were fever, fatigue, depression and vomiting. Rhipicephalus sanguineus Dermacentor variabilis and Haemaphysalis leporis-palustris ticks were positive for E. canis. Phylogenetic analysis showed that the sequences of dogs and ticks in Mexico form a third group diverging of sequences from South America and USA. Conclusions. This is the first phylogenetic analysis of E. canis in Mexico. There are differences in the sequences of Mexico with those reported in South America and USA. This research lays the foundation for further study of genetic variability.
Schultz, Sharon M; Nicholson, William L; Comer, James A; Childs, James E; Humphreys, Jan G
Serum samples from 381 black bears (Ursus americanus) killed in Pennsylvania (USA) on 24 November 1997 were analyzed for antibodies reactive to the agent of human granulocytic ehrlichiosis (HGE; Ehrlichia sp.) by indirect immunofluorescence assay. Antibody reactivity to HGE antigen was detected in 21% (81/381) of the samples collected. Reactive samples were reported from 56% (14/25) of the counties where bear samples were collected. Endpoint antibody titer ranged from 1:8 to 1:16, 192, with a geometric mean titer of 1:582. There was no significant difference in antibody prevalence between male and female bears (P bears were significantly more likely to have reactive antibodies than juvenile bears (P bear blood clots (n = 181) through nested polymerase chain reaction assays were unsuccessful. Further studies are needed for identification of the pathogen-responsible for induction of HGE-reactive. This is the first description of antibodies reactive to the HGE agent in black bears and suggests these mammals are infected with the agent of HGE or an antigenically related ehrlichial species.
Vargas-Hernández, G; André, M R; Faria, J L M; Munhoz, T D; Hernandez-Rodriguez, M; Machado, R Z; Tinucci-Costa, M
Ehrlichiosis and babesiosis are tick-borne diseases, caused mainly by Ehrlichia canis and Babesia canis, respectively, with a worldwide occurrence in dogs, whose main vector is the brown-dog tick, Rhipicephalus sanguineus. The present work aimed to detect the presence of E. canis and Babesia sp. in 91 dog blood samples in Colombia, by molecular and serological techniques. We also performed sequence alignment to indicate the identity of the parasite species infecting these animals. The present work shows the first molecular detection of E. canis and B. vogeli in dogs from Colombia. Immunoglobulin-G (IgG) antibodies to E. canis and Babesia vogeli were found in 75 (82.4%) and 47 (51.6%) sampled dogs, respectively. Thirty-seven (40.6%) and 5 (5.5%) dogs were positive in PCR for E. canis and Babesia sp., respectively. After sequencing, amplicons showed 99% of identity with isolates of E. canis and B. vogeli. The phylogenetic trees based on 16S rRNA-Anaplasmataceae sequences and 18S rRNA-piroplasmid sequences supported the identity of the found E. canis and B. vogeli DNAs, respectively. The present work shows the first molecular detection of E. canis and B. vogeli in dogs in Colombia. Copyright © 2011 Elsevier B.V. All rights reserved.
Mavromatis, K.; Kuyler Doyle, C.; Lykidis, A.; Ivanova, N.; Francino, P.; Chain, P.; Shin, M.; Malfatti, S.; Larimer, F.; Copeland,A.; Detter, J.C.; Land, M.; Richardson, P.M.; Yu, X.J.; Walker, D.H.; McBride, J.W.; Kyrpides, N.C.
Ehrlichia canis, a small obligately intracellular, tick-transmitted, gram-negative, a-proteobacterium is the primary etiologic agent of globally distributed canine monocytic ehrlichiosis. Complete genome sequencing revealed that the E. canis genome consists of a single circular chromosome of 1,315,030 bp predicted to encode 925 proteins, 40 stable RNA species, and 17 putative pseudogenes, and a substantial proportion of non-coding sequence (27 percent). Interesting genome features include a large set of proteins with transmembrane helices and/or signal sequences, and a unique serine-threonine bias associated with the potential for O-glycosylation that was prominent in proteins associated with pathogen-host interactions. Furthermore, two paralogous protein families associated with immune evasion were identified, one of which contains poly G:C tracts, suggesting that they may play a role in phase variation and facilitation of persistent infections. Proteins associated with pathogen-host interactions were identified including a small group of proteins (12) with tandem repeats and another with eukaryotic-like ankyrin domains (7).
Full Text Available Ehrlichia ruminantium is an obligatory intracellular bacterium that causes heartwater, a fatal disease in ruminants. Due to its intracellular nature, E. ruminantium requires a set of specific virulence factors, such as the type IV secretion system (T4SS, and outer membrane proteins (Map proteins in order to avoid and subvert the host's immune response. Several studies have been conducted to understand the regulation of the T4SS or outer membrane proteins, in Ehrlichia, but no integrated approach has been used to understand the regulation of Ehrlichia pathogenicity determinants in response to environmental cues. Iron is known to be a key nutrient for bacterial growth both in the environment and within hosts. In this study, we experimentally demonstrated the regulation of virB, map1, and tr1 genes by the newly identified master regulator ErxR (for Ehrlichia ruminantium expression regulator. We also analyzed the effect of iron depletion on the expression of erxR gene, tr1 transcription factor, T4SS and map1 genes clusters in E. ruminantium. We show that exposure of E. ruminantium to iron starvation induces erxR and subsequently tr1, virB, and map1 genes. Our results reveal tight co-regulation of T4SS and map1 genes via the ErxR regulatory protein at the transcriptional level, and, for the first time link map genes to the virulence function sensu stricto, thereby advancing our understanding of Ehrlichia's infection process. These results suggest that Ehrlichia is able to sense changes in iron concentrations in the environment and to regulate the expression of virulence factors accordingly.
Jafar Bekloo, Ahmad; Ramzgouyan, Maryam Roya; Shirian, Sadegh; Faghihi, Faezeh; Bakhshi, Hassan; Naseri, Fatemeh; Sedaghat, Mehdi; Telmadarraiy, Zakkyeh
Anaplasma/Ehrlichia species are tick-transmitted pathogens that cause infections in humans and numerous domestic and wild animal species. There is no information available on the molecular characteristics and phylogenetic position of Anaplasma/Ehrlichia spp. isolated from tick species from different geographic locations in Iran. The aim of this study was to determine the prevalence, molecular characteristics, and phylogenetic relationship of both Anaplasma spp. and Ehrlichia spp. in tick species isolated from different domestic animals from two different geographical locations of Iran. A total of 930 ticks were collected from 93 cattle, 250 sheep, and 587 goats inhabiting the study areas. The collected ticks were then investigated for the presence of Anaplasma/Ehrlichia spp. using nested PCR based on the 16S rRNA gene, followed by sequencing. Sequence analysis was done based on the data published in the GenBank on Anaplasma/Ehrlichia spp. isolates using bioinformatic tools such as the standard nucleotide BLAST. Genome of Anaplasma or Ehrlichia spp. was detected in 14 ticks collected in Heris, including 5 Dermacentor marginatus, 1 Haemaphysalis erinacei, 3 Hyalomma anatolicum, and 4 Rhipicephalus sanguineus, also in 29 ticks collected in Chabahar, including 14 R. sanguineus, 8 D. marginatus, 3 Hyalomma Anatolicum, and 4 Hyalomma dromedarii. Partial analysis of the 16S rRNA gene sequence of positive samples collected from goats and sheep showed that they were infected with Anaplasma/Ehrlichia spp. that were 94-98% identical to ovine Anaplasma and 91-96% identical to Neoehrlichia and Ehrlichia spp. The various ticks identified in this study suggest the possible emergence of tick-borne diseases in animals and humans in these regions. R. sanguineus and D. marginatus seem to be predominant vectors responsible for anaplasmosis in these regions. Partial sequence analysis of the 16S rRNA gene showed that A. ovis is genetically polymorphic in these regions. Furthermore, an
Kledmanee, Kan; Suwanpakdee, Sarin; Krajangwong, Sakranmanee; Chatsiriwech, Jarin; Suksai, Parut; Suwannachat, Pongpun; Sariya, Ladawan; Buddhirongawatr, Ruangrat; Charoonrut, Phingphol; Chaichoun, Kridsada
A multiplex polymerase chain reaction (PCR) has been developed for simultaneous detection of canine blood parasites, Ehrlichia canis, Babesia spp and Hepatozoon canis, from blood samples in a single reaction. The multiplex PCR primers were specific to E. canis VirB9, Babesia spp 16S rRNA and H. canis 16S rRNA genes. Specificity of the amplicons was confirmed by DNA sequencing. The assay was evaluated using normal canine and infected blood samples, which were detected by microscopic examination. This multiplex PCR offers scope for simultaneous detection of three important canine blood parasites and should be valuable in monitoring parasite infections in dogs and ticks.
Aroch, Itamar; Baneth, Gad; Salant, Harold; Nachum-Biala, Yaarit; Berkowitz, Asaf; Shamir, Merav; Chai, Orit
An 8-year-old mixed-breed dog was presented for acute, progressive weakness and ataxia, inappetence, and weight loss. The patient was mentally normal, but nonambulatory, with a right head tilt, right positional ventral strabismus, and slight head tremors. A neurologic lesion was localized to the cerebellum and right brainstem. Cerebrospinal fluid (CSF) analysis showed a markedly increased protein concentration and mixed pleocytosis, with eosinophil predominance (44%), intracytoplasmic inclusions within eosinophils, consistent with Ehrlichia canis (E canis) morulae, and Toxoplasma gondii (T gondii) or Neospora caninum (N caninum) tachyzoites within eosinophils and monocytes. A serum indirect immunofluorescent antibody test was positive for N caninum (titer 1:12 800) and negative for T gondii. Both blood and CSF PCR results were N caninum- and E canis-positive and T gondii- and Anaplasma phagocytophilum-negative, and blood PCR, but not CSF PCR, was Hepatozoon canis-positive. The dog was treated for 30 days with clindamycin, sulfamethoxazole-trimethoprim, doxycycline, prednisone, and cephalosporin, but did not improve neurologically, and was euthanized. Brain histopathology showed moderate multifocal, subacute meningoencephalitis with necrosis and gliosis. The neurologic disease was mostly attributed to central nervous system (CNS) neosporosis, with the possible contribution of ehrlichiosis, which was likely a manifestation of blood-brain barrier disruption. Hepatozoonosis was probably a result or cause of underlying immunosuppression. To our knowledge, this is the first report of CNSN caninum and E canis co-infection detected by both CSF PCR and cytology and E canis morulae identified within CSF eosinophils. © 2018 American Society for Veterinary Clinical Pathology.
Cangi, Nídia; Gordon, Jonathan L; Bournez, Laure; Pinarello, Valérie; Aprelon, Rosalie; Huber, Karine; Lefrançois, Thierry; Neves, Luís; Meyer, Damien F; Vachiéry, Nathalie
The disease, Heartwater, caused by the Anaplasmataceae E. ruminantium , represents a major problem for tropical livestock and wild ruminants. Up to now, no effective vaccine has been available due to a limited cross protection of vaccinal strains on field strains and a high genetic diversity of Ehrlichia ruminantium within geographical locations. To address this issue, we inferred the genetic diversity and population structure of 194 E. ruminantium isolates circulating worldwide using Multilocus Sequence Typing based on lipA, lipB, secY, sodB , and sucA genes . Phylogenetic trees and networks were generated using BEAST and SplitsTree, respectively, and recombination between the different genetic groups was tested using the PHI test for recombination. Our study reveals the repeated occurrence of recombination between E. ruminantium strains, suggesting that it may occur frequently in the genome and has likely played an important role in the maintenance of genetic diversity and the evolution of E. ruminantium . Despite the unclear phylogeny and phylogeography, E. ruminantium isolates are clustered into two main groups: Group 1 (West Africa) and a Group 2 (worldwide) which is represented by West, East, and Southern Africa, Indian Ocean, and Caribbean strains. Some sequence types are common between West Africa and Caribbean and between Southern Africa and Indian Ocean strains. These common sequence types highlight two main introduction events due to the movement of cattle: from West Africa to Caribbean and from Southern Africa to the Indian Ocean islands. Due to the long branch lengths between Group 1 and Group 2, and the propensity for recombination between these groups, it seems that the West African clusters of Subgroup 2 arrived there more recently than the original divergence of the two groups, possibly with the original waves of domesticated ruminants that spread across the African continent several thousand years ago.
Lanza-Perea, M; Zieger, U; Qurollo, B A; Hegarty, B C; Pultorak, E L; Kumthekar, S; Bruhl-Day, R; Breitschwerdt, E B
Frequent exposure of Grenadian dogs to Rhipicephalus sanguineus results in Anaplasma platys, and Ehrlichia canis seroreactivity. During elective surgeries, substantial intraoperative hemorrhage occurs in some seroreactive dogs. To assess hemostatic parameters and bleeding tendencies as well as prevalence of PCR positivity in apparently healthy A. platys and E. canis seroreactive and seronegative free-roaming dogs from Grenada. Forty-seven elective surgery dogs allocated to 4 groups: Seronegative control (n = 12), A. platys (n = 10), E. canis (n = 14) and A. platys, and E. canis (n = 11) seroreactive. Preoperatively, hemostasis was assessed by platelet count, prothrombin time, activated partial thromboplastin time, and buccal mucosal bleeding time. Intra- and postoperative bleeding scores were subjectively assigned. Blood, spleen, bone marrow, and lymph node aspirates were tested by PCR. Bleeding scores in dogs coseroreactive for A. platys and E. canis were higher (P = .015) than those of seronegative dogs. A. platys DNA was amplified from 7/21 (33%) A. platys seroreactive dogs and from 1 E. canis seroreactive dog; E. canis DNA was amplified from 21/25 (84%) E. canis seroreactive dogs. E. canis DNA was amplified most often from blood, whereas A. platys DNA was amplified most often from bone marrow. Apparently healthy, free-roaming dogs coseropositive for A. platys and E. canis may have increased intraoperative bleeding tendencies despite normal hemostatic parameters. Future investigations should explore the potential for vascular injury as a cause for bleeding in these dogs. Improved tick control is needed for dogs in Grenada. Copyright © 2014 by the American College of Veterinary Internal Medicine.
Tereza Emmanuelle de Farias Rotondano
Full Text Available This study assessed the occurrence of Ehrlichia spp., Babesia spp. and Hepatozoon spp. infections in 100 tick-harboring dogs from a semiarid region of the State of Paraíba, Northeastern Brazil. Blood samples and ticks were collected from the animals, and a questionnaire was submitted to dog owners to obtain general data. Blood samples were used to perform hemogram, direct blood smear and immunological and molecular hemoparasite detection. The 1,151 ticks collected were identified as Rhipicephalus sanguineus; direct smears revealed E. canis-like morulae in the monocytes of 4% (4/100 of the non-vaccinated female dogs, and 34% and 25% of the dogs tested positive for Ehrlichia canis by indirect immunofluorescence assay (IFA and polymerase chain reaction (PCR, respectively. Blood smear examination revealed Babesia-suggestive merozoites in the erythrocytes of 2% (2/100 of the animals. Babesia vogeli was detected by PCR in ten animals (10% and was correlated with young age (p = 0.007 and thrombocytopenia (p = 0.01. None of the animals showed Hepatozoon spp. positivity. These results indicate that E. canis is the main tick-borne canine pathogen in the study area and provide the first report of B. vogeli infection in dogs from Paraiba State.
Rotondano, Tereza Emmanuelle de Farias; Almeida, Herta Karyanne Araújo; Krawczak, Felipe da Silva; Santana, Vanessa Lira; Vidal, Ivana Fernandes; Labruna, Marcelo Bahia; de Azevedo, Sérgio Santos; Ade lmeida, Alzira Maria Paiva; de Melo, Marcia Almeida
This study assessed the occurrence of Ehrlichia spp., Babesia spp. and Hepatozoon spp. infections in 100 tick-harboring dogs from a semiarid region of the State of Paraíba, Northeastern Brazil. Blood samples and ticks were collected from the animals, and a questionnaire was submitted to dog owners to obtain general data. Blood samples were used to perform hemogram, direct blood smear and immunological and molecular hemoparasite detection. The 1,151 ticks collected were identified as Rhipicephalus sanguineus; direct smears revealed E. canis-like morulae in the monocytes of 4% (4/100) of the non-vaccinated female dogs, and 34% and 25% of the dogs tested positive for Ehrlichia canis by indirect immunofluorescence assay (IFA) and polymerase chain reaction (PCR), respectively. Blood smear examination revealed Babesia-suggestive merozoites in the erythrocytes of 2% (2/100) of the animals. Babesia vogeli was detected by PCR in ten animals (10%) and was correlated with young age (p = 0.007) and thrombocytopenia (p = 0.01). None of the animals showed Hepatozoon spp. positivity. These results indicate that E. canis is the main tick-borne canine pathogen in the study area and provide the first report of B. vogeli infection in dogs from Paraiba State.
Herrin, Brian H; Peregrine, Andrew S; Goring, Jonas; Beall, Melissa J; Little, Susan E
Canine test results generated by veterinarians throughout Canada from 2013-2014 were evaluated to assess the geographical distribution of canine infection with Borrelia burgdorferi, Dirofilaria immitis, Ehrlichia spp., and Anaplasma spp. The percent positive test results of 115,636 SNAP® 4Dx® Plus tests from dogs tested were collated by province and municipality to determine the distribution of these vector-borne infections in Canada. A total of 2,844/115,636 (2.5%) dogs tested positive for antibody to B. burgdorferi. In contrast, positive test results for D. immitis antigen and antibodies to Ehrlichia spp. and Anaplasma spp. were low, with less than 0.5% of dogs testing positive for any one of these three agents nationwide. Provincial seroprevalence for antibodies to B. burgdorferi ranged from 0.5% (Saskatchewan)-15.7% (Nova Scotia); the areas of highest percent positive test results were in proximity to regions in the USA considered endemic for Lyme borreliosis, including Nova Scotia (15.7%) and Eastern Ontario (5.1%). These high endemic foci, which had significantly higher percent positive test results than the rest of the nation (P Canada. Using dogs as sentinels for these pathogens can aid in recognition of the public and veterinary health threat that each pose.
Full Text Available In recent years, some tick-borne diseases such as anaplasmosis and ehrlichiosis became widespread worldwide, threatening the health of humans, domestic animals and wildlife. The aims of this study were to determine the presence of Anaplasma phagocytophilum, Ehrlichia canis, and Ehrlichia chaffeensis in 102 opossums (Didelphis spp. and 44 owned free-ranging dogs in southeastern Mexico using a specific polymerase chain reaction (PCR. A. phagocytophilum was detected in opossums and dogs with a prevalence of 3 and 27%, respectively. E. canis was only present in 7% of dogs, while we didn't detect E. chaffeensis in any host. We report the first evidence of infections of A. phagocytophilum in Didelphis virginiana and D. marsupialis in Mexico. The infection rates and patterns we found of A. phagocytophilum suggest that dogs are more directly involved in the ecology of this pathogen than opossums. Despite the small prevalence found, our results are of public health concern because of the zoonotic capabilities of A. phagocytophilum, the high tick infestation rates found and because both opossums and free-ranging dogs can achieve high population densities in the region.
Williams, Brianna M; Berentsen, Are; Shock, Barbara C; Teixiera, Maria; Dunbar, Michael R; Becker, Matthew S; Yabsley, Michael J
A molecular survey was conducted for several hemoparasites of domestic dogs and three species of wild carnivores from two sites in Zambia. Three Babesia spp. were detected including Babesia felis and Babesia leo in lions (Panthera leo) and a Babesia sp. (similar to Babesia lengau) in spotted hyenas (Crocuta crocuta) and a single lion. All wild dogs (Lycaon pictus) and domestic dogs were negative for Babesia. High prevalences for Hepatozoon were noted in all three wild carnivores (38-61%) and in domestic dogs (13%). Significantly higher prevalences were noted in hyenas and wild dogs compared with domestic dogs and lions. All carnivores were PCR negative for Ehrlichia canis, Ehrlichia ewingii, and Bartonella spp. Overall, high prevalences and diversity of Babesia and Hepatozoon were noted in wild carnivores from Zambia. This study is the first molecular characterization of Babesia from any hyena species and is the first report of a Babesia sp. closely related to B. lengau, a parasite previously only reported from cheetahs (Acinonyx jubatus), in lions and hyenas. Although usually benign in wild carnivores, these hemoparasites can be pathogenic under certain circumstances. Importantly, data on vectors for these parasites are lacking, so studies are needed to identify vectors as well as determine transmission routes, infection dynamics, and host specificity of these hemoparasites in wildlife in Africa and also the risk of transmission between domestic animals and wildlife.
Mdladla, Khanyisile; Dzomba, Edgar F; Muchadeyi, Farai C
The present study investigated the seroprevalence of antibodies to Ehrlichia ruminantium and the associated risk factors in goats from five different farming provinces of South Africa. Sera collected from 686 goats of the commercial meat type (n=179), mohair type (n=9), non-descript indigenous goats from Eastern Cape (n=56), KwaZulu-Natal (n=209), Limpopo (n=111), North West (n=61) and Northern Cape (n=11) provinces and a feral Tankwa goat (n=50) were tested for the presence of immunoglobulin G (IgG) antibodies to antigens of E. ruminantium using the indirect fluorescent-antibody test (IFAT). Fifty two percent of these goats had ticks. The overall seroprevalence of antibodies to E. ruminantium was 64.87% (445/686) with the highest seroprevalence reported for Limpopo (95.50%) and lowest for Northern Cape (20.29%). Highest seroprevalence for antibodies to E. ruminantium was observed in goats from endemic regions (76.09%), and from smallholder production systems (89.54%). High seroprevalence was also observed in non-descript indigenous goats (85.04%), adult goat (69.62%), in does (67.46%) and goats infested with ticks (85.79%). The logistic model showed a gradient of increasing risk for commercial meat type Savanna (OR=3.681; CI=1.335-10.149) and non-descript indigenous (OR=3.466; CI=1.57-7.645) compared to Boer goats and for goats from the smallholder production system (OR=2.582; CI=1.182-5.639) and those with ticks (OR=3.587; CI=2.105-6.112). Results from this study showed that E. ruminantium infections were prevalent but were widely and unevenly distributed throughout South Africa. Findings from the study facilitate identification and mapping of risk areas for heartwater and its endeminicity in South Africa and should be taken into consideration for future disease control strategies and local goat improvement programs. Copyright © 2016 Elsevier B.V. All rights reserved.
Barrantes-González, Alexander V; Jiménez-Rocha, Ana E; Romero-Zuñiga, Juan José; Dolz, Gaby
A cross-sectional study combining different serological and molecular techniques for the detection of Ehrlichia species in dogs and their ticks was carried out with data from all regions of Costa Rica. A seroprevalence of 32.1% (131/408), and infection with E. canis of 3.2% (13/407) was found, whereas 6.9% (9/130) of ticks attached to the dogs were PCR positive to E. canis. Higher prevalences were found outside the Greater Metropolitan Area (GMA). Risk factors associated with E. canis seropositivity were age, between 2 and 7 years (RR: 1.6, 95% CI: 1.2-2.2) and 8-15 years (RR: 1.8, 95% CI: 1.2-3.0), number of dogs/total of households [Dogs per Household Ratio (DHR) ≥3.1 (RR: 2.0; 95% CI: 1.4-3.0)], number of dogs infested with at least one tick/total of dogs sampled [Tick Infestation Prevalence (TIP)≥31% (RR: 2.1; 95% CI:1.3-3.3)] and living outside the GMA (RR: 1.7; 95% CI: 1.2-2.4) and being a mixed-breed dog (RR: 1.5; 95% CI: 1.1-2.1). Risk factors for E. canis PCR positive dogs were a depressive attitude (OR: 11.2; 95% CI: 1.1-115.9), fever (OR:4.8; 95% CI:1.2-19.3), DHR≥3.1 (OR: 5.7; 95% CI:1.7-19.2)], number of ticks/total of dogs sampled [Tick Distribution Ratio (TDR) ≥2.1 (OR: 6.5; 95% CI: 1.3-31.8)], and TIP≥40% (OR: 5.7; 95% CI: 1.7-19.2). This paper describes E. canis seroprevalence, PCR prevalence and tick analysis in dogs from Costa Rica, with associated clinical signs and owner perceptions. In summary, most of the E. canis infections in dogs in our country seemed to pass unnoticed by owners. Since most of the seropositive dogs (97.7%, 131/134) were negative for E. canis DNA in their blood, it is important to determine in future studies if these dogs recovered from the E. canis infection without any medication, or are persistently infected, and will develop chronic disease. Copyright © 2016 Elsevier GmbH. All rights reserved.
Andoh, Masako; Sakata, Akiko; Takano, Ai; Kawabata, Hiroki; Fujita, Hiromi; Une, Yumi; Goka, Koichi; Kishimoto, Toshio; Ando, Shuji
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
Full Text Available One of the major routes of transmission of rickettsial and ehrlichial diseases is via ticks that infest numerous host species, including humans. Besides mammals, reptiles and amphibians also carry ticks that may harbor Rickettsia and Ehrlichia strains that are pathogenic to humans. Furthermore, reptiles and amphibians are exempt from quarantine in Japan, thus facilitating the entry of parasites and pathogens to the country through import. Accordingly, in the current study, we examined the presence of Rickettsia and Ehrlichia spp. genes in ticks associated with reptiles and amphibians originating from outside Japan. Ninety-three ticks representing nine tick species (genera Amblyomma and Hyalomma were isolated from at least 28 animals spanning 10 species and originating from 12 countries (Ghana, Jordan, Madagascar, Panama, Russia, Sri Lanka, Sudan, Suriname, Tanzania, Togo, Uzbekistan, and Zambia. None of the nine tick species are indigenous in Japan. The genes encoding the common rickettsial 17-kDa antigen, citrate synthase (gltA, and outer membrane protein A (ompA were positively detected in 45.2% (42/93, 40.9% (38/93, and 23.7% (22/93 of the ticks, respectively, by polymerase chain reaction (PCR. The genes encoding ehrlichial heat shock protein (groEL and major outer membrane protein (omp-1 were PCR-positive in 7.5% (7/93 and 2.2% (2/93 of the ticks, respectively. The p44 gene, which encodes the Anaplasma outer membrane protein, was not detected. Phylogenetic analysis showed that several of the rickettsial and ehrlichial sequences isolated in this study were highly similar to human pathogen genes, including agents not previously detected in Japan. These data demonstrate the global transportation of pathogenic Rickettsia and Ehrlichia through reptile- and amphibian-associated ticks. These imported animals have potential to transfer pathogens into human life. These results highlight the need to control the international transportation of known
Shpynov, Stanislav; Fournier, Pierre-Edouard; Rudakov, Nikolay; Tankibaev, Marat; Tarasevich, Irina; Raoult, Didier
Using PCR, we screened 411 ticks from four genera collected in Russia and Kazakhstan for the presence of rickettsiae and ehrlichiae. In Russia, we detected “Rickettsia heilongjiangensis,” Rickettsia sp. strain RpA4, and Ehrlichia muris. In Kazakhstan, we detected Rickettsia sp. strain RpA4 and a rickettsia closely related to Rickettsia aeschlimannii. These agents should be considered in a differential diagnosis of tick-borne infections in these areas.
In Vitro Activities of Telithromycin (HMR 3647) against Rickettsia rickettsii, Rickettsia conorii, Rickettsia africae, Rickettsia typhi, Rickettsia prowazekii, Coxiella burnetii, Bartonella henselae, Bartonella quintana, Bartonella bacilliformis, and Ehrlichia chaffeensis
Rolain, Jean-Marc; Maurin, Max; Bryskier, André; Raoult, Didier
In vitro activities of telithromycin compared to those of erythromycin against Rickettsia spp., Bartonella spp., Coxiella burnetii, and Ehrlichia chaffeensis were determined. Telithromycin was more active than erythromycin against Rickettsia, Bartonella, and Coxiella burnetii, with MICs of 0.5 μg/ml, 0.003 to 0.015 μg/ml, and 1 μg/ml, respectively, but was inactive against Ehrlichia chaffeensis.
Baneth, Gad; Harrus, Shimon; Gal, Arnon; Aroch, Itamar
The protozoon Hepatozoon canis and the rickettsia Ehrlichia canis are tick-borne pathogens, transmitted by Rhipicephalus sanguineus, which cause canine hepatozoonosis and canine monocytic ehrlichiosis, respectively. Co-infection of the same host monocytes with H. canis and E. canis confirmed by molecular characterization of the infecting agents and quantitative assessment of co-infected cells is described for the first time in three naturally-infected dogs. Blood smear evaluation indicated that at least 50% of the leukocytes infected with H. canis gamonts contained E. canis morulae. Co-infection of the same host cell demonstrated in this report suggests that infection with one pathogen may permit or enhance invasion or prolonged cellular survival of the other. Copyright © 2014 Elsevier B.V. All rights reserved.
Mixson, T.R.; Ginsberg, H.S.; Campbell, S.R.; Sumner, J.W.; Paddock, C.D.
The lone star tick, Amblyomma americanum (L.), has increased in abundance in several regions of the northeastern United States, including areas of Long Island, NY. Adult and nymphal stage A. americanum collected from several sites on Long Island were evaluated for infection with Ehrlichia chaffeensis, the causative agent of human monocytic ehrlichiosis (HME), by using a nested polymerase chain reaction assay. Fifty-nine (12.5%) of ,17.3 adults and eight of 11.3 pools of five nymphs each (estimated minimum prevalence of infection 1.4%) contained DNA of E. chaffeensis. These data, coupled with the documented expansion of lone star tick populations in the northeastern United States, confirm that E. chaffeensis is endemic to many areas of Long Island and that HME should be considered among the differential diagnoses of the many distinct tick-borne diseases that occur in this region.
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.
José Nivaldo da Silva
Full Text Available A erliquiose canina é uma doença transmitida por carrapatos Rhipicephalus sanguineus e ocasionada pela Ehrlichia canis, bactéria intracelular obrigatória. O presente estudo verificou a prevalência de anticorpos anti-E. canis em 254 cães de quatro regiões administrativas de Cuiabá, Estado de Mato Grosso, por imunofluorescência indireta, observando-se uma prevalência de 42,5% (108/254 sem diferença significativa entre as regiões. As variáveis idade, raça, sexo, hábitat, acesso à zona rural e presença de carrapatos foram analisadas. Os títulos de anticorpos variaram entre 1:40 a 1:2.560. Somente 32 (29,63% cães soropositivos estavam infestados por carrapatos, todos R. sanguineus. O resultado encontrado confirma que não há predisposição racial, sexual ou etária, enquanto a menor ocorrência de cães reagentes no intradomicílio provavelmente está relacionada à baixa infestação por carrapato, apesar de não ter sido observada diferença significativa entre os cães com ou sem a infestação com o carrapato vetor.Canine ehrlichiosis is a disease transmitted by ticks Rhipicephalus sanguineus and caused by Ehrlichia canis, obligatory intracellular bacteria. The present study examined the prevalence of anti-E. canis in 254 dogs from four administrative regions of Cuiabá, Mato Grosso, by indirect immunofluorescence assay. There was a prevalence of 42.5% (108/254 without significant difference between the studied regions. The variables age, breed, sex, habitat, access to rural and ticks were analyzed. The antibody titers ranged from 1:40 to 1:2,560. Only 32 (29.63% seropositive dogs were infested with ticks, all R. sanguineus. The results confirm that do not have breed, sex or age predisposition to ehrlichiosis due E. canis, while the lowest occurrence of reactive dogs indoors probably related to low tick infestation, although no significant difference between dogs with or without infestation with the tick vector.
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.
Luana Gabriela Ferreira dos Santos
Full Text Available The present study evaluated the presence of Ehrlichia DNA in the blood samples of 320 dogs from the urban and rural areas of the municipality of Poconé, Pantanal region, Mato Grosso state, by Polymerase Chain Reaction (PCR, targeting the ehrlichial dsb gene. Risk factors for infection in dogs were also evaluated. Forty-eight (15%, 95% CI: 11.4-19.5% dogs were positive: 25 (15.6%, 95% CI: 10.4-22.2% from the urban area and 23 (14.4%, 95% CI: 9.3-20.8% from the rural area (P > 0.05. Partial DNA sequence obtained from PCR products of 18 samples from the urban area and 16 samples from the rural area were 100% identical to E. canis from Brazil and the USA. This study reports the first E. canis molecular detection in dogs from the northern Pantanal region.O presente estudo avaliou a presença de DNA de Ehrlichia spp. em 320 cães das áreas urbana e rural do município de Poconé, região do Pantanal de Mato Grosso, pela PCR visando o gene dsb. Os fatores de risco para a infecção em cães também foram avaliados. Quarenta e oito (15%, IC 95%: 11,4-19,5% cães foram positivos, 25 (15,6%, IC 95%: 10,4-22,2% da área urbana e 23 (14,37%, 95% CI: 9,3-20,8% da área rural (P > 0,05. Sequências parciais de DNAs obtidos a partir de produtos da PCR de 18 amostras da área urbana e 16 da área rural foram 100% idênticas a E. canis do Brasil e EUA. Este estudo relata a primeira detecção molecular de E. canis em cães da região norte do Pantanal.
