New paradigms in the prevention of canine vector-borne diseases.

The prevention of canine vector-borne diseases (CVBDs) is pivotal for the health and welfare of dogs as well as for reducing their zoonotic risk to humans. Scientific knowledge gained in recent years contributed to the development of new strategies for the control of these diseases in different social and cultural contexts. Here, we discuss recent advances in the prevention of vector-borne pathogens (VBPs) affecting dogs with a focus on those of zoonotic relevance.

Geographical analysis of seroprevalence of Ehrlichia spp., Anaplasma spp., Borrelia burgdorferi and Dirofilaria immitis, in clinics and dog shelters in different Mexican states.

This study aimed to determine the seroprevalence and geographical distribution of Ehrlichia spp., Anaplasma spp., Borrelia burgdorferi and Dirofilaria immitis in dogs in Mexico, including owned dogs from veterinary clinics with regular medical care and shelter dogs. The Mexican territory was divided into eight geographical regions; 22 out of 32 states were included; 110 veterinary clinics and 53 dog shelters participated. SNAP® 4Dx Plus® (IDEXX® Laboratories) was used to detect antibodies against Ehrlichia spp., Anaplasma spp., Borrelia burgdorferi and Dirofilaria immitis antigens. A total of 3522 apparently healthy dogs were tested, 1648 from clinics and 1874 from shelters. The highest seroprevalence of infection/exposure was found for Ehrlichia spp. (30.9%), followed by Anaplasma spp. (14.6%), D. immitis (5.3%) and B. burgdorferi (0.1%). Significantly more positive dogs were older than 3 years. Regarding differences between facility types, there were only differences for D. immitis which was more prevalent in clinics than in shelters (OR â€‹= â€‹1.97; 95% CI: 1.45-2.69; P â€‹< â€‹0.0001). Co-infections were detected in 38.4% of the positive samples. Dogs from Mexican states located on the Atlantic and the Pacific coast were significantly more at risk for Ehrlichia spp. and Anaplasma spp. infections than dogs from interior states. Dogs in Atlantic coastal states were more at risk for Dirofilaria immitis infection.

The Relevance of Screening for Vector-Borne Diseases in Dogs with Proteinuria Living in an Endemic Region: A Retrospective Study.

This study aims to assess the main causes of proteinuria in dogs from the region of Lisbon (Portugal), estimating the relevance of screening for canine vector-borne diseases (CVBDs). A cross-sectional retrospective study was conducted. Medical records from proteinuric dogs (urinary protein−creatinine ratio > 0.5) presented to a Veterinary Teaching Hospital over a two-year period were reviewed for signalment, established diagnosis, proteinuria origin, and CVBD screening results. A total of 106 dogs were included. The median age was 9.5 years old (IQR: 7−12). Proteinuria was considered of renal origin in 76% of cases (46% of them had a presumptive diagnosis of glomerulonephritis secondary to CVBD, 27% chronic kidney disease, 26% systemic disease possible to induce proteinuria, and 1% leptospirosis). Proteinuria was classified as post-renal or mixed-origin in 17% and 7% of cases, respectively. About 35% of proteinuric dogs were positive for at least one CVBD. Of them, 84% were seropositive for one CVBD, while 16% tested positive for two or more. Among dogs testing positive for CVBD, 89% were seropositive for Leishmania infantum. This study showed that about one-third of proteinuric dogs tested positive for CVBDs, highlighting the relevance of their screening in dogs with proteinuria living in endemic regions.

Vector-borne pathogens in clinically healthy military working dogs in eastern Austria.

Military working dogs have an increased risk of acquiring an infection with vector-borne pathogens due to kennel housing and regular exposure to wildlife and vectors. To evaluate the level of infections in clinically healthy dogs of the Austrian Armed Forces, 94 individuals of the Military Working Dog Training Centre (MWDTC) Kaisersteinbruch/eastern Austria were examined in August 2016, February 2019 and August 2019. A modified Knott test was used to determine the presence of microfilariae, PCR for DNA detection of filarioid nematodes (incl. Dirofilaria), Leishmania spp., piroplasms, Borrelia spp., Bartonella spp. and Anaplasmataceae, and serological examination for antibodies against Borrelia burgdoferi s. l. and Leishmania infantum in all dogs. Two dogs were positive for Dirofilaria repens in the Knott test, and one of them also by PCR. Six clinically healthy dogs (4.2%) were positive for Babesia canis (PCR). In serology, 10 (10.6%) of the dogs were positive for specific antibodies against Borrelia burgdoferi s. l. The results suggest that the current measures against arthropod vector exposure and the pathogens they can transmit are not fully sufficient for these dogs. Further investigations of the tick and mosquito fauna in this area will shed more light on the risk of exposure for both the dogs and the staff of the MWDTC.

Risk of transmittable blood-borne pathogens in blood units from blood donor dogs in Canada.

