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In August 2023, ehrlichiosis was confirmed in a patient in Italy with a Haemaphysalis punctata tick attached to his neck. Gene sequences of Ehrlichia canis from the tick and the patient were identical, indicating a potential risk for this uncommon infection for persons participating in outdoor activities.

Haemoparasitic infections are frequently observed in dogs from tropical regions, including India. The present investigation combined microscopic blood smear examination and PCR assays to assess the occurrence of canine tick-borne diseases (CTBD) from suspected dogs in and around Hisar, Haryana. Using the Giemsa-stained peripheral thin blood smear examination, 15 (12.5%) of the 120 dogs were infected with CTBD, with 5.8%, 3.3%, 2.5%, and 0.8% dogs testing positive for Hepatozoon canis , Ehrlichia canis , Babesia vogeli , and Babesia gibsoni , respectively. Using the PCR assay, CTBD was found to be 64.16% (77/120) in examined dogs. Of the 77 PCR-positive canines, 56 were infected with a single haemoparasite, while 21 were infected with two or more species. H. canis was the most abundant tick-borne pathogen, representing 35%, followed by E. canis 25.8%, B. vogeli 20%, and B. gibsoni 2.5%. The most common co-infection was with H. canis along with E. canis (7.5%). The PCR assay was proven to be more efficient for detecting haemoparasites in dogs compared to blood smear examinations. The study suggests that canine tick-borne diseases are common in Haryana and recommends using PCR-based molecular tests in addition to conventional microscopic examination to diagnose these infections for effective treatment and management of infected canines.

Background Dogs in the US are commonly infected with vector-borne pathogens, including heartworm and tick-borne disease agents. The geographic distribution of both arthropod vectors and the pathogens they transmit continues to expand. Methods To describe the current geographic distribution and prevalence of antigen of Dirofilaria immitis and antibody to Borrelia burgdorferi , Ehrlichia spp., and Anaplasma spp. in dogs, we summarized over 144 million test results from 2013 to 2019, inclusive, by county, state, and region. Canine seroprevalence by state was compared to population-adjusted human reports of tick-borne diseases. Results Results varied regionally, with D. immitis antigen and Ehrlichia spp. antibodies more frequently detected in the Southeast (2.6% and 5.2%, respectively) and antibody to B. burgdorferi and Anaplasma spp. most common in the Northeast (12.1% and 7.3%, respectively). Overall, percent positive test results to D. immitis decreased in the Southeast by 33.3% when compared to earlier summaries using the same strategy (from 3.9 to 2.6%). Geographic expansion of areas where dogs commonly test positive for Ehrlichia spp. was evident, likely because of a change in the test made in 2012 to allow detection of antibodies to E. ewingii concomitant with expansion of vector tick populations. Percent positive test results to Ehrlichia spp. increased in every region; this shift was particularly pronounced in the Southeast, where percent positive test results increased fourfold (from 1.3 to 5.2%). Continued geographic expansion of B. burgdorferi and A. phagocytophilum was apparent in the Northeast, Midwest, and Upper South, although canine seroprevalence of antibody to B. burgdorferi was much lower than prior surveys in many Lyme-endemic areas. Annual reports of human cases of Lyme disease, ehrlichiosis, and anaplasmosis were associated with percent positive canine results by state for the three tick-borne disease agents ( R 2  = 0.812, 0.521, and 0.546, respectively). Within endemic areas, percent positive test results for all three tick-borne agents demonstrated evidence of geographic expansion. Conclusions Large scale analysis of results from screening dogs in practice for evidence of vector-borne infections, including those with zoonotic importance, continues to be a valuable strategy for understanding geographic trends in infection risk over time. Graphical Abstract

Ehrlichia canis is a tick-borne bacterium that causes a potentially fatal disease in dogs called canine monocytic ehrlichiosis. In this cross-sectional study, we used a One Health framework to identify statistical associations between E. canis infection in dogs and multiple dog-related, human and environmental factors in Iquitos, Peru. Due to the lack of consensus regarding the positivity threshold for E. canis qPCR assays, we also evaluated if the factors associated with infection remained conserved regardless of the Ct value cut-off used: Ct < 35, a conservative but commonly accepted Ct cut-off for bacterial screening, or Ct ≤ 40, which has been used in several E. canis studies. Under the more conservative scenario, we found that the prevalence of E. canis among dogs was 19.6% (95% CI 15.8–23.9%). Additionally, we showed that risk factor analyses utilizing a qPCR Ct cut-off of 35 or 40 (with conventional PCR confirmation for samples with a Ct > 35) yield comparable results in statistical models, although some differences should be considered. Our findings suggest that in Iquitos, Peru, interventions to prevent E. canis infection should prioritize dogs living in houses with corrugated iron walls. Additionally, comprehensive strategies targeting dogs that have recently traveled and incorporating neutering/spaying and widespread acaricide programs may also prove beneficial. We also discuss the challenges encountered during molecular testing for E. canis detection, highlighting the broader difficulties of studying poorly understood intracellular pathogens in Global South countries.

Background The severity of canine leishmaniosis (CanL) due to Leishmania infantum might be affected by other vector-borne organisms that mimic its clinical signs and clinicopathological abnormalities. The aim of this study was to determine co-infections with other vector-borne pathogens based on serological and molecular techniques in dogs with clinical leishmaniosis living in Spain and to associate them with clinical signs and clinicopathological abnormalities as well as disease severity. Methods Sixty-one dogs with clinical leishmaniosis and 16 apparently healthy dogs were tested for Rickettsia conorii , Ehrlichia canis , Anaplasma phagocytophilum and Bartonella henselae antigens by the immunofluorescence antibody test (IFAT) and for E. canis , Anaplasma spp., Hepatozoon spp., Babesia spp. and filarioid DNA by polymerase chain reaction (PCR). Results Among the dogs examined by IFAT, the seroprevalences were: 69% for R. conorii , 57% for E. canis , 44% for A. phagocytophilum and 37% for B. henselae ; while the prevalences found by PCR were: 8% for Ehrlichia / Anaplasma , 3% for Anaplasma platys and 1% for H. canis . No other pathogen DNA was detected. Statistical association was found between dogs with clinical leishmaniosis and seroreactivity to R. conorii antigen (Fisher’s exact test: P  = 0.025, OR = 4.1, 95% CI = 1–17) and A. phagocytophilum antigen (Fisher’s exact test: P  = 0.002, OR = 14.3, 95% CI = 2–626) and being positive to more than one serological or molecular tests (co-infections) (Mann-Whitney test: U  = 243, Z  = -2.6, n 1  = 14, n 2  = 61, P  = 0.01) when compared with healthy dogs. Interestingly, a statistical association was found between the presence of R. conorii , E. canis , A. phagocytophilum and B. henselae antibodies in sick dogs and some clinicopathological abnormalities such as albumin and albumin/globulin ratio decrease and increase in serum globulins. Furthermore, seroreactivity with A. phagocytophilum antigens was statistically associated with CanL clinical stages III and IV. Conclusions This study demonstrates that dogs with clinical leishmaniosis from Catalonia (Spain) have a higher rate of co-infections with other vector-borne pathogens when compared with healthy controls. Furthermore, positivity to some vector-borne pathogens was associated with more marked clinicopathological abnormalities as well as disease severity with CanL.

Ehrlichia canis, a rickettsial organism, is responsible for causing ehrlichiosis, a tick-borne disease affecting dogs. This study aimed to estimate ehrlichiosis prevalence and identify associated risk factors in pet dogs. A total of 246 peripheral blood samples were purposively collected from pet dogs in Dhaka, Mymensingh, and Rajshahi districts between December 2018 and December 2020. Risk factor data were obtained through face-to-face interviews with dog owners using a pre-structured questionnaire. Multivariable logistic regression analysis identified risk factors. Polymerase chain reaction targeting the 16S rRNA gene confirmed Ehrlichia spp. PCR results were further validated by sequencing. The prevalence and case fatality of ehrlichiosis were 6.9% and 47.1%, respectively. Dogs in rural areas had 5.8 times higher odds of ehrlichiosis (odd ratio, OR: 5.84; 95% CI: 1.72-19.89) compared to urban areas. Dogs with access to other dogs had 5.14 times higher odds of ehrlichiosis (OR: 5.14; 95% CI: 1.63-16.27) than those without such access. Similarly, irregularly treated dogs with ectoparasitic drugs had 4.01 times higher odds of ehrlichiosis (OR: 4.01; 95% CI: 1.17-14.14) compared to regularly treated dogs. The presence of ticks on dogs increased ehrlichiosis odds nearly by 3 times (OR: 3.02; 95% CI: 1.02-8.97). Phylogenetic analysis, based on 17 commercially sequenced isolates, showed different clusters of aggregation, however, BAUMAH-13 (PP321265) perfectly settled with a China isolate (OK667945), similarly, BAUMAH-05 (PP321257) with Greece isolate (MN922610), BAUMAH-16 (PP321268) with Italian isolate (KX180945), and BAUMAH-07 (PP321259) with Thailand isolate (OP164610). Pet owners and veterinarians in rural areas should be vigilant in monitoring dogs for ticks and ensuring proper preventive care. Limiting access to other dogs in high-risk areas can help mitigate disease spread. Tick prevention measures and regular treatment with ectoparasitic drugs will reduce the risk of ehrlichiosis in dogs. The observed genetic similarity of the Bangladeshi Ehrlichia canis strain highlights the need for ongoing surveillance and research to develop effective control and prevention strategies, both within Bangladesh and globally.