Wei, Lanjing; Kelly, Patrick; Ackerson, Kate; El-Mahallawy, Heba S; Kaltenboeck, Bernhard; Wang, Chengming
Although vector-borne diseases are important causes of morbidity and mortality in dogs in tropical areas, there is little information on these conditions in Costa Rica. In PCRs of blood from dogs in Costa Rica, we did not detect DNAs of Rickettsia (R.) felis and Coxiella (C.) burnetii but we did find evidence of infection with Dirofilaria (D.) immitis (9/40, 22.5%), Hepatozoon (H.) canis (15/40, 37.5%), Babesia spp. (10/40, 25%; 2 with B. gibsoni and 8 with B. vogeli), Anaplasma (A.) platys (3/40, 7.5%) and Ehrlichia (E.) canis (20/40, 50%). Nine dogs (22.5%) were free of any vector-borne pathogens while 14 (35%) were infected with a single pathogen, 11 (27.5%) with two, 4 (10%) with three, 1 (2.5%) with four, and 1 (2.5%) with five pathogens. Dogs in Costa Rica are commonly infected with vector-borne agents.
Piratae, Supawadee; Pimpjong, Kiattisak; Vaisusuk, Kotchaphon; Chatan, Wasupon
Canine tick borne diseases showing distribution worldwide have caused morbidity and mortality in dogs. This study observed the mainly tick borne pathogens described for dogs in Thailand, Ehrlichia canis, Hepatozoon canis and Babesia canis vogeli. From May to July 2014, blood samples were collected from 79 stray dogs from 7 districts of Mahasarakham province to molecular surveyed for 16s rRNA gene of E. canis and 18s rRNA gene of H. canis and B. canis vogeli. Twenty eight (35.44%) of stray dogs showed the infection with tick borne pathogens. The prevalence of E. canis infection was the highest with 21.5% (17/79). DNA of H. canis and B. canis vogeli were detected at the prevalence of 10.1% (8/79) and 6.3% (5/79), respectively. Co-infection between E. canis and B. canis vogeli were identified in 2 (2.5%) dogs. The results indicated that a wide range of tick borne pathogens are circulation in the canine population in Mahasarakham province. This study is the first report on prevalence of E. canis, H. canis and B. canis vogeli in stray dogs in Mahasarakham, a province in northern part of Thailand. This data providing is important to understand the prevalence of E. canis, H. canis and B. canis vogeli infection in stray dogs in this region, which will assist in the management of these blood parasite.
Gisele Braziliano Andrade
Full Text Available Different parasites that commonly occur concomitantly can influence one another, sometimes with unpredictable effects. We evaluated pathological aspects of dogs naturally co-infected with Leishmania infantum and Ehrlichia canis. The health status of the dogs was investigated based on histopathological, hematological and biochemical analyses of 21 animals infected solely with L. infantum and 22 dogs co- infected with L. infantum and E. canis. The skin of both groups showed chronic, predominantly lymphohistioplasmacytic inflammatory reaction. The plasmacytosis in the lymphoid tissues was likely related with the hypergammaglobulinemia detected in all the dogs. The disorganization of extracellular matrix found in the reticular dermis of the inguinal region and ear, characterized by the substitution of thick collagen fibers for thin fibers, was attributed to the degree of inflammatory reaction, irrespective of the presence of parasites. In addition, the histopathological analysis revealed that twice as many dogs in the co-infected group presented Leishmania amastigotes in the ear skin than those infected solely with Leishmania, increasing the possibility of becoming infected through sand fly vectors. Our findings highlight the fact that the health of dogs infected concomitantly with L. infantum and E. canis is severely compromised due to their high levels of total plasma protein, globulins, alkaline phosphatase and creatine kinase, and severe anemia.
Buchko, Garry W.; Hewitt, Stephen N.; Van Voorhis, Wesley C.; Myler, Peter J.
Thioredoxins (Trxs) are small ubiquitous proteins that participate in a diverse variety of redox reactions via the reversible oxidation of two cysteine thiol groups in a structurally conserved active site, CGPC. Here, we describe the NMR solution structures of a Trx from Ehrlichia chaffeensis (Ec-Trx, ECH_0218), the etiological agent responsible for human monocytic ehrlichiosis, in both the oxidized and reduced states. The overall topology of the calculated structures is similar in both redox states and similar to other Trx structures, a five-strand, mixed -sheet (1:3:2:4:5) surrounded by four -helices. Unlike other Trxs studied by NMR in both redox states, the 1H-15N HSQC spectra of reduced Ec-Trx was missing eight amide cross peaks relative to the spectra of oxidized Ec-Trx. These missing amides correspond to residues C32-E39 in the active site containing helix (2) and S72-I75 in a loop near the active site and suggest a substantial change in the backbone dynamics associated with the formation of an intramolecular C32-C35 disulfide bond.
Claudia M. Ribeiro
Full Text Available ABSTRACT: Hemoparasitic infections are tick-borne diseases, which affect animals and humans. Considering the importance of canine hemoparasitic infections in veterinary clinics, this study aimed to determine the occurrence of Anaplasma platys, Ehrlichia canis and Babesia vogeli in blood samples from 182 dogs not domiciled in the city of Pato Branco, southwestern region of Paraná State, Brazil, using polymerase chain reaction (PCR. The prevalence of A. platys and B. vogeli was 32.9% and 10.9% respectively, and A. platys infection prevailed (p<0.001. The number of dogs positive for A. platys was larger in Winter (p<0.05. All blood samples were negative for E. canis. In the dogs, infestation by Amblyomma cajennense predominated over that by Rhipicephalus sanguineus (p<0.001; but there was no significant association between PCR and the variables presence of ticks, sex and age. Dogs infected by A. platys and B. vogeli showed thrombocytopenia, lymphopenia and leukocytosis; but there was no correlation between such hematological changes and infection by hemoparasites. This appears to be the first molecular study that demonstrates the existence of A. platys and B. vogeli in dogs from the southwestern region of Paraná.
Identificación morfológica de Ehrlichia sp. en las plaquetas de pacientes con infección por virus de la inmunodeficiencia humana, en Venezuela Morphologic identification of Ehrlichia sp. in the platelets of patients infected with the human immunodeficiency virus in Venezuela
Irene del Carmen de Tamí
Full Text Available OBJETIVO: Determinar la frecuencia de infección por bacterias del género Ehrlichia en una población de pacientes con infección por virus de la inmunodeficiencia humana (VIH. Como se sabe, la infección por VIH suele culminar en el síndrome de inmunodeficiencia adquirida (SIDA después de un período variable en que la persona queda expuesta a distintos tipos de infecciones oportunistas, cuya gravedad varía según el deterioro de su sistema inmunitario. La ehrliquiosis humana es una enfermedad infecciosa de reconocimiento reciente que se transmite por garrapatas y que es causada por especies de Ehrlichia. Se trata de microorganismos gramnegativos, intracelulares obligados, que muestran tropismo por células sanguíneas (leucocitos y plaquetas. En Venezuela se identificó por primera vez en seres humanos en 1994, con la detección de microcolonias (mórulas en el citoplasma de plaquetas. MATERIAL Y MÉTODOS: Para el estudio se reunieron muestras de sangre periférica de 87 pacientes con seropositividad a VIH recibidos en el Laboratorio de Inmunología del Instituto de Oncología y Hematología, en Caracas, Venezuela, que es uno de los centros de referencia nacionales para casos de infección por VIH. Se revisaron retrospectivamente, al microscopio óptico, frotis elaborados con capa blanca sanguínea concentrada, teñidos con colorante de Wright y sellados con resina para su conservación. RESULTADO: Se identificó Ehrlichia trombocítica en 12 de los 87 sujetos del estudio, es decir, en 13,8%. CONCLUSIÓN: La identificación de Ehrlichia trombocítica en pacientes seropositivos sugiere la necesidad de tener en cuenta la ehrliquiosis como posible diagnóstico en la población con infección por VIH.OBJECTIVE: To determine the frequency of infection by bacteria of the genus Ehrlichia in a population of patients infected with the human immunodeficiency virus (HIV. As is widely recognized, HIV infection is usually followed by the acquired
Kawahara, M; Ito, T; Suto, C; Shibata, S; Rikihisa, Y; Hata, K; Hirai, K
In metropolitan Tokyo, the Ehrlichia muris seropositivity rate of 24 wild mice was 63% in Hinohara Village, but in the surrounding areas, it was 0 to 5%. This finding suggests that the reservoir of E. muris is focal. Among the 15 seropositive mice, ehrlichiae were isolated from 9 Apodemus speciosus mice and 1 A. argenteus mouse, respectively. Five ehrlichial isolates were obtained from 10 ticks (Haemaphysalis flava) collected in Asuke Town, Aichi Prefecture, where the E. muris type strain had been isolated. These new isolates were compared with the E. muris type strain. The mouse virulence and ultrastructure of the new isolates were similar to those of the type strain, and all of them were cross-reactive with each other, as well as with the type strain, by indirect immunofluorescent-antibody test. The levels of similarity of the base sequences of the 16S rRNA gene of one of the A. speciosus isolates and one of the tick isolates to that of the E. muris type strain were 99.79 and 99.93%, respectively. We suggest that all of these isolates are E. muris; that E. muris is not limited to Eothenomys kageus but infects other species of mice; and that E. muris is present at locations other than Aichi Prefecture. It appears that H. flava is a potential vector of E. muris. Twenty (1%) of 1803 humans from metropolitan Tokyo were found to be seropositive for E. muris antibodies. A serological survey revealed that exposure to E. muris or organisms antigenically cross-reactive to E. muris occurred among dogs, wild mice, monkeys, bears, deer, and wild boars in Gifu Prefecture, nearby prefectures, and Nagoya City, central Japan. However, human beings and Rattus norvegicus rats in this area were seronegative. These results indicate broader geographic distribution of and human and animal species exposure to E. muris or related Ehrlichia spp. in Japan.
Parola, Philippe; Cornet, Jean-Paul; Sanogo, Yibayiri Osée; Miller, R. Scott; Thien, Huynh Van; Gonzalez, Jean-Paul; Raoult, Didier; Telford III, Sam R.; Wongsrichanalai, Chansuda
A total of 650 ticks, including 13 species from five genera, were collected from animals, from people, or by flagging of the vegetation at sites on the Thai-Myanmar border and in Vietnam. They were tested by PCR to detect DNA of bacteria of the order Rickettsiales. Three Anaplasma spp. were detected in ticks collected in Thailand, including (i) Anaplasma sp. strain AnDa465, which was considered a genotype of Anaplasma platys (formerly Ehrlichia platys) and which was obtained from Dermacentor ...
Full Text Available Survival of Ehrlichia chaffeensis depends on obligatory intracellular infection. One of the barriers to E. chaffeensis research progress has been the inability, using conventional techniques, to generate knock-out mutants for genes essential for intracellular infection. This study examined the use of Peptide Nucleic Acids (PNAs technology to interrupt type IV secretion system (T4SS effector protein expression in E. chaffeensis followed by intracellular complementation of the effector to determine its requirement for infection. Successful E. chaffeensis infection depends on the E. chaffeensis-specific T4SS protein effector, ehrlichial translocated factor-1 (Etf-1, which induces Rab5-regulated autophagy to provide host cytosolic nutrients required for E. chaffeensis proliferation. Etf-1 is also imported by host cell mitochondria where it inhibits host cell apoptosis to prolong its infection. We designed a PNA specific to Etf-1 and showed that the PNA bound to the target region of single-stranded Etf-1 RNA using a competitive binding assay. Electroporation of E. chaffeensis with this PNA significantly reduced Etf-1 mRNA and protein, and the bacteria's ability to induce host cell autophagy and infect host cells. Etf-1 PNA-mediated inhibition of ehrlichial Etf-1 expression and E. chaffeensis infection could be intracellularly trans-complemented by ectopic expression of Etf-1-GFP in host cells. These data affirmed the critical role of bacterial T4SS effector in host cell autophagy and E. chaffeensis infection, and demonstrated the use of PNA to analyze the gene functions of obligate intracellular bacteria.
Taslima T. Lina
Full Text Available Ehrlichia chaffeensis preferentially targets mononuclear phagocytes and survives through a strategy of subverting innate immune defenses, but the mechanisms are unknown. We have shown E. chaffeensis type 1 secreted tandem repeat protein (TRP effectors are involved in diverse molecular pathogen-host interactions, such as the TRP120 interaction with the Notch receptor-cleaving metalloprotease ADAM17. In the present study, we demonstrate E. chaffeensis, via the TRP120 effector, activates the canonical Notch signaling pathway to promote intracellular survival. We found that nuclear translocation of the transcriptionally active Notch intracellular domain (NICD occurs in response to E. chaffeensis or recombinant TRP120, resulting in upregulation of Notch signaling pathway components and target genes notch1, adam17, hes, and hey. Significant differences in canonical Notch signaling gene expression levels (>40% were observed during early and late stages of infection, indicating activation of the Notch pathway. We linked Notch pathway activation specifically to the TRP120 effector, which directly interacts with the Notch metalloprotease ADAM17. Using pharmacological inhibitors and small interfering RNAs (siRNAs against γ-secretase enzyme, Notch transcription factor complex, Notch1, and ADAM17, we demonstrated that Notch signaling is required for ehrlichial survival. We studied the downstream effects and found that E. chaffeensis TRP120-mediated activation of the Notch pathway causes inhibition of the extracellular signal-regulated kinase 1/2 (ERK1/2 and p38 mitogen-activated protein kinase (MAPK pathways required for PU.1 and subsequent Toll-like receptor 2/4 (TLR2/4 expression. This investigation reveals a novel mechanism whereby E. chaffeensis exploits the Notch pathway to evade the host innate immune response for intracellular survival.
Huber, Doroteja; Reil, Irena; Duvnjak, Sanja; Jurković, Daria; Lukačević, Damir; Pilat, Miroslav; Beck, Ana; Mihaljević, Željko; Vojta, Lea; Polkinghorne, Adam; Beck, Relja
The bacteria Anaplasma platys, Anaplasma phagocytophilum and Ehrlichia canis are tick-borne agents that cause canine vector-borne disease. The prevalence of these pathogens in South Eastern Europe is unknown with the exception of an isolated case of A. platys detected in a dog imported into Germany from Croatia. To gain a better insight into their presence and prevalence, PCR-based screening for these bacterial pathogens was performed on domesticated dogs from different regions of Croatia. Blood samples from 1080 apparently healthy dogs from coastal and continental parts of Croatia as well as tissue samples collected from 63 deceased dogs with a history of anaemia and thrombocytopenia were collected for molecular screening by an Anaplasmataceae-specific 16S rRNA conventional PCR. Positive samples were confirmed using a second Anaplasmataceae-specific PCR assay with the PCR product sequenced for the purpose of bacterial species identification. All sequenced isolates were georeferenced and a kernel intensity estimator was used to identify clusters of greater case intensity. 42/1080 (3.8%; CI 2.7-5.0) of the healthy dogs were PCR positive for bacteria in the Anaplasmataceae. Sequencing of the 16S rRNA gene amplified from these positive samples revealed the presence of A. platys in 2.5% (CI 1.6-3.4%, 27 dogs), A. phagocytophilum in 0.3% (CI 0-0.6%, 3 dogs) and a Wolbachia endosymbiont in 1.1% (CI 0.4-1.6%, 12 dogs) of dogs screened in this study. Necropsied dogs were free from infection. Notably, no evidence of E. canis infection was found in any animal. This survey represents a rare molecular study of Anaplasmataceae in dogs in South Eastern Europe, confirming the presence of A. platys and A. phagocytophilum but not E. canis. The absence of E. canis was surprising given it has been described in all other Mediterranean countries surveyed and raises questions over the regional vector capacity of the Rhipicephalus sanguineus tick.
Maurício Franco Zanette
Full Text Available Introduction: The aim of this study was to evaluate the serological cross-reactivity between Leishmania sp. and other canine pathogens. Methods: Positive serum samples for Ehrlichia canis, Babesia canis, Toxoplasma gondii, Neospora caninum and Trypanosoma cruzi were tested using three serological methods enzyme linked immunosorbent assay (ELISA, indirect immunofluorescent antibody test (IFAT and Kalazar Detect™, for canine visceral leishmaniasis. Results: Of the 57 dog samples tested, 24 (42.1% tested positive using one of the three serological methods: 10/57 (17.5% for ELISA, 11/57 (19.3% for IFAT and 3/57 (5.3% for Kalazar Detect™. Conclusions: Our results demonstrated that the presence of other infectious agents may lead to cross-reactivity on leishmaniasis serological tests.
Lin, Mingqun; Liu, Hongyan; Xiong, Qingming; Niu, Hua; Cheng, Zhihui; Yamamoto, Akitsugu; Rikihisa, Yasuko
Ehrlichia chaffeensis is an obligatory intracellular bacterium that causes a potentially fatal emerging zoonosis, human monocytic ehrlichiosis. E. chaffeensis has a limited capacity for biosynthesis and metabolism and thus depends mostly on host-synthesized nutrients for growth. Although the host cell cytoplasm is rich with these nutrients, as E. chaffeensis is confined within the early endosome-like membrane-bound compartment, only host nutrients that enter the compartment can be used by this bacterium. How this occurs is unknown. We found that ehrlichial replication depended on autophagy induction involving class III phosphatidylinositol 3-kinase (PtdIns3K) activity, BECN1 (Beclin 1), and ATG5 (autophagy-related 5). Ehrlichia acquired host cell preincorporated amino acids in a class III PtdIns3K-dependent manner and ehrlichial growth was enhanced by treatment with rapamycin, an autophagy inducer. Moreover, ATG5 and RAB5A/B/C were routed to ehrlichial inclusions. RAB5A/B/C siRNA knockdown, or overexpression of a RAB5-specific GTPase-activating protein or dominant-negative RAB5A inhibited ehrlichial infection, indicating the critical role of GTP-bound RAB5 during infection. Both native and ectopically expressed ehrlichial type IV secretion effector protein, Etf-1, bound RAB5 and the autophagy-initiating class III PtdIns3K complex, PIK3C3/VPS34, and BECN1, and homed to ehrlichial inclusions. Ectopically expressed Etf-1 activated class III PtdIns3K as in E. chaffeensis infection and induced autophagosome formation, cleared an aggregation-prone mutant huntingtin protein in a class III PtdIns3K-dependent manner, and enhanced ehrlichial proliferation. These data support the notion that E. chaffeensis secretes Etf-1 to induce autophagy to repurpose the host cytoplasm and capture nutrients for its growth through RAB5 and class III PtdIns3K, while avoiding autolysosomal killing.
Maylin Gonzalez Navarrete
Full Text Available Os carrapatos (Acari: Ixodidae são de importância médica e veterinária relevantes em todo o mundo por causa da variedade de agentes patogênicos que podem transmitir. No presente trabalho, foi realizada uma pesquisa para identificar Babesia spp. e Ehrlichia spp. em carrapatos coletados de cães de Cuba. Foram coletados 431 carrapatos de 378 cães, tendo sido identificados como pertencentes às espécies de Ripicephalus sanguineus sensu lato (s. 1. O DNA genômico foi extraído com protocolo usando fenol/clorofórmio. Os carrapatos foram organizados em “pools” e o DNA extraído foi testado pela reação em cadeia da polimerase (nPCR para amplificar 398 pares de bases (pb do DNA ribossômico 16S (rDNA de Ehrlichia canis e PCR para amplificar aproximadamente 560 pb do DNA ribossômico 18S (rDNA. Dos 49 pools testados, 8,16% (n = 4/49 foram positivos para o E. canis por nPCR visando o gene do 16S rDNA e apenas um pool (n = 1/49; 2,04% foi positivo para o gene 18S rDNA para Babesia canis. As quatro sequências obtidas para o fragmento de 16S rDNA foram idênticas entre si e resultaram em 100% de identidade com E. canis de diferentes países. A sequência obtida do gene do 18S rDNA para Babesia spp. apresentou semelhança de 100% com Babesia canis vogeli quando comparada às sequências depositadas no Genbank. Esta foi a primeira detecção molecular desses agentes no carrapato R. sanguineus s. l. em Cuba.
Elisabete S Correa
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
Mara Rúbia Rocha Pereira Sales
Full Text Available ABSTRACT. Sales M.R.R.P., Ignacchiti M.D.C., Mendes Junior A.F., Suhett W.G., Porfírio L.C., Marins M., Aptekmann K.P. & Pereira Júnior O.S. [Prevalence of Ehrlichia canis using the nested-PCR, correlation with the presence of morulae and thrombocytopenia in dogs treated in Veterinary Hospital of the Federal University of Espirito Santo.] Prevalência de Ehrlichia canis pela Nested- -PCR, correlação com a presença de mórula e trombocitopenia em cães atendidos no Hospital Veterinário da Universidade Federal do Espírito Santo. Revista Brasileira de Medicina Veterinária, 37(1:47-51, 2015. Centro de Ciências Agrárias, Universidade Federal do Espírito Santo, Rua Projetada s/nº, Caixa Postal 25, Pontal, Marataízes, ES 29349-000, Brasil. E-mail: email@example.com Ehrlichia canis, is the primary etiologic agent of canine monocytic ehrlichiosis. The disease is mainly transmitted by the brown dog ticks Rhipicephalus sanguineus in different endemic regions of Brazil. The purpose of this study was determinated using the Nested Polymerase Chain Reaction (nested-PCR the prevalence of Ehrlichia canis in 85 dogs, regardless of race, age, sex or health status, treated at the Veterinary Hospital of Federal University of Espirito Santo, in Alegre-ES and evaluate its correlation with the presence of morulae and thrombocytopenia. It was observed that 1.17% of the samples were positive by blood smear, for the presence of morulae. However, the nested-PCR showed 5.88% positivity of samples. And 17.64% samples showed thrombocytopenia. By analyzing all the techniques, it was concluded that the introduction of diagnostic techniques such as nested-PCR is an important method for aid in early diagnosis of pathologies.
Aspectos epidemiológicos, clínicos e hematológicos de 251 cães portadores de mórula de Ehrlichia spp. naturalmente infectados Epidemiological, clinical, and hematological aspects of 251 dogs naturally infected with Ehrlichia spp. morulae
Full Text Available Realizou-se um estudo retrospectivo dos aspectos epidemiológicos, sinais clínicos, dados de exame físico e alterações hematológicas da erliquiose em 251 cães naturalmente infectados por Ehrlichia spp. Dos 4407 casos atendidos em hospital veterinário no período de janeiro de 2002 a dezembro de 2003, verificou-se que 251 cães eram portadores de mórula de Ehrlichia spp. em leucócitos de sangue periférico. Destes, 48 foram eliminados das avaliações por apresentarem patologias concomitantes. Nos 203 cães restantes, verificou-se que houve maior ocorrência em fêmeas (61,1% e que a doença manteve-se constante durante todo o período avaliado. Observou-se que 38% encontravam-se na faixa etária entre um e 23 meses e 58,6% eram de raça definida. As principais alterações clínicas observadas foram apatia, anorexia/hiporexia, vômito, secreção oculonasal e esplenomegalia. Cento e cinco cães apresentaram temperatura retal entre 38 e 39,5°C. As alterações observadas com maior frequência no hemograma foram anemia, predominando o tipo normocítica normocrômica (58,2%; desvio nuclear de neutrófilos para a esquerda (67% e eosinopenia (58,1%.A study of epidemiological and clinical aspects, alterations of physical exams, and hematological changes of canine ehrlichiosis was performed. A retrospective study was performed in 4,407 dogs referred to a Veterinary Hospital from January 2002 to December 2003. Of all cases, 251 dogs showed Ehrlichia spp. morulae. Among these, 48 were excluded from the study due to other co-infection by other pathologies. In the other 203 evaluated dogs, females (61.1% were more infected than males. The dogs aged from one to 23 months (68.6% and 58.6% were definite breed. Emesis, apathy, anorexia/hypoxeria, spleenomegaly, and nasal discharge were the most common signs presented. Rectal temperature was 38 - 39.5ÚC in 105 dogs. The most usual changes seen during the hematological tests were normochromic and
Kaewkong, Worasak; Intapan, Pewpan M; Sanpool, Oranuch; Janwan, Penchom; Thanchomnang, Tongjit; Kongklieng, Amornmas; Tantrawatpan, Chairat; Boonmars, Thidarut; Lulitanond, Viraphong; Taweethavonsawat, Piyanan; Chungpivat, Sudchit; Maleewong, Wanchai
Canine babesiosis, hepatozoonosis, ehrlichiosis, and anaplasmosis are tick-borne diseases caused by different hemopathogens. These diseases are causes of morbidity and mortality in dogs. The classic method for parasite detection and differentiation is based on microscopic observation of blood smears. The limitations of the microscopic method are that its performance requires a specially qualified person with professional competence, and it is ineffective in differentiating closely related species. This study applied PCR amplification with high throughput pyrosequencing for molecular differential detection of the following 4 hemoparasites common to tropical areas in dog blood samples: Babesia vogeli, Hepatozoon canis, Ehrlichia canis, and Anaplasma platys. PCR was initially used to amplify specific target regions of the ribosomal RNA genes of each parasite using 2 primer pairs that included 18S rRNA for protozoa (B. vogeli and H. canis) and 16S rRNA for rickettsia (E. canis and A. platys). Babesia vogeli and H. canis were discriminated using 9 nucleotide positions out of 30 base pairs, whereas E. canis and A. platys were differentiated using 15 nucleotide positions out of 34 base pairs that were determined from regions adjacent to 3' ends of the sequencing primers. This method provides a challenging alternative for a rapid diagnosis and surveillance of these tick-borne diseases in canines. Copyright © 2014 Elsevier GmbH. All rights reserved.
Singla, Lachhman Das; Sumbria, Deepak; Mandhotra, Ajay; Bal, M S; Kaur, Paramjit
There are few published studies on various vector borne diseases of dogs in India and most depict clinical infection in dogs, diagnosed by observation of the haemopathogens in stained blood smears. This study provides the first report regarding molecular confirmation and ancestral relationship analysis of blood smears positive cases of assorted haemopathogens in Punjab province of India. On blood smear examination, haemopathogens were observed in 124 out of 778 (15.95%, 95% CI: 13.53- 18.68) blood smears. Further polymerase chain reactions (PCR) was used on bloods smear positive cases to validate the results. Out of 778 blood samples, Babesia gibsoni was most common parasite infecting dogs (15.04%, 95% CI: 12.7-17.72), followed by Ehrlichia canis (0.39%, 95% CI: 0.0-1.13), infection of Babesia vogeli and Hepatozoon canis was same (0.26%, 95% CI: 0.0-0.9). Among various risk factors studied (age, sex, season), prevalence of infection was non-significantly higher in 1-2 year of age group (19.88%, 95% CI: 14.45-26.71), regarding sex same prevalence was recorded (15.94%), and chances of infection was highest in pre-monsoon i.e. summer (18.26%, 95% CI: 14.49-22.76). Phylogenetic analysis revealed ancestral background of Ludhiana isolates of B. vogeli, B. gibsoni, H. canis, and E. canis with the isolates of Philippines, Mongolia and Tunisia.
Sérgio Santos de Azevedo
Full Text Available Aiming to determine the seroprevalence of Ehrlichia canis infection, as well as to identify risk factors associated to the seropositivity, a serological survey was conducted in 109 dogs assisted at the Hospital Veterinário/Centro de Saúde e Tecnologia Rural (CSTR/Universidade Federal de Campina Grande (UFCG, Campus de Patos, Paraíba State, Northeastern Brazil. Serological diagnosis of ehrlichiosis was performed by the indirect fluorescent antibody test (IFAT and sera presenting antibody titers >; 40 were considered positive. Of the 109 samples, 72.5% were positive (95% CI = 63.1% - 80.6%. Animals that had contact with other dogs (odds ratio = 3.59; 95% CI = 1.41 - 9.12, contact with ponds (odds ratio = 8.39; 95% CI = 1.01 - 69.87 or with free access to the street (odds ratio = 6.24; 95% CI = 1.28 - 30.39 were more exposed to the risk of infection.
Full Text Available An Ehrlichia ruminantium culture system was utilized for the anti-rickettsial evaluation of two ethnoveterinary plants, Elephantorrhiza elephantina and Aloe marlothii. Well-established E. ruminantium cultures were incubated with the plant leaf acetone extracts and compared to oxytetracycline and untreated controls. Effectivity was established by comparing the percentage parasitised cells and the calculation of both EC50 and extrapolated EC90 in µg/ml. The plant extracts were also screened for antibacterial activity using bioautography. Elephantorrhiza elephantina and A. marlothii demonstrated anti-ehrlichial activity with an EC50 of 111.4 and 64.5 µg/ml and EC 90 of 228.9 and 129.9 µg/ml, respectively. The corresponding EC50 and EC90 for oxytetracycline was 0.29 and 0.08 µg/ml. Both plants appeared to produce their inhibitory activity by a similar mechanism, unrelated to that of the tetracyclines. Both the plant acetone extracts demonstrated antibacterial activity against Escherichia coli and Staphylococcus aureus (ATCC strains.
Andréa Pereira da Costa
Full Text Available This study evaluated exposure and infection by tick-borne agents (Babesia vogeli, Ehrlichia canis and Rickettsia spp. in 172 dogs in rural areas and 150 dogs in urban areas of the municipality of Chapadinha, state of Maranhão, northeastern Brazil, using molecular and serological methods. Overall, 16.1% of the sampled dogs (52/322 were seroreactive to B. vogeli, with endpoint titers ranging from 40 to 640. For E. canis, 14.6% of the dogs (47/322 were seroreactive, with endpoint titers from 80 to 163,840. Antibodies reactive to at least one of the five species of Rickettsia were detected in 18.9% of the dogs (61/322, with endpoint titers ranging from 64 to 4,096. High endpoint titers were observed for Rickettsia amblyommii. Three (0.9% and nine (2.8% canine blood samples were PCR-positive for Babesia spp. and E. canis. The ticks collected from urban dogs were all Rhipicephalus sanguineus sensu lato, whereas the rural dogs were infested by R. sanguineus s.l, Amblyomma cajennense sensu lato and Amblyomma ovale. One A. ovale tick was found to be infected by Rickettsia bellii. This study provides an epidemiological background for controlling and preventing canine tick-borne diseases in a neglected region of Brazil.
Arathy D S Nair
Full Text Available Ehrlichia chaffeensis, transmitted from Amblyomma americanum ticks, causes human monocytic ehrlichiosis. It also infects white-tailed deer, dogs and several other vertebrates. Deer are its reservoir hosts, while humans and dogs are incidental hosts. E. chaffeensis protein expression is influenced by its growth in macrophages and tick cells. We report here infection progression in deer or dogs infected intravenously with macrophage- or tick cell-grown E. chaffeensis or by tick transmission in deer. Deer and dogs developed mild fever and persistent rickettsemia; the infection was detected more frequently in the blood of infected animals with macrophage inoculum compared to tick cell inoculum or tick transmission. Tick cell inoculum and tick transmission caused a drop in tick infection acquisition rates compared to infection rates in ticks fed on deer receiving macrophage inoculum. Independent of deer or dogs, IgG antibody response was higher in animals receiving macrophage inoculum against macrophage-derived Ehrlichia antigens, while it was significantly lower in the same animals against tick cell-derived Ehrlichia antigens. Deer infected with tick cell inoculum and tick transmission caused a higher antibody response to tick cell cultured bacterial antigens compared to the antibody response for macrophage cultured antigens for the same animals. The data demonstrate that the host cell-specific E. chaffeensis protein expression influences rickettsemia in a host and its acquisition by ticks. The data also reveal that tick cell-derived inoculum is similar to tick transmission with reduced rickettsemia, IgG response and tick acquisition of E. chaffeensis.