BACKGROUND: Canine blood donors can be infected by various vector-borne or other pathogens that could be an important cause of morbidity and death in transfusion recipients. HYPOTHESIS/OBJECTIVES: To estimate and predict positivity to transmittable blood-borne pathogens in blood units collected from blood donor dogs in Canada. ANIMALS: Six thousand one hundred and fifty blood units from 1914 active blood donors registered to the Canadian Animal Blood Bank (CABB) between March 2010 and December 2016. METHODS: A registry-based retrospective study. Blood units were screened by SNAP 4Dx/4Dx Plus and PCR panel tests. Information on blood donors and test results were extracted from multiple databases and collated. Logistic regressions were used to predict blood unit positivity. RESULTS: Of 1779 blood units, 0.56% were antibody-positive for Anaplasma phagocytophilum/platys and 0% for Ehrlichia canis/ewingii. After exclusion of antibody-positive units to Anaplasma spp., 1.1% of 6140 blood units were PCR-positive to Anaplasma phagocytophilum, Bartonella spp., Brucella canis, "Candidatus Mycoplasma haematoparvum," Mycoplasma haemocanis, or a combination of these pathogens. Babesia spp., Ehrlichia spp., and Leishmania spp. were not detected. Units from the first blood collection from a dog had higher odds of testing PCR-positive (P < .001) for at least 1 pathogen than units from subsequent collections. CONCLUSIONS AND CLINICAL IMPORTANCE: Although our study indicates a low probability of detecting blood-borne pathogen in blood units collected by this Canadian blood bank, the presence of positive units highlights the importance of the preemptive identification and screening of blood units from healthy blood donors for safe blood banking, especially in first-time donors.

Exposure to Major Vector-Borne Diseases in Dogs Subjected to Different Preventative Regimens in Endemic Areas of Italy.

Vector-borne diseases (VBDs) are globally widespread arthropod-transmitted diseases with a significant impact on animal and human health. Many drivers have recently spurred the geographic spread of VBDs in dogs. This study has evaluated the exposure to most important VBDs in dogs under different preventative treatments in different regions of Italy, i.e., Veneto, Friuli Venezia-Giulia, Umbria, Giglio Island (Tuscany), Abruzzo and Latium. Serological analyses were performed to detect antibodies against Leishmania infantum, Babesia canis, Anaplasma phagocytophilum/Anaplasma platys, Ehrlichia canis/Ehrlichia ewingii, Borrelia burgdorferi, Rickettsia conorii and the circulating antigen of Dirofilaria immitis. Dogs were categorized according to the treatment schedule usually received, and the association between seropositivity and possible risk factors was statistically evaluated. Overall, 124/242 (51.2%) dogs tested positive for at least one pathogen, while 34 (14.0%) were exposed to two or more pathogens. The most detected seropositivity was against R. conorii, followed by Anaplasma spp., L. infantum, B. canis, and the other pathogens under study. Significant statistical associations were found according to geographical provenance, history of tick infestation, lifestyle and inadequate prophylactic treatments. Random/irregular treatments have been identified as a clear risk factor. These results show that adequate prophylactic treatment protocols are overlooked by dog owners, despite the availability of several effective products, with possible implications in veterinary medicine and on public health.

Molecular survey of vector-borne diseases in two groups of domestic dogs from Lisbon, Portugal.

BACKGROUND: Canine vector-borne diseases (CVBDs) are caused by a wide range of pathogens transmitted by arthropods. They have been an issue of growing importance in recent years; however, there is limited information about the vector-borne pathogens circulating in Portugal. The aim of the present study was to detect canine vector-borne bacteria and protozoa of veterinary and zoonotic importance using molecular methods. METHODS: One hundred and forty-two dogs from Lisbon, southern Portugal, were tested: 48 dogs from a veterinary hospital clinically suspected of vector-borne diseases and 94 apparently healthy dogs from shelters. Anaplasma spp./Ehrlichia spp., Babesia/Theileria spp., Hepatozoon spp., and Mycoplasma spp. infections were detected by PCR from blood samples and examined under light microscopy. Other information including clinical status and diagnostic test results were collected for each animal. RESULTS: Infections were detected by PCR in 48 (33.80%) dogs. Single infections were found in 35 dogs (24.64%), and co-infections were found in 13 (9.15%) dogs. Twenty-nine (20.42%) dogs were positive for Hepatozoon spp., 15 (10.56%) for Mycoplasma spp., 11 (7.75%) for Anaplasma spp./Ehrlichia spp., and six (4.21%) for Babesia spp. DNA sequencing was used to identify Babesia vogeli (2.81%), Babesia canis (1.40%), Hepatozoon canis (20.42%), Mycoplasma haematoparvum (2.11%), Mycoplasma haemocanis (8.45%), Anaplasma platys (7.04%), and Ehrlichia canis (0.70%). CONCLUSIONS: This is the first molecular identification of B. canis and M. haematoparvum in dogs from southern Portugal. This study highlights the importance of molecular methods to identify CVBD pathogens in endemic areas and helps to guide the clinical approach of veterinarians in practice.

Seroprevalence of Ehrlichia spp., Anaplasma spp., Borrelia burgdorferi sensu lato, and Dirofilaria immitis in Stray Dogs, from 2016 to 2019, in Southern Italy.

Canine vector-borne diseases (CVBD) are an important and emerging health concern for humans and animals worldwide. The purpose of the presented study was to assess, from 2016 to 2019, the seroprevalence of CVBD agents and clarify the epidemiology of tick-borne disease in stray dogs living in the Campania Region, Southern Italy. For this purpose, blood samples were collected from January 2016 to December 2019 from 1023 dogs in authorized kennels located in the five municipalities of the Campania Region. SNAP® 4DX® from IDEXX® Laboratories was used for detection of Ehrlichia spp., Anaplasma spp., Borrelia burgdorferi sensu lato (s.l.), and Dirofilaria immitis antibodies. The overall seroprevalence of CVBD in stray dogs was 19.6% (95% Confidence Intervals (CI): 17.2-22.8%; 201/1023). The most common pathogen was Ehrlichia spp., with a percentage of positivity of 16.03%, followed by Anaplasma spp. with 7.8%. B. burgdorferi s.l. and D. immitis were detected in only 0.2% of dogs; co-infection was detected in 4.5% of stray dogs tested. No link was detected between the gender, age, location, and CVBD seropositivity, except for Ehrlichia spp. for which location (Avellino Province; p = 0.007) and gender (male, p = 0.002) were risk factors for seropositivity. Our results demonstrated that animals are exposed to at least one of the four etiological agents (Ehrlichia spp., Anaplasma spp. Borrelia burgdorferi s.l., and Dirofilaria immitis) transmitted by vectors. Finally, this study highlighted the utility of serological monitoring in stray dogs, housed in kennels, given the threat posed by CVBD to animals and the zoonotic implications of these etiological agents and their vectors on human health.

Molecular Diagnosis, Prevalence and Importance of Zoonotic Vector-Borne Pathogens in Cuban Shelter Dogs-A Preliminary Study.

The present study aimed to determine the prevalence of zoonotic vector-borne pathogens, including Anaplasma platys, Anaplasma phagocytophilum, Borrelia burgdorferi sensu lato, Ehrlichia canis and Rickettsia spp. in shelter dogs from Cuba. Blood samples were collected from 100 shelter dogs and examined by molecular methods. Overall, 85 (85%; 95% CI: 77.88-92.12) dogs tested positive for at least one vector-borne pathogen using species-specific qPCR assays. Among the positive samples, E. canis was the most prevalent 62% (95% CI: 52.32-71.68), followed by A. platys 40% (95% CI: 30.23-49.77) and Rickettsia felis 27% (95% CI: 18.15-35.85), whereas 36% (95% CI: 26.43-45.57) showed co-infections. All samples were negative for A. phagocytophilum and B. burgdorferi s.l. The presence of 248 Rhipicephalus sanguineus ticks collected from the dogs was not statistically associated with the occurrence of infections. Thrombocytopenia was the most frequent haematological alteration found in PCR-positive dogs; it was statistically associated with the presence of E. canis, as well as co-infections (p < 0.05). The phylogenetic analyses of A. platys and E. canis based on 16S rRNA, groEL and gltA genes showed a low genetic diversity between Cuban strains. The present study demonstrates the high prevalence of vector-borne pathogens with zoonotic potential in shelter dogs from Cuba.

Detection of Leishmania infantum DNA and antibodies against Anaplasma spp., Borrelia burgdorferi s.l. and Ehrlichia canis in a dog kennel in South-Central Romania.

Canine vector-borne diseases are caused by pathogens transmitted by arthropods including ticks, mosquitoes and sand flies. Many canine vector-borne diseases are of zoonotic importance. This study aimed to assess the prevalence of vector-borne infections caused by Dirofilaria immitis, Ehrlichia canis, Borrelia burgdorferi sensu lato, Anaplasma spp. and Leishmania infantum in a dog kennel in Argeș County, Romania. Dog kennels are shelters for stray dogs with no officially registered owners that are gathered to be neutered and/or boarded for national/international adoptions by various public or private organizations. The international dog adoptions might represent a risk in the transmission of pathogens into new regions. In this context, a total number of 149 blood samples and 149 conjunctival swabs from asymptomatic kennel dogs were assessed using serology and quantitative real-time polymerase chain reaction. Antibodies against B. burgdorferi s.l. were detected in one dog (0.6%), anti-Anaplasma antibodies were found in five dogs (3.3%), while ten dogs (6.7%) tested positive for D. immitis antigen. