Frequency of co-seropositivities for certain pathogens and their relationship with clinical and histopathological changes and parasite load in dogs infected with Leishmania infantum.

In canine leishmaniosis caused by the protozoan Leishmania infantum, little is known about how co-infections with or co-seropositivities for other pathogens can influence aggravation of this disease. Therefore, the objectives of this study were to evaluate the frequency of co-infections with or co-seropositivities for certain pathogens in dogs seropositive for L. infantum and their relationship with clinical signs, histological changes and L. infantum load. Sixty-six L. infantum-seropositive dogs were submitted to clinical examination, collection of blood and bone marrow, culling, and necropsy. Antibodies against Anaplasma spp., Borrelia burgdorferi sensu lato, Ehrlichia spp. and Toxoplasma gondii and Dirofilaria immitis antigens were investigated in serum. Samples from different tissues were submitted to histopathology and immunohistochemistry for the detection of Leishmania spp. and T. gondii. Quantitative real-time PCR was used to assess the L. infantum load in spleen samples. For detection of Coxiella burnetii, conventional PCR and nested PCR were performed using bone marrow samples. All 66 dogs tested positive for L. infantum by qPCR and/or culture. Fifty dogs (76%) were co-seropositive for at least one pathogen: T. gondii (59%), Ehrlichia spp., (41%), and Anaplasma spp. (18%). Clinical signs were observed in 15 (94%) dogs monoinfected with L. infantum and in 45 (90%) dogs co-seropositive for certain pathogens. The L. infantum load in spleen and skin did not differ significantly between monoinfected and co-seropositive dogs. The number of inflammatory cells was higher in the spleen, lung and mammary gland of co-seropositive dogs and in the mitral valve of monoinfected dogs. These results suggest that dogs infected with L. infantum and co-seropositive for certain pathogens are common in the region studied. However, co-seropositivities for certain pathogens did not aggravate clinical signs or L. infantum load, although they were associated with a more intense inflammatory reaction in some organs.

Diversity unearthed by the estimated molecular phylogeny and ecologically quantitative characteristics of uncultured Ehrlichia bacteria in Haemaphysalis ticks, Japan.

Ehrlichia species are obligatory intracellular bacteria transmitted by arthropods, and some of these species cause febrile diseases in humans and livestock. Genome sequencing has only been performed with cultured Ehrlichia species, and the taxonomic status of such ehrlichiae has been estimated by core genome-based phylogenetic analysis. However, many uncultured ehrlichiae exist in nature throughout the world, including Japan. This study aimed to conduct a molecular-based taxonomic and ecological characterization of uncultured Ehrlichia species or genotypes from ticks in Japan. We first surveyed 616 Haemaphysalis ticks by p28-PCR screening and analyzed five additional housekeeping genes (16S rRNA, groEL, gltA, ftsZ, and rpoB) from 11 p28-PCR-positive ticks. Phylogenetic analyses of the respective genes showed similar trees but with some differences. Furthermore, we found that V1 in the V1-V9 regions of Ehrlichia 16S rRNA exhibited the greatest variability. From an ecological viewpoint, the amounts of ehrlichiae in a single tick were found to equal approx. 6.3E+3 to 2.0E+6. Subsequently, core-partial-RGGFR-based phylogenetic analysis based on the concatenated sequences of the five housekeeping loci revealed six Ehrlichia genotypes, which included potentially new Ehrlichia species. Thus, our approach contributes to the taxonomic profiling and ecological quantitative analysis of uncultured or unidentified Ehrlichia species or genotypes worldwide.

Epidemiological link between canine monocytic ehrlichiosis caused by Ehrlichia canis and the presence of Rhipicephalus sanguineus sensu stricto in Argentina.

In this work, we analyze data that support an epidemiological link between cases of canine monocytic ehrlichiosis (CME) by Ehrlichia canis and the presence of Rhipicephalus sanguineus sensu stricto as vector in an endemic area for this tick in Argentina. In a blood sample of a 1-year-old toy poodle with CME compatible clinical signs, which showed CME typical morulae in monocytes in Giemsa-stained blood smear, DNA of E. canis was detected by PCR. Further, DNA of E. canis was also detected in a female of R. sanguineus s.s. collected on the infected dog. Rhipicephalus sanguineus s.s. is the only member of the R. sanguineus group that prevails in the study area. The results of this study suggest that R. sanguineus s.s. may play a more important role in the transmission of E. canis than it was assumed so far. The epidemiological link between CME cases and R. sanguineus s.s. as vector in temperate areas of Argentina described in this work contrast previous studies which found that R. sanguineus sensu lato "tropical lineage" (which is absent in the study area) is competent to transmit E. canis but not R. sanguineus s.s.