Yabsley, Michael J; McKibben, John; Macpherson, Calum N; Cattan, Peggy F; Cherry, Natalie A; Hegarty, Barbara C; Breitschwerdt, Edward B; O'Connor, Tom; Chandrashekar, Ramaswamy; Paterson, Tara; Perea, Marta Lanza; Ball, Geoffrey; Friesen, Stanley; Goedde, Jill; Henderson, Brooke; Sylvester, Wayne
To identify the tick-borne pathogens in dogs from Grenada, we conducted a serologic survey for Ehrlichia canis in 2004 (104 dogs) and a comprehensive serologic and molecular survey for a variety of tick-borne pathogens in 2006 (73 dogs). In 2004 and 2006, 44 and 32 dogs (42.3% and 43.8%) were seropositive for E. canis, respectively. In 2006, several tick-borne pathogens were identified by serology and PCR. DNA of E. canis, Anaplasma platys, Babesia canis vogeli, Hepatozoon canis, and Bartonella sp. were identified in 18 (24.7%), 14 (19.2%), 5 (7%), 5 (7%), and 1 (1.4%) dogs, respectively. Six (8.2%) dogs were seropositive for Bartonella vinsonii subsp. berkhoffii. All dogs were seronegative and PCR-negative for Rickettsia spp. Coinfection with two or three pathogens was observed in eight dogs. Partial 16S rRNA E. canis and A. platys sequences were identical to sequences in GenBank. Partial 18S rRNA gene sequences from the Grenadian H. canis were identical to each other and had one possible mismatch (ambiguous base) from H. canis detected from Spain and Brazil. Grenadian B. c. vogeli sequences were identical to B. c. vogeli from Brazil and Japan. All of the detected pathogens are transmitted, or suspected to be transmitted, by Rhipicephalus sanguineus. Results of this study indicate that dogs from Grenada are infected with multiple tick-borne pathogens; therefore, tick-borne diseases should be included as differentials for dogs exhibiting thrombocytopenia, leukopenia, fever, or lethargy. One pathogen, E. canis, is also of potential public health significance.
Piantedosi, Diego; Neola, Benedetto; D'Alessio, Nicola; Di Prisco, Francesca; Santoro, Mario; Pacifico, Laura; Sgroi, Giovanni; Auletta, Luigi; Buch, Jesse; Chandrashekar, Ramaswamy; Breitschwerdt, Edward B; Veneziano, Vincenzo
Canine vector-borne diseases (CVBDs) are caused by a range of pathogens transmitted to dogs by arthropods. The present study investigates Ehrlichia canis, Anaplasma spp., Borrelia burgdorferi sensu lato, and Dirofilaria immitis seroprevalences in hunting dogs from southern Italy. Dogs (no. 1335) were tested using a commercial in-clinic enzyme-linked immunosorbent assay kit. Odds ratios (ORs) were calculated by logistic regression analysis to identify risk factors. Overall, 138/1335 dogs (10.3%) were seroreactive to at least one CVBD pathogen. E. canis, Anaplasma spp., B. burgdorferi s.l., and D. immitis seroprevalences were 7.6, 4.4, 0.3, and 0.2%, respectively. E. canis and Anaplasma spp. co-exposures were found in 30 dogs (2.2%), compared with Anaplasma spp. and B. burgdorferi s.l. co-exposures in 2 dogs (0.1%). Adult age was a risk factor for E. canis (OR 2.35) seroreactivity whereas hunting fur-bearing animals for E. canis (OR 4.75) and Anaplasma spp. (OR 1.87), respectively. The historical presence of tick infestation was identified as a risk factor for positivity to E. canis (OR 2.08) and Anaplasma spp. (OR 2.15). Finally, larger dog pack size was significantly associated with E. canis (OR 1.85) and Anaplasma spp. (OR 2.42) exposures. The results of the present survey indicated that hunting dog populations are at relative risk of CVBDs in southern Italy. Further studies are needed to evaluate the role of hunting dogs in the epidemiology of vector-borne organisms due to sharing common environments with wild, sympatric animal populations.
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
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. Copyright © 2016
Valentina Virginia Ebani
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
Study of cross-reactivity in serum samples from dogs positive for Leishmania sp., Babesia canis and Ehrlichia canis in enzyme-linked immunosorbent assay and indirect fluorescent antibody test Estudo da reatividade cruzada em amostras de soro de cães positivos para Leishmania sp., Babesia canis e Ehrlichia canis, pelo ensaio imunoenzimático indireto e pela reação de imunofluorescência indireta
Trícia Maria F. de Sousa Oliveira
Full Text Available To verify the presence of cross-reaction among leishmaniosis, ehrlichiosis and babesiosis in serological diagnostics used in human visceral leishmaniasis control programs, serum samples from leishmaniasis endemic and non-endemic areas were collected and tested by Indirect Fluorescent Antibody (IFAT and Enzyme-linked immunosorbent assay (ELISA. All serum samples from endemic areas were positive for Leishmania sp., by ELISA and IFAT, 51% positive for Babesia canis and 43% for Ehrlichia canis by IFAT. None of the serum samples from non-endemic areas were positive for Leishmania sp., by IFAT, but 67% were positive for B. canis and 78% for E. canis using the same test. When tested by ELISA for Leishmania sp., four samples from non-endemic area were positive. These dogs were then located and no clinical signs, parasites or antibody was detected in new tests for a six month period. Only one of these 4 samples was positive for B. canis by IFAT and ELISA and three for E. canis by IFAT. The results of the work suggest a co-infection in the endemic area and no serological cross-reaction among these parasites by IFAT and ELISA.Para verificar a existência de reação cruzada entre leishmaniose visceral, erliquiose e babesiose, nos testes sorológicos utilizados em programas de controle da leishmaniose visceral humana, amostras de soro canino provenientes de áreas endêmicas e não endêmicas para essa enfermidade, foram testadas pela Reação de Imunofluorescência (RIFI e Ensaio imunoenzimático (ELISA. Todos os soros provenientes de área endêmica foram positivos para Leishmania sp pelo ELISA e RIFI, 51% para Babesia canis e 43% para Ehrlichia canis pela RIFI. Pela RIFI, nenhum dos soros provenientes de área não endêmica foi positivo para Leishmania sp, sendo 67% positivos para B. canis e 78% para E. canis pelo mesmo teste. Quando testados pelo ELISA para Leishmania sp., quatro soros da área não endêmica foram positivos. Os cães foram localizados
Presence of anti-Toxoplasma gondii, -Neospora caninum, -Leishmania spp. and -Ehrlichia canis antibodies in free-ranging maned wolves (Chrysocyon brachyurus in the northeastern region of the state of São Paulo, Brazil
Full Text Available O lobo-guará (Chrysocyon brachyurus habita o ecossistema de Cerrado e é considerado o maior canídeo da América do Sul e uma espécie ameaçada de extinção pela "International Union for Conservation of Nature" (IUNC. O objetivo desse estudo foi investigar a presença de anticorpos anti-Toxoplasma gondii, -Neospora caninum, -Leishmania spp. e -Ehrlichia canis em lobos-guará da região nordeste do estado de São Paulo, Brasil. Das 17 amostras de soro testadas por meio da reação de imunofluorescência indireta (RIFI, 88,2% (15/17, 17,6% (3/17 e 52,9% (9/17 apresentaram anticorpos anti-T. gondii, -Leishmania spp. e -E. canis, respectivamente. Todos os animais testados foram soronegativos para N. caninum. Esses resultados indicam a exposição dos lobos-guará dessa região aos agentes pesquisados. A presença de um complexo industrial, agricultura extensiva e fragmentação de habitat na região nordeste do estado de São Paulo, favorece a proximidade desses animais silvestres a ambientes urbanos o que pode contribuir para a transmissão de doenças entre os animais silvestres, domésticos e o homem.
Jodi L McGill
Full Text Available Ehrlichia chaffeensis is a tick-borne rickettsial pathogen and the causative agent of human monocytic ehrlichiosis. Transmitted by the Amblyomma americanum tick, E. chaffeensis also causes disease in several other vertebrate species including white-tailed deer and dogs. We have recently described the generation of an attenuated mutant strain of E. chaffeensis, with a mutation in the Ech_0660 gene, which is able to confer protection from secondary, intravenous-administered, wild-type E. chaffeensis infection in dogs. Here, we extend our previous results, demonstrating that vaccination with the Ech_0660 mutant protects dogs from physiologic, tick-transmitted, secondary challenge with wild-type E. chaffeensis; and describing, for the first time, the cellular and humoral immune responses induced by Ech_0660 mutant vaccination and wild-type E. chaffeensis infection in the canine host. Both vaccination and infection induced a rise in E. chaffeensis-specific antibody titers and a significant Th1 response in peripheral blood as measured by E. chaffeensis antigen-dependent CD4+ T cell proliferation and IFNγ production. Further, we describe for the first time significant IL-17 production by peripheral blood leukocytes from both Ech_0660 mutant vaccinated animals and control animals infected with wild-type E. chaffeensis, suggesting a previously unrecognized role for IL-17 and Th17 cells in the immune response to rickettsial pathogens. Our results are a critical first step towards defining the role of the immune system in vaccine-induced protection from E. chaffeensis infection in an incidental host; and confirm the potential of the attenuated mutant clone, Ech_0660, to be used as a vaccine candidate for protection against tick-transmitted E. chaffeensis infection.
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
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
The effect of the parasite on body weight, packed cell volume, total protein and daily ... However, LSD test showed no differences between the samples. ... The goat died after 15 days and brain squash smears prepared were positive for ...
Cabezas-Cruz, A.; Zweygarth, E.; Vancová, Marie; Broniszewska, M.; Grubhoffer, Libor; Friche Passos, L.M.; Barbosa Ribeiro, M.F.; Alberdi, P.; de la Fuente, J.
Roč. 66, č. 3 (2016), s. 1426-1430 ISSN 1466-5026 EU Projects: European Commission(XE) 278976 - ANTIGONE Institutional support: RVO:60077344 Keywords : cell lines * Anaplasma * genotype * pathogen * Brazil Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 2.134, year: 2016
Full Text Available Canine Ehrlichiosis is an important immunosuppressive tick borne disease in dogs. The geographical distribution and transmission is mostly related with Rhipicephalus sanguineus which acts as a vector. There is no predilection of age or sex; all breeds may be infected with Canine Monocytic Ehrlichiosis (CME. The primary targets are monocytic cells. Platelet disorders and serum protein alterations are the principal hematological and biochemical consequences of infections. Clinical signs are almost non-specific. A definitive diagnosis requires: visualization of morulae within monocytes on cytology, detection of serum antibodies with E. canis, the IFA test, or the PCR. The objective of this study was to present information about haematological and biochemical tests of E. canis infected dogs in Lesvos island in Greece, which is an endemic area.
Brunker, Jill D; Hoover, John P
A mixed breed dog treated for ehrlichiosis and systemic histoplasmosis developed a refractory thrombocytopenia. When an abdominal mass was detected, exploratory laparotomy and biopsies confirmed lymphoma, which on immunohistochemical stains was determined to be of B-cell origin. Conceivably, the B-cell lymphoma in this dog was associated with chronic inflammation from ehrlichiosis, histoplasmosis, or both.
Nair, Arathy D S; Cheng, Chuanmin; Ganta, Chanran K; Sanderson, Michael W; Alleman, Arthur R; Munderloh, Ulrike G; Ganta, Roman R
Dogs acquire infections with the Anaplasmataceae family pathogens, E. canis, E. chaffeensis, E. ewingii, A. platys and A. phagocytophilum mostly during summer months when ticks are actively feeding on animals. These pathogens are also identified as causing diseases in people. Despite the long history of tick-borne diseases in dogs, much remains to be defined pertaining to the clinical and pathological outcomes of infections with these pathogens. In the current study, we performed experimental infections in dogs with E. canis, E. chaffeensis, A. platys and A. phagocytophilum. Animals were monitored for 42 days to evaluate infection-specific clinical, hematological and pathological differences. All four pathogens caused systemic persistent infections detectible throughout the 6 weeks of infection assessment. Fever was frequently detected in animals infected with E. canis, E. chaffeensis, and A. platys, but not in dogs infected with A. phagocytophilum. Hematological differences were evident in all four infected groups, although significant overlap existed between the groups. A marked reduction in packed cell volume that correlated with reduced erythrocytes and hemoglobin was observed only in E. canis infected animals. A decline in platelet numbers was common with E. canis, A. platys and A. phagocytophilum infections. Histopathological lesions in lung, liver and spleen were observed in all four groups of infected dogs; infection with E. canis had the highest pathological scores, followed by E. chaffeensis, then A. platys and A. phagocytophilum. All four pathogens induced IgG responses starting on day 7 post infection, which was predominantly comprised of IgG2 subclass antibodies. This is the first detailed investigation comparing the infection progression and host responses in dogs after inoculation with four pathogens belonging to the Anaplasmataceae family. The study revealed a significant overlap in clinical, hematological and pathological changes resulting from the infections.
Harrus, Shimon; Kenny, Martin; Miara, Limor; Aizenberg, Itzhak; Waner, Trevor; Shaw, Susan
This report presents evidence that dogs recover from acute canine monocytic ehrlichiosis (CME) after 16 days of doxycycline treatment (10 mg/kg of body weight every 24 h). Blood PCR was as valuable as splenic aspirate PCR for early diagnosis of acute CME. Splenic aspirate PCR was, however, superior to blood PCR for the evaluation of ehrlichial elimination.
Full Text Available H01142 Ehrlichia ewingii infection Human ehrlichiosis is a recently recognized tic...k-borne infection. Ehrlichia ewingii has been identified as a cause of human disease in addition to formerly... known pathogenic Ehrlichia species. Infectious disease ... Ehrlichia ewingii ... 16S rRNA [KO:K01977] ... Other dis... Manian FA, Liddell AM, Schmulewitz N, Storch GA ... TITLE ... Ehrlichia ewingii, a
Full Text Available obligate intracellular bacteria closely related to Ehrlichia and Anaplasma, causes an infectious mononucleosis...tis, anorexia, generalized lymphadenopathy, and peripheral blood mononucleosis. Infectious disease ... Neoricke
Full Text Available CBRC-AGAM-04-0115 ref|YP_507138.1| thymidylate synthase, flavin-dependent [Ehrlichia chaffee...nsis str. Arkansas] gb|ABD45587.1| thymidylate synthase, flavin-dependent [Ehrlichia chaffeensis str. Arkansas] YP_507138.1 2.8 29% ...
Full Text Available CBRC-MDOM-01-0247 ref|ZP_00545283.1| 60 kDa inner membrane protein [Ehrlichia chaffee...nsis str. Sapulpa] gb|EAM85346.1| 60 kDa inner membrane protein [Ehrlichia chaffeensis str. Sapulpa] ZP_00545283.1 2.1 30% ...
Julie C Dunning Hotopp
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.
... Ehrlichia bacteria can be carried by the: American dog tick Deer tick ( Ixodes scapularis ), which can also ... your vital signs, including: Blood pressure Heart rate Temperature Other tests include: Complete blood count ( CBC ) Granulocyte ...
Full Text Available Ehrlichiae are gram-negative obligate intracellular bacteria that cause potentially fatal human monocytic ehrlichiosis. We previously showed that natural killer (NK cells play a critical role in host defense against Ehrlichia during primary infection. However, the contribution of NK cells to the memory response against Ehrlichia remains elusive. Primary infection of C57BL/6 mice with Ehrlichia muris provides long-term protection against a second challenge with the highly virulent Ixodes ovatus Ehrlichia (IOE, which ordinarily causes fatal disease in naïve mice. Here, we show that the depletion of NK cells in E. muris-primed mice abrogates the protective memory response against IOE. Approximately, 80% of NK cell-depleted E. muris-primed mice succumbed to lethal IOE infection on days 8-10 after IOE infection, similar to naïve mice infected with the same dose of IOE. The lack of a recall response in NK cell-depleted mice correlated with an increased bacterial burden, extensive liver injury, decreased frequency of Ehrlichia-specific IFN-γ-producing memory CD4+ and CD8+ T-cells, and a low titer of Ehrlichia-specific antibodies. Intraperitoneal infection of mice with E. muris resulted in the production of IL-15, IL-12, and IFN-γ as well as an expansion of activated NKG2D+ NK cells. The adoptive transfer of purified E. muris-primed hepatic and splenic NK cells into Rag2-/-Il2rg-/- recipient mice provided protective immunity against challenge with E. muris. Together, these data suggest that E. muris-induced memory-like NK cells, which contribute to the protective, recall response against Ehrlichia.
Full Text Available Introduction. Ehrlichiosis is a bacterial zoonosis transmitted by hematophagous arthropods - ticks. In humans, it occurs as monocytic, granulocytic, and ewingii ehrlichiosis. Pathological process is based on parasitic presence of Ehrlichia organisms within peripheral blood cells - monocytes and granulocytes. Case Outline. Fifty-two year old patient was admitted to hospital due to high fever of over 40°C that lasted two days, accompanied with chills, muscle aches, malaise, loss of appetite, headache, confusion, breathing difficulties, and mild dry cough. The history suggested tick bite that occurred seven days before the onset of disease. Doxycycline was introduced and administered for 14 days, causing the disease to subside. Indirect immunofluorescence assay was used to analyze three serum samples obtained from this patient for Ehrlichia chaffeensis antibodies, and peripheral blood smear was evaluated for the presence of Ehrlichia and Ehrlichia aggregation into morulae. Conclusion. Ehrlichiosis should be considered in each case where there is a history of tick bite together with the clinical picture (high fever, chills, muscle aches, headache, generalized weakness and malaise, and possible maculopapular rash. The presence of Ehrlichia chaffeensis antibodies was confirmed in a patient with the history of tick bite, appropriate clinical picture and indirect immunofluorescence assay. This confirmed the presence of human monocytotropic ehrlichiosis, a disease that is uncommonly identified in our country.
Anderson Cort L
Full Text Available Abstract Background Although closely related, the alpha-proteobacteria Wolbachia and the Rickettsiacae (Rickettsia and Ehrlichia, employ different evolutionary life history strategies. Wolbachia are obligate endocellular symbionts that infect an extraordinary host range and, in contrast to the infectious and pathogenic Rickettsia and Ehrlichia, profoundly influence host reproductive biology. Results Phylogenies of the Rickettsia, Ehrlichia, and Wolbachia were independently inferred from 16S rDNA sequences and GroEL amino acid sequences. Topologies inferred from both sets of sequence data were consistent with one another, and both indicate the genus Wolbachia shared a common ancestor most recently with Ehrlichia. These two genera are a sister group to the genus Rickettsia. Mapping biological properties onto this phylogeny reveals that manipulation of host reproduction, characteristic of Wolbachia strains, is a derived characteristic. This evolutionary novelty is accompanied by the loss of the ability to infect vertebrate hosts. Conclusions Because of the contrasting transmission strategies employed by each, Wolbachia is expected to maximize efficiency of vertical transmission, while Ehrlichia and Rickettsia will optimize horizontal transfer of infection. Wolbachia manipulation of host reproduction could thus be viewed as strategy employed by this bacterium to foster its own propagation via vertical transmission.
Lorelei L. Clarke
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.
Full Text Available haliana PE=4 SV=1 33 5.4 tr|B1N6C1|B1N6C1_9RICK Omp-1-18 OS=Ehrlichia ewingii GN=omp118 P... 33 7.1 tr|B3EJD...+ YSC H ++LCS P TKYF Sbjct: 637 DLHLSAAGCRCSPEKYSCLTHVKELCSC----PWVTKYF 671 >tr|B1N6C1|B1N6C1_9RICK Omp-1-18 OS=Ehrlichia ewing
Paula Elisa Brandão Guedes
Full Text Available Ehrlichiosis is a zoonotic disease that is caused by bacteria of the genus Ehrlichia. The aims of this study were to detect the presence of Ehrlichia spp. in the blood of dogs in Ituberá, Bahia, and to compare the sensitivities and specificities of blood smear, serological, and molecular examinations. Furthermore, this study identified factors associated with exposure to the agent in dogs in this locality. Blood samples were collected from 379 dogs and submitted for indirect immunofluorescent assay and polymerase chain reaction testing for the detection of Ehrlichia spp. antibodies and DNA, respectively. Additionally, a peripheral blood smear was obtained from the ear tip for parasite identification. Of the 379 animals, 12.4%, 32.7%, and 25.6% were identified as positive on the blood smear, serological, and molecular tests, respectively. The dogs positive in one of the three techniques were considered exposed (46.9%. Younger dogs and rural habitat were protective factors and presence of ticks and contact with other dogs were the risk factors associated with exposure to the agent. It was concluded that dogs of Ituberá have high positivity for Ehrlichia spp. and that the diagnostic methods used for detection are complementary.
Department of Veterinary Microbiology and Parasitology, Faculty of Veterinaiy Medicine. ... bariierii, Ehrlichia coats, Rickerrsia conorii, .... (TAG) by an enzymatic method coupled with spectrophotometry using assay kit (Randox. Lab. Ltd., Co. Antrim, UK). Statistical analysis. Statistical analysis was performed with the use.
Spitalska, E.; Literák, I.; Sparagano, O. A .E.; Golovchenko, Maryna; Kocianová, E.
Roč. 118, 23/24 (2006), 759-764 ISSN 0043-5325 Institutional research plan: CEZ:AV0Z60220518 Keywords : Passerine birds * Ixodes ricinus * Borrelia * Rickettsia * Ehrlichia * Carpathians Subject RIV: EC - Immunology Impact factor: 0.804, year: 2006
Westmoreland, Lori S H; Stoskopf, Michael K; Maggi, Ricardo G
We detected Anaplasma phagocytophilum by DNA amplification in whole blood from free-ranging, hunter-killed American black bears ( Ursus americanus ) from the east coast of North Carolina, US. Molecular prevalence for Anaplasma phagocytophilum was 3% from 68 black bears. No DNA of other Anaplasma or Ehrlichia spp. was identified.
Ramsauer, Sandra; Bay, Gert; Meli, Marina; Hofmann-Lehmann, Regina; Lutz, Hans
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.
Ramsauer, Sandra; Bay, Gert; Meli, Marina; Hofmann-Lehmann, Regina; Lutz, Hans
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.
Ioannidis, Panagiotis; Dunning Hotopp, Julie C; Sapountzis, Panagiotis
, 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...
Full Text Available Objectives The objective of the current study was to investigate the prevalence rates of the following infectious agents in 116 stray cats in the Barcelona area of Spain: Anaplasma phagocytophilum, Bartonella species, Borrelia burgdorferi, Chlamydia felis, Dirofilaria immitis, Ehrlichia species, feline calicivirus (FCV, feline herpesvirus-1 (FHV-1, feline leukaemia virus (FeLV, feline immunodeficiency virus (FIV, haemoplasmas, Mycoplasma species and Rickettsia species. Methods Serum antibodies were used to estimate the prevalence of exposure to A phagocytophilum, Bartonella species, B burgdorferi, Ehrlichia species and FIV; serum antigens were used to assess for infection by D immitis and FeLV; and molecular assays were used to amplify nucleic acids of Anaplasma species, Bartonella species, C felis, D immitis, Ehrlichia species, FCV, FHV-1, haemoplasmas, Mycoplasma species and Rickettsia species from blood and nasal or oral swabs. Results Of the 116 cats, 63 (54.3% had evidence of infection by Bartonella species, FeLV, FIV or a haemoplasma. Anaplasma species, Ehrlichia species or Rickettsia species DNA was not amplified from these cats. A total of 18/116 cats (15.5% were positive for FCV RNA (six cats, Mycoplasma species DNA (six cats, FHV-1 DNA (three cats or C felis DNA (three cats. Conclusions and relevance This study documents that shelter cats in Catalonia are exposed to many infectious agents with clinical and zoonotic significance, and that flea control is indicated for cats in the region.
R. sanguinens ticks are known vectors of pathogens like Babesia aunts and Ehrlichia cents, the etiological agents of canine babesiosis and canine monocytic ehrlichiosis, respectively. Interestingly, it has been suspected that R, sanguineus is involved in the transmission of other major pathogens such as. Leishmania ...
Verheijden, J.H.M.; Miert, A. S. J. P. A. M. Van; Duin, C.T.M. van; Schotman, A.J.H.; Nieuwenhuis, J.
In goats with trypanosomiasis (T. vivax or T. congolense) no marked fall in plasma zinc concentration was seen despite high temperature peaks, whereas plasma concentrations of iron tended to undergo some decline. In goats infected with Ehrlichia phagocytophila, there was a marked decline in plasma
Miert, A.S.J.P.A.M. van; Duin, C.T.M. van; Schotman, A.J.H.; Franssen, F.F.
Tick-borne fever in goats caused by Ehrlichia (Cytoecetes) phagocytophila was characterised by high fever, dullness, anorexia, tachycardia and a slight to moderate inhibition of rumen motility. The animals developed a gradual decline in the total number of circulating white blood cells. There was a
Wei, Feng; Song, Mingxin; Liu, Huanhuan; Wang, Bo; Wang, Shuchao; Wang, Zedong; Ma, Hongyu; Li, Zhongyu; Zeng, Zheng; Qian, Jun; Liu, Quan
Tick-borne diseases are considered as emerging infectious diseases in humans and animals in China. In this study, Ixodes persulcatus (n = 1699), Haemaphysalis concinna (n = 412), Haemaphysalis longicornis (n = 390), Dermacentor nuttalli (n = 253), and Dermacentor silvarum (n = 204) ticks were collected by flagging from northeastern China, and detected for infection with Anaplasma, Ehrlichia, Babesia, and Hepatozoon spp. by using nested polymerase chain reaction assays and sequencing analysis. Anaplasma phagocytophilum was detected in all tick species, i.e., I. persulcatus (9.4%), H. longicornis (1.9%), H. concinna (6.5%), D. nuttalli (1.7%), and D. silvarum (2.3%); Anaplasma bovis was detected in H. longicornis (0.3%) and H. concinna (0.2%); Ehrlichia muris was detected in I. persulcatus (2.5%) and H. concinna (0.2%); Candidatus Neoehrlichia mikurensis was only detected in I. persulcatus (0.4%). The Ehrlichia variant (GenBank access number KU921424), closely related to Ehrlichia ewingii, was found in H. longicornis (0.8%) and H. concinna (0.2%). I. persulcatus was infected with Babesia venatorum (1.2%), Babesia microti (0.6%), and Babesia divergens (0.6%). Additionally, four Babesia sequence variants (GenBank access numbers 862303–862306) were detected in I. persulcatus, H. longicornis, and H. concinna, which belonged to the clusters formed by the parasites of dogs, sheep, and cattle (B. gibsoni, B. motasi, and B. crassa). Two Hepatozoon spp. (GenBank access numbers KX016028 and KX016029) associated with hepatozoonosis in Japanese martens were found in the collected ticks (0.1–3.1%). These findings showed the genetic variability of Anaplasma, Ehrlichia, Babesia, and Hepatozoon spp. circulating in ticks in northeastern China, highlighting the necessity for further research of these tick-associated pathogens and their role in human and animal diseases. PMID:27965644
Mohan Kumar, Dipu; Lin, Mingqun; Xiong, Qingming; Webber, Mathew James; Kural, Comert; Rikihisa, Yasuko
Obligate intracellular bacteria, such as Ehrlichia chaffeensis, perish unless they can enter eukaryotic cells. E. chaffeensis is the etiological agent of human monocytic ehrlichiosis, an emerging infectious disease. To infect cells, Ehrlichia uses the C terminus of the outer membrane invasin entry-triggering protein (EtpE) of Ehrlichia (EtpE-C), which directly binds the mammalian cell surface glycosylphosphatidyl inositol-anchored protein, DNase X. How this binding drives Ehrlichia entry is unknown. Here, using affinity pulldown of host cell lysates with recombinant EtpE-C (rEtpE-C), we identified two new human proteins that interact with EtpE-C: CD147 and heterogeneous nuclear ribonucleoprotein K (hnRNP-K). The interaction of CD147 with rEtpE-C was validated by far-Western blotting and coimmunoprecipitation of native EtpE with endogenous CD147. CD147 was ubiquitous on the cell surface and also present around foci of rEtpE-C-coated-bead entry. Functional neutralization of surface-exposed CD147 with a specific antibody inhibited Ehrlichia internalization and infection but not binding. Downregulation of CD147 by short hairpin RNA (shRNA) impaired E. chaffeensis infection. Functional ablation of cytoplasmic hnRNP-K by a nanoscale intracellular antibody markedly attenuated bacterial entry and infection but not binding. EtpE-C also interacted with neuronal Wiskott-Aldrich syndrome protein (N-WASP), which is activated by hnRNP-K. Wiskostatin, which inhibits N-WASP activation, and cytochalasin D, which inhibits actin polymerization, inhibited Ehrlichia entry. Upon incubation with host cell lysate, EtpE-C but not an EtpE N-terminal fragment stimulated in vitro actin polymerization in an N-WASP- and DNase X-dependent manner. Time-lapse video images revealed N-WASP recruitment at EtpE-C-coated bead entry foci. Thus, EtpE-C binding to DNase X drives Ehrlichia entry by engaging CD147 and hnRNP-K and activating N-WASP-dependent actin polymerization. Ehrlichia chaffeensis, an
Valéria Régia Franco Sousa
Full Text Available A erliquiose monocítica canina é uma doença cosmopolita causada por Ehrlichia canis e transmitida pelo carrapato Rhipicephalus sanguineus, sendo frequentemente diagnosticada em cães em todo o Brasil. Este trabalho teve por objetivo investigar citológica e molecularmente a infecção por Ehrlichia em 195 cães atendidos no Hospital Veterinário da Universidade Federal de Mato Grosso, analisando os achados clínicos e laboratoriais. Nos 48 cães atendidos com citologia positiva para Ehrlichia sp., foi possível verificar a diversidade de sinais, com predominância estatisticamente significativa de palidez de mucosas (P≤0,05, assim como variados achados hematológicos, ocorrendo tanto anemia, leucopenia e trombocitopenia, quanto normalidade ou aumento dessas células. Ocorreu aumento das proteínas plasmáticas, com hiperglobulinemia, sem, no entanto, haver diferença significativa (P≥0,05, apesar de esse achado ser frequente nessa afecção. Por meio do PCR nested, confirmou-se a infecção por E. canis em cães da cidade de Cuiabá.The canine monocytic ehrlichiosis is a cosmopolitan disease, caused by Ehrlichia canis, transmitted by ticks Rhipicephalus sanguineus that has been frequently diagnosed in dogs throughout the country. This study aimed to investigate the cytological and molecular Ehrlichia infection in 195 dogs examined at the University Veterinary Hospital of Mato Grosso, by analyzing the clinical and laboratory findings. In 48 dogs with positive cytology for Ehrlichia sp it was possible to detect the diversity of signs, with predominance statistically significant of pallor of mucous membranes (P≤0.05 as well as several hematological findings, occurring anemia, leukopenia and thrombocytopenia, or increased as normal cells. There was increased of plasma proteins, with hyperglobulinemia, however without any significant difference (P≥0.05, although this finding is common in that infection. Through the nested PCR technique it
Moro, Pedro L; Shah, Jyotsna; Li, Olga; Gilman, Robert H; Harris, Nick; Moro, Manuel H
A serosurvey for human ehrlichiosis caused by Ehrlichia chaffeensis and Anaplasma phagocytophilum was performed in different regions of Peru by using indirect immunofluorescence assays (IFAs). Regions included an urban community in a shantytown in Lima (Pampas) and three rural communities located on the northern coast of Peru (Cura Mori), in the southern Peruvian Andes (Cochapata), and in the Peruvian jungle region (Santo Tomas). An overall E. chaffeensis seroprevalence of 13% (21 of 160) was found by IFA. Seroprevalences in females and males was 15% (16 of 106) and 9% (5 of 53), respectively. Seroprevalences in Cura Mori, Cochapata, Pampas, and Santo Tomas were 25% (10 of 40), 23% (9 of 40), 3% (1 of 40), and 3% (1 of 40), respectively. Seroprevalences in Cura Mori and Cochapata were significantly higher than in Santo Tomas or Pampas (P Peru. Further studies are needed to characterize Ehrlichia species in Peru, their vectors and their clinical significance.
André, Marcos Rogério; Filgueira, Kilder Dantas; Calchi, Ana Cláudia; Sousa, Keyla Carstens Marques de; Gonçalves, Luiz Ricardo; Medeiros, Vitor Brasil; Ximenes, Poliana Araújo; Lelis, Ivana Cristina Nunes Gadelha; Meireles, Maria Vanuza Nunes de; Machado, Rosangela Zacarias
The role of several feline vector-borne pathogens (FVBP) as a cause of disease in cats has not been clearly determined. In fact, with the exception of Bartonella spp. and hemoplasmas, FVBP in cats has not been clearly determined in Brazil yet. The present study aimed at identifying, by using molecular methods, the presence of FVBP in three cats showing non-specific clinical signs and inclusions suggestive of hemoparasites in blood smears. Cytauxzoon felis, 'Candidatus Mycoplasma haemominutum', Ehrlichia sp. closely related to Ehrlichia canis, and Anaplasma sp. closely related to Anaplasma phagocytophilum were detected in blood samples from two out of three sampled cats. Both cats positive for multiple FVBP did not show hematological and biochemical abnormalities. The present work emphasizes the need for molecular confirmation of co-infection by multiple FVBP in cats presenting non-specific clinical signs and inclusions resembling hemoparasites in blood smears.
Marcos Rogério André
Full Text Available Abstract The role of several feline vector-borne pathogens (FVBP as a cause of disease in cats has not been clearly determined. In fact, with the exception of Bartonella spp. and hemoplasmas, FVBP in cats has not been clearly determined in Brazil yet. The present study aimed at identifying, by using molecular methods, the presence of FVBP in three cats showing non-specific clinical signs and inclusions suggestive of hemoparasites in blood smears. Cytauxzoon felis, ‘Candidatus Mycoplasma haemominutum’, Ehrlichia sp. closely related to Ehrlichia canis, and Anaplasma sp. closely related to Anaplasma phagocytophilum were detected in blood samples from two out of three sampled cats. Both cats positive for multiple FVBP did not show hematological and biochemical abnormalities. The present work emphasizes the need for molecular confirmation of co-infection by multiple FVBP in cats presenting non-specific clinical signs and inclusions resembling hemoparasites in blood smears.
Gutierrez, Clara Nancy; Perez Yabarra, Luis; agrela, Irma Fatima
La ehrlichiosis canina es una enfermedad infecciosa emergente transmitida por garrapatas, producida por Ehrlichia spp. (Proteobacteria: Ricketsiales), la cual afecta a miembros de la familia Canidae. Los agentes etiológicos son bacterias Gram negativas, intracelulares obligatorias, redondeadas y pleomórficas, esto último especialmente en cultivos celulares. Estas bacterias se localizan en vacuolas rodeadas de membranas (mórulas) en el citoplasma de células sanguíneas y dependiendo de la espec...
Antonio Vicente Mundim
Full Text Available Hepatozoon canis gametocytes, measuring 9,56 µm x 5,60 µm were identified in circulating leukocytes of two dogs admitted to the Veterinary Hospital of the Universidade Federal de Uberlândia. Morulae of Ehrlichia canis were also found in circulating monocytes. The authors report the first occurrence of H. canis in Uberlândia, Minas Gerais state.
Kelly, Patrick John; Köster, Liza; Li, Jing; Zhang, Jilei; Huang, Ke; Branford, Gillian Carmichael; Marchi, Silvia; Vandenplas, Michel; Wang, Chengming
As there is little data on vector-borne diseases of cats in the Caribbean region and even around the world, we tested feral cats from St Kitts by PCR to detect infections with Babesia, Ehrlichia and spotted fever group Rickettsia (SFGR) and surveyed them for antibodies to Rickettsia rickettsii and Ehrlichia canis. Whole blood was collected from apparently healthy feral cats during spay/ neuter campaigns on St Kitts in 2011 (N = 68) and 2014 (N = 52). Sera from the 52 cats from 2014 were used to detect antibodies to Ehrlichia canis and Rickettsia rickettsii using indirect fluorescent antibody tests and DNA extracted from whole blood of a total of 119 cats (68 from 2011, and 51 from 2014) was used for PCRs for Babesia, Ehrlichia and Rickettsia. We could not amplify DNA of SFG Rickettsia in any of the samples but found DNA of E. canis in 5% (6/119), Babesia vogeli in 13% (15/119), Babesia gibsoni in 4% (5/119), mixed infections with B. gibsoni and B. vogeli in 3% (3/119), and a poorly characterized Babesia sp. in 1% (1/119). Overall, 10% of the 52 cats we tested by IFA for E. canis were positive while 42% we tested by indirect fluorescent antibody (IFA) for R. rickettsii antigens were positive. Our study provides the first evidence that cats can be infected with B. gibsoni and also indicates that cats in the Caribbean may be commonly exposed to other vector-borne agents including SFGR, E. canis and B. vogeli. Animal health workers should be alerted to the possibility of clinical infections in their patients while public health workers should be alerted to the possibility that zoonotic SFGR are likely circulating in the region.
Michelet, Lorraine; Delannoy, Sabine; Devillers, Elodie
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...
Stephanie L. Richards
Full Text Available Improvements to risk assessments are needed to enhance our understanding of tick-borne disease epidemiology. We review tick vectors and duration of tick attachment required for pathogen transmission for the following pathogens/toxins and diseases: (1 Anaplasma phagocytophilum (anaplasmosis; (2 Babesia microti (babesiosis; (3 Borrelia burgdorferi (Lyme disease; (4 Southern tick-associated rash illness; (5 Borrelia hermsii (tick-borne relapsing fever; (6 Borrelia parkeri (tick-borne relapsing fever; (7 Borrelia turicatae (tick-borne relapsing fever; (8 Borrelia mayonii; (9 Borrelia miyamotoi; (10 Coxiella burnetii (Query fever; (11 Ehrlichia chaffeensis (ehrlichiosis; (12 Ehrlichia ewingii (ehrlichiosis; (13 Ehrlichia muris; (14 Francisella tularensis (tularemia; (15 Rickettsia 364D; (16 Rickettsia montanensis; (17 Rickettsia parkeri (American boutonneuse fever, American tick bite fever; (18 Rickettsia ricketsii (Rocky Mountain spotted fever; (19 Colorado tick fever virus (Colorado tick fever; (20 Heartland virus; (21 Powassan virus (Powassan disease; (22 tick paralysis neurotoxin; and (23 Galactose-α-1,3-galactose (Mammalian Meat Allergy-alpha-gal syndrome. Published studies for 12 of the 23 pathogens/diseases showed tick attachment times. Reported tick attachment times varied (<1 h to seven days between pathogen/toxin type and tick vector. Not all studies were designed to detect the duration of attachment required for transmission. Knowledge of this important aspect of vector competence is lacking and impairs risk assessment for some tick-borne pathogens.
Campos-Calderón, Liliana; Ábrego-Sánchez, Leyda; Solórzano-Morales, Antony; Alberti, Alberto; Tore, Gessica; Zobba, Rosanna; Jiménez-Rocha, Ana E; Dolz, Gaby
Although vector-borne diseases are globally widespread with considerable impact on animal production and on public health, few reports document their presence in Central America. This study focuses on the detection and molecular identification of species belonging to selected bacterial genera (Ehrlichia, Anaplasma and Rickettsia) in ticks sampled from dogs in Costa Rica by targeting several genes: 16S rRNA/dsb genes for Ehrlichia; 16S rRNA/groEL genes for Anaplasma, and ompA/gltA/groEL genes for Rickettsia. PCR and sequence analyses provides evidences of Ehrlichia canis, Anaplasma platys, and Anaplasma phagocytophilum infection in Rhipicephalus sanguineus s.l ticks, and allow establishing the presence of Rickettsia monacensis in Ixodes boliviensis. Furthermore, the presence of recently discovered Mediterranean A. platys-like strains is reported for the first time in Central America. Results provide new background on geographical distribution of selected tick-transmitted bacterial pathogens in Costa Rica and on their molecular epidemiology, and are pivotal to the development of effective and reliable diagnostic tools in Central America. Copyright © 2016 Elsevier GmbH. All rights reserved.
Background Feline vector-borne diseases (FVBD) have emerged in recent years, showing a wider geographic distribution and increased global prevalence. In addition to their veterinary importance, domestic cats play a central role in the transmission cycles of some FVBD agents by acting as reservoirs and sentinels, a circumstance that requires a One Health approach. The aim of the present work was to molecularly detect feline vector-borne bacteria and protozoa with veterinary and zoonotic importance, and to assess associated risk factors in cats from southern Portugal. Methods Six hundred and forty-nine cats (320 domestic and 329 stray), from veterinary medical centres and animal shelters in southern Portugal, were studied. Anaplasma spp./Ehrlichia spp., Babesia spp., Bartonella spp., Borrelia burgdorferi sensu lato, Hepatozoon spp. and Leishmania spp. infections were evaluated by polymerase chain reaction (PCR) in blood samples. Results One hundred and ninety-four (29.9%) cats were PCR-positive to at least one of the tested genera or complex of FVBD agents. Sixty-four (9.9%) cats were positive to Leishmania spp., 56 (8.6%) to Hepatozoon spp., 43 (6.6%) to Babesia spp., 35 (5.4%) to Anaplasma spp./Ehrlichia spp., 19 (2.9%) to Bartonella spp. and 14 (2.2%) to B. burgdorferi s.l. Thirty-three (5.1%) cats were positive to two (n = 29) or three (n = 4) genera/complex. Babesia vogeli, Bartonella clarridgeiae, Bartonella henselae, Ehrlichia canis, Hepatozoon felis and Leishmania infantum were identified by DNA sequencing. Conclusions The occurrence of FVBD agents in southern Portugal, some of them with zoonotic character, emphasizes the need to alert the veterinary community, owners and public health authorities for the risk of infection. Control measures should be implemented to prevent the infection of cats, other vertebrate hosts and people. PMID:24655431
Full Text Available Tick-borne diseases are considered as emerging infectious diseases in humans and animals in China. In this study, Ixodes persulcatus (n=1699, Haemaphysalis concinna (n=412, Haemaphysalis longicornis (n=390, Dermacentor nuttalli (n=253, and Dermacentor silvarum (n=204 ticks were collected by flagging from northeastern China, and detected for infection with Anaplasma, Ehrlichia, Babesia, and Hepatozoon spp. by using nested polymerase chain reaction assays and sequencing analysis. A. phagocytophilum was detected in all tick species, i.e., I. persulcatus (9.4%, H. longicornis (1.9%, H. concinna (6.5%, D. nuttalli (1.7%, and D. silvarum (2.3%; A. bovis was detected in H. longicornis (0.3% and H. concinna (0.2%; E. muris was detected in I. persulcatus (2.5% and H. concinna (0.2%; Candidatus Neoehrlichia mikurensis was only detected in I. persulcatus (0.4%. The Ehrlichia variant (GenBank access number KU921424, closely related to E. ewingii, was found in H. longicornis (0.8% and H. concinna (0.2%. I. persulcatus was infected with B. venatorum (1.2%, B. microti (0.6%, and B. divergens (0.6%. Additionally, four Babesia sequence variants (GenBank access numbers 862303–862306 were detected in I. persulcatus, H. longicornis, and H. concinna, which belonged to the clusters formed by the parasites of dogs, sheep and cattle (B. gibsoni, B. motasi, and B. crassa. Two Hepatozoon spp. (GenBank access numbers KX016028 and KX016029 associated with hepatozoonosis in Japanese martens were found in the collected ticks (0.1–3.1%. These findings showed the genetic variability of Anaplasma, Ehrlichia, Babesia, and Hepatozoon spp. circulating in ticks in northeastern China, highlighting the necessity for further research of these tick-associated pathogens and their role in human and animal diseases.
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
García-Acosta, J; Aguilar-García, CR; Aguilar-Arce, IE
Resumen El género Rickettsia está compuesto por dos grupos definidos antigenicamente: el grupo tifus, que incluye a Rickettsiaprowazekii, causante del tifus epidémico o exantemático, y a R. typhi, causante del tifus murino o endémico; el otro grupo es el de las fiebres manchadas. El género Rickettsia está constituido por diferentes especies de bacterias gramnegativas; a su vez, forma parte de la familia Rickettsiaceae (junto a Coxiella, Ehrlichia y Bartonella). Todas las especies del género t...
Full Text Available t WO0168911. 40 0.31 3 ( FG284490 ) 1108770671670 New World Screwworm Egg 9261 ES...Ts C... 40 0.36 3 ( FG285211 ) 1108770694495 New World Screwworm Egg 9261 ESTs C... 40 0.40 3 ( FG284535 ) 1108770671722 New World...7821 ) Ehrlichia ruminantium strain Welgevonden, complet... 38 0.42 12 ( FG286862 ) 1108770727001 New World ...1 ( CP000083 ) Colwellia psychrerythraea 34H, complete genome. 48 0.45 1 ( FG284489 ) 1108770671669 New World...e 3 clone RP11-84J20, WORKI... 38 0.48 5 ( FG287745 ) 1108770753631 New World Scr
Márcio A. M. Galvão
Full Text Available O trabalho descreve um inquérito sorológico para rickettsioses em escolares e cães de Novo Cruzeiro, Minas Gerais, Brasil, em 1998. Trezentos e trinta e um escolares pertenciam a uma área endêmica e 142 a uma área não endêmica do município. Trinta e nove (10,1% soros foram reativos à Reação de Imunofluorescência Indireta (RIFI para Rickettsia rickettsiino título de 1:64, sendo que dentre esses reativos, 35 eram de estudantes de escolas de área endêmica. Dentre os 73 cães analisados quanto à presença de anticorpos anti R. rickettsii, anti Ehrlichia chaffeensise anti Ehrlichia canisà RIFI no título de 1:64, 3 (4,11%, 11 (15,07% e 13 (17,81% desses animais foram reativos respectivamente aos antígenos testados. Conclui-se que, a sororeatividade para R. rickettsiiem indivíduos sadios sem história prévia de febre maculosa brasileira, uma doença marcante por sua alta letalidade, e a presença de sororeatividade para Ehrlichiacom potencial patogênico para o homem em cães, nos leva a indagar sobre a transmissão ao homem de outras espécies da família Rickettsiae na área estudada.This article describes a serological survey for rickettsiosis in the county of Novo Cruzeiro, Minas Gerais State, Brazil, in 1998, testing schoolchildren and dogs. Sera included 331 samples from schoolchildren from an endemic area and 142 samples from schoolchildren from a non-endemic area in the county. All children examined were healthy and had not reported clinical symptoms of Brazilian spotted fever prior to the serological survey. Some 35 children in the endemic area were reactive to Rickettsia rickettsiiby indirect fluorescent antibody (IFA with a titer of 1:64, corresponding to 10.6%. Sera from 73 dogs were tested, showing seroreactivity (IFA 1:64 to Rickettsia rickettsi, Ehrlichia chaffeensis, and Ehrlichia canisin 3 (4.11%, 11 (15.07%, and 13 (17.81%, respectively. The results in schoolchildren and the presence of canine seroreactivity to
Keyla Carstens Marques de Sousa
Full Text Available Tick-borne pathogens affect a wide range of vertebrate hosts. To identify tick-borne pathogens among dogs from Campo Grande, MS, Brazil testing seropositive for Leishmania infantum (syn. L. chagasi, a serological and molecular study was conducted to detectEhrlichia canis, Anaplasma platys and Babesia vogeli in 60 serum and spleen samples. A confirmatory diagnosis ofL. infantum based on serological and molecular assays was also performed, as was sequence alignment and phylogenetic analysis to assess the identity of the parasite species infecting these animals. IgG antibodies toEhrlichia spp., B. vogeli and L. infantum were found, respectively, in 39 (65%, 49 (81.6% and 60 (100% of the sampled dogs. Twenty-seven (45%, fifty-four (90%, fifty-three (88.3%, two (3.3% and one (1.6% dog were positive, respectively, forE. canis, Leishmania spp., Leishmania donovani complex, Babesia sp. and Anaplasma sp. in PCR assays. After sequencing, the amplicons showed 99% of identity with E. canis, B. vogeli, A. platys andLeishmania chagasi isolates. The findings of this study indicate that L. infantum-seropositive dogs from Campo Grande are exposed to multiple tick-borne pathogens, which should therefore be included in the differential diagnosis of dogs with clinical suspicion of leishmaniasis.
Herbert Sousa Soares
Full Text Available Blood samples were collected from 99 domestic dogs from the urban and rural areas of the Lábrea municipality, state of Amazonas, Brazil. Canine serum samples were tested by immunofluorescence assay against Rickettsia spp., which revealed that only 3.0% (1/33 and 7.6% (5/66 of the dogs from urban and rural areas, respectively, reacted positively to at least one Rickettsia species. DNA was extracted from canine blood and tested by a battery of PCR assays targeting protozoa of the genera Babesia and Hepatozoon, and bacteria of the genera Rickettsia and Ehrlichia and family Anaplasmataceae. All samples were negative in the PCR assays targeting the genera Babesia, Hepatozoon, Ehrlichia and Rickettsia. For Anaplasmataceae, 3% (1/33 and 39.4% (26/66 of the urban and rural dogs, respectively, yielded amplicons that generated DNA sequences 100% identical to the corresponding sequence of Wolbachia endosymbiont of Dirofilaria immitis. Because of these results, all canine DNA samples were further tested in a PCR assay targeting filarial nematodes, which was positive for 18.2% (6/33 and 57.6% (38/66 urban and rural dogs, respectively. Filarial-PCR products generated DNA sequences 100% identical to D. immitis. While tick-borne infections were rare in Lábrea, D. immitis infection rates were among the highest reported in South America.
A. Valeria Scorza
Full Text Available The prevalence of intestinal parasites and vector-borne agents of dogs and cats in the Pine Ridge Reservation, South Dakota were determined. Fecal samples (84 dogs, 9 cats were examined by centrifugal floatation and by immunofluorescence assay (FA for Giardia and Cryptosporidium. PCR was performed on Giardia [beta-giardin (bg, triose phosphate isomerase (tpi, glutamate dehydrogenase genes (gdh] and Cryptosporidium [heat shock protein-70 gene (hsp] FA positive samples. Cat sera (n = 32 were tested for antibodies against Bartonella spp., Toxoplasma gondii, and FIV, and antigens of FeLV and Dirofilaria immitis. Dog sera (n = 82 were tested for antibodies against T. gondii, Borrelia burgdorferi, Ehrlichia canis, and Anaplasma phagocytophilum and D. immitis antigen. Blood samples (92 dogs, 39 cats were assessed by PCR for amplification of DNA of Bartonella spp., Ehrlichia spp., Anaplasma spp., haemoplasmas, and Babesia spp. (dogs only. The most significant results were Giardia spp. (32% by FA, Taenia spp. (17.8% and Cryptosporidium spp. (7.1%. The Giardia isolates typed as the dog-specific assemblages C or D and four Cryptosporidium isolates typed as C. canis. Antibodies against T. gondii were detected in 15% of the dogs. Antibodies against Bartonella spp. and against T. gondii were detected in 37.5% and 6% of the cats respectively. FeLV antigen was detected in 10% of the cats.
Tereza Emmanuelle de Farias Rotondano
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.
Tadin, Ante; Tokarz, Rafal; Markotić, Alemka; Margaletić, Josip; Turk, Nenad; Habuš, Josipa; Svoboda, Petra; Vucelja, Marko; Desai, Aaloki; Jain, Komal; Lipkin, W Ian
Croatia is a focus for many rodent-borne zoonosis. Here, we report a survey of 242 rodents and small mammals, including 43 Myodes glareolus, 131 Apodemus flavicollis, 53 Apodemus agrarius, three Apodemus sylvaticus, six Sorex araneus, four Microtus arvalis, one Microtus agrestis, and one Muscardinus avellanarius, collected at eight sites in Croatia over an 8-year period. Multiplex MassTag polymerase chain reaction (PCR) was used for detection of Borrelia, Rickettsia, Bartonella, Babesia, Ehrlichia, Anaplasma, Francisella tularensis, and Coxiella burnetii. Individual PCR assays were used for detection of Leptospira, lymphocytic choriomeningitis virus, orthopoxviruses, flaviviruses, hantaviruses, and Toxoplasma gondii. Of the rodents, 52 (21.5%) were infected with Leptospira, 9 (3.7%) with Borrelia miyamotoi, 5 (2%) with Borrelia afzelii, 29 (12.0%) with Bartonella, 8 (3.3%) with Babesia microti, 2 (0.8%) with Ehrlichia, 4 (1.7%) with Anaplasma, 2 (0.8%) with F. tularensis, 43 (17.8%) with hantaviruses, and 1 (0.4%) with an orthopoxvirus. Other agents were not detected. Multiple infections were found in 32 rodents (13.2%): dual infections in 26 rodents (10.7%), triple infections in four rodents (2.9%), and quadruple infections in two rodents (0.8%). Our findings indicate that rodents in Croatia harbor a wide range of bacteria and viruses that are pathogenic to humans. © The American Society of Tropical Medicine and Hygiene.
Yabsley, Michael J; Nims, Todd N; Savage, Mason Y; Durden, Lance A
Ticks were collected from 38 black bears (Ursus americanus floridanus) from northwestern Florida (n = 18) from 2003 to 2005 and southern Georgia (n = 20) in 2006. Five species (Amblyomma americanum, A. maculatum, Dermacentor variabilis, Ixodes scapularis, and I. affinis) were collected from Florida bears, and 4 species (A. americanum, A. maculatum, D. variabilis, I. scapularis) were collected from bears in Georgia. Ixodes scapularis was the most frequently collected tick, followed by D. variabilis, A. americanum, A. maculatum, and I. affinis. The collection of I. affinis from a Florida bear represents a new host record. A subset of ticks was screened for pathogens and putative symbionts by polymerase chain reaction (PCR). The zoonotic tick-borne pathogens Ehrlichia chaffeensis and Rickettsia parkeri were detected in 1 of 23 (4.3%) A. americanum and 1 of 12 (8.3%) A. maculatum, respectively. The putative zoonotic pathogen "Rickettsia amblyommii" was detected in 4 (17.4%) A. americanum and 1 (8.3%) A. maculatum. Other putative symbiotic rickettsiae detected included R. bellii and R. montanensis in D. variabilis, a Rickettsia cooleyi-like sp. and Rickettsia sp. Is-1 in I. scapularis, and Rickettsia TR39-like sp. in I. scapularis and A. americanum. All ticks were PCR-negative for Anaplasma phagocytophilum, Panola Mountain Ehrlichia sp., E. ewingii, Francisella tularensis, and Borrelia spp.
Christensen, Steen; Serbus, Laura Renee
Two-component regulatory systems are commonly used by bacteria to coordinate intracellular responses with environmental cues. These systems are composed of functional protein pairs consisting of a sensor histidine kinase and cognate response regulator. In contrast to the well-studied Caulobacter crescentus system, which carries dozens of these pairs, the streamlined bacterial endosymbiont Wolbachia pipientis encodes only two pairs: CckA/CtrA and PleC/PleD. Here, we used bioinformatic tools to compare characterized two-component system relays from C. crescentus, the related Anaplasmataceae species Anaplasma phagocytophilum and Ehrlichia chaffeensis, and 12 sequenced Wolbachia strains. We found the core protein pairs and a subset of interacting partners to be highly conserved within Wolbachia and these other Anaplasmataceae. Genes involved in two-component signaling were positioned differently within the various Wolbachia genomes, whereas the local context of each gene was conserved. Unlike Anaplasma and Ehrlichia, Wolbachia two-component genes were more consistently found clustered with metabolic genes. The domain architecture and key functional residues standard for two-component system proteins were well-conserved in Wolbachia, although residues that specify cognate pairing diverged substantially from other Anaplasmataceae. These findings indicate that Wolbachia two-component signaling pairs share considerable functional overlap with other α-proteobacterial systems, whereas their divergence suggests the potential for regulatory differences and cross-talk. PMID:25809075
Silvana de Cássia Paulan
Full Text Available Canine visceral leishmaniasis (CVL is caused by the protozoan Leishmania infantum, which infects dogs and humans in many regions of Brazil. The present study involved an indirect fluorescent antibody test (IFAT to analyze L. infantum,Ehrlichia spp., Babesia canis,Toxoplasma gondii and Neospora caninuminfection rates in serum samples from 93 dogs in a rural settlement in Ilha Solteira, SP, Brazil. The seroprevalence rates of anti-L. infantum, anti-Ehrlichia, anti-B. canis, anti-T. gondii and anti-N. caninum antibodies were 37.6%, 75.3%, 72%, 47.3% and 6.4%, respectively. In addition to IFAT, direct microscopic examination of popliteal lymph node aspirates revealed 26.9% of CVL positive dogs. Serological tests revealed that 17.2% of the dogs were seropositive for a single parasite, 29% for two parasites, 33% for three, 16.1% for four, and 1.1% for five parasites, while 3.2% were seronegative for five parasites. The presence of antibodies against these parasites in serum samples from dogs confirmed their exposure to these parasites in this rural area. Because of the potential zoonotic risk of these diseases, mainly leishmaniasis, ehrlichiosis and toxoplasmosis, special attention should focus on programs for the improvement of diagnostic assays and control measures against these parasites.
Cardoso, Luís; Oliveira, Ana Cristina; Granada, Sara; Nachum-Biala, Yaarit; Gilad, Matan; Lopes, Ana Patrícia; Sousa, Sérgio Ramalho; Vilhena, Hugo; Baneth, Gad
No molecular data have been available on tick-borne pathogens that infect dogs from Angola. The occurrence of agents from the genera Anaplasma, Babesia, Ehrlichia and Hepatozoon was assessed in 103 domestic dogs from Luanda, by means of the polymerase chain reaction (PCR) and DNA sequence analysis. Forty-six dogs (44.7 %) were positive for at least one pathogen. Twenty-one animals (20.4 %) were found infected with Anaplasma platys, 18 (17.5 %) with Hepatozoon canis, six (5.8 %) with Ehrlichia canis, six (5.8 %) with Babesia vogeli, one (1.0 %) with Babesia gibsoni and one (1.0 %) with an unnamed Babesia sp. The molecular frequency of single infections taken together was 37.9 % and that of co-infections with several combinations of two pathogens accounted for 6.8 % of the animals. This is the first report of A. platys, B. vogeli, B. gibsoni, E. canis and H. canis infections diagnosed by PCR in domestic dogs from Angola. The present study provides evidence that dogs in Luanda are widely exposed to, and at risk of becoming infected with, tick-borne pathogens. Further investigation is needed, including a larger number of animals, canine populations from other cities and provinces of the country, as well as potential vector ticks, aiming at better characterizing and controlling canine vector-borne diseases in Angola.
Malheiros, J; Costa, M M; do Amaral, R B; de Sousa, K C M; André, M R; Machado, R Z; Vieira, M I B
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. Copyright © 2016 Elsevier GmbH. All rights reserved.
Amoli, Aa Rahmani; Khoshnegah, J; Razmi, Ghr
We attempted to determine the prevalence of Hepatozoon spp. infection in Mashhad, northeast of Iran, via blood smear parasitology. The prevalence was investigated by examination of blood smear parasitology, using blood samples collected from 254 dogs (51 strays and 203 privately owned-dogs). Two stray dogs (2/51; 3.92%) and two privately-owned dogs (2/203; 0.98%) were infected with Hepatozoon spp. Therefore, as per blood smear parasitology, the prevalence of Hepatozoon spp. infection was 1.57% (4/254). Sixteen out of 254 dogs (6.29%) were infested with ticks; all of which were Rhipicephalus sanguineus. One of the dogs infected with Hepatozoon spp. exhibited ticks at the time of examination. Concurrent infection with Ehrlichia canis and Leishmania infantum was not detected in the four Hepatozoon spp. infected dogs. This is the first epidemiological study on the prevalence of Hepatozoon spp. infection in dogs in Iran.
Luiz Ricardo Gonçalves
Full Text Available Since dogs presenting several vector borne diseases can show none or nonspecific clinical signs depending on the phase of infection, the assessment of the particular agents involved is mandatory. The present study aimed to investigate the presence of Babesia spp., Ehrlichia spp., Anaplasma spp., Hepatozoon spp. and Leishmania spp. in blood samples and ticks, collected from two dogs from Rio Grande do Norte showing suggestive tick-borne disease by using molecular techniques. DNA of E. canis, H. canis and L. infantum were detected in blood samples and R. sanguineus ticks collected from dogs. Among all samples analyzed, two showed the presence of multiple infections with E. canis, H. canis and L. infantum chagasi. Here we highlighted the need for molecular differential diagnosis in dogs showing nonspecific clinical signs.
Mylonakis, Mathios E; Leontides, Leonidas; Gonen, Liat; Billinis, Charalambos; Koutinas, Alexander F; Baneth, Gad
The prevalence of IgG antibodies to Hepatozoon canis and the presence of gamonts in the blood and hemolymphatic tissues were studied in dogs with canine monocytic ehrlichiosis (CME) caused by Ehrlichia canis. Both pathogens are transmitted by the tick Rhipicephalus sanguineus. Forty-five out of 69 (65.2%) dogs with CME were seropositive to H. canis by an enzyme-linked immunosorbent assay (ELISA). Intra-neutrophilic gamonts of H. canis were found in 2 out of 69 dogs (2.9%) comprising 4.5% of the seropositive dogs. The present study indicated that the prevalence of antibodies to H. canis was high among dogs with CME in an area where both infections are endemic. However, previous exposure to H. canis was not found as an important contributor to clinical or clinicopathologic abnormalities found in dogs with CME.
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. PMID:18691408
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.
Tago, Damian; Meyer, Damien F
Diseases induced by obligate intracellular pathogens have a large burden on global human and animal health. Understanding the factors involved in the virulence and fitness of these pathogens contributes to the development of control strategies against these diseases. Based on biological observations, a theoretical model using game theory is proposed to explain how obligate intracellular bacteria interact with their host. The equilibrium in such a game shows that the virulence and fitness of the bacterium is host-triggered and by changing the host's defense system to which the bacterium is confronted, an evolutionary process leads to an attenuated strain. Although, the attenuation procedure has already been conducted in practice in order to develop an attenuated vaccine (e.g., with Ehrlichia ruminantium), there was a lack of understanding of the theoretical basis behind this process. Our work provides a model to better comprehend the existence of different phenotypes and some underlying evolutionary mechanisms for the virulence of obligate intracellular bacteria.
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Keysary, Avi; Amram, Lili; Keren, Gershon; Sthoeger, Zev; Potasman, Israel; Jacob, Amir; Strenger, Carmella; Dawson, Jacqueline E.
We conducted a retrospective serosurvey of 1,000 persons in Israel who had fever of undetermined cause to look for Ehrlichia chaffeensis antibodies. Four of five cases with antibodies reactive to E. chaffeensis were diagnosed in the summer, when ticks are more active. All patients had influenzalike symptoms with high fever. None of the cases was fatal. Three serum samples were also seroreactive for antibodies to E. canis, and one was also reactive to the human granulocytic ehrlichiosis (HGE) agent. The titer to the HGE agent in this patient was higher than the serum titer to E. chaffeensis, and the Western blot analysis also indicated that the HGE agent was the primary cause of infection. We present the first serologic evidence that the agents of human monocytic ehrlichiosis (HME) and HGE are present in Israel. Therefore, human ehrlichiosis should be included in the differential diagnoses for persons in Israel who have been exposed to ticks and have influenzalike symptoms. PMID:10603210
Full Text Available A total of 344 dogs belonging to people in resource-poor communities in North West Province, South Africa, was examined for ectoparasites, and all visible arthropods were collected from the left side of each dog. By doubling these numbers it was estimated that the dogs harboured 14 724 ixodid ticks, belonging to 6 species, 1028 fleas, belonging to 2 species, and 26 lice. Haemaphysalis leachi accounted for 420 and Rhipicephalus sanguineus for 14 226 of the ticks. Pure infestations of H. leachi were present on 14 dogs and of R. sanguineus on 172 dogs. Small numbers of Amblyomma hebraeum, R. appendiculatus, R. evertsi evertsi and R. simus were also collected. The predominance of R. sanguineus accounts for the high prevalence of canine ehrlichiosis (Ehrlichia canis within the survey region, compared to canine babesiosis (Babesia canis, which is transmitted by H. leachi, and is a much rarer disease.
Liu, Lijuan; Chen, Qian; Yang, Yu; Wang, Jiancheng; Cao, Xiaomei; Zhang, Sheng; Li, Hong; Hou, Yong; Wang, Fuxiang; Xu, Baoliang
To describe the prevalence of Rickettsia in ticks at the Sino-Russian and Sino-Mongolian borders, a total of 292 ticks were collected and tested by conventional PCR assays. The prevalence of Rickettsia was 53.4%, and phylogenetic analysis showed that they belonged to R. raoultii species after alignment for the ompA, ompB, and gltA genes, respectively. Coxiella burnetii DNA was detected for 14%, and no Ehrlichia, Borrelia burgdorferi, and Babesia species were found. Co-infection of two pathogens was 9.9%, and no co-infection with three or more pathogens was found. This study suggested Rickettsia was the most common pathogen in the ticks and co-infection was found. The findings might be helpful to provide advice on the prevention and control of tick-borne disease potential for tourists and residents.
Full Text Available The present work describes a retrospective study of clinical cases of ehrlichiosis in dogs examined from March 1998 to September 2001. From the clinical records with laboratorial confirmation of Ehrlichia canis or E. platys infections, the following parameters were analyzed: demographic aspects (age, race, sex, period of the year and origin, clinical characteristics (body temperature, exposure to ticks and clinical signs, and hematological characteristics (blood cell counts and type of infected cell. A total of 194 clinical records were analyzed, from which 31 animals were infected with E. canis and 21 animals with E. platys. The number of cases of canine ehrlichiosis increased considerably from the year 2000 onwards, and 24.4% of the cases occurred in 13- to 24-month-old animals, in different urban and per-urban regions of the municipality of Belo Horizonte. The most frequent symptoms were fever, anorexia, apathy, abdominal pain, lymphadenopathy and dispnea. Regarding hematological alterations, 70.3% of the animals presented anemia, 50% presented thrombocytopenia and 30% leukopenia, and most E. canis morulae were seen in monocytes. The results point to the importance of canine ehrlichiosis, as 35.9% of the dogs with suspected hemoparasitic diseases were infected with Ehrlichia canis or E. platys.O presente trabalho descreve um estudo retrospectivo da casuística clínica de erliquiose em cães atendidos entre março de 1998 e setembro de 2001. Foram analisadas 194 fichas clínicas de animais com suspeita de hemoparasitoses, nas quais 31 cães foram diagnosticados com Ehrlichia canis e 21 com Ehrlichia platys, por meio de exame parasitológico direto de esfregaços sangüíneos. Foram considerados alguns aspectos demográficos (idade, raça, sexo, época do ano e região de origem, características clínicas (temperatura corporal, presença e/ou histórico de carrapatos e sinais clínicos e hematológicas (hemograma completo e célula parasitada
Seroprevalence rates of antibodies against Leishmania infantum and other protozoan and rickettsial parasites in dogs Soroprevalência de anticorpos contra Leishmania infantum e outras espécies de protozoários e rickettsia em cães
Silvana de Cássia Paulan
Full Text Available Canine visceral leishmaniasis (CVL is caused by the protozoan Leishmania infantum, which infects dogs and humans in many regions of Brazil. The present study involved an indirect fluorescent antibody test (IFAT to analyze L. infantum, Ehrlichia spp., Babesia canis, Toxoplasma gondii and Neospora caninum infection rates in serum samples from 93 dogs in a rural settlement in Ilha Solteira, SP, Brazil. The seroprevalence rates of anti-L. infantum, anti-Ehrlichia, anti-B. canis, anti-T. gondii and anti-N. caninum antibodies were 37.6%, 75.3%, 72%, 47.3% and 6.4%, respectively. In addition to IFAT, direct microscopic examination of popliteal lymph node aspirates revealed 26.9% of CVL positive dogs. Serological tests revealed that 17.2% of the dogs were seropositive for a single parasite, 29% for two parasites, 33% for three, 16.1% for four, and 1.1% for five parasites, while 3.2% were seronegative for five parasites. The presence of antibodies against these parasites in serum samples from dogs confirmed their exposure to these parasites in this rural area. Because of the potential zoonotic risk of these diseases, mainly leishmaniasis, ehrlichiosis and toxoplasmosis, special attention should focus on programs for the improvement of diagnostic assays and control measures against these parasites.Leishmaniose Visceral Canina (LVC é causada pelo protozoário Leishmania infantum, podendo infectar cães e humanos em várias regiões do Brasil. O presente estudo teve por objetivo realizar a reação de imunofluorescência indireta (RIFI para analisar os índices de infecção parasitária para L. infantum, Ehrlichia spp., Babesia canis, Toxoplasma gondii e Neospora caninum, em 93 amostras de soro de cães de um assentamento rural no município de Ilha Solteira, SP, Brasil. A taxa de soroprevalência de cães com anticorpos anti-L. infantum, anti-Ehrlichia, anti-B. canis, anti-T. gondii e anti-N. caninum foi de 37,6%, 75,3%, 72%, 47,3% e 6
Full Text Available Objective: To evaluate the prevalence of blood parasite infection in stray dogs by PCR technique and the association between levels of packed cell volume (PCV and blood parasitic infection in stray dogs. Methods: A total of 65 blood samples were collected from stray dogs in animal quarantine station from Mahasarakham, Thailand to evaluate the levels of PCV before molecular screening for tick-borne pathogens infection. Results: Stray dogs were positive with one or more pathogens in 44 (67.69% out of 65 blood samples. Ehrlichia canis [43.1%, 95% confidence interval (CI: 38.1–48.1] was the most common blood pathogen found infecting in stray dogs in Mahasarakham Province, followed by Anaplasma platys (29.2%, 95% CI: 24.2–34.2, Hepatozoon canis (12.3%, 95% CI: 7.3–17.3 and Babesia canis vogeli (6.2%, 95% CI: 1.2–11.2, respectively. Moreover, co-infections with two pathogens were identified in 11 (16.9% of dogs examined and two (2.9% dogs were coinfections with three pathogens. Statistically significant relationship between the PCV levels and Ehrlichia canis infection was found (P < 0.05. Conclusions: This study indicated that blood pathogens are spreading in stray dogs and they are potentially high risk of agent transmission to human via exposure with tick vectors. It was also the first report of Anaplasma platys infection in dogs in north-eastern part of Thailand.
Wolf, Rafael William; Aragona, Mônica; Muñoz-Leal, Sebastián; Pinto, Leticia Borges; Melo, Andréia Lima Tomé; Braga, Isis Assis; Costa, Jackeliny dos Santos; Martins, Thiago Fernandes; Marcili, Arlei; Pacheco, Richard de Campos; Labruna, Marcelo B; Aguiar, Daniel Moura
Taking into account the diversity of small terrestrial mammals of the Pantanal, the present study aimed to verify the occurrence of infection by Ehrlichia spp., Anaplasma spp., Rickettsia spp., Hepatozoon spp., Babesia spp. and parasitism by ticks in non-volant small mammals collected in the Brazilian Pantanal. Samples of blood, liver and spleen were collected from 64 captured animals, 22 marsupials and 42 rodents. Pathogen detection was performed by the use of genus-specific Polymerase Chain Reaction (PCR) assays. Ticks collected from the animals consisted of Amblyomma sculptum and Amblyomma triste nymphs, and Ornithodoros guaporensis larvae. None of the vertebrate samples (blood, liver, or spleen) yielded detectable DNA of Rickettsia spp. or Ehrlichia spp. The blood of the rodent Hylaeamys megacephalus yielded an Anaplasma sp. genotype (partial 16S rRNA gene) 99% similar to multiple Anaplasma spp. genotypes around the world. The blood of three rodents of the species Calomys callosus were positive for a novel Hepatozoon sp. agent, phylogenetically related (18S rDNA gene) to distinct Hepatozoon genotypes that have been detected in rodents from different parts of the world. One marsupial (Monodelphis domestica) and three rodents (Thrichomys pachyurus) were positive to novel piroplasmid genotypes, phylogenetically (18S rDNA gene) related to Theileria bicornis, Cytauxzoon manul, and Cytauxzoon felis. The present study provides the first molecular detection of Hepatozoon sp. and piroplasmids in small mammals in Brazil. Additionally, we expanded the distribution of O. guaporensis to Brazil, since this tick species was previously known to occur only in Bolivia. Copyright © 2016 Elsevier GmbH. All rights reserved.
Alho, Ana Margarida; Lima, Clara; Latrofa, Maria Stefania; Colella, Vito; Ravagnan, Silvia; Capelli, Gioia; Madeira de Carvalho, Luís; Cardoso, Luís; Otranto, Domenico
Vector-borne diseases (VBDs) have been increasingly reported in dogs and cats worldwide. However, no data are currently available regarding canine and feline VBDs in Qatar and limited information is available from other Persian Gulf countries. Blood samples from 98 client-owned animals (i.e. 64 dogs and 34 cats) living in Doha (Qatar) were collected and the presence of genomic DNA of Anaplasma spp., Babesia spp., Dirofilaria spp., Ehrlichia spp., Hepatozoon spp., Mycoplasma spp. and Rickettsia spp. was assessed by polymerase chain reaction (PCR), real time-PCR (rt-PCR) and sequence analysis. Of the 64 dogs, 12 (18.8%) were infected with at least one pathogen (i.e. 7.8% with Mycoplasma spp., 4.7% with Babesia vogeli, 3.1% with Ehrlichia canis, and 1.6% with Anaplasma platys, Babesia gibsoni and Hepatozoon canis, each). One of the 12 dogs was co-infected with B. vogeli and E. canis. Of the 34 cats, seven (20.6%) animals were infected with at least one pathogen (i.e. 5.9% were positive for Mycoplasma spp., and 2.9% for Babesia felis, B. vogeli, E. canis, "Candidatus Mycoplasma haemominutum" and Mycoplasma haemofelis, each). No dogs or cats were positive for Dirofilaria spp. or Rickettsia spp. Although the sample sizes of dogs and cats herein analysed was moderately small, data from this study report the occurrence of A. platys, B. vogeli, B. gibsoni, E. canis, H. canis and Mycoplasma spp. in domestic dogs and of B. felis, B. vogeli, "Candidatus M. haemominutum", E. canis and M. haemofelis in domestic cats from Qatar. Further investigations along with prophylactic measures are strongly recommended in order to reduce the risk of dogs and cats acquiring VBDs in Qatar.
Pereira, André; Parreira, Ricardo; Nunes, Mónica; Casadinho, Afonso; Vieira, Maria Luísa; Campino, Lenea; Maia, Carla
Wildlife can act as reservoir of different tick-borne pathogens, such as bacteria, parasites and viruses. The aim of the present study was to assess the presence of tick-borne bacteria and protozoa with veterinary and zoonotic importance in cervids and wild boars from the Centre and South of Portugal. One hundred and forty one blood samples from free-ranging ungulates including 73 red deer (Cervus elaphus), 65 wild boars (Sus scrofa) and three fallow deer (Dama dama) were tested for the presence of Anaplasma marginale/A. ovis, A. phagocytophilum, Anaplasma/Ehrlichia spp., Babesia/Theileria spp., Borrelia burgdorferi (sensu lato) (s.l.), and Rickettsia spp. DNA by PCR. Anaplasma spp. DNA was detected in 33 (43.4 %) cervids (31 red deer and two fallow deer) and in two (3.1 %) wild boars while Theileria spp. were found in 34 (44.7 %) cervids (32 red deer and two fallow deer) and in three (4.6 %) wild boar blood samples. Sequence analysis of msp4 sequences identified A. marginale, A. ovis, while the analysis of rDNA sequence data disclosed the presence of A. platys and A. phagocytophilum and T. capreoli and Theileria sp. OT3. Anaplasma spp./Theileria spp. mixed infections were found in 17 cervids (22.4 %) and in two wild boars (3.1 %). All samples were negative for Babesia sp., B. burgdorferi (s.l.), Ehrlichia sp. or Rickettsia sp. This is the first detection of Anaplasma marginale, A. ovis, A. phagocytophilum, A. platys, Theileria capreoli and Theileria sp. OT3 in cervids and wild boars from Portugal. Further studies concerning the potential pathogenicity of the different species of Anaplasma and Theileria infecting wild ungulates, the identification of their vector range, and their putative infectivity to domestic livestock and humans should be undertaken.
Galvão Márcio A. M.
Full Text Available O trabalho descreve um inquérito sorológico para rickettsioses em escolares e cães de Novo Cruzeiro, Minas Gerais, Brasil, em 1998. Trezentos e trinta e um escolares pertenciam a uma área endêmica e 142 a uma área não endêmica do município. Trinta e nove (10,1% soros foram reativos à Reação de Imunofluorescência Indireta (RIFI para Rickettsia rickettsiino título de 1:64, sendo que dentre esses reativos, 35 eram de estudantes de escolas de área endêmica. Dentre os 73 cães analisados quanto à presença de anticorpos anti R. rickettsii, anti Ehrlichia chaffeensise anti Ehrlichia canisà RIFI no título de 1:64, 3 (4,11%, 11 (15,07% e 13 (17,81% desses animais foram reativos respectivamente aos antígenos testados. Conclui-se que, a sororeatividade para R. rickettsiiem indivíduos sadios sem história prévia de febre maculosa brasileira, uma doença marcante por sua alta letalidade, e a presença de sororeatividade para Ehrlichiacom potencial patogênico para o homem em cães, nos leva a indagar sobre a transmissão ao homem de outras espécies da família Rickettsiae na área estudada.
René-Martellet, Magalie; Lebert, Isabelle; Chêne, Jeanne; Massot, Raphaël; Leon, Marta; Leal, Ana; Badavelli, Stefania; Chalvet-Monfray, Karine; Ducrot, Christian; Abrial, David; Chabanne, Luc; Halos, Lénaïg
Canine Monocytic Ehrlichiosis (CME), due to the bacterium Ehrlichia canis and transmitted by the brown dog tick Rhipicephalus sanguineus, is a major tick-borne disease in southern Europe. In this area, infections with other vector-borne pathogens (VBP) are also described and result in similar clinical expression. The aim of the present study was to evaluate the incidence risk of clinical CME in those endemic areas and to assess the potential involvement of other VBP in the occurrence of clinical and/or biological signs evocative of the disease. The study was conducted from April to November 2011 in veterinary clinics across Italy, Spain and Portugal. Sick animals were included when fitting at least three clinical and/or biological criteria compatible with ehrlichiosis. Serological tests (SNAP®4Dx, SNAP®Leish tests, Idexx, USA) and diagnostic PCR for E. canis, Anaplasma platys, Anaplasma phagocytophilum, Babesia spp, Hepatozoon canis and Leishmania infantum detection were performed to identify the etiological agents. Ehrlichiosis was considered when three clinical and/or biological suggestive signs were associated with at least one positive paraclinical test (serology or PCR). The annual incidence risk was calculated and data were geo-referenced for map construction. The probabilities of CME and other vector-borne diseases when facing clinical and/or biological signs suggestive of CME were then evaluated. A total of 366 dogs from 78 veterinary clinics were enrolled in the survey. Among them, 99 (27%) were confirmed CME cases, which allowed an estimation of the average annual incidence risk of CME amongst the investigated dog population to be 0.08%. Maps showed an increasing gradient of CME incidence risk from northern towards southern areas, in particular in Italy. It also suggested the existence of hot-spots of infections by VBP in Portugal. In addition, the detection of other VBP in the samples was common and the study demonstrated that a dog with clinical signs
Daniel C. L. Linhares
Full Text Available Este trabalho foi desenvolvido com o objetivo de padronizar um protocolo para a reação em cadeia da polimerase (PCR e selecionar oligonucleotídeos iniciadores para a detecção específica de Ehrlichia canis, em uma única etapa de reação. Inicialmente foram obtidas seqüências depositadas no Genbank, referentes ao gene que codifica o 16S rRNA das espécies E. canis (número de acesso = AF162860, E. ewingii (U96436, E. platys (AF1567844, E. chaffeensis, (U86665, E. phagocytophila genogrupo (U02521, E. bovis (AF294789 e E. risticii (M21290, as quais foram submetidas ao alinhamento genético para a construção dos iniciadores. Do alinhamento foi selecionado, a partir de uma região semiconservada, um iniciador específico para E. canis, designado EBR1 (5’-cctctggctataggaaattg- 3’ e, de uma região conservada, um iniciador genérico EBR5 (5’-ggagtgcttaacgcgttag- 3’. Paralelamente foram obtidas amostras de sangue de dez cães que apresentavam infecção aguda por E. canis, confirmado pela presença de mórulas intracitoplasmáticas, características da riquétsia, em células mononucleares sangüíneas. O DNA genômico extraído dessas amostras foi utilizado para a avaliação da reação de PCR, empregando-se um protocolo adaptado de outros autores e o par de oligos EBR1/EBR5, selecionado neste trabalho. A reação de PCR apresentou resultados positivos para os 10 isolados de E. canis, amplificando o fragmento esperado de 765 pares de bases do gene 16S rRNA. Resultados negativos verificados nas reações de PCR para amostras de DNA genômico de Babesia canis, Hepatozoon canis, Haemobartonella sp., Trypanosoma evansi e do hospedeiro (Canis familiaris livre de infecção indicaram a segurança do método quanto à especificidade para a discriminação de E. canis. PALAVRAS-CHAVE: Cães, Ehrlichia canis, erliquiose canina, hemoparasitose, reação em cadeia da polimerase
Background Protozoa and bacteria transmitted by arthropods, including ticks and phlebotomine sand flies, may cause a wide range of canine vector-borne diseases. Dogs can be simultaneously or sequentially infected with multiple pathogens. Canine babesiosis caused by Babesia canis canis and Babesia canis vogeli is known to occur in Portugal. This study assessed, by means of blood smear examination, PCR and DNA nucleotide sequencing, the presence of Babesia spp. and co-infecting agents Leishmania, Anaplasma/Ehrlichia and Hepatozoon in 45 dogs from northern Portugal clinically suspected of babesiosis. Results Forty-four dogs (98%) had infection with B. canis canis and one with B. canis vogeli. Co-infections were detected in nine animals (20%). Eight dogs were found infected with two vector-borne agents: six with B. canis canis and Leishmania infantum; one with B. canis canis and Ehrlichia canis; and one with B. canis canis and Hepatozoon canis. Another dog was infected with three vector-borne pathogens: B. canis vogeli, E. canis and L. infantum. Overall, L. infantum was found in seven (16%), E. canis in two (4%), and H. canis in one (2%) out of the 45 dogs with babesiosis. Almost 90% of the 45 cases of canine babesiosis were diagnosed in the colder months of October (18%), November (27%), December (20%), February (13%) and March (9%). Co-infections were detected in February, March, April, May, October and November. Twenty-two (50%) out of 44 dogs infected with B. canis were found infested by ticks including Dermacentor spp., Ixodes spp. and Rhipicephalus sanguineus. Mortality (9%) included two co-infected dogs that died spontaneously and two with single infections that were euthanized. Conclusions Babesia canis canis is the main etiological agent of canine babesiosis in northern Portugal. A higher sensitivity of Babesia spp. detection was obtained with PCR assays, compared to the observation of blood smears. Twenty percent of the dogs were co-infected with L. infantum
Full Text Available Tick-borne diseases are increasing all over the word, including Turkey. The aim of this study was to determine the bacterial and protozoan vector-borne pathogens in ticks infesting humans in the Corum province of Turkey.From March to November 2014 a total of 322 ticks were collected from patients who attended the local hospitals with tick bites. Ticks were screened by real time-PCR and PCR, and obtained amplicons were sequenced. The dedected tick was belonging to the genus Hyalomma, Haemaphysalis, Rhipicephalus, Dermacentor and Ixodes. A total of 17 microorganism species were identified in ticks. The most prevalent Rickettsia spp. were: R. aeschlimannii (19.5%, R. slovaca (4.5%, R. raoultii (2.2%, R. hoogstraalii (1.9%, R. sibirica subsp. mongolitimonae (1.2%, R. monacensis (0.31%, and Rickettsia spp. (1.2%. In addition, the following pathogens were identified: Borrelia afzelii (0.31%, Anaplasma spp. (0.31%, Ehrlichia spp. (0.93%, Babesia microti (0.93%, Babesia ovis (0.31%, Babesia occultans (3.4%, Theileria spp. (1.6%, Hepatozoon felis (0.31%, Hepatozoon canis (0.31%, and Hemolivia mauritanica (2.1%. All samples were negative for Francisella tularensis, Coxiella burnetii, Bartonella spp., Toxoplasma gondii and Leishmania spp.Ticks in Corum carry a large variety of human and zoonotic pathogens that were detected not only in known vectors, but showed a wider vector diversity. There is an increase in the prevalence of ticks infected with the spotted fever group and lymphangitis-associated rickettsiosis, while Ehrlichia spp. and Anaplasma spp. were reported for the first time from this region. B. microti was detected for the first time in Hyalomma marginatum infesting humans. The detection of B. occultans, B. ovis, Hepatozoon spp., Theileria spp. and Hemolivia mauritanica indicate the importance of these ticks as vectors of pathogens of veterinary importance, therefore patients with a tick infestation should be followed for a variety of pathogens
Penzhorn, Barend L; Netherlands, Edward C; Cook, Courtney A; Smit, Nico J; Vorster, Ilse; Harrison-White, Robert F; Oosthuizen, Marinda C
Domestic dogs are not native to sub-Saharan Africa, which may account for their susceptibility to Babesia rossi, of which endemic black-backed jackals (Canis mesomelas) are natural reservoirs. There is virtually no information on the occurrence of potentially pathogenic haemogregarines (e.g. Hepatozoon canis) or even rickettsial bacteria (e.g. Ehrlichia spp. and Anaplasma spp.) in indigenous canids in sub-Saharan Africa. Such organisms could pose a risk to domestic dogs, as well as to populations of endangered indigenous canid species. Genomic DNA extracted from blood samples taken from 126 free-ranging and 16 captive black-backed jackals was subjected to reverse line blot (RLB) hybridization assay; 82 (57.8%) specimens reacted only with the Ehrlichia/Anaplasma genera-specific probe. Full-length bacterial 16S rRNA gene of five of these specimens was cloned and the recombinants sequenced. The ten 16S rDNA sequences obtained were most closely related, with approximately 99% identity, to Anaplasma sp. South African Dog, various uncultured Anaplasma spp., as well as various Anaplasma phagocytophilum genotypes. Ninety-one specimens were screened for haemogregarines through PCR amplification using the 18S rRNA gene; 20 (21.9%) specimens reacted positively, of which 14 (15.4%) were confirmed positive for Hepatozoon genotypes from within H. canis. Two (2.2%) specimens were found positive for two different Hepatozoon genotypes. Sequence analyses confirmed the presence of 16S rDNA sequences closely related to A. phagocytophilum and Anaplasma sp. South African Dog as well as two H. canis genotypes in both free-ranging and captive black-backed jackals. Distinguishing between closely related lineages may provide insight into differences in pathogenicity and virulence of various Anaplasma and H. canis genotypes. By building up a more comprehensive understanding of the range and diversity of the bacteria and eukaryotic organisms (piroplasms and haemogregarines) in the blood of
Coleman Glen T
Full Text Available Abstract Background There are few published reports on canine Babesia, Ehrlichia, Anaplasma, Hepatozoon and haemotropic Mycoplasma infections in India and most describe clinical disease in individual dogs, diagnosed by morphological observation of the microorganisms in stained blood smears. This study investigated the occurrence and distribution of canine tick-borne disease (TBD pathogens using a combination of conventional and molecular diagnostic techniques in four cities in India. Results On microscopy examination, only Hepatozoon gamonts were observed in twelve out of 525 (2.3%; 95% CI: 1.2, 4 blood smears. Using polymerase chain reaction (PCR, a total of 261 from 525 dogs (49.7%; 95% CI: 45.4, 54.1 in this study were infected with one or more canine tick-borne pathogen. Hepatozoon canis (30%; 95% CI: 26.0, 34.0 was the most common TBD pathogen found infecting dogs in India followed by Ehrlichia canis (20.6%; 95% CI: 17.2, 24.3, Mycoplasma haemocanis (12.2%; 95% CI: 9.5, 15.3, Anaplasma platys (6.5%; 95% CI: 4.5, 8.9, Babesia vogeli (5.5%, 95% CI: 3.7, 7.8 and Babesia gibsoni (0.2%, 95% CI: 0.01, 1.06. Concurrent infection with more than one TBD pathogen occurred in 39% of cases. Potential tick vectors, Rhipicephalus (most commonly and/or Haemaphysalis ticks were found on 278 (53% of dogs examined. Conclusions At least 6 species of canine tick-borne pathogens are present in India. Hepatozoon canis was the most common pathogen and ticks belonging to the genus Rhipicephalus were encountered most frequently. Polymerase chain reaction was more sensitive in detecting circulating pathogens compared with peripheral blood smear examination. As co-infections with canine TBD pathogens were common, Indian veterinary practitioners should be cognisant that the discovery of one such pathogen raises the potential for multiple infections which may warrant different clinical management strategies.
Full Text Available Abstract Background Protozoa and bacteria transmitted by arthropods, including ticks and phlebotomine sand flies, may cause a wide range of canine vector-borne diseases. Dogs can be simultaneously or sequentially infected with multiple pathogens. Canine babesiosis caused by Babesia canis canis and Babesia canis vogeli is known to occur in Portugal. This study assessed, by means of blood smear examination, PCR and DNA nucleotide sequencing, the presence of Babesia spp. and co-infecting agents Leishmania, Anaplasma/Ehrlichia and Hepatozoon in 45 dogs from northern Portugal clinically suspected of babesiosis. Results Forty-four dogs (98% had infection with B. canis canis and one with B. canis vogeli. Co-infections were detected in nine animals (20%. Eight dogs were found infected with two vector-borne agents: six with B. canis canis and Leishmania infantum; one with B. canis canis and Ehrlichia canis; and one with B. canis canis and Hepatozoon canis. Another dog was infected with three vector-borne pathogens: B. canis vogeli, E. canis and L. infantum. Overall, L. infantum was found in seven (16%, E. canis in two (4%, and H. canis in one (2% out of the 45 dogs with babesiosis. Almost 90% of the 45 cases of canine babesiosis were diagnosed in the colder months of October (18%, November (27%, December (20%, February (13% and March (9%. Co-infections were detected in February, March, April, May, October and November. Twenty-two (50% out of 44 dogs infected with B. canis were found infested by ticks including Dermacentor spp., Ixodes spp. and Rhipicephalus sanguineus. Mortality (9% included two co-infected dogs that died spontaneously and two with single infections that were euthanized. Conclusions Babesia canis canis is the main etiological agent of canine babesiosis in northern Portugal. A higher sensitivity of Babesia spp. detection was obtained with PCR assays, compared to the observation of blood smears. Twenty percent of the dogs were co
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.
Full Text Available Although diverse tick-borne pathogens (TBPs are endemic to East Africa, with recognized impact on human and livestock health, their diversity and specific interactions with tick and vertebrate host species remain poorly understood in the region. In particular, the role of reptiles in TBP epidemiology remains unknown, despite having been implicated with TBPs of livestock among exported tortoises and lizards. Understanding TBP ecologies, and the potential role of common reptiles, is critical for the development of targeted transmission control strategies for these neglected tropical disease agents. During the wet months (April–May; October–December of 2012–2013, we surveyed TBP diversity among 4,126 ticks parasitizing livestock and reptiles at homesteads along the shores and islands of Lake Baringo and Lake Victoria in Kenya, regions endemic to diverse neglected tick-borne diseases. After morphological identification of 13 distinct Rhipicephalus, Amblyomma, and Hyalomma tick species, ticks were pooled (≤8 individuals by species, host, sampling site, and collection date into 585 tick pools. By supplementing previously established molecular assays for TBP detection with high-resolution melting analysis of PCR products before sequencing, we identified high frequencies of potential disease agents of ehrlichiosis (12.48% Ehrlichia ruminantium, 9.06% Ehrlichia canis, anaplasmosis (6.32% Anaplasma ovis, 14.36% Anaplasma platys, and 3.08% Anaplasma bovis,, and rickettsiosis (6.15% Rickettsia africae, 2.22% Rickettsia aeschlimannii, 4.27% Rickettsia rhipicephali, and 4.95% Rickettsia spp., as well as Paracoccus sp. and apicomplexan hemoparasites (0.51% Theileria sp., 2.56% Hepatozoon fitzsimonsi, and 1.37% Babesia caballi among tick pools. Notably, we identified E. ruminantium in both Amblyomma and Rhipicephalus pools of ticks sampled from livestock in both study areas as well as in Amblyomma falsomarmoreum (66.7% and Amblyomma nuttalli (100
Movilla, Rebeca; Altet, Laura; Serrano, Lorena; Tabar, María-Dolores; Roura, Xavier
The spleen is a highly perfused organ involved in the immunological control and elimination of vector-borne pathogens (VBP), which could have a fundamental role in the pathogenesis of splenic disease. This study aimed to evaluate certain VBP in samples from dogs with splenic lesions. Seventy-seven EDTA-blood and 64 splenic tissue samples were collected from 78 dogs with splenic disease in a Mediterranean area. Babesia spp., Bartonella spp., Ehrlichia/Anaplasma spp., Hepatozoon canis, Leishmania infantum, hemotropic Mycoplasma spp. and Rickettsia spp. were targeted using PCR assays. Sixty EDTA-blood samples from dogs without evidence of splenic lesions were included as a control group. More than half (51.56%) of the biopsies (33/64) were consistent with benign lesions and 48.43% (31/64) with malignancy, mostly hemangiosarcoma (25/31). PCR yielded positive results in 13 dogs with spleen alterations (16.67%), for Babesia canis (n = 3), Babesia gibsoni (n = 2), hemotropic Mycoplasma spp. (n = 2), Rickettsia massiliae (n = 1) and "Babesia vulpes" (n = 1), in blood; and for B. canis, B. gibsoni, Ehrlichia canis and L. infantum (n = 1 each), in spleen. Two control dogs (3.3%) were positive for B. gibsoni and H. canis (n = 1 each). Benign lesions were detected in the 61.54% of infected dogs (8/13); the remaining 38.46% were diagnosed with malignancies (5/13). Infection was significantly associated to the presence of splenic disease (P = 0.013). There was no difference in the prevalence of infection between dogs with benign and malignant splenic lesions (P = 0.69); however B. canis was more prevalent in dogs with hemangiosarcoma (P = 0.006). VBP infection could be involved in the pathogenesis of splenic disease. The immunological role of the spleen could predispose to alterations of this organ in infected dogs. Interestingly, all dogs with B. canis infection were diagnosed with hemangiosarcoma in the present survey. As previously
Full Text Available Tick-borne diseases (TBDs cause significant losses among livestock and impact the livelihoods of resource-poor farming communities worldwide. In Ethiopia, detailed studies on the epidemiology of tick-borne pathogens (TBPs in cattle using sensitive molecular detection methods are scarce. The objective of this study was to determine the prevalence and species composition of bovine TBPs of veterinary significance in local cattle populations. A comprehensive cross-sectional epidemiological study was conducted in cattle populations of Illubabor zone in Southwestern Ethiopia from June to August 2013. For this purpose, blood samples were collected from 392 cattle. A combination of polymerase chain reaction (PCR and a Reverse Line Blot (RLB hybridization assay was employed for the detection of TBPs in these samples. The PCR/RLB results of the 392 blood samples indicated a high overall prevalence of 96.9% for TBPs, including Theileria mutans (66.1%, Theileria orientalis (51.8%, Anaplasma sp. Omatjenne (25.5%, Anaplasma marginale (14.5%, Babesia bigemina (14.0% and Theileria velifera (13.0% and minor occurrences of Ehrlichia ruminantium (0.5% and Ehrlichia minasensis (0.26%. Moreover, three novel Anaplasma genotypes were detected in bovine blood samples. A phylogenetic analysis revealed that they most likely represent three, but at least two, new species. The prevalence of the three novel Anaplasma species, preliminary designated as Anaplasma sp. Hadesa, Anaplasma sp. Saso and Anaplasma sp. Dedessa, was 12.5%, 14.3% and 5.6%, respectively. Overall, a total of 227 cattle (57.9% were found to be co-infected with two or more TBPs simultaneously and 86 different species combinations were observed. The findings show a very high burden of infection of cattle with TBPs in Ethiopia. The high frequency of co-infections suggests that clinical manifestations might be complex. Further research is required to determine the pathogenicity, host cell types and vector of
Full Text Available Abstract Background A variety of human diseases transmitted by arthropod vectors, including ticks, are emerging around the globe. Birds are known to be hosts of ticks and can disperse exotic ticks and tick-borne pathogens. In Taiwan, previous studies have focused predominantly on mammals, leaving the role of birds in the maintenance of ticks and dissemination of tick-borne pathogens undetermined. Methods Ticks were collected opportunistically when birds were studied from 1995 to 2013. Furthermore, to improve knowledge on the prevalence and mean load of tick infestation on birds in Taiwan, ticks were thoroughly searched for when birds were mist-netted at seven sites between September 2014 and April 2016 in eastern Taiwan. Ticks were identified based on both morphological and molecular information and were screened for potential tick-borne pathogens, including the genera Anaplasma, Babesia, Borrelia, Ehrlichia and Rickettsia. Finally, a list of hard tick species collected from birds in Taiwan was compiled based on past work and the current study. Results Nineteen ticks (all larvae were recovered from four of the 3096 unique mist-netted bird individuals, yielding a mean load of 0.006 ticks/individual and an overall prevalence of 0.13%. A total of 139 ticks from birds, comprising 48 larvae, 35 nymphs, 55 adults and one individual of unknown life stage, were collected from 1995 to 2016, and 11 species of four genera were identified, including three newly recorded species (Haemaphysalis wellingtoni, Ixodes columnae and Ixodes turdus. A total of eight tick-borne pathogens were detected, with five species (Borrelia turdi, Anaplasma sp. clone BJ01, Ehrlichia sp. BL157-9, Rickettsia helvetica and Rickettsia monacensis not previously isolated in Taiwan. Overall, 16 tick species of five genera have been recorded feeding on birds, including nine species first discovered in this study. Conclusion Our study demonstrates the paucity of information on ticks of
Omondi, David; Masiga, Daniel K; Fielding, Burtram C; Kariuki, Edward; Ajamma, Yvonne Ukamaka; Mwamuye, Micky M; Ouso, Daniel O; Villinger, Jandouwe
Although diverse tick-borne pathogens (TBPs) are endemic to East Africa, with recognized impact on human and livestock health, their diversity and specific interactions with tick and vertebrate host species remain poorly understood in the region. In particular, the role of reptiles in TBP epidemiology remains unknown, despite having been implicated with TBPs of livestock among exported tortoises and lizards. Understanding TBP ecologies, and the potential role of common reptiles, is critical for the development of targeted transmission control strategies for these neglected tropical disease agents. During the wet months (April-May; October-December) of 2012-2013, we surveyed TBP diversity among 4,126 ticks parasitizing livestock and reptiles at homesteads along the shores and islands of Lake Baringo and Lake Victoria in Kenya, regions endemic to diverse neglected tick-borne diseases. After morphological identification of 13 distinct Rhipicephalus, Amblyomma , and Hyalomma tick species, ticks were pooled (≤8 individuals) by species, host, sampling site, and collection date into 585 tick pools. By supplementing previously established molecular assays for TBP detection with high-resolution melting analysis of PCR products before sequencing, we identified high frequencies of potential disease agents of ehrlichiosis (12.48% Ehrlichia ruminantium , 9.06% Ehrlichia canis ), anaplasmosis (6.32% Anaplasma ovis , 14.36% Anaplasma platys , and 3.08% Anaplasma bovis ,), and rickettsiosis (6.15% Rickettsia africae , 2.22% Rickettsia aeschlimannii , 4.27% Rickettsia rhipicephali , and 4.95% Rickettsia spp.), as well as Paracoccus sp. and apicomplexan hemoparasites (0.51% Theileria sp., 2.56% Hepatozoon fitzsimonsi , and 1.37% Babesia caballi ) among tick pools. Notably, we identified E. ruminantium in both Amblyomma and Rhipicephalus pools of ticks sampled from livestock in both study areas as well as in Amblyomma falsomarmoreum (66.7%) and Amblyomma nuttalli (100%) sampled
Cecília Braga Laposy
Full Text Available In order to study the biochemical changes in naturally infected dogs with Ehrlichia canis, blood samples from 57 animals were collected. Thirty-five healthy dogs were used as control group. Dosage of serum protein, albumin, globulin, albumin:globulin ratio (A:G, urea, creatinine, alanine aminotransferase (ALT e alkaline phosphatase (AP were carried out. The comparison of mean in infected and control group as well as the distribution of frequencies were investigated in order to associate the disease to the findings. Hypoproteinemia, hypoalbuminemia, decreased A:G ratio, hyperglobulinemia, and uremia were associated to ehrlichiosis (p Com o objetivo de estudar os parâmetros bioquímicos associados às funções renal e hepática em cães naturalmente infectados por Ehrlichia canis e associá-los à ehrlichiose, foram colhidas 57 amostras de sangue, com realização de dosagem de proteína plasmática total, albumina, globulina, relação albumina/ globulina (A/G, uréia, creatinina, alanino aminotransferase (ALT e fosfatase alcalina (FA. Outros 35 cães em higidez foram utilizados como controle. Foram comparadas as médias dos animais infectados e controle, bem como a distribuição de freqüências, para associação dos resultados com a doença. Hipoproteinemia, hipoalbuminemia, redução da A/G, hiperglobulinemia e aumento de uréia estiveram associados à doença (p < 0,05. Foram comparados também os achados entre animais não pancitopênicos (NPan e em pancitopenia (Pan, verificando-se que no primeiro grupo, houve aumento de ALT, e nos animais pancitopênicos, a creatinina foi o principal indicador de ehrlichiose. No caso do perfil protéico, não houve diferença que pudesse servir como marcador da fase da enfermidade nos animais infectados.
Woodroffe, Rosie; Prager, Katherine C.; Conrad, Patricia A.; Dubovi, Edward J.; Mazet, Jonna A. K.
Background Infectious diseases have contributed to the decline and local extinction of several wildlife species, including African wild dogs (Lycaon pictus). Mitigating such disease threats is challenging, partly because uncertainty about disease dynamics makes it difficult to identify the best management approaches. Serious impacts on susceptible populations most frequently occur when generalist pathogens are maintained within populations of abundant (often domestic) “reservoir” hosts, and spill over into less abundant host species. If this is the case, disease control directed at the reservoir host might be most appropriate. However, pathogen transmission within threatened host populations may also be important, and may not be controllable by managing another host species. Methodology/Principal Findings We investigated interspecific and intraspecific transmission routes, by comparing African wild dogs' exposure to six canine pathogens with behavioural measures of their opportunities for contact with domestic dogs and with other wild dogs. Domestic dog contact was associated with exposure to canine parvovirus, Ehrlichia canis, Neospora caninum and perhaps rabies virus, but not with exposure to canine distemper virus or canine coronavirus. Contact with other wild dogs appeared not to increase the risk of exposure to any of the pathogens. Conclusions/Significance These findings, combined with other data, suggest that management directed at domestic dogs might help to protect wild dog populations from rabies virus, but not from canine distemper virus. However, further analyses are needed to determine the management approaches – including no intervention – which are most appropriate for each pathogen. PMID:22238695
Bronson, Ellen; Spiker, Harry; Driscoll, Cindy P
American black bears (Ursus americanus) in Maryland, USA, live in forested areas in close proximity to humans and their domestic pets. From 1999 to 2011, we collected 84 serum samples from 63 black bears (18 males; 45 females) in five Maryland counties and tested them for exposure to infectious, including zoonotic, pathogens. A large portion of the bears had antibody to canine distemper virus and Toxoplasma gondii, many at high titers. Prevalences of antibodies to zoonotic agents such as rabies virus and to infectious agents of carnivores including canine adenovirus and canine parvovirus were lower. Bears also had antibodies to vector-borne pathogens common to bears and humans such as West Nile virus, Borrelia burgdorferi, Rickettsia rickettsii, and Anaplasma phagocytophilum. Antibodies were detected to Leptospira interrogans serovars Pomona, Icterohaemorrhagiae, Canicola, Grippotyphosa, and Bratislava. We did not detect antibodies to Brucella canis or Ehrlichia canis. Although this population of Maryland black bears demonstrated exposure to multiple pathogens of concern for humans and domesticated animals, the low levels of clinical disease in this and other free-ranging black bear populations indicate the black bear is likely a spillover host for the majority of pathogens studied. Nevertheless, bear populations living at the human-domestic-wildlife interface with increasing human and domestic animal exposure should continue to be monitored because this population likely serves as a useful sentinel of ecosystem health.
Fesler, Melissa C; Middelveen, Marianne J; Stricker, Raphael B
Morgellons disease (MD) is a dermatological condition characterized by aberrant production of keratin and collagen fibers in skin. Although infection with Borrelia burgdorferi , the causative agent of Lyme disease (LD), has been associated with MD, relatively few studies have hitherto provided epidemiological evidence regarding this association. A cohort of 1000 seropositive North American LD patients was evaluated for the presence of MD. Patients were diagnosed with MD based on detection of microscopic fibers in skin lesions or under unbroken skin. Demographic and clinical features of MD patients were analyzed, and laboratory testing for tickborne coinfections and other infectious agents, was performed. Subjective and objective features of MD were analyzed using statistical methods. Of 1000 seropositive LD patients, 60 (6%) were diagnosed with MD. Of these 60 patients, 75% were female and 78% presented in the late disseminated stage of MD. All 60 patients (100%) were seropositive for B. burgdorferi infection. Tickborne coinfections in these patients included Babesia spp (62%), Bartonella and Rickettsia (25% each), Ehrlichia (15%) and Anaplasma (10%). Helicobacter pylori was detected in 12% of MD patients. In all, 77% of MD patients had one or more coinfections. This study confirms recent findings that MD occurs in a limited subset of LD patients. The clinical and genetic determinants of MD in LD patients require further study.
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.
Bruce H. Noden
Full Text Available Namibia has a rich history in veterinary health but little is known about the vector-borne diseases that affect companion dogs and cats. The aim of this review is to summarise the existing published and available unpublished literature, put it into a wider geographical context, and explore some significant knowledge gaps. To date, only two filarial pathogens (Dirofilaria repens and Acanthocheilonema dracunculoides and three tick-borne pathogens (Babesia canis vogeli, Hepatozoon canis and Ehrlichia canis have been reported. Most studies have focused solely on dogs and cats in the urban Windhoek and surrounding areas, with almost nothing reported in rural farming areas, in either the populous northern regions or the low-income urban areas where animal owners have limited access to veterinary services. With the development of several biomedical training programmes in the country, there is now an excellent opportunity to address zoonotic vector-borne diseases through a One Health approach so as to assess the risks to small companion animals as well as diseases of public health importance.
Full Text Available Background: We attempted to determine the prevalence of Hepatozoon spp. infection in Mashhad, northeast of Iran, via blood smear parasitology.Methods: The prevalence was investigated by examination of blood smear parasitology, using blood samples collected from 254 dogs (51 strays and 203 privately owned-dogs.Results: Two stray dogs (2/51; 3.92% and two privately-owned dogs (2/203; 0.98% were infected with Hepatozoon spp. Therefore, as per blood smear parasitology, the prevalence of Hepatozoon spp. infection was 1.57% (4/254. Sixteen out of 254 dogs (6.29% were infested with ticks; all of which were Rhipicephalus sanguineus. One of the dogs infected with Hepatozoon spp. exhibited ticks at the time of examination. Concurrent infection with Ehrlichia canis and Leishmania infantum was not detected in the four Hepatozoon spp. infected dogs.Conclusion: This is the first epidemiological study on the prevalence of Hepatozoon spp. infection in dogs in Iran.
Lakshmanan, Bindu; Jose, K Jain; George, Arun; Usha, N P; Devada, K
India has a wide range of agro-climatic zones which is highly conducive for a diverse range of vectors and canines are continuously exposed to the risk of spectrum of tick borne protozoan diseases. The brown dog tick, Rhipicephalus sanguineus is widely prevalent among dogs in Kerala and there is a high prevalence of this tick transmitted Babesia and Ehrlichia spp. infection. However, the incidence of Hepatozoon canis transmitted by the same tick species had not been reported in the state since 2004. Preliminary screening of client owned dogs revealed six dogs to be positive for typical gelatin capsule shaped gamonts of H. canis within neutrophils in blood smear by microscopic examination. A PCR assay was standardized to amplify a specific 737 bp fragment of 18S rRNA gene of H. canis. Phylogenetic analysis revealed closest relationship with West Indies isolate deposited at GenBank database. The present study records the molecular detection of this haemoparasite in the state, for the first time.
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.
Full Text Available Objective: To explore haemato-biochemical and oxidative stress indices due to concomitant tick borne intracellular diseases in dogs presented at Referral Veterinary Polyclinic, Indian Veterinary Research Institute, Bareilly during May 2010 to May 2012. Methods: Microscopy of Giemsa blood smear and ELISA test (SNAP 4D伊 were carried out in suspected cases to confirm haemo-parasitic infection. Blood and serum samples were analyzed for oxidative stress indices and haemato-biochemical changes. All the ailing conditions were recorded to investigate the clinical pattern of concomitant tick borne diseases. Ultrasonographic study was carried out to obtain the hepatic involvement. Results: Examination of 3 650 dogs revealed that 2.77% dog were positive for various tick borne diseases, out of which 21.78% were with concomitant infection. Clinical symptoms were noted with overall mean clinical score of 9.95依0.30. Ultrasonographic examination revealed hepatomegaly, distension of gall bladder, and ascites. Haemato-biochemical evaluation confirmed anaemia, leucopenia, thrombocytopenia, hypoproteinemia, hypoalbuminemia, hyperglobulinemia and hyperbilirubinemia with increased serum alanine amino transferase, alkaline phosphatase and gamma-glutamyl transpeptidase in concomitant infected dogs. The lipid peroxidation level of concomitant infection was significantly higher (P<0.05 than healthy group whereas superoxide dismutase, glutathione-reduced and catalase activity in concomitant infected group were decreased. Conclusions: The severity of infection was more pronounced in dogs harboring Ehrlichia, Babesia and Hepatozoon and the oxidative stress may have a pathophysiological role in concomitant infection in dogs.
Chhabra, Sushma; Uppal, Sanjeev Kumar; Singla, Lachhman Das
Objective To evaluate clinical and hematological aspects of dogs naturally infected with Hepatozoon canis (H. canis) presented at the Small Animal Clinics of Guru Angad Dev Veterinary and Animal Sciences University, Ludhiana. Methods Blood films of 34 naturally infected dogs were examined for haematological alterations and parasitaemia. Signalment and clinical signs were recorded from the animals. Clinical histories were filled out during the consultation. Results Of the 34 positive dogs by Giemsa stained peripheral blood films, 88.23% presented parasitaemia by H. canis only, while 11.77% had the combination of H. canis, Babesia sp. and Ehrlichia sp. Young male dogs less than one-year-old, of non-descript breed, were the most commonly affected. And 26.47% were presented with anorexia/inappetence as the only clinical symptom. Other clinical symptoms were mild to moderate fever, pale mucosae and lethargy; a few were also showing the signs of vomiting and diarrhoea. Haematological alterations showed mainly normochromic-normocytic anaemia, leukocytosis and neutrophilia. Conclusions The findings of this study substantiate that H. canis caused clinical and haematological alterations of the varied intensity in dogs, even with low parasitaemia, should be taken into consideration. PMID:23730562
Chhabra, Sushma; Uppal, Sanjeev Kumar; Singla, Lachhman Das
To evaluate clinical and hematological aspects of dogs naturally infected with Hepatozoon canis (H. canis) presented at the Small Animal Clinics of Guru Angad Dev Veterinary and Animal Sciences University, Ludhiana. Blood films of 34 naturally infected dogs were examined for haematological alterations and parasitaemia. Signalment and clinical signs were recorded from the animals. Clinical histories were filled out during the consultation. Of the 34 positive dogs by Giemsa stained peripheral blood films, 88.23% presented parasitaemia by H. canis only, while 11.77% had the combination of H. canis, Babesia sp. and Ehrlichia sp. Young male dogs less than one-year-old, of non-descript breed, were the most commonly affected. And 26.47% were presented with anorexia/inappetence as the only clinical symptom. Other clinical symptoms were mild to moderate fever, pale mucosae and lethargy; a few were also showing the signs of vomiting and diarrhoea. Haematological alterations showed mainly normochromic-normocytic anaemia, leukocytosis and neutrophilia. The findings of this study substantiate that H. canis caused clinical and haematological alterations of the varied intensity in dogs, even with low parasitaemia, should be taken into consideration.
Collett, M G; Doyle, A S; Reyers, F; Kruse, T; Fabian, B
Laboratory findings in an adult bull terrier presented with a history of anorexia and weight loss included the following: severe anaemia, leukocytosis, neutrophilia, lymphopaenia, thrombocytopaenia, Ehrlichia canis morulae in monocytes, hypergammaglo-bulinaemia, a bleeding tendency, icterus and proteinuria. In addition, a high Haemobartonella canis parasitaemia, non-encapsulated yeasts on urinalysis and a localised Demodex canis infestation were present. Treatment for ehrlichiosis was initiated but the dog died. Lesions found were a severe cryptococcal granulomatous pneumonia and cryptococcal colonies in the lungs, bronchial lymph nodes, kidneys, liver, spleen, heart, meninges, eyes and thoracic cavity. In addition, hyphal forms resembling Filobasidiella neoformans, the teleomorph of Cryptococcus neoformans, were seen in lung fine needle aspiration smears, impression smears and lung sections. C. neoformans was cultured from urine, lung and liver. Lung and kidney also yielded Salmonella typhimureum. Cortical atrophy with T-cell depletion of lymph nodes as well as splenic lymphoid follicular atrophy, typical of chronic ehrlichiosis-induced cell mediated immunosuppression, could have predisposed to the fatal disseminated cryptococcis.
Barrett, Anne W; Little, Susan E
To determine the prevalence of infection with vector-borne agents in a cross-section of dogs from Oklahoma, where canine vector-borne diseases are common, blood samples were evaluated through serology and molecular analysis. Antibodies reactive to Ehrlichia spp., Rickettsia rickettsii, R. montanensis, and "R. amblyommii" were detected in 10.5% (11/105), 74.3% (78/105), 58.1% (61/105), and 55.2% (58/105) of dogs, respectively. Presence of spotted fever group Rickettsia spp. DNA was identified in 13.1% (8/61) of shelter dogs but not in any pet dogs (0/44). DNA of "R. amblyommii" was confirmed by sequencing, constituting the first report of this agent in a naturally infected dog. Antigen of Dirofilaria immitis was detected in 10.5% (11/105) and 16.2% (17/105) of samples before and after heat treatment, respectively. In total, 87.6% (92/105) of the dogs had evidence of infection with at least one vector-borne disease agent, confirming high risk of exposure to multiple vector-borne disease agents, several of which are zoonotic.
Lv, Jizhou; Fernández de Marco, Maria Del Mar; Goharriz, Hooman; Phipps, L Paul; McElhinney, Lorraine M; Hernández-Triana, Luis M; Wu, Shaoqiang; Lin, Xiangmei; Fooks, Anthony R; Johnson, Nicholas
Ticks host a wide range of zoonotic pathogens and are a significant source of diseases that affect humans and livestock. However, little is known about the pathogens associated with bat ticks. We have collected ectoparasites from bat carcasses over a seven year period. Nucleic acids (DNA and RNA) were extracted from 296 ticks removed from bats and the species designation was confirmed in all ticks as Argas (Carios) vespertilionis. A subset of these samples (n = 120) were tested for the presence of zoonotic pathogens by molecular methods. Babesia species, Rickettsia spp., within the spotted fever group (SFG), and Ehrlichia spp. were detected in ticks removed from 26 bats submitted from 14 counties across England. The prevalence of Rickettsia spp. was found to be highest in Pipistrellus pipistrellus from southern England. This study suggests that the tick species that host B. venatorum may include the genus Argas in addition to the genus Ixodes. As A. vespertilionis has been reported to feed on humans, detection of B. venatorum and SFG Rickettsia spp. could present a risk of disease transmission in England. No evidence for the presence of flaviviruses or Issyk-Kul virus (nairovirus) was found in these tick samples.
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.
Spolidorio, Mariana G.; Labruna, Marcelo B.; Machado, Rosangela Z.; Moraes-Filho, Jonas; Zago, Augusto M.; Donatele, Dirlei M.; Pinheiro, Sônia R.; Silveira, Iara; Caliari, Késia M.; Yoshinari, Natalino H.
Blood samples collected from 201 humans, 92 dogs, and 27 horses in the state of Espirito Santo, Brazil, were tested by polymerase chain reaction, indirect immunofluorescence assays, and indirect enzyme-linked immunosorbent assay for tick-borne diseases (rickettsiosis, ehrlichiosis, anaplasmosis, borreliosis, babesiosis). Our results indicated that the surveyed counties are endemic for spotted fever group rickettsiosis because sera from 70 (34.8%) humans, 7 (7.6%) dogs, and 7 (25.9%) horses were reactive to at least one of the six Rickettsia species tested. Although there was evidence of ehrlichiosis (Ehrlichia canis) and babesiosis (Babesia canis vogeli, Theileria equi) in domestic animals, no human was positive for babesiosis and only four individuals were serologically positive for E. canis. Borrelia burgdorferi-serologic reactive sera were rare among humans and horses, but encompassed 51% of the canine samples, suggesting that dogs and their ticks can be part of the epidemiological cycle of the causative agent of the Brazilian zoonosis, named Baggio-Yoshinari Syndrome. PMID:20595502
Chapes, Stephen Keith; Ganta, Roman Reddy; Chapers, S. K. (Principal Investigator)
Several immunological processes can be affected by space flight. However, there is little evidence to suggest that flight-induced immunological deficits lead to illness. Therefore, one of our goals has been to define models to examine host resistance during space flight. Our working hypothesis is that space flight crews will come from a heterogeneous population; the immune response gene make-up will be quite varied. It is unknown how much the immune response gene variation contributes to the potential threat from infectious organisms, allergic responses or other long term health problems (e.g. cancer). This article details recent efforts of the Kansas State University gravitational immunology group to assess how population heterogeneity impacts host health, either in laboratory experimental situations and/or using the skeletal unloading model of space-flight stress. This paper details our use of several mouse strains with several different genotypes. In particular, mice with varying MHCII allotypes and mice on the C57BL background with different genetic defects have been particularly useful tools with which to study infections by Staphylococcus aureus, Salmonella typhimurium, Pasteurella pneumotropica and Ehrlichia chaffeensis. We propose that some of these experimental challenge models will be useful to assess the effects of space flight on host resistance to infection.
Coelho, Eduardo Antonio Ferraz; Costa, Lourena Emanuele; Lage, Daniela Pagliara; Martins, Vívian Tamietti; Garde, Esther; de Jesus Pereira, Nathália Cristina; Lopes, Eliane Gonçalves Paiva; Borges, Luiz Felipe Nunes Menezes; Duarte, Mariana Costa; Menezes-Souza, Daniel; de Magalhães-Soares, Danielle Ferreira; Chávez-Fumagalli, Miguel Angel; Soto, Manuel; Tavares, Carlos Alberto Pereira
Serological diagnostic tests for canine and human leishmaniasis present problems related with their sensitivity and/or specificity. Recently, an immunoproteomic approach performed with Leishmania infantum proteins identified new parasite antigens. In the present study, the diagnostic properties of two of these proteins, cytochrome c oxidase and IgE-dependent histamine-releasing factor, were evaluated for the serodiagnosis of canine visceral (CVL) and human tegumentary (HTL) leishmaniasis. For the CVL diagnosis, sera samples from non-infected dogs living in an endemic or non-endemic area of leishmaniasis, sera from asymptomatic or symptomatic visceral leishmaniasis (VL) dogs, from Leish-Tec(®)-vaccinated dogs, and sera from animals experimentally infected by Trypanosoma cruzi or Ehrlichia canis were used. For the HTL diagnosis, sera from non-infected subjects living in an endemic area of leishmaniasis, sera from active cutaneous or mucosal leishmaniasis patients, as well as those from T. cruzi-infected patients were employed. ELISA assays using the recombinant proteins showed both sensitivity and specificity values of 100% for the serodiagnosis of both forms of disease, with high positive and negative predictive values, showing better diagnostic properties than the parasite recombinant A2 protein or a soluble Leishmania antigen extract. In this context, the two new recombinant proteins could be considered to be used in the serodiagnosis of CVL and HTL. Copyright © 2015 Elsevier B.V. All rights reserved.
Full Text Available Cholesterol is a multifunctional lipid that plays important metabolic and structural roles in the eukaryotic cell. Despite having diverse lifestyles, the obligate intracellular bacterial pathogens Chlamydia, Coxiella, Anaplasma, Ehrlichia, and Rickettsia all target cholesterol during host cell colonization as a potential source of membrane, as well as a means to manipulate host cell signaling and trafficking. To promote host cell entry, these pathogens utilize cholesterol-rich microdomains known as lipid rafts, which serve as organizational and functional platforms for host signaling pathways involved in phagocytosis. Once a pathogen gains entrance to the intracellular space, it can manipulate host cholesterol trafficking pathways to access nutrient-rich vesicles or acquire membrane components for the bacteria or bacteria-containing vacuole. To acquire cholesterol, these pathogens specifically target host cholesterol metabolism, uptake, efflux, and storage. In this review, we examine the strategies obligate intracellular bacterial pathogens employ to manipulate cholesterol during host cell colonization. Understanding how obligate intracellular pathogens target and use host cholesterol provides critical insight into the host-pathogen relationship.
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.
Full Text Available BACKGROUND: Infectious diseases have contributed to the decline and local extinction of several wildlife species, including African wild dogs (Lycaon pictus. Mitigating such disease threats is challenging, partly because uncertainty about disease dynamics makes it difficult to identify the best management approaches. Serious impacts on susceptible populations most frequently occur when generalist pathogens are maintained within populations of abundant (often domestic "reservoir" hosts, and spill over into less abundant host species. If this is the case, disease control directed at the reservoir host might be most appropriate. However, pathogen transmission within threatened host populations may also be important, and may not be controllable by managing another host species. METHODOLOGY/PRINCIPAL FINDINGS: We investigated interspecific and intraspecific transmission routes, by comparing African wild dogs' exposure to six canine pathogens with behavioural measures of their opportunities for contact with domestic dogs and with other wild dogs. Domestic dog contact was associated with exposure to canine parvovirus, Ehrlichia canis, Neospora caninum and perhaps rabies virus, but not with exposure to canine distemper virus or canine coronavirus. Contact with other wild dogs appeared not to increase the risk of exposure to any of the pathogens. CONCLUSIONS/SIGNIFICANCE: These findings, combined with other data, suggest that management directed at domestic dogs might help to protect wild dog populations from rabies virus, but not from canine distemper virus. However, further analyses are needed to determine the management approaches--including no intervention--which are most appropriate for each pathogen.
Carolina Guadalupe Sosa-Gutierrez
Full Text Available Ehrlichia canis is a rickettsial intracellular obligate bacterial pathogen and agent of canine monocytic ehrlichiosis. The prevalence of this disease in veterinary medicine can vary depending on the diagnostic method used and the geographic location. One hundred and fifty-two canine blood samples from six veterinary clinics and two shelters from Sinaloa State (Mexico were analyzed in this study. All animals were suspected of having Canine Monocytic Ehrlichiosis (CME. The diagnostic methods used were the ELISA (Snap4Dx, IDEXX together with blood smear and platelet count. From all dogs blood samples analyzed, 74.3% were positive to E. canis by ELISA and 40.1% were positive by blood smear. The sensitivity and specificity observed in the ELISA test were 78.8% and 86.7%. In addition, thrombocytopenia was presented in 87.6% of positive dogs. The predominant clinical manifestations observed were fever, anorexia, depression, lethargy, and petechiae. Consequently, this is the first report in which the morulae were visualized in the blood samples, and E. canis-specific antibodies were detected in dogs from Sinaloa, Northwest of Mexico.
Escobedo-Hinojosa, Wendy; Pardo-López, Liliana
Little is known about the diversity of bacteria in the Southwestern Gulf of Mexico. The aim of the study illustrated in this perspective was to search for the presence of bacterial pathogens in this ecosystem, using metagenomic data recently generated by the Mexican research group known as the Gulf of Mexico Research Consortium. Several genera of bacteria annotated as pathogens were detected in water and sediment marine samples. As expected, native and ubiquitous pathogenic bacteria genera such as Burkolderia, Halomonas, Pseudomonas, Shewanella and Vibrio were highly represented. Surprisingly, non-native genera of public health concern were also detected, including Borrelia, Ehrlichia, Leptospira, Mycobacterium, Mycoplasma, Salmonella, Staphylococcus, Streptococcus and Treponema. While there are no previous metagenomics studies of this environment, the potential influences of natural, anthropogenic and ecological factors on the diversity of putative pathogenic bacteria found in it are reviewed. The taxonomic annotation herein reported provides a starting point for an improved understanding of bacterial biodiversity in the Southwestern Gulf of Mexico. It also represents a useful tool in public health as it may help identify infectious diseases associated with exposure to marine water and ingestion of fish or shellfish, and thus may be useful in predicting and preventing waterborne disease outbreaks. © FEMS 2017. All rights reserved. For permissions, please e-mail: firstname.lastname@example.org.
Schroeder, H; Jardine, J E; Davis, V
An 8-year-old, Maltese-cross bitch presented with chronic neck and back pain and an acute onset of circling, hyperaesthesia and constant crying. Clinical examination revealed temporal muscle atrophy, an abnormal hanging reflex, cervical rigidity and severe hepatomegaly. Ultrasonography of the liver showed several disseminated, poorly demarcated, hypoechoic areas which on fine needle aspirates, contained large numbers of pigmented fungal hyphae. Cerebrospinal fluid examination revealed fungal hyphae and numerous Ehrlichia canis morulae. A diagnosis of systemic phaeohyphomycosis secondary to ehrlichiosis was proposed. Treatment was unsuccessful and the dog was euthanased. At necropsy, multiple yellowish-green to black, necro-granulomatous foci were found throughout the liver parenchyma and similar foci were present in the spleen, renal cortices and adrenal glands. Irregular, multifocal, grey to black foci of malacia were present in both the grey and the white matter of the brain. On histopathological examination pigmented fungal hyphae were demonstrated in the liver, spleen, kidneys, portal lymph node and adrenals, as well as in the brain. Cultures of various organs yielded a fungal organism identified as Xylohypha bantiana.
Horak, Ivan G; Heyne, Heloise; Halajian, Ali; Booysen, Shalaine; Smit, Willem J
The aim of the study was to determine the species spectrum of ixodid ticks that infest horses and donkeys in South Africa and to identify those species that act as vectors of disease to domestic livestock. Ticks were collected opportunistically from 391 horses countrywide by their owners or grooms, or by veterinary students and staff at the Faculty of Veterinary Science, University of Pretoria. Ticks were also collected from 76 donkeys in Limpopo Province, 2 in Gauteng Province and 1 in North West province. All the ticks were identified by means of a stereoscopic microscope. Horses were infested with 17 tick species, 72.1% with Rhipicephalus evertsi evertsi, 19.4% with Amblyomma hebraeum and 15.6% with Rhipicephalus decoloratus. Rhipicephalus evertsi evertsi was recovered from horses in all nine provinces of South Africa and R. decoloratus in eight provinces. Donkeys were infested with eight tick species, and 81.6% were infested with R. evertsi evertsi, 23.7% with A. hebraeum and 10.5% with R. decoloratus. Several tick species collected from the horses and donkeys are the vectors of economically important diseases of livestock. Rhipicephalus evertsi evertsi is the vector of Theileria equi, the causative organism of equine piroplasmosis. It also transmits Anaplasma marginale, the causative organism of anaplasmosis in cattle. Amblyomma hebraeum is the vector of Ehrlichia ruminantium, the causative organism of heartwater in cattle, sheep and goats, whereas R. decoloratus transmits Babesia bigemina, the causative organism of babesiosis in cattle.
Biggs, Holly M; Behravesh, Casey Barton; Bradley, Kristy K; Dahlgren, F Scott; Drexler, Naomi A; Dumler, J Stephen; Folk, Scott M; Kato, Cecilia Y; Lash, R Ryan; Levin, Michael L; Massung, Robert F; Nadelman, Robert B; Nicholson, William L; Paddock, Christopher D; Pritt, Bobbi S; Traeger, Marc S
Tickborne rickettsial diseases continue to cause severe illness and death in otherwise healthy adults and children, despite the availability of low-cost, effective antibacterial therapy. Recognition early in the clinical course is critical because this is the period when antibacterial therapy is most effective. Early signs and symptoms of these illnesses are nonspecific or mimic other illnesses, which can make diagnosis challenging. Previously undescribed tickborne rickettsial diseases continue to be recognized, and since 2004, three additional agents have been described as causes of human disease in the United States: Rickettsia parkeri, Ehrlichia muris-like agent, and Rickettsia species 364D. This report updates the 2006 CDC recommendations on the diagnosis and management of tickborne rickettsial diseases in the United States and includes information on the practical aspects of epidemiology, clinical assessment, treatment, laboratory diagnosis, and prevention of tickborne rickettsial diseases. The CDC Rickettsial Zoonoses Branch, in consultation with external clinical and academic specialists and public health professionals, developed this report to assist health care providers and public health professionals to 1) recognize key epidemiologic features and clinical manifestations of tickborne rickettsial diseases, 2) recognize that doxycycline is the treatment of choice for suspected tickborne rickettsial diseases in adults and children, 3) understand that early empiric antibacterial therapy can prevent severe disease and death, 4) request the appropriate confirmatory diagnostic tests and understand their usefulness and limitations, and 5) report probable and confirmed cases of tickborne rickettsial diseases to public health authorities.
Dumler J Stephen
Full Text Available Abstract Background Human monocytic ehrlichiosis (HME and Rocky Mountain spotted fever (RMSF are caused by Ehrlichia chaffeensis and Rickettsia rickettsii, respectively. The pathogenesis of RMSF relates to rickettsia-mediated vascular injury, but it is unclear in HME. Methods To study histopathologic responses in the lymphatic system for correlates of immune injury, lymph nodes from patients with HME (n = 6 and RMSF (n = 5 were examined. H&E-stained lymph node tissues were examined for five histopathologic features, including hemophagocytosis, cellularity, necrosis, and vascular congestion and edema. The relative proportions of CD68 macrophages, CD8 and CD4 T lymphocytes, and CD20 B lymphocytes were evaluated by immunohistochemical staining. Results Hemophagocytosis was similar in HME and RMSF, and was greater than in control cases (p = .015. Cellularity in HME was not different from controls, whereas RMSF lymph nodes were markedly less cellular (p E. chaffeensis-infected mononuclear phagocytes were infrequent compared to R. rickettsii-infected endothelial cells. More CD8 cells in lymph nodes were observed with HME (p Conclusion Hemophagocytosis, CD8 T cell expansion, and the paucity of infected cells in HME, suggest that E. chaffeensis infection leads to macrophage activation and immune-mediated injury.
Hailemariam, Zerihun; Ahmed, Jabbar Sabir; Clausen, Peter-Henning; Nijhof, Ard Menzo
An essential step in the molecular detection of tick-borne pathogens (TBPs) in blood is the extraction of DNA. When cooled storage of blood under field conditions prior to DNA extraction in a dedicated laboratory is not possible, the storage of blood on filter paper forms a promising alternative. We evaluated six DNA extraction methods from blood spotted on FTA Classic ® cards (FTA cards), to determine the optimal protocol for the subsequent molecular detection of TBPs by PCR and the Reverse Line Blot hybridization assay (RLB). Ten-fold serial dilutions of bovine blood infected with Babesia bovis, Theileria mutans, Anaplasma marginale or Ehrlichia ruminantium were made by dilution with uninfected blood and spotted on FTA cards. Subsequently, DNA was extracted from FTA cards using six different DNA extraction protocols. DNA was also isolated from whole blood dilutions using a commercial kit. PCR/RLB results showed that washing of 3mm discs punched from FTA cards with FTA purification reagent followed by DNA extraction using Chelex ® resin was the most sensitive procedure. The detection limit could be improved when more discs were used as starting material for the DNA extraction, whereby the use of sixteen 3mm discs proved to be most practical. The presented best practice method for the extraction of DNA from blood spotted on FTA cards will facilitate epidemiological studies on TBPs. It may be particularly useful for field studies where a cold chain is absent. Copyright Â© 2016 Elsevier GmbH. All rights reserved.
Full Text Available The Republic of Cuba is a popular destination for Russian tourists and about 30-50 thousands of Russian citizens visit Cuba annually. However, the recreational activity is often associated with the risk of Ixodid ticks bites and infection with tick-borne pathogens. According to published literature, the fauna of the hard tick in Cuba is represented by nine species including Ixodes capromydis, Amblyomma albopictum, A. cajennense, A. dissimile, A. quadricavum, A. torrei, Dermacentor nitens, Rhipicephalus sanguineus and R. (Boophilus microplus. Five of these species, i.e. A. cajennense, A. dissimile, D. nitens, R. sanguineus and R. (Boophilus microplus, were reported as human parasites. Ticks are spread over the most part of the island territory. Several tick-borne pathogens should be considered as a potential threat for the bitten humans in Cuba, including Borrelia burgdorferi sensu lato, Rickettsia sp., Anaplasma sp., Ehrlichia sp., Coxiella sp., thogotovirus and Crimean-Congo hemorrhagic fever virus. Implications for the epidemiology of tick-borne infections in the Russian Federation are discussed.
Hecht, Joy A; Allerdice, Michelle E J; Krawczak, Felipe S; Labruna, Marcelo B; Paddock, Christopher D; Karpathy, Sandor E
Rickettsia bellii is a rickettsial species of unknown pathogenicity that infects argasid and ixodid ticks throughout the Americas. Many molecular assays used to detect spotted fever group (SFG) Rickettsia species do not detect R. bellii, so that infection with this bacterium may be concealed in tick populations when assays are used that screen specifically for SFG rickettsiae. We describe the development and validation of a R. bellii-specific, quantitative, real-time PCR TaqMan assay that targets a segment of the citrate synthase (gltA) gene. The specificity of this assay was validated against a panel of DNA samples that included 26 species of Rickettsia, Orientia, Ehrlichia, Anaplasma, and Bartonella, five samples of tick and human DNA, and DNA from 20 isolates of R. bellii, including 11 from North America and nine from South America. A R. bellii control plasmid was constructed, and serial dilutions of the plasmid were used to determine the limit of detection of the assay to be one copy per 4 µl of template DNA. This assay can be used to better determine the role of R. bellii in the epidemiology of tick-borne rickettsioses in the Western Hemisphere. Published by Oxford University Press on behalf of Entomological Society of America 2016. This work is written by US Government employees and is in the public domain in the US.
Scorza, Andrea V; Duncan, Colleen; Miles, Laura; Lappin, Michael R
To estimate the prevalence of enteric parasites and selected vector-borne agents of dogs and cats in San Isidro de El General, Costa Rica, fecal and serum samples were collected from animals voluntarily undergoing sterilization. Each fecal sample was examined for parasites by microscopic examination after fecal flotation and for Giardia and Cryptosporidium using an immunofluorescence assay (IFA). Giardia and Cryptosporidium IFA positive samples were genotyped after PCR amplification of specific DNA if possible. The seroprevalence rates for the vector-borne agents (Dirofilaria immitis, Borrelia burgdorferi, Ehrlichia canis, and Anaplasma phagocytophilum) were estimated based on results from a commercially available ELISA. Enteric parasites were detected in samples from 75% of the dogs; Ancylostoma caninum, Trichuris vulpis, Giardia, and Toxocara canis were detected. Of the cats, 67.5% harbored Giardia spp., Cryptosporidium spp., Ancylostoma tubaeforme, or Toxocara cati. Both Cryptosporidium spp. isolates that could be sequenced were Cryptosporidium parvum (one dog isolate and one cat isolate). Of the Giardia spp. isolates that were successfully sequenced, the 2 cat isolates were assemblage A and the 2 dog isolates were assemblage D. D. immitis antigen and E. canis antibodies were identified in 2.3% and 3.5% of the serum samples, respectively. The prevalence of enteric zoonotic parasites in San Isidro de El General in Costa Rica is high in companion animals and this information should be used to mitigate public health risks. Copyright © 2011. Published by Elsevier B.V.
Bush, Larry M; Vazquez-Pertejo, Maria T
Lyme disease is the most commonly reported tick-borneillness in the United States. Thecausative spirochete, Borrelia burgdorferi is transmitted by 4 species of Ixodes tick species. Over 90% of US cases occur in northeasternstates from Maine to Virginia, and in Wisconsin, Minnesota, and Michigan. Infection also takes place in northern California and Oregon. Lyme borreliosis is also diagnosed in parts of Europe, China, and Japan. The white-footed mouse is the primary animal reservoir for B. burgdorferi in the U.S. and the preferred host for nymphal and larval forms of the deer tick. Deer are hosts for the adult ticks but do not carry the spirochete. Signs and symptomsof infection occur in 3 stages; early localized, typified by erythema migrans; early disseminated with a flu-like syndrome, neurologic, and cardiac manifestations; and late, characteristically with arthritis. Although, the term 'Chronic Lyme Disease' has been assigned to many patients with a variety of unexplained symptoms, experts in the field question the validity of this diagnosis and warn against prolonged unproven antimicrobial therapies. Diagnosis relies upon clinical evaluation and is supported by serologic testing using a 2-step process which requires careful interpretation. Treatmentvaries with stage of disease, but normally includes doxycycline, amoxicillin,and ceftriaxone. Currently, no preventative vaccine is available. In some geographic areas, patients may be confected with Babesia, Ehrlichia, and Anaplasma since the same Ixodes ticks transmit these pathogens. Copyright © 2018 Mosby, Inc. All rights reserved.
Raphael B Stricker
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
Stricker, Raphael B; Johnson, Lorraine
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. PMID:21694904
Dittrich, Sabine; Phuklia, Weerawat; Turner, Gareth D H; Rattanavong, Sayaphet; Chansamouth, Vilada; Dumler, Stephen J; Ferguson, David J P; Paris, Daniel H; Newton, Paul N
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.
Bovine petechial fever is a Rickettsial disease of cattle, which has been diagnosed, only in Kenya, East Africa. Other countries in the region share some of the biotopes in which the disease occurs, and may well have the infection. The disease is characterised by widespread petechial and ecchymotic haemorrhages on the mucosal surfaces, and throughout the serosal and subserosal surfaces of the body organs and cavities. It may be fatal in up to 50% of untreated cases. The causal organism may be demonstrated most readily in the cytoplasm of polymorphonuclear granulocytes of the peripheral blood, as well as other leucocytes, and has been classified as Cytoecetes ondirii, a member of the tribe Ehrlichiae. Circumstantial and other evidence suggests that the disease is transmitted by an arthropod vector, which has yet to be identified. The blood of a naturally infected wild ruminant, the bushbuck, Tragelaphus scriptus has been shown to remain infective for at least 2 years, and other species such as the African buffalo, Syncercus caffer for at least 5 weeks. These and possibly other species, may serve as the amplifying and reservoir hosts.
Soares, Aline O; Souza, Aline D; Feliciano, Eveline A; Rodrigues, André F S F; D'Agosto, Marta; Daemon, Erik
Fleas and ticks transmit various pathogens while feeding on the blood of dogs. This study sought to verify the occurrence of ectoparasitism and hemoparasitism in dogs from two urban areas in the city of Juiz de Fora, Minas Gerais, Brazil. Between February and August 2003, 101 dogs were studied: 50 came from apartments in the downtown region and 51 from houses with grassy yards. The ectoparasites were collected and conserved in etanol 70%. The occurrence of hemoparasites was verified by examining blood smears from sample taken from the dogs'ears. The blood smears were stained with Giemsa and 100 fields per slide were examined, studying the erythrocytes to determine parasitism. From among the dogs living in apartments, we found (with respective prevalence and mean intensity): Ctenocephalides felis (12%), (3.3+/-2.0); Rhipicephalus sanguineus (2%); and ixodid nymphs (2%). In this environment in the dogs were not found hemoparasites. From the houses with grassy yards, we observed the following prevalence levels and mean intensities: C. felis (14%), (2.28+/-1.9); R. sanguineus (35%), (7.8+/-9.8); ixodid nymph (18%), (1.4+/-0.7); and ixodid larvae (4%), (12+/-14.4). The hemoparasites found were: Ehrlichia canis (16%) and Babesia canis (2%).
Ghai, Ria R; Mutinda, Mathew; Ezenwa, Vanessa O
Tick-borne hemoparasites (TBHs) are a group of pathogens of concern in animal management because they are associated with a diversity of hosts, including both wild and domestic species. However, little is known about how frequently TBHs are shared across the wildlife-livestock interface in natural settings. Here, we compared the TBHs of wild Grant's gazelle (Nanger granti) and domestic sheep (Ovis aries) in a region of Kenya where these species extensively overlap. Blood samples collected from each species were screened for piroplasm and rickettsial TBHs by PCR-based amplification of 18S/16S ribosomal DNA, respectively. Overall, 99% of gazelle and 66% of sheep were positive for Babesia/Theileria, and 32% of gazelle and 47% sheep were positive for Anaplasma/Ehrlichia. Sequencing a subset of positive samples revealed infections of Theileria and Anaplasma. Sequences sorted into seven phylogenetically distinct genotypes-two Theileria, and five Anaplasma. With the exception of a putatively novel Anaplasma lineage from Grant's gazelle, these genotypes appeared to be divergent forms of previously described species, including T. ovis, A. ovis, A. bovis, and A. platys. Only one genotype, which clustered within the A. platys clade, contained sequences from both gazelle and sheep. This suggests that despite niche, habitat, and phylogenetic overlap, the majority of circulating tick-borne diseases may not be shared between these two focal species. Copyright © 2016 Elsevier B.V. All rights reserved.
Erwin, John A.; Fitak, Robert R.; Dwyer, James F.; Morrison, Joan L.; Culver, Melanie
Bacterial pathogens of the families Anaplasmataceae and Rickettsiaceae are often spread to humans or other animals from bites from infected arthropod hosts. Recently, an increasing number of studies have implicated migratory birds in the circulation of these pathogens through the spread of arthropod vectors. However, few studies have examined the potential for resident bird populations to serve as reservoirs for these zoonoses. In this study, we used nested PCRs of the GroESL and 17 kDa genes to screen for Anaplasmataceae and Rickettsiaceae, respectively, in a resident population of the northern crested caracara (Caracara cheriway) from Florida (n = 55). Additionally, a small number (n = 6) of captive individuals from Texas were included. We identified one individual (1.64%) positive for Rickettsia felis and one (1.64%) positive for Ehrlichia chaffeensis; both these individuals were from Florida. Presence of these pathogens demonstrates that these birds are potential hosts; however, the low prevalence of infections suggests that these populations likely do not function as an ecological reservoir.
Fernando de Freitas Fernandes
Full Text Available The action of fenthion was studied in a 15% epicutaneous formulation upon Rhipicephalus sanguineus, which may transmit pathogens to men and other animals, such as Ehrlichia, Babesia and Ricketsia. Dogs were artificially infected for the trial. The fenthion bioassays were begun four months after artificial infestation. The test group, having a mean of 186 ticks per dog, received the formulation dosage according to body weight on the neck region. Tick counts were performed, considering diameters > or = 2mm, during 11 days of treatment, in the most affected body areas: back, ears and paws. Before the application of fenthion in the dogs, it were observed an average 43.3% ticks in the ears, 38.1% in the back area and 17.6% in the paws. The number of ticks in dogs decreased by 36.2%, 63.8%, 82.7%, 67%, 40% and 4.9%, respectively on days 1, 2, 3, 5, 7, 9 and 11 after treatment. R. sanguineus anti-tick activity, lower than that officially recommended, was verified. The number of ticks increased progressively after the 5th day, demonstrating residual insecticide inefficacy. The results obtained did not indicate the use of this formulation, at the tested dosage, as an elective measure for R. sanguineus control.Investigou-se a atividade de fenthion em formulação epicutânea a 15% sobre Rhipicephalus sanguineus, transmissor de patógenos ao homem e animais, tais como Ehrlichia, Babesia e Ricketsia. Infestou-se artificialmente cães com larvas deste carrapato. Os bioensaios com o fenthion iniciaram-se 4 meses após a infestação artificial. Constatada a média de 186 ixodídeos/cão, cães do grupo teste receberam na região da nuca a dosagem correspondente ao seu peso. Avaliaram-se a eficiência e a atividade residual através de contagens dos carrapatos com diâmetro > ou = 2mm, durante 11 dias, nas áreas corpóreas mais parasitadas: dorso, orelhas e patas. Anteriormente, à aplicação do fenthion, 44,3% dos carrapatos format observados nas orelhas
Mariana Granziera Spolidorio
Full Text Available Canine ehrlichiosis and babesiosis are the most prevalent tick-borne diseases in Brazilian dogs. Few studies have focused attention in surveying tick-borne diseases in the Brazilian Amazon region. A total of 129 blood samples were collected from dogs living in the Brazilian eastern Amazon. Seventy-two samples from dogs from rural areas of 19 municipalities and 57 samples from urban stray dogs from Santarém municipality were collected. Serum samples were submitted to Indirect Immunofluorescence Assay (IFA with antigens of Babesia canis vogeli, Ehrlichia canis, and six Rickettsia species. The frequency of dogs containing anti-B. canis vogeli, anti-E. canis, and anti-Rickettsia spp. antibodies was 42.6%, 16.2%, and 31.7%, respectively. Anti-B. canis vogeli antibodies were detected in 59.6% of the urban dogs, and in 29.1% of the rural dogs (P Ehrliquiose canina e babesiose canina são as doenças parasitárias transmitidas por carrapatos de maior prevalência em cães do Brasil. Poucos estudos pesquisaram doenças transmitidas por carrapatos na região da Amazônia brasileira. Um total de 129 amostras de sangue foram colhidas de cães da Amazônia oriental brasileira. Setenta e dois cães eram de áreas rurais de 19 municípios do Estado do Pará, e 57 amostras foram colhidas de cães errantes vadios da área urbana do município de Santarém-PA. As amostras de soro foram submetidas ao ensaio de imunofluorescência indireta, com antígenos de Babesia canis vogeli, Ehrlichia canis, e seis espécies de Rickettsia. A frequência de cães com anticorpos anti-B. canis vogeli, anti-E. canis, e anti-Rickettsia spp. foi de 42,6%, 16,2% e 31,7%, respectivamente. Anticorpos anti-B. canis vogeli foram detectados em 59,6% dos cães urbanos, e em 29,1% dos cães rurais (P < 0.05. Para E. canis, a soroprevalência foi parecida entre os cães urbanos (15,7% e rurais (16,6%. Para Rickettsia spp., cães rurais apresentaram prevalência (P < 0.05 significativamente
Full Text Available Canine distemper virus is a member of the genus Morbillivirus of the family Paramyxoviridaethat causes severe disease in dogs and a range of wild mammals. The clinical signs relate essentially to the respiratory, gastrointestinal and central nervous systems. In South Africa, infection with Ehrlichia canis and canine parvovirus may present similarly. Many dogs will initially present with a wide range of central nervous system signs without any history of systemic disease. A recent South African study evaluating ante mortem diagnosis highlighted the importance of recognising clinical signs, cerebrospinal fluid IgG titres, serum IgM titres and immunocytochemistry of epithelial tissue. A 2-year retrospective evaluation of cerebrospinal fluid samples collected from dogs presented to the Onderstepoort Veterinary Academic Hospital indicates that distemper infection is common, and this disease should routinely be suspected in cases of diverse neurological disease in dogs. The South African dog population is specifically at high risk for the disease because of the large pool of unvaccinated, reproductively-active dogs that expose the wildlife resources of the country to risk of fatal disease. Outbreaks of disease in dogs continue to occur in developed and developing communities in both vaccinated and unvaccinated dogs worldwide, and have also been described in a wide range of free-ranging wildlife, including seals, dolphins and lions, and in endangered zoo animals. Modified live virus vaccines have contributed markedly to disease control in the dog population but have caused mortality in some wild carnivores. New recombinant vaccines are being developed that will be safe in wild animals. The pathogenesis of CNS demyelination has been compared to various important demyelinating diseases in humans and, amongst other things, relates to down-regulation of the oligodendrocyte gene coding for myelin synthesis and non-immunocyte CNS cell expression of type II
Carslake, R J; Hill, K E; Sjölander, K; Hii, S F; Prattley, D; Acke, E
To determine the prevalence of selected canine vector-borne diseases (Leishmania infantum, Anaplasma spp., Ehrlichia canis, Borrelia burgdorferi and Dirofilaria immitis) and endo- and ectoparasites in Samoan dogs presenting for surgical sterilisation and to report on the general health management of the dogs. This study was a prospective serological cross-sectional survey. Management data were obtained for 242 dogs by interview with their owners. Blood samples were collected from 237 dogs and screened for the canine vector-borne diseases using point-of-care qualitative ELISA assays. Anaplasma spp. positive samples were screened by PCR and sequenced for species identification. Rectal faecal samples were collected from 204 dogs for faecal flotation and immunofluorescent antibody tests were performed for Giardia and Cryptosporidium spp. on a subset of 93 faecal samples. The skin and coat of 221 dogs were examined for presence of ectoparasites. The D. immitis antigen was detected in 46.8% (111/237) of dogs. Seroprevalence of Anaplasma spp. was 8.4% (20/237); A. platys was confirmed by PCR. Prevalence of hookworm was 92.6% (185/205) and Giardia was 29.0% (27/93). Ectoparasites were detected on 210/221 (95.0%) of dogs examined and 228/242 dogs (94.2%) had previously never received any preventative medication. There was a very high prevalence of D. immitis, hookworm and external parasites in Samoan dogs, and prophylactic medication is rarely administered. This is the first report confirming A. platys in Samoa and the South Pacific islands. The public health implications of poor management of the dogs should be considered and investigated further. © 2017 Australian Veterinary Association.
Ker, Henrique Gama; Coura-Vital, Wendel; Aguiar-Soares, Rodrigo Dian de Oliveira; Roatt, Bruno Mendes; das Dores Moreira, Nádia; Carneiro, Cláudia Martins; Machado, Evandro Marques de Menezes; Teixeira-Carvalho, Andréa; Martins-Filho, Olindo Assis; Giunchetti, Rodolfo Cordeiro; Araújo, Márcio Sobreira Silva; Coelho, Eduardo Antonio Ferraz; da Silveira-Lemos, Denise; Reis, Alexandre Barbosa
Diagnosing canine visceral leishmaniasis (CVL) is a critical challenge since conventional immunoserological tests still present some deficiencies. The current study evaluated a prototype flow cytometry serology test, using antigens and fluorescent antibodies that had been stored for 1 year at 4°C, on a broad range of serum samples. Noninfected control dogs and Leishmania infantum-infected dogs were tested, and the prototype test showed excellent performance in differentiating these groups with high sensitivity, specificity, positive and negative predictive values, and accuracy (100% in all analyses). When the CVL group was evaluated according to the dogs' clinical status, the prototype test showed outstanding accuracy in all groups with positive serology (asymptomatic II, oligosymptomatic, and symptomatic). However, in dogs which had positive results by PCR-restriction fragment length polymorphism (RFLP) but negative results by conventional serology (asymptomatic I), serological reactivity was not observed. Additionally, sera from 40 dogs immunized with different vaccines (Leishmune, Leish-Tec, or LBSap) did not present serological reactivity in the prototype test. Eighty-eight dogs infected with other pathogens (Trypanosoma cruzi, Leishmania braziliensis, Ehrlichia canis, and Babesia canis) were used to determine cross-reactivity and specificity, and the prototype test performed well, particularly in dogs infected with B. canis and E. canis (100% and 93.3% specificities, respectively). In conclusion, our data reinforce the potential of the prototype test for use as a commercial kit and highlight its outstanding performance even after storage for 1 year at 4°C. Moreover, the prototype test efficiently provided accurate CVL serodiagnosis with an absence of false-positive results in vaccinated dogs and minor cross-reactivity against other canine pathogens.
Tolnai, Z; Sréter-Lancz, Z; Sréter, T
In recent years, Ehrlichia canis and Hepatozoon canis transmitted by Rhipicephalus sanguineus were reported from Hungary. The aim of the present study was to reveal the spatial distribution pattern of pathogens transmitted by R. sanguineus in a sentinel species, red fox (Vulpes vulpes) in Hungary and to analyse the relationship of these patterns with landscape and climate by geographical information systems. Fox carcasses, representing 0.5% of the total fox population were randomly selected out of all the foxes of Hungary. The spleen samples of the animals were tested by real-time PCR for Anaplasma platys, Babesia vogeli, E. canis and H. canis infection. Positive results were confirmed by conventional PCR followed by sequencing. The prevalence of H. canis infection was 22.2% (95% CI=18.4-26.4%), and this parasite was detected in all areas including the mountain regions of Hungary. These findings indicate that other tick species or other transmission routes (oral and transplacental) might be in the background of the countrywide distribution of H. canis. Anaplasma platys was not found; nevertheless, the prevalence of Anaplasma phagocytophilum infection transmitted by Ixodes ricinus was 12.5% (95% CI=9.7-16.1%) in foxes. B. vogeli and E. canis infection was not detected. There was no correlation between environmental parameter values in the home range of foxes and A. phagocytophilum or H. canis infection, which is in line with that observed in the case of tick species infesting foxes in Hungary. The results of this study indicate that R. sanguineus, if present, might be rare in Hungary. Our baseline study can be used for future evaluation of the effect of climate change on the spreading and emergence of R. sanguineus transmitted pathogens in Hungary. Copyright © 2015 Elsevier GmbH. All rights reserved.
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.
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.
Rojas, Alicia; Rojas, Diana; Montenegro, Víctor; Gutiérrez, Ricardo; Yasur-Landau, Daniel; Baneth, Gad
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 (pcanis were significantly associated with R. sanguineus s.l. infestation (pcanis 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. Copyright © 2013 Elsevier B.V. All rights reserved.
Kelly, Patrick J.; Xu, Chuanling; Lucas, Helene; Loftis, Amanda; Abete, Jamie; Zeoli, Frank; Stevens, Audrey; Jaegersen, Kirsten; Ackerson, Kate; Gessner, April; Kaltenboeck, Bernhard; Wang, Chengming
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. PMID:23335965
Kamani, Joshua; Baneth, Gad; Mumcuoglu, Kosta Y.; Waziri, Ndadilnasiya E.; Eyal, Osnat; Guthmann, Yifat; Harrus, Shimon
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. PMID:23505591
McDermid, Kimberly R; Snyman, Andrei; Verreynne, Frederick J; Carroll, John P; Penzhorn, Banie L; Yabsley, Michael J
African lion ( Panthera leo ) numbers are decreasing rapidly and populations are becoming smaller and more fragmented. Infectious diseases are one of numerous issues threatening free-ranging lion populations, and low-density populations are particularly at risk. We collected data on the prevalence and diversity of viral and parasitic pathogens in a small lion population in eastern Botswana. During 2012 and 2014, blood samples were collected from 59% (n=13) of the adult-subadult lions in the Northern Tuli Game Reserve in eastern Botswana. One lion had antibodies to feline panleukopenia virus, two had antibodies to canine distemper virus, and two had feline calicivirus antibodies. Ten of the 13 had antibodies to feline immunodeficiency virus and 11 had feline herpesvirus antibodies. All lions were negative for antibodies to feline coronavirus. Blood samples from all lions were negative for Trypanosoma, Anaplasma, Theileria, and Ehrlichia spp. by molecular testing; however, all lions were positive for Babesia spp. by reverse line blot hybridization assay. Sequencing of amplicons from four lions revealed four groups of Babesia spp. including several genetic variants of Babesia felis , Babesia lengau, and Babesia canis and a group of novel Babesia sequences which were only 96% similar to other Babesia spp. Six lions were infested with four species of ticks (Rhipicentor nuttalli, Rhipicephalus simus, Rhipicephalus sulcatus, and Rhipicephalus appendiculatus). These data provide the first health assessment of this population and can be used to identify management and conservation strategies to decrease the impact of pathogens on this population. This is particularly important as there is an initiative to incorporate this population into a larger metapopulation of lions from adjacent South Africa and Zimbabwe.
S. K. Borthakur
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.
Mrljak, Vladimir; Kuleš, Josipa; Mihaljević, Željko; Torti, Marin; Gotić, Jelena; Crnogaj, Martina; Živičnjak, Tatjana; Mayer, Iva; Šmit, Iva; Bhide, Mangesh; Barić Rafaj, Renata
Vector-borne pathogens (VBPs) are a group of globally extended and quickly spreading pathogens that are transmitted by various arthropod vectors. The aim of the present study was to investigate the seroprevalence against Babesia canis, Anaplasma phagocytophilum, Borrelia burgdorferi sensu lato, Leishmania infantum, Dirofilaria immitis, and Ehrlichia canis in dogs in Croatia. We investigated 435 randomly selected apparently healthy dogs in 13 different locations of Croatia for antibodies to B. canis by indirect immunofluorescence using a commercial IFA IgG Antibody Kit. All samples were also tested for qualitative detection of D. immitis antigen and for antibodies to A. phagocytophilum, B. burgdorferi sensu lato, L. infantum, and E. canis with two point-of-care assays. Overall, 112 dogs (25.74%, 95% confidence interval [CI] 21.70-30.12) were serologically positive for one or more of the pathogens. B. canis was the most prevalent pathogen (20.00%, 95% CI 16.34-24.07), followed by A. phagocytophilum (6.21%, 95% CI 4.12-8.90), L. infantum, (1.38%, 95% CI 0.51-2.97), and B. burgdorferi sensu lato (0.69%, 95% CI 0.01-2.00). The lowest seroprevalence was for D. immitis and E. canis (0.46%, 95% CI 0.01-1.65). Coinfection was determined in 12 dogs (2.76%, 95% CI 1.43-4.77), of which 10 were positive to two pathogens (7 with B. canis and A. phagocytophilum and 1 B. canis with B. burgdorferi sensu lato or L. infantum or E. canis). One dog was positive to three pathogens and another dog to four pathogens. Seroprevalence for babesia was age, breed, and lifestyle/use dependent. Purebred dogs had almost half the chance of developing disease than crossbred (OR = 0.58, p Croatia. Some of these VBPs are zoonotic and represent a potential risk to public health.
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
Clara Nancy Gutiérrez
Full Text Available La ehrlichiosis canina es una enfermedad infecciosa emergente transmitida por garrapatas, producida por Ehrlichia spp. (Proteobacteria: Ricketsiales, la cual afecta a miembros de la familia Canidae. Los agentes etiológicos son bacterias Gram negativas, intracelulares obligatorias, redondeadas y pleomórficas, esto último especialmente en cultivos celulares. Estas bacterias se localizan en vacuolas rodeadas de membranas (mórulas en el citoplasma de células sanguíneas y dependiendo de la especie, tienen tropismo por linfocitos, monocitos y granulocitos. Históricamente la enfermedad es endémica en regiones tropicales y subtropicales, pero se reporta cada vez más en regiones de clima templado. Ello puede atribuirse a varios factores, los cuales incluyen el mejoramiento en las herramientas de diagnóstico, los cambios ambientales y climáticos (calentamiento global que influyen directamente en la distribución de las garrapatas y la gran cantidad de viajes con mascotas de un lugar a otro del planeta, lo cual ha contribuido al establecimiento de esta enfermedad en áreas no endémicas. Es común la presencia de coinfección con otros patógenos transmitidos por garrapatas y esto complica la patogénesis, las manifestaciones clínicas, el diagnóstico y el tratamiento. Frecuentemente, el patógeno no puede ser eliminado por completo a pesar de la terapia con antibióticos y la resolución de los signos clínicos. Actualmente, no se dispone de una vacuna, por lo que el uso de ectoparasiticidas resulta ser una buena opción para la prevención de la enfermedad. Esta enfermedad constituye un problema en medicina veterinaria y el potencial zoonótico de estos agentes es una consideración de gran relevancia para la salud humana.
Leydet, Brian F; Liang, Fang-Ting
There are 4 major human-biting tick species in the northeastern United States, which include: Amblyomma americanum, Amblyomma maculatum, Dermacentor variabilis, and Ixodes scapularis. The black bear is a large mammal that has been shown to be parasitized by all the aforementioned ticks. We investigated the bacterial infections in ticks collected from Louisiana black bears (Ursus americanus subspecies luteolus). Eighty-six ticks were collected from 17 black bears in Louisiana from June 2010 to March 2011. All 4 common human-biting tick species were represented. Each tick was subjected to polymerase chain reaction (PCR) targeting select bacterial pathogens and symbionts. Bacterial DNA was detected in 62% of ticks (n=53). Rickettsia parkeri, the causative agent of an emerging spotted fever group rickettsiosis, was identified in 66% of A. maculatum, 28% of D. variabilis, and 11% of I. scapularis. The Lyme disease bacterium, Borrelia burgdorferi, was detected in 2 I. scapularis, while one A. americanum was positive for Borrelia bissettii, a putative human pathogen. The rickettsial endosymbionts Candidatus Rickettsia andeanae, rickettsial endosymbiont of I. scapularis, and Rickettsia amblyommii were detected in their common tick hosts at 21%, 39%, and 60%, respectively. All ticks were PCR-negative for Anaplasma phagocytophilum, Ehrlichia spp., and Babesia microti. This is the first reported detection of R. parkeri in vector ticks in Louisiana; we also report the novel association of R. parkeri with I. scapularis. Detection of both R. parkeri and B. burgdorferi in their respective vectors in Louisiana demands further investigation to determine potential for human exposure to these pathogens. Copyright © 2013 Elsevier GmbH. All rights reserved.
Khoo, Jing-Jing; Chen, Fezshin; Kho, Kai Ling; Ahmad Shanizza, Azzy Iyzati; Lim, Fang-Shiang; Tan, Kim-Kee; Chang, Li-Yen; AbuBakar, Sazaly
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. Copyright © 2016 The Authors. Published by Elsevier GmbH.. All rights reserved.
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.
Diaz, Nicole M; Mendez, Gabriella S; Grijalva, C Jaime; Walden, Heather S; Cruz, Marilyn; Aragon, Eduardo; Hernandez, Jorge A
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%) (pdog 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. Copyright © 2015 Elsevier B.V. All rights reserved.
Guo, Huanping; Sevinc, Ferda; Ceylan, Onur; Sevinc, Mutlu; Ince, Ege; Gao, Yang; Moumouni, Paul Franck Adjou; Liu, Mingming; Efstratiou, Artemis; Wang, Guanbo; Cao, Shinuo; Zhou, Mo; Jirapattharasate, Charoonluk; Ringo, Aaron Edmond; Zheng, Weiqing; Xuan, Xuenan
In the present study, a total of 192 blood samples were collected from pet dogs, kennel dogs and shepherd dogs in Konya district, Turkey, and tested by specific PCR for the presence of vector-borne pathogens. Several pathogens were identified, most of which can cause substantial morbidity in dogs. PCR results revealed that 54 (28.1%) dogs were infected with one or more pathogens. Positive results were obtained for Babesia spp. in 4 dogs (2.1%), Hepatozoon spp. in 8 dogs (4.2%) and Mycoplasma spp. in 46 dogs (24%). Three dogs (1.6%) were infected with two or three pathogens. The sequence analysis of the positive DNA samples revealed the presence of Babesia canis vogeli, Hepatozoon canis, Hepatozoon sp. MF, Mycoplasma haemocanis and Candidatus Mycoplasma haematoparvum. Ehrlichia canis and Anaplasma platys were not detected. Regardless of ownership status, vector-borne diseases were common in these dog populations. There was significant difference of pathogen prevalence among the different dog populations. Mycoplasma spp. was more frequent in the kennel dogs (31.9%) than in the pet (21.4%) and shepherd dogs (13.8%). Additionally, the frequency of Babesia spp. and Hepatozoon spp. was higher in the shepherd dogs which account for three quarters and half of the total number of Babesia spp. and Hepatozoon spp., respectively. To our knowledge, this is the first report of Mycoplasma infection in dogs in Turkey. The results of the present study provide a foundation for understanding the epidemiology of canine vector-borne diseases (CVBDs), and for strategies to control these diseases in Turkey.
Oliveira, Valéria da Costa; Boechat, Viviane Cardoso; Mendes Junior, Artur Augusto Velho; Madeira, Maria de Fátima; Ferreira, Luiz Claudio; Figueiredo, Fabiano Borges; Campos, Monique Paiva; de Carvalho Rodrigues, Francisco das Chagas; Carvalhaes de Oliveira, Raquel de Vasconcellos; Amendoeira, Maria Regina Reis; Menezes, Rodrigo Caldas
Zoonotic visceral leishmaniasis is caused by the protozoan Leishmania infantum and little is known about the occurrence and pathogenesis of this parasite in the CNS. The aims of this study were to evaluate the occurrence, viability and load of L. infantum in the CNS, and to identify the neurological histological alterations associated with this protozoan and its co-infections in naturally infected dogs. Forty-eight Leishmania-seropositive dogs from which L. infantum was isolated after necropsy were examined. Cerebrospinal fluid (CSF) samples were analyzed by parasitological culture, quantitative real-time PCR (qPCR) and the rapid immunochromatographic Dual Path Platform test. Brain, spinal cord and spleen samples were submitted to parasitological culture, qPCR, and histological techniques. Additionally, anti-Toxoplasma gondii and anti-Ehrlichia canis antibodies in serum and distemper virus antigens in CSF were investigated. None of the dogs showed neurological signs. All dogs tested positive for L. infantum in the CNS. Viable forms of L. infantum were isolated from CSF, brain and spinal cord in 25% of the dogs. Anti-L. infantum antibodies were detected in CSF in 61% of 36 dogs. Inflammatory histological alterations were observed in the CNS of 31% of the animals; of these, 66% were seropositive for E. canis and/or T. gondii. Amastigote forms were associated with granulomatous non-suppurative encephalomyelitis in a dog without evidence of co-infections. The highest frequency of L. infantum DNA was observed in the brain (98%), followed by the spinal cord (96%), spleen (95%), and CSF (50%). The highest L. infantum load in CNS was found in the spinal cord. These results demonstrate that L. infantum can cross the blood-brain barrier, spread through CSF, and cause active infection in the entire CNS of dogs. Additionally, L. infantum can cause inflammation in the CNS that can lead to neurological signs with progression of the disease.
Valéria da Costa Oliveira
Full Text Available Zoonotic visceral leishmaniasis is caused by the protozoan Leishmania infantum and little is known about the occurrence and pathogenesis of this parasite in the CNS. The aims of this study were to evaluate the occurrence, viability and load of L. infantum in the CNS, and to identify the neurological histological alterations associated with this protozoan and its co-infections in naturally infected dogs. Forty-eight Leishmania-seropositive dogs from which L. infantum was isolated after necropsy were examined. Cerebrospinal fluid (CSF samples were analyzed by parasitological culture, quantitative real-time PCR (qPCR and the rapid immunochromatographic Dual Path Platform test. Brain, spinal cord and spleen samples were submitted to parasitological culture, qPCR, and histological techniques. Additionally, anti-Toxoplasma gondii and anti-Ehrlichia canis antibodies in serum and distemper virus antigens in CSF were investigated. None of the dogs showed neurological signs. All dogs tested positive for L. infantum in the CNS. Viable forms of L. infantum were isolated from CSF, brain and spinal cord in 25% of the dogs. Anti-L. infantum antibodies were detected in CSF in 61% of 36 dogs. Inflammatory histological alterations were observed in the CNS of 31% of the animals; of these, 66% were seropositive for E. canis and/or T. gondii. Amastigote forms were associated with granulomatous non-suppurative encephalomyelitis in a dog without evidence of co-infections. The highest frequency of L. infantum DNA was observed in the brain (98%, followed by the spinal cord (96%, spleen (95%, and CSF (50%. The highest L. infantum load in CNS was found in the spinal cord. These results demonstrate that L. infantum can cross the blood-brain barrier, spread through CSF, and cause active infection in the entire CNS of dogs. Additionally, L. infantum can cause inflammation in the CNS that can lead to neurological signs with progression of the disease.
Ionita, Mariana; Silaghi, Cornelia; Mitrea, Ioan Liviu; Edouard, Sophie; Parola, Philippe; Pfister, Kurt
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. Copyright © 2015 Elsevier GmbH. All rights reserved.
Ionita, Mariana; Mitrea, Ioan Liviu; Pfister, Kurt; Hamel, Dietmar; Silaghi, Cornelia
The aim of the present study was to provide a preliminary insight into the diversity of tick-borne pathogens circulating at the domestic host-tick interface in Romania. For this, feeding and questing ticks were analyzed by real-time polymerase chain reaction (PCR) for the presence of Anaplasma phagocytophilum, Anaplasma platys, Ehrlichia canis, Borrelia burgdorferi sensu latu, and by PCR and subsequent sequencing for Rickettsia spp., Babesia spp. and Theileria spp. A total of 382 ticks, encompassing 5 species from 4 genera, were collected in April-July 2010 from different areas of Romania; of them, 40 were questing ticks and the remainder was collected from naturally infested cattle, sheep, goats, horses or dogs. Tick species analyzed included Ixodes ricinus, Dermacentor marginatus, Hyalomma marginatum, Rhipicephalus bursa, and Rhipicephalus sanguineus. Four rickettsiae of the spotted fever group of zoonotic concern were identified for the first time in Romania: Rickettsia monacensis and Rickettsia helvetica in I. ricinus, and Rickettsia slovaca and Rickettsia raoultii in D. marginatus. Other zoonotic pathogens such as A. phagocytophilum, Borrelia afzelii, and Babesia microti were found in I. ricinus. Pathogens of veterinary importance were also identified, including Theileria equi in H. marginatum, Babesia occultans in D. marginatus and H. marginatum, Theileria orientalis/sergenti/buffeli-group in I. ricinus and in H. marginatum and E. canis in R. sanguineus. These findings show a wide distribution of very diverse bacterial and protozoan pathogens at the domestic host-tick interface in Romania, with the potential of causing both animal and human diseases. Copyright © 2013 Elsevier B.V. All rights reserved.
Williams-Newkirk, Amanda Jo; Rowe, Lori A; Mixson-Hayden, Tonya R; Dasch, Gregory A
The lone star tick (Amblyomma americanum) is an abundant and aggressive biter of humans, domestic animals, and wildlife in the southeastern-central USA and an important vector of several known and suspected zoonotic bacterial pathogens. However, the biological drivers of bacterial community variation in this tick are still poorly defined. Knowing the community context in which tick-borne bacterial pathogens exist and evolve is required to fully understand the ecology and immunobiology of the ticks and to design effective public health and veterinary interventions. We performed a metagenomic survey of the bacterial communities of questing A. americanum and tested 131 individuals (66 nymphs, 24 males, and 41 females) from five sites in three states. Pyrosequencing was performed with barcoded eubacterial primers targeting variable 16S rRNA gene regions 5-3. The bacterial communities were dominated by Rickettsia (likely R. amblyommii) and an obligate Coxiella symbiont, together accounting for 6.7-100% of sequences per tick. DNAs from Midichloria, Borrelia, Wolbachia, Ehrlichia, Pseudomonas, or unidentified Bacillales, Enterobacteriaceae, or Rhizobiales groups were also detected frequently. Wolbachia and Midichloria significantly co-occurred in Georgia (pmales containing more Rickettsia and females containing more Coxiella. Comparisons among adult ticks collected in New York and North Carolina supported the findings from the Georgia collection despite differences in geography, collection date, and sample handling, implying that the differences detected are consistent attributes. The data also suggest that some members of the bacterial community change during the tick life cycle and that some sex-specific attributes may be detectable in nymphs.
Marcondes, Mary; Hirata, Karina Y; Vides, Juliana P; Sobrinho, Ludmila S V; Azevedo, Jaqueline S; Vieira, Thállitha S W J; Vieira, Rafael F C
Visceral leishmaniasis (VL) has been increasingly recognized in cats living in areas endemic for the disease. Co-infection with Leishmania infantum and other infectious agents is well established in dogs. However, for cats, data on co-infections with L. infantum and other infectious agents are still sparse. The aim of this study was to identify the prevalence of vector-borne pathogens, Mycoplasma spp., feline immunodeficiency virus (FIV) and feline leukaemia virus (FeLV) in cats from an area endemic for VL in southeastern Brazil. Of the 90 cats, eight (8.9%) were infected with Mycoplasma spp., five (5.5%) were FIV- positive and one (1.1%) was FeLV-positive. Co-infection with L. infantum and at least one other infectious agent was found in 9/50 (18.0%; CI: 8.6-31.4%) cats. In Group 1 (cats infected naturally by L. infantum), 4/50 (8.0%) cats were positive for FIV, 4/50 (8%) for Mycoplasma spp. and 1/50 (2.0%) was co-infected with FeLV and Mycoplasma spp. In Group 2 (cats non-infected with L. infantum), 2/40 (5.0%) cats were infected with Mycoplasma spp. and 1/40 (2.5%) was co-infected with FIV and Mycoplasma spp. All cats were negative for Ehrlichia spp., Babesia spp. and Anaplasma platys. A low prevalence of co-infection in Leishmania-infected and non-infected cats was found. Co-infections with Leishmania and vector-borne diseases in cats are not common in this area endemic for VL in Brazil.
Apperson, Charles S; Engber, Barry; Nicholson, William L; Mead, Daniel G; Engel, Jeffrey; Yabsley, Michael J; Dail, Kathy; Johnson, Joey; Watson, D Wesley
Cases of Rocky Mountain spotted fever (RMSF) in North Carolina have escalated markedly since 2000. In 2005, we identified a county in the Piedmont region with high case numbers of RMSF. We collected ticks and examined them for bacterial pathogens using molecular methods to determine if a novel tick vector or spotted fever group rickettsiae (SFGR) might be emerging. Amblyomma americanum, the lone star tick, comprised 99.6% of 6,502 specimens collected in suburban landscapes. In contrast, Dermacentor variabilis, the American dog tick, a principal vector of Rickettsia rickettsii, comprised < 1% of the ticks collected. Eleven of 25 lone star tick pools tested were infected with "Rickettsia amblyommii," an informally named SFGR. Sera from patients from the same county who were presumptively diagnosed by local physicians with a tick-borne illness were tested by an indirect immunofluorescence antibody (IFA) assay to confirm clinical diagnoses. Three of six patients classified as probable RMSF cases demonstrated a fourfold or greater rise in IgG class antibody titers between paired acute and convalescent sera to "R. amblyommii" antigens, but not to R. rickettsii antigens. White-tailed deer, Odocoileus virginianus, are preferred hosts of lone star ticks. Blood samples collected from hunter-killed deer from the same county were tested by IFA test for antibodies to Ehrlichia chaffeensis and "R. amblyommii." Twenty-eight (87%) of 32 deer were positive for antibodies to E. chaffeensis, but only 1 (3%) of the deer exhibited antibodies to "R. amblyommii," suggesting that deer are not the source of "R. amblyommii" infection for lone star ticks. We propose that some cases of rickettsiosis reported as RMSF may have been caused by "R. amblyommii" transmitted through the bite of A. americanum.
Cicuttin, Gabriel L; De Salvo, María N; La Rosa, Isabel; Dohmen, Federico E Gury
Bats are potential reservoirs of many vector-borne bacterial pathogens. The aim of the present study was to detect species of Anaplasma, Ehrlichia, Neorickettsia, Rickettsia, Borrelia and Bartonella in Brazilian free-tailed bats (Tadarida brasiliensis, Molossidae) from Buenos Aires city, Argentina. Between 2012 and 2013, 61 T. brasiliensis from urban areas of Buenos Aires city were studied. The samples were molecularly screened by PCR and sequencing. Five bats (8.2%) were positive to Neorickettsia risticii, one (1.6%) was positive to Rickettsia sp. and three bats (4.9%) to Bartonella sp. For molecular characterization, the positive samples were subjected to amplification and sequencing of a fragment of p51 gene for N. risticii, a fragment of citrate synthase gene (gltA) for Rickettsia genus and a fragment of gltA for Bartonella genus. Phylogenetic tree was constructed using the maximum-likelihood method. Phylogenetic analysis of N. risticii detect in our study revealed that it relates to findings in the USA West Coast; Rickettsia sp. detected is phylogenetically within R. bellii group, which also includes many other Rickettsia endosymbionts of insects; and Bartonella sp. found is related to various Bartonella spp. described in Vespertilionidae bats, which are phylogenetically related to Molossidae. Our results are in accordance to previous findings, which demonstrate that insectivorous bats could be infected with vector-borne bacteria representing a potential risk to public health. Future research is necessary to clarify the circulation of these pathogens in bats from Buenos Aires. Copyright © 2017 Elsevier Ltd. All rights reserved.
Mariana Cristina Hoeppner Rondelli
Full Text Available This study aimed to compare the effects of two doses of doxycycline hyclate on red blood cells, hemoglobin, hematocrit, white blood cells and platelets of dogs with ehrlichiosis. Group I, comprised of healthy dogs (n=6, negative on serology for Ehrlichia canis and Leptospira spp., real time PCR for E. canis and Anaplasma platys, and on semi–nested PCR for Babesia canis; Groups II (n=6 and III (n=6, comprised of dogs with suggestive clinical history, positive serology and/or real time PCR for E. canis, negative on research for anti-Leptospira spp. antibodies and real time PCR for A. platys, and on semi–nested PCR for B. canis were studied. Sick dogs were treated with doxycycline hyclate every 12 hours, by mouth, for 30 days (5 mg/kg, group II; 10 mg/kg, group III. Complete blood counts were performed before, after 15 days, and 10 days after period of treatment was complete. No difference between groups at the studied time points were noticed for red blood cells, hemoglobin, haematocrit and white blood cells. Difference was observed for platelets between group I and groups II and III (p<0.0001 at the study onset. After 15 days of treatment, the mean platelet for group III was lower than groups I (p=0.008 and II (p=0.0007, indicative of persistent thrombocytopenia, already absent in group II. No difference between groups was noticed at final time point, which suggests that both treatments increased platelets in dogs naturally infected with E. canis.
Chantal T. Rosa
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.
Tabar, M-D; Movilla, R; Serrano, L; Altet, L; Francino, O; Roura, X
To investigate evidence for selected vector-borne pathogen infections in dogs with pericardial effusion living in a Mediterranean area in which several canine vector-borne diseases are endemic. Archived EDTA blood (n=68) and pericardial fluid samples (n=58) from dogs with pericardial effusion (n=68) were included. Dogs without pericardial effusion examined for other reasons were included as controls (n=60). Pericardial effusion was classified as neoplastic in 40 dogs, idiopathic in 23 dogs and of unknown aetiology in 5 dogs. Real-time PCR was performed for Leishmania infantum, Ehrlichia/Anaplasma species, Hepatozoon canis, Babesia species, Rickettsia species and Bartonella species, and sequencing of PCR products from positive samples was used to confirm species specificity. Vector-borne pathogens were found in 18 dogs: 16 of 68 dogs with pericardial effusion (23·5%) and two of 60 control dogs (3·3%). Positive dogs demonstrated DNA of Leishmania infantum (n=7), Anaplasma platys (n=2, one dog coinfected with Leishmania infantum), Babesia canis (n=5), Babesia gibsoni (n=3) and Hepatozoon canis (n=2). Vector-borne pathogens were more commonly detected among dogs with pericardial effusion than controls (P=0·001). There was no relationship between aetiology of the pericardial effusion and evidence of vector-borne pathogens (P=0·932). Vector-borne pathogens are often detected in dogs with pericardial effusion and require further investigation, especially in dogs with idiopathic pericardial effusion. PCR can provide additional information about the potential role of vector-borne pathogens in dogs with pericardial effusion living in endemic areas. © 2018 British Small Animal Veterinary Association.
Gubbels, J. M.; de Vos, A. P.; van der Weide, M.; Viseras, J.; Schouls, L. M.; de Vries, E.; Jongejan, F.
A reverse line blot (RLB) assay was developed for the identification of cattle carrying different species of Theileria and Babesia simultaneously. We included Theileria annulata, T. parva, T. mutans, T. taurotragi, and T. velifera in the assay, as well as parasites belonging to the T. sergenti-T. buffeli-T. orientalis group. The Babesia species included were Babesia bovis, B. bigemina, and B. divergens. The assay employs one set of primers for specific amplification of the rRNA gene V4 hypervariable regions of all Theileria and Babesia species. PCR products obtained from blood samples were hybridized to a membrane onto which nine species-specific oligonucleotides were covalently linked. Cross-reactions were not observed between any of the tested species. No DNA sequences from Bos taurus or other hemoparasites (Trypanosoma species, Cowdria ruminantium, Anaplasma marginale, and Ehrlichia species) were amplified. The sensitivity of the assay was determined at 0.000001% parasitemia, enabling detection of the carrier state of most parasites. Mixed DNAs from five different parasites were correctly identified. Moreover, blood samples from cattle experimentally infected with two different parasites reacted only with the corresponding species-specific oligonucleotides. Finally, RLB was used to screen blood samples collected from carrier cattle in two regions of Spain. T. annulata, T. orientalis, and B. bigemina were identified in these samples. In conclusion, the RLB is a versatile technique for simultaneous detection of all bovine tick-borne protozoan parasites. We recommend its use for integrated epidemiological monitoring of tick-borne disease, since RLB can also be used for screening ticks and can easily be expanded to include additional hemoparasite species. PMID:10325324
Samara Machuca Figueroa
Full Text Available La línea celular RF/6A ha sido utilizada en estudios de corto plazo evaluando fármacos o infecciones experimentales con Anaplasma marginale; en contraste, DH82 es utilizada para la multiplicación de Ehrlichia canis. No obstante, se desconocen condiciones específicas de su crecimiento, por lo que se diseñaron varios experimentos para resolver interrogantes de su propagación. Ambas líneas, se adquirieron de la American Type Culture Collection, mantenidas en Medio Mínimo Esencial suplementado con suero fetal bovino, piruvato de Na y NaHCO3 e incubadas en atmósfera de 5% de CO2 en aire, a 37 °C. Los primeros ensayos, en placas de 24 pozos, esclarecieron los valores de dosis mínima inicial, que fueron 62,500 y 8,836 células/pozo para DH82 y RF/6A; así como los de densidad de siembra; cultivos con concentraciones de 5, 10, 20 y 40 células por mm2, cosechados con solución Tripsina-EDTA al alcanzar >95% de confluencia. Los índices estimados fueron: 3,319.32, 1,956.70, 870.73 y 422.14 para DH82 y 62.38, 63.51, 25.31 y 12.16 veces con RF/6A. La cinética del crecimiento, en cajas de Petri de 35 mm Ø, incluyó la siembra de 20 células/mm2, cambio del medio cada 63 h y cosecha cada 21 h para DH82; para RF/6A; la siembra fue 10 células/ mm2, cambio de medio cada 45 h y cosecha cada 15 h. El máximo crecimiento se observó hasta las 336 y 315 h con tiempos de duplicación de 42.9 y 36.9 h respectivamente para DH82 y RF/6A. Los datos permitieron proponer un modelo patrón de cultivo, para estudios futuros.
Full Text Available Rosalie Greenberg Medical Arts Psychotherapy Associates PA, Summit, NJ, USAI read with great interest the article by Bransfield,1 wherein the author reviewed potential contributors to and manifestations of heightened loss of control in Lyme disease patients. As a child psychiatrist living in a Lyme-endemic state, New Jersey, in the USA, I have seen a number of children and adolescents who exhibit significant acute or gradual onset of highly oppositional behaviors often both at home and at school. Refusal to do classwork or homework, heightened paranoia or feeling rejected by others is often present. In addition, they often demonstrate increased irritability, extreme temper tantrums, problems concentrating, elevated impulsivity, sensory hypersensitivity (sound, touch, smell, taste and/or light and intense emotional lability. Parents describe these behaviors as either a sudden change or a period of worsening of a previous condition, such as attention-deficit hyperactivity disorder or mood disorder. During periods of intense anger over minimal issues, they can appear menacing and threaten to kill a sibling, parent or a friend or state that they want to die themselves. In a recent record review of 69 youth evaluated in my private psychopharmacology practice, 49 of these new patients, without a known history of Lyme disease or other tick-borne disorders, demonstrated evidence of exposure to one or more of the pathogens Borrelia burgdorferi, Bartonella, Babesia, Ehrlichia and Anaplasma on serologic testing performed by multiple laboratories. In addition, 6 of the 69 patients were initially referred because of psychiatric difficulties observed by other physicians during the active treatment of tick-borne infections. Supporting this connection of childhood neuropsychiatric symptoms and tick-borne illness is a recent article looking at 27 youth diagnosed with bipolar disorder I or II, which found that 74% or 20/27 were diagnosed with at least one tick
Díaz-Regañón, David; Villaescusa, Alejandra; Ayllón, Tania; Rodríguez-Franco, Fernando; Baneth, Gad; Calleja-Bueno, Lydia; García-Sancho, Mercedes; Agulla, Beatriz; Sainz, Ángel
-borne pathogens, such as Ehrlichia canis and Bartonella henselae. Our results indicate that cats from Madrid, central Spain, are infected with Hepatozoon spp. and Cytauxzoon sp., although with a low prevalence. Further studies are needed to determine the virulence of these agents in Spanish cats.
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
Annoscia, Giada; Latrofa, Maria Stefania; Cantacessi, Cinzia; Olivieri, Emanuela; Manfredi, Maria Teresa; Dantas-Torres, Filipe; Otranto, Domenico
Babesia spp. are globally distributed tick-borne protozoan parasites that infect the red blood cells of a wide range of vertebrate hosts, including humans. Diagnosis of babesiosis is often impeded by the transient presence of the parasites in peripheral blood, as well as by their pleomorphic nature. Given the reports of an expanding and, in some cases, sympatric geographical distribution of Babesia canis and Babesia vogeli in dogs and associated vectors, in Europe, the development of time-efficient and cost-effective diagnostic tools to detect and differentiate these two species is warranted. In this study, we designed and developed a novel polymerase chain reaction (PCR) assay targeting the parasite cytochrome c oxidase subunit 1 (cox1) gene, for the simultaneous detection and differentiation of B. canis and B. vogeli. The analytical sensitivity of the PCR was evaluated using serial dilutions of genomic DNA extracted from individual and artificially mixed canine blood samples infected by B. canis (3×10 2 infected erythrocytes/ml, ie/ml) and B. vogeli (2.1×10 1 ie/ml). The analytical specificity of the assay was assessed using blood samples positive for Hepatozoon canis, Ehrlichia canis, Anaplasma platys, Babesia microti, Babesia rossi and Theileria annae (syn. Babesia vulpes). The clinical specificity of the PCR assay was evaluated on 147 blood samples from dogs and 128 tick specimens (Dermacentor reticulatus and Rhipicephalus sanguineus sensu lato). Species-specific bands of the expected sizes (i.e., 750bp for B. canis and 450bp for B. vogeli), and two bands in the mixed blood samples were obtained. The PCR assay developed herein detected a low number of infected erythrocytes (i.e., 3×10 -2 B. canis, 2.1×10 -2 B. vogeli ie/ml). Of the 147 blood samples, nine (6.1%) were positive for B. canis and six (4.1%) for B. vogeli, whereas only one tick (D. reticulatus) was positive for B. canis. This PCR assay represents a rapid and reliable tool for the diagnosis of B
Full Text Available Mosquito-borne diseases are common high-impact diseases in tropical and subtropical areas. However, other non-mosquito vector-borne pathogens (VBPs may share their geographic distribution, seasonality, and clinical manifestations, thereby contributing their share to the morbidity and mortality caused by febrile illnesses in these regions. The purpose of this work was to collect and review existing information and identify knowledge gaps about tick, flea-, and louse-borne diseases of veterinary and public health significance in Nigeria. Full-length articles about VBPs were reviewed and relevant information about the vectors, their hosts, geographic distribution, seasonality, and association(s with human or veterinary diseases was extracted. Specific laboratory tools used for detection and identification of VBPs in Nigeria were also identified. A total of 62 original publications were examined. Substantial information about the prevalence and impacts of ticks and fleas on pet and service dogs (18 articles, and livestock animals (23 articles were available; however, information about their association with and potential for causing human illnesses was largely absent despite the zoonotic nature of many of these peri-domestic veterinary diseases. Recent publications that employed molecular methods of detection demonstrated the occurrence of several classic (Ehrlichia canis, Rickettsia africae, Bartonella sp. and emerging human pathogens (R. aeschlimannii, Neoehrlichia mikurensis in ticks and fleas. However, information about other pathogens often found in association with ticks (R. conorii and fleas (R. typhi, R. felis across the African continent was lacking. Records of louse-borne epidemic typhus in Nigeria date to 1947; however, its current status is not known. This review provides an essential baseline summary of the current knowledge in Nigeria of non-mosquito VBPs, and should stimulate improvements in the surveillance of the veterinary and
Ghatalia, Pooja; Porter, Joanne; Wroblewski, Danielle; Carlson, John Andrew
Primary cutaneous marginal zone lymphoma (PCMZL) has rarely been reported in teenagers and is occasionally associated with Borrelia burgdorferi infection. Juxta-articular fibrotic nodules represent a unique, localized fibrosing response to spirochete infections, namely Borreliosis. Herein, we report a 15-year-old healthy boy who presented with a 4-year history of progressive acquisition of asymptomatic, erythematous nodules, ≤ 3 cm, beginning with his right forearm (3), then right arm (1) and lastly his right inner thigh (1). Biopsy showed PCMZL in three of five samples, and inflamed, fibrotic nodules, near the elbow in two. The bottom heavy lymphomatous nodules consisted of mostly small CD20+ CD43+ lymphocytes, some with plasmacytoid features. Mature plasma cells were lambda light chain restricted by in situ hybridization. The juxta-articular fibrotic nodules were located in the deep dermis and subcutis, had peripheral plasma cell-rich infiltrates, and showed nodular sclerosis (morphea profunda-like) in one, and lamellar and angiocentric sclerosis in the other reminiscent of quiescent lesions of chronic localized fibrosing leukocytoclastic vasculitis. Immunohistochemistry for B. burgdorferi revealed rare positive organisms; however, polymerase chain reaction (PCR) and serology were negative for B. burgdorferi as were serologic and/or PCR assays for Bartonella henselae, Ba. quintana, Ehrlichia chaffeensis, Treponema pallidum, Helicobacter pylori and Babesia microti. No evidence of extracutaneous disease was found by the review of systems and imaging studies. A 4-week trial of doxycycline therapy failed, whereas intralesional (IL) corticosteroid therapy induced rapid regression of his nodules. After two local recurrences, also treated with IL corticosteroids, he is well, without cutaneous disease, 20 months later. A literature review of 19 pediatric cases PCMZL reveals a similar natural history as adult PCMZL. Despite negative serology and PCR for B. burgdorferi
Schötta, Anna-Margarita; Wijnveld, Michiel; Stockinger, Hannes; Stanek, Gerold
Ticks transmit a large number of pathogens capable of causing human disease. In this study, the PCR-reverse line blot (RLB) method was used to screen for pathogens in a total of 554 Ixodes ricinus ticks collected from all provinces of Austria. These pathogens belong to the genera Borrelia , Rickettsiae , Anaplasma / Ehrlichia (including " Candidatus Neoehrlichia"), Babesia , and Coxiella The pathogens with the highest detected prevalence were spirochetes of the Borrelia burgdorferi sensu lato complex, in 142 ticks (25.6%). Borrelia afzelii (80/142) was the most frequently detected species, followed by Borrelia burgdorferi sensu stricto (38/142) and Borrelia valaisiana (36/142). Borrelia garinii/Borrelia bavariensis , Borrelia lusitaniae , and Borrelia spielmanii were found in 28 ticks, 5 ticks, and 1 tick, respectively. Rickettsia spp. were detected in 93 ticks (16.8%): R. helvetica (39/93), R. raoultii (38/93), R. monacensis (2/93), and R. slovaca (1/93). Thirteen Rickettsia samples remain uncharacterized. " Candidatus Neoehrlichia mikurensis," Babesia spp. ( B. venatorum , B. divergens , B. microti ), and Anaplasma phagocytophilum were found in 4.5%, 2.7%, and 0.7%, respectively. Coxiella burnetii was not detected. Multiple microorganisms were detected in 40 ticks (7.2%), and the cooccurrence of Babesia spp. and " Candidatus Neoehrlichia mikurensis" showed a significant positive correlation. We also compared different PCR-RLBs for detection of Borrelia burgdorferi sensu lato and Rickettsia spp. and showed that different detection approaches provide highly diverse results, indicating that analysis of environmental samples remains challenging. IMPORTANCE This study determined the wide spectrum of tick-borne bacterial and protozoal pathogens that can be encountered in Austria. Surveillance of (putative) pathogenic microorganisms occurring in the environment is of medical importance, especially when those agents can be transmitted by ticks and cause disease. The
van Vuuren, M; Penzhorn, B L
The role of African wildlife in the occurrence of vector-borne infections in domestic animals has gained renewed interest as emerging and re-emerging infections occur worldwide at an increasing rate. In Africa, biodiversity conservation and the expansion of livestock production have increased the risk of transmitting vector-borne infections between wildlife and livestock. The indigenous African pathogens with transboundary potential, such as Rift Valley fever virus, African horse sickness virus, bluetongue virus, lumpy skin disease virus, African swine fever virus, and blood-borne parasites have received the most attention. There is no evidence for persistent vector-borne viral infections in African wildlife. For some viral infections, wildlife may act as a reservoir through the inter-epidemic circulation of viruses with mild or subclinical manifestations. Wildlife may also act as introductory or transporting hosts when moved to new regions, e.g. for lumpy skin disease virus, Rift Valley fever virus and West Nile virus. Wildlife may also act as amplifying hosts when exposed to viruses in the early part of the warm season when vectors are active, with spillover to domestic animals later in the season, e.g. with bluetongue and African horse sickness. Some tick species found on domestic animals are more abundant on wildlife hosts; some depend on wildlife hosts to complete their life cycle. Since the endemic stability of a disease depends on a sufficiently large tick population to ensure that domestic animals become infected at an early age, the presence of wildlife hosts that augment tick numbers may be beneficial. Many wild ungulate species are reservoirs of Anaplasma spp., while the role of wildlife in the epidemiology of heartwater (Ehrlichia ruminantium infection) has not been elucidated. Wild ungulates are not usually reservoirs of piroplasms that affect livestock; however, there are two exceptions: zebra, which are reservoirs of Babesia caballi and Theileria
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
Solcà, Manuela da Silva; Guedes, Carlos Eduardo Sampaio; Nascimento, Eliane Gomes; Oliveira, Geraldo Gileno de Sá; dos Santos, Washington Luis Conrado; Fraga, Deborah Bittencourt Mothé; Veras, Patrícia Sampaio Tavares
Because infected dogs are widely considered to be the main domestic reservoir for Leishmania infantum (syn Leishmania chagasi) parasites in Brazil, the diagnosis of canine visceral leishmaniasis (CVL) must be made both accurately and promptly. The present study attempted to standardize a conventional polymerase chain reaction (cPCR) protocol for the detection of L. infantum DNA in canine spleen samples. Quantitative PCR (qPCR) technique was used to confirm the presence of Leishmania DNA in the canine spleen fragments. A comparison was made between the efficacies of these molecular diagnostic techniques and conventional parasitological and serological methods. cPCR protocols for spleen samples were standardized using primers that amplify a 145 bp fragment, located at the parasite kinetoplast minicircle. The genus specificity of the cPCR protocol was assessed by its inability to amplify the DNA of other common canine pathogens, such as Ehrlichia canis, Babesia canis, Toxoplasma gondii and Trypanosoma cruzi. cPCR protocol sensitivity was tested by assessing the reaction detection limit, determined to be 10 fg of L. infantum reference strain DNA, which corresponds to a range of 0.03-0.1 parasites per fragment. Standardized cPCR protocol was used to detect the presence of Leishmania in 45 dog spleen samples. Our results showed that 40% of the spleen fragment cultures were positive for Leishmania parasites, 58% of the dog serum samples tested positive using ELISA, and parasite DNA was detected in 44% using qPCR, while 47% of the spleen samples using cPCR. Diagnostic methods performance was assessed and revealed a better degree of ascertainment for cPCR when compared to other diagnostic methods. The sensitivity of ELISA was 83.3%, qPCR was 83.3%, and cPCR was 88.9%; PPV for ELISA was 57.7%, qPCR was 75% and cPCR was 76.2%; the Kappa coefficients were found to be 0.40 (fair) for ELISA, 0.64 (substantial) for qPCR and 0.68 (substantial) for cPCR. In both oligosymptomatic