Background Cats can be carriers of infected arthropods and be infected with several vector-borne pathogens (VBP) but there is limited knowledge about their pathogenic role in cats. Results A cross-sectional controlled study investigated the clinical status and antibody ( Bartonella henselae , Rickettsia conorii , Ehrlichia canis , Anaplasma phagocytophilum , Babesia microti and Leishmania infantum ) and/or blood PCR ( Mycoplasma spp., Bartonella spp., Rickettsia spp., Ehrlichia/Anaplasma spp., piroplasmids, L. infantum , Hepatozoon felis ) prevalence in 197 cats. Outdoor cats lacking ectoparasiticide treatment or hosting ectoparasites (study group [SG], n = 134) and indoor cats treated against ectoparasites (control group [CG], n = 63) were enrolled. Clinical data and retroviral co-infections were compared between the two groups. Multivariable analysis tested associations between variables and VBP exposure. Lymphadenia, stomatitis, and various haematological abnormalities were statistically more frequent in SG. Antibodies against R. conorii , B. henselae , A. phagocytophylum , B. microti , E. canis and L. infantum were detected. Bartonella henselae , Bartonella clarridgeiae , Mycoplasma haemofelis , “ Candidatus Mycoplasma haemominutum” and “ Candidatus Mycoplasma turicensis” DNA were identified. Very high antibody (87.8%) and PCR (40.1%) positivity to at least one pathogen were detected and were significantly higher in SG. Co-infections were confirmed in about one-third of the cats and were more frequent in SG cats. Molecular and overall (antibody and PCR) positivity to Bartonella and antibody positivity to R. conorii were higher in SG. Multivariable analysis found significant associations of Bartonella spp. infection with Feline Immunodeficiency Virus (FIV) infection and increased globulins, and of Mycoplasma spp. infection with adult age, FIV infection, anaemia, and increased creatinine. Conclusions A very high prevalence of exposure to zoonotic VBP was found in cats, with Rickettsia and Bartonella infections being most prevalent. Some risk factors were documented namely for Mycoplasma spp. and Bartonella spp. The lifestyle of cats is clinically relevant and requires specific preventative measures to protect their health.

Despite the high global impacts of canine vector-borne diseases (CVBD) due to their wide distribution and zoonotic potential, the current epidemiological situation of CVBD in many tropical and subtropical regions remains unknown. This study examines the seroprevalence and molecular prevalence of Ehrlichia canis and other pathogens causing CVBDs ( Leishmania infantum , Dirofilaria immitis, Babesia spp., Anaplasma spp. and Hepatozoon canis ) in dogs living on the island of Boa Vista (Cape Verde Republic). Blood samples and infesting ticks were taken from 150 dogs across the island (stray, shelter, and pet dogs). Serum samples were tested using a rapid immunochromatographic test (Uranotest ® Quattro) that detects antibodies against E. canis , L. infantum , Anaplasma spp. and D. immitis antigen. Levels of serum antibodies against E. canis were measured using the immunofluorescence antibody test (IFAT). In addition, tick-borne pathogens in blood samples ( Anaplasma spp., Babesia spp., Hepatozoon spp., and Ehrlichia canis ) were detected by microscopy observation and/or PCR plus sequencing. The seroprevalence of E. canis was extremely high at 82% (123/150), as revealed by both immunochromatography and IFAT. Most dogs returning a seropositive test result (82.92%; 102/123) had antibody titres > 1:1280 but showed no clinical signs or notable laboratory abnormalities. Of the 123 animals testing seropositive for E. canis , 67 (54.47%) also presented antibodies against Anaplasma spp ., and 13 (10.56%) showed the presence of Hepatozoon spp. gamonts in the blood smear. Ehrlichia canis infection was detected in 17.1% (25/146) of dogs tested by direct sequencing of polymerase chain reaction (PCR) products. Co-infections were detected in seven of these dogs: four dogs tested PCR-positive for both E. canis and A. platys , two dogs tested positive for E. canis and Hepatozoon spp., and one dog tested positive for E. canis , A. platys and Hepatozoon spp. Rhipicephalus sanguineus sensu lato was the only tick species found infesting the canine study population. The high prevalence of tick-borne pathogens detected in dogs from Boa Vista Island highlights a need for improved control measures designed to prevent the transmission of these pathogens. Graphical Abstract

BACKGROUND: Red foxes (Vulpes vulpes) have recently been recognized as potential reservoirs of several vector-borne pathogens and a source of infection for domestic dogs and humans, mostly due to their close vicinity to urban areas and frequent exposure to different arthropod vectors. The aim of this study was to investigate the presence and distribution of Babesia spp., Hepatozoon canis, Anaplasma spp., Bartonella spp., ‘Candidatus Neoehrlichia mikurensis’, Ehrlichia canis, Rickettsia spp. and blood filaroid nematodes in free-ranging red foxes from Bosnia and Herzegovina. METHODS: Spleen samples from a total of 119 red foxes, shot during the hunting season between October 2013 and April 2014 throughout Bosnia and Herzegovina, were examined for the presence of blood vector-borne pathogens by conventional PCRs and sequencing. RESULTS: In the present study, three species of apicomplexan parasites were molecularly identified in 73 red foxes from the entire sample area, with an overall prevalence of 60.8%. The DNA of B. canis, B. cf. microti and H. canis was found in 1 (0.8%), 38 (31.9%) and 46 (38.6%) spleen samples, respectively. In 11 samples (9.2%) co-infections with B. cf. microti and H. canis were detected and one fox harboured all three parasites (0.8%). There were no statistically significant differences between geographical region, sex or age of the host in the infection prevalence of B. cf. microti, although females (52.9%; 18/34) were significantly more infected with H. canis than males (32.9%; 28/85). The presence of vector-borne bacteria and filaroid nematodes was not detected in our study. CONCLUSION: This is the first report of B. canis, B. cf. microti and H. canis parasites in foxes from Bosnia and Herzegovina and the data presented here provide a first insight into the distribution of these pathogens among the red fox population. Moreover, the relatively high prevalence of B. cf. microti and H. canis reinforces the assumption that this wild canid species might be a possible reservoir and source of infection for domestic dogs.

Background Due to limited documentation on vector-borne pathogens of companion animals in Namibia, a country-wide, multi-site field study was conducted to estimate the prevalence of these pathogens in domestic dogs. Methods Samples of whole blood and serum from 375 dogs in 15 towns across eight regions were analysed. Vector-borne pathogens were screened by light microscopic examination of blood smears, point-of-care serology, and quantitative real-time polymerase chain reaction (qPCR). Haematology and serum biochemistry analyses were also performed. Results Collectively, the SNAP ® 4Dx ® Plus Test provided 64% seropositive results, comprising Ehrlichia species (59%), Anaplasma species (45%), Dirofilaria immitis (2%), and Borrelia burgdorferi (< 1%). Altogether, prevalence as determined by probe-based qPCR assays was 54%, comprising Ehrlichia canis (27%), Hepatozoon canis (25%), Anaplasma species (13%), and Babesia vogeli (8%). Light microscopy yielded the least number of positives, indicating a collective positive result of only 11% in screening for Ehrlichia , Anaplasma , Hepatozoon , Babesia , and microfilaria species. On the whole, Kunene and Otjozondjupa regions showed the highest pathogen prevalence (75%), and the lowest was from Erongo region (38%), on qPCR testing. Significant associations between tick presence and infection by E. canis ( P  = 0.001), Anaplasma species ( P  = 0.006), and B. vogeli ( P  = 0.008) were demonstrated. Likewise, relevant associations between haemoparasite infection and variables of patient signalment, history, and various disease manifestations were shown. Finally, significant associations were found between pathogen infection and numerous clinical pathology abnormalities of the erythron, leukon, and thrombon, including thrombocytopenia ( P  = 0.022). Conclusions Diagnostic modalities should be used contextually to test for canine pathogens, with due consideration of the limitations. Appropriate diagnostic testing such as qPCR, guided by relevant known associations with disease manifestation, should guide responsible treatment strategies and identify potential zoonotic risks in pets. Graphical Abstract