Molecular identification and characterization of Rickettsia spp. and other tick-borne pathogens in cattle and their ticks from Huambo, Angola.

Ticks are one of the most common vectors of a broad variety of pathogenic agents that significantly affects cattle production causing reduced productivity and important economic losses, while simultaneously having an impact on human health due to the zoonotic risk. In much of the territory of Angola urban population has grown rapidly in recent decades, sharing today close contact with large farms that are generally owned by city residents, providing the ideal conditions for vector-borne pathogens (VBP) transmission between animals and humans. Here we studied the occurrence of Rickettsia, Anaplasma, Ehrlichia, Babesia and Theileria in domestic cattle (n = 98) from Huambo, Angola, and their ticks (n = 116) to obtain a more detailed knowledge into the spectrum of tick-borne agents circulating in this population. We morphologically identified Amblyomma variegatum, Rhipicephalus decoloratus and R. evertsi mimeticus ticks, further confirmed by molecular analysis of the 12S rDNA and 16S rDNA genes. Although none of the bovine blood showed to be positive for R. africae by the ompB, ompA and gltA assays, five ticks showed to be positive for R. africae by the ompB, ompA and the gltA PCRs. Two 18S rRNA sequences were retrieved from bovine blood and one sequence from A. variegatum tick, showing 100% identity with Theileria mutans. By using a PCR targeting the 16S rRNA gene of Anaplama spp. we have also obtained six bovine blood samples showing 99-100 % nucleotide sequence identity with A. capra, two showing 98 % nucleotide sequence identity with A. phagocytophilum and three showing 98-100 % nucleotide sequence identity with A. platys. None of the ticks were positive. The present study shows the presence of a wide range of vector-borne diseases in domestic cattle and their ticks in Huambo province, Angola. Given the lack of both animal and human health infrastructures in this rural region, swift diagnosis and treatment is hampered which could produce a more severe impact on health.

Serological survey of vector-borne pathogens in stray dogs from Sofia area, Bulgaria.

Vector-borne diseases represent a significant part of canine infectious pathology posing serious zoonotic potential. Stray dogs are found to be an important health and ecological factor in urban areas. The objectives of the present study were to update the current knowledge on the seroprevalence of Anaplasma phagocytophilum/ Anaplasma platys, Ehrlichia canis, Borrelia burgdorferi and Dirofilaria immitis in stray dogs from Sofia, Bulgaria. A total of 448 apparently healthy and randomly selected dogs were tested with Anigen Rapid CaniV-4 Test Kit. Overall 28.57% (128/448) of all tested animals were positive for one or more of the pathogenic species. Total percentage was distributed as follows (single plus mix infections): Anaplasma spp. - 16.29% (73/448), D. immitis - 13.39% (60/448), Ehrlichia spp. - 1.34% (6/448) and B.burgdorferi - 0.67% (3/448). The current study demonstrated the circulation of Anaplasma spp. and D. immitis among stray dogs in Sofia, Bulgaria.

Parasites of veterinary importance from domestic animals in uMkhanyakude district of KwaZulu-Natal province.

This study investigated the occurrence and phylogenetic relationship of protozoan parasites and Ehrlichia infecting domestic animals from three municipalities in uMkhanyakude district of KwaZulu-Natal province, South Africa. A total of 208 blood samples collected from clinically healthy cattle, sheep, goats and dogs from uMkhanyakude district were examined by polymerase chain reaction (PCR) assays, using either genus or species-specific primers to determine the occurrence and phylogenetic relationship of various protozoan parasites and Ehrlichia of veterinary importance. A total of 5/109 (4.6%) cattle were PCR-positive for the presence of Toxoplasma gondii, 33/109 (30.3%) for Babesia bovis, 24/109 (22.02%) for Babesia bigemina and 20/109 (18.3%) for Trypanosoma sp., while 3/10 (30%) of sheep were PCR-positive for Theileria ovis and none of the goats were positive for any of the detected pathogens. The co-infection of 4/109 (3.7%) B. bovis and B. bigemina was detected in cattle. Only Ehrlichia canis was detected in dogs with infection rate of 20/48 (41.7%). Sequences of PCR-positive isolates (B. bovis, B. bigemina, E. canis, T. ovis and T. gondii) showed that they were closely related to their relevant species from various countries. These findings have expanded our knowledge about the prevalence and phylogenetic similarity between protozoan parasites and Ehrlichia isolates of South African origin. To date, this is the first study in South Africa to detect T. gondii infections from cattle blood using PCR.

Transmission of Anaplasma phagocytophilum (Foggie, 1949) by Ixodes ricinus (Linnaeus, 1758) ticks feeding on dogs and artificial membranes.

BACKGROUND: The interplay of speed of activity of acaricidal products and tick-borne pathogen transmission time is the major driver for disease prevention. This study aimed to investigate the time required for transmission of Anaplasma phagocytophilum by adult Ixodes ricinus ticks in vivo on dogs, and to confirm the time required for transmission observed in vivo, in vitro. METHODS: Nymphs of I. ricinus were experimentally infected with an A. phagocytophilum strain of canine origin. Dogs were allocated to 6 groups of 3 dogs each. Groups 1-5 were infested with 50 A. phagocytophilum-infected female adult ticks on Day 0. Ticks were removed post-infestation at 3, 6, 12, 24 and 48 h. Dogs in Group 6 were infested with 60 A. phagocytophilum-infected female adult ticks (left on dogs until engorged). Dogs were observed daily for general health and clinically examined on Day 0, and weekly from Day 14. Blood was collected for qPCR and serological analysis on Day 0 (pre-challenge) and weekly thereafter. In the in vitro study each artificial feeding chamber was seeded with 10 adult ticks (5 male/5 female), attachment assessed, and blood pools sampled for qPCR at 6 h intervals up to 72 h after first tick attachment. RESULTS: Anaplasma phagocytophilum specific antibodies and DNA were detected in all 3 dogs in Group 6. No A. phagocytophilum-specific antibodies or DNA were detected in any dogs in Groups 1-5. All dogs remained healthy. Female tick attachment in 60 artificial feeding chambers over 72 h ranged between 20-60%. Anaplasma phagocytophilum DNA was detected in the blood collected from 5% of chambers sampled at 6 h, with the highest number of positive samples (16.3%) observed at 36 h. CONCLUSIONS: Transmission of A. phagocytophilum by I. ricinus ticks starts within a few hours after attachment but establishment of infections in dogs is apparently dependent on a minimum inoculation dose that was only observed when ticks attached for greater than 48 h. These findings highlight the need for acaricidal products to exert a repellent and/or rapid killing effect on ticks to forestall transmission and subsequent disease.

Prevalence of Borrelia burgdorferi, Anaplasma spp., Ehrlichia spp. and Dirofilaria immitis in Canadian dogs, 2008 to 2015: a repeat cross-sectional study.

BACKGROUND: Vector-borne pathogens are emerging concerns in multiple regions of Canada. Determining regional prevalence of canine vector-borne pathogens and documenting change will improve clinician awareness, enable targeted prevention, enhance diagnosis and ideally reduce the risk of disease. Study objectives were to: (i) estimate the prevalence of positive canine vector-borne test results from samples submitted in Canada; (ii) assess change in prevalence over time, from baseline (2008) to 2015; and (iii) estimate the prevalence of pathogen co-infections. METHODS: This repeat cross-sectional study evaluated 753,468 test results for D. immitis antigen and B. burgdorferi, Ehrlichia canis/ewingii/muris serology, and 753,208 test results for Anaplasma phagocytophilum/platys serology using the SNAP® 4Dx®Test and SNAP 4Dx® Plus Test. RESULTS: Based on all submitted samples from Canada (2008-2015), the period seroprevalence of B. burgdorferi, Ehrlichia spp., Anaplasma spp. and D. immitis antigen were 2.0%, 0.5%, 0.4% and 0.2%, respectively. Over the 7 years (2008 compared to 2015) we observed a significant increase in seroprevalence for B. burgdorferi (144.4%) and Ehrlichia spp. (150%). Co-infections (positive for two or more pathogens on a single 4 pathogen test kit) were estimated at 5.4% (1162/21,612) of total positive tests. CONCLUSIONS: The temporal rise and geographical differences in prevalence detected for these pathogens (notably B. burgdorferi) are consistent with anecdotal information on canine illness related to tick-borne pathogen exposure in multiple regions of Canada, particularly canine Lyme disease.

Repeated cross-sectional study of Trypanosoma cruzi in shelter dogs in Texas, in the context of Dirofilaria immitis and tick-borne pathogen prevalence.

BACKGROUND: Vector-borne diseases have an adverse impact on health of dogs, and infected dogs can be sentinels for human infection. Infection with Trypanosoma cruzi, an agent of Chagas disease, causes fatal heart disease in dogs across the southern United States but has been neglected from wide-scale prevalence studies. OBJECTIVES: To determine the prevalence of exposure to T. cruzi, Ehrlichia spp., Anaplasma spp., Borrelia burgdorferi, and infection with Dirofilaria immitis among dogs in shelters across Texas and to identify risk factors for T. cruzi seropositivity. ANIMALS: Six hundred and eight dogs. METHODS: This repeated cross-sectional study was performed by collecting blood from ~30 dogs during each of the 3 visits to 7 shelters. We tested serum for antibodies to T. cruzi using 2 tests in series and for antibodies to Ehrlichia spp., Anaplasma spp., and B. burgdorferi and D. immitis antigen using the IDEXX SNAP 4DX Plus point-of-care test. DNA was extracted from blood clots and tested for T. cruzi DNA and strain type via quantitative polymerase chain reactions (qPCR). We used logistic regression to assess risk factors. RESULTS: One hundred ten (18.1%) of 608 dogs were seropositive for T. cruzi. Prevalence of exposure to the other vector-borne agents was: Ehrlichia spp. 3.6%; Anaplasma spp. 6.9%; B. burgdorferi 0.2%; and D. immitis infection 16.0%. Six of 559 (1.1%) dogs were qPCR-positive for T. cruzi. CONCLUSIONS AND CLINICAL IMPORTANCE: T. cruzi seroprevalence was comparable to D. immitis prevalence and higher than seroprevalence of the tick-borne pathogens. T. cruzi is an underrecognized health threat to dogs across Texas and possibly other southern states where triatomine vectors are endemic.

Association between canine leishmaniosis and Ehrlichia canis co-infection: a prospective case-control study.

BACKGROUND: 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. METHODS: 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. RESULTS: 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 be co-infected with M. haemocanis. CONCLUSIONS: Dogs with ClinL are at a higher risk of co-infection with E. canis than clinically healthy dogs. We recommend that dogs diagnosed with ClinL should be tested for E. canis co-infection using PCR.

Evaluating acarological risk for exposure to Ixodes scapularis and Ixodes scapularis-borne pathogens in recreational and residential settings in Washington County, Minnesota.

The distribution of I. scapularis, the tick vector of the bacteria that cause Lyme disease, has been expanding over the last two decades in the north-central United States in parallel with increasing incidence of human cases of Lyme disease in that region. However, assessments of residential risk for exposure to ticks are lacking from this region. Here, we measured the density of host-seeking I. scapularis nymphs in two suburban and two rural public recreational sites located in Washington County, Minnesota as well as in nearby residential properties. We sought to compare tick densities across land use types and to identify environmental factors that might impact nymphal density. We also assessed the prevalence of infection in the collected ticks with Lyme disease spirochetes (Borrelia burgdorferi sensu stricto, B. mayonii), and other I. scapularis-borne pathogens including B. miyamotoi, Babesia microti and Anaplasma phagocytophilum. Similar to studies from the eastern United States, on residential properties, I. scapularis nymphal densities were highest in the ecotonal areas between the forest edge and the lawn. Residences with the highest densities of nymphs were more likely to have a higher percentage of forest cover, log piles, and signs of deer on their property. In recreational areas, we found the highest nymphal densities both in the wooded areas next to trails as well as on mowed trails. Among the 303 host-seeking I. scapularis nymphs tested for pathogens, B. burgdorferi sensu stricto, A. phagocytophilum and B. miyamotoi were detected in 42 (13.8%), 14 (4.6%), and 2 (0.6%) nymphs, respectively.

Molecular Detection and Phylogenetic Analysis of Anaplasma phagocytophilum in Horses in Korea.

The identification and characterization of pathogenic and zoonotic tick-borne diseases like granulocytic anaplasmosis are essential for developing effective control programs. The differential diagnosis of pathogenic Anaplasma phagocytophilum and non-pathogenic A. phagocytophilum-like Anaplasma spp. is important for implementing effective treatment from control programs. The objective of the present study was to investigate the prevalence of Anaplasma spp. in horses in Korea by nucleotide sequencing and restriction enzyme fragment length polymorphism assay. Of the 627 horses included in the study, only 1 (0.2%) was infected with A. phagocytophilum. Co-infection with A. phagocytophilum- like Anaplasma spp. was not detected in the study. The 16S rRNA sequence of A. phagocytophilum was similar (99.5- 100%) to A. phagocytophilum 16S rRNA isolated from horses in other countries. PCR adapted to amplify A. phagocytophilum groEL and msp2 genes failed to generate amplicons, suggesting genetic diversity in these genes. This study is the first molecular detection of A. phagocytophilum in horses in Korea. Human granulocytic anaplasmosis and animal infection of A. phagocytophilum have been reported in Korea recently. Because of vector tick distribution, global warming, and the increase of the horse industry, horses should be considered as a potential reservoir for A. phagocytophilum, and cross infectivity should be evaluated even though a low prevalence of infection was detected in this study. Furthermore, continuous surveillance and effective control measures for A. phagocytophilum should be established to prevent disease distribution and possible transmission to humans.

Serum antioxidant capacity and oxidative damage in clinical and subclinical canine ehrlichiosis.

The objective of this study was to evaluate and compare the antioxidant response and the products of oxidative damage analysed by various assays in clinical and subclinical canine monocytic ehrlichiosis (CME). For this purpose, four assays to measure the total serum antioxidant capacity (TAC), such as the cupric reducing antioxidant capacity (CUPRAC), ferric reducing ability of plasma (FRAP), trolox equivalent antioxidant capacity (TEAC) using acidic medium (TEACA), and the TEAC using the horseradish peroxidase (TEACH) were used. In addition, the serum thiol concentrations were analysed. Reactive oxygen species (ROS), thiobarbituric acid reactive substances (TBARS), and ferrous oxidation-xylenol orange (FOX) were measured to determine the concentrations of free radical and the products of oxidative damage as result of the disease. All antioxidant markers were significantly lower in the dogs on clinical ehrlichiosis when compared with healthy dogs; however only the CUPRAC, FRAP and thiol were significantly lower in subclinical CME compared with healthy dogs. TBARS and FOX showed no significant differences between dogs with CME and healthy dogs; however, a significant increased ROS concentration was observed in dogs with clinical and subclinical CME when compared with healthy dogs. Results showed that in CME there is a state of oxidative stress with significant changes in markers of antioxidant defence and in concentrations of free radicals. However, the detection of these changes would depend of the assay used.

A Bayesian spatio-temporal model for forecasting the prevalence of antibodies to Ehrlichia species in domestic dogs within the contiguous United States.

BACKGROUND: Dogs in the United States are hosts to a diverse range of vector-borne pathogens, several of which are important zoonoses. This paper describes factors deemed to be significantly related to the prevalence of antibodies to Ehrlichia spp. in domestic dogs, including climatic conditions, geographical factors, and societal factors. These factors are used in concert with a spatio-temporal model to construct an annual seroprevalence forecast. The proposed method of forecasting and an assessment of its fidelity are described. METHODS: Approximately twelve million serological test results for canine exposure to Ehrlichia spp. were used in the development of a Bayesian approach to forecast canine infection. Data used were collected on the county level across the contiguous United States from routine veterinary diagnostic tests between 2011-2015. Maps depicting the spatial baseline Ehrlichia spp. prevalence were constructed using Kriging and head-banging smoothing methods. Data were statistically analyzed to identify factors related to antibody prevalence via a Bayesian spatio-temporal conditional autoregressive (CAR) model. Finally, a forecast of future Ehrlichia seroprevalence was constructed based on the proposed model using county-level data on five predictive factors identified at a workshop hosted by the Companion Animal Parasite Council and published in 2014: annual temperature, percentage forest coverage, percentage surface water coverage, population density and median household income. Data were statistically analyzed to identify factors related to disease prevalence via a Bayesian spatio-temporal model. The fitted model and factor extrapolations were then used to forecast the regional seroprevalence for 2016. RESULTS: The correlation between the observed and model-estimated county-by-county Ehrlichia seroprevalence for the five-year period 2011-2015 is 0.842, demonstrating reasonable model accuracy. The weighted correlation (acknowledging unequal sample sizes) between 2015 observed and forecasted county-by-county Ehrlichia seroprevalence is 0.970, demonstrating that Ehrlichia seroprevalence can be forecasted accurately. CONCLUSIONS: The forecast presented herein can be an a priori alert to veterinarians regarding areas expected to see expansion of Ehrlichia beyond the accepted endemic range, or in some regions a dynamic change from historical average prevalence. Moreover, this forecast could potentially serve as a surveillance tool for human health and prove useful for forecasting other vector-borne diseases.

A Loop-Mediated Isothermal Amplification Assay Targeting 16S rRNA Gene for Rapid Detection of Anaplasma phagocytophilum Infection in Sheep and Goats.

Anaplasma phagocytophilum is a zoonotic pathogen and the causative agent of human granulocytic anaplasmosis in humans and tick-borne fever in various kinds of animals. In the present study, a loop-mediated isothermal amplification (LAMP) assay for rapid detection of A. phagocytophilum was developed using primers specific to 16S rRNA gene of this organism. The LAMP assay was performed at 65 C for 60 min and terminated at 80 C for 10 min. The optimal reaction conditions under which no cross-reaction was observed with other closely related tick-borne parasites ( Anaplasma bovis , Anaplasma ovis , Theileria luwenshuni, Babesia motasi, and Schistosoma japonicum ) were established. The assay exhibited much higher sensitivity compared with conventional polymerase chain reaction (PCR) (1 copy vs. 1,000 copies). To evaluate the applicability of the LAMP assay, 94 field samples of sheep blood were analyzed for A. phagocytophilum infection by using LAMP, nested PCR, and conventional PCR assays at the same time. A positive LAMP result was obtained from 53 (56.4%) of the 94 samples, whereas only 12 (12.8%) and 3 (3.2%) tested positive by nested and conventional PCR, respectively. In conclusion, this LAMP assay is a specific, sensitive, and rapid method for the detection of A. phagocytophilum in sheep/goats.

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

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.

Limited sharing of tick-borne hemoparasites between sympatric wild and domestic ungulates.

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.

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

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.

Rangelia vitalii, Babesia spp. and Ehrlichia spp. in dogs in Passo Fundo, state of Rio Grande do Sul, Brazil / Rangelia vitalii, Babesia spp. e Ehrlichia spp. em cães de Passo Fundo, estado do Rio Grande do Sul, Brasil

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.

Resumo Patógenos transmitidos por carrapatos são um problema emergente em todo o mundo, o trabalho objetivou diagnosticar os agentes causais da infecção em cães com suspeita de hemoparasitoses. Cinquenta e oito caninos com sinais clínicos como depressão, diáteses hemorrágicas e febre foram avaliados quanto à apresentação clínica, hemograma, esfregaço sanguíneo, sorologia pelo método de Imunofluorescência Indireta para os agentes Babesia vogeli e Ehrlichia canis e na PCR convencional para Babesia spp. (gene 18S rRNA), Rangelia vitalii (gene 18S rRNA) e Ehrlichia spp. (gene dsb). Cinco (8,6%) dos 58 cães apresentaram sorologia positiva para Babesia spp. e três (5,1%) para E. canis. Quatro (6,8%) animais mostraram-se positivos para R. vitalii no diagnóstico molecular. Os produtos da PCR foram sequenciados e o DNA encontrado de R. vitalii mostrou 99% de identidade genética com amostras de R. vitalii isoladas no Brasil. Não foi observada a presença de Babesia spp. e E. canis na PCR dos cães avaliados. Os resultados indicaram a presença de R. vitalii e exposição a Babesia spp. e Ehrlichia spp. entre os cães analisados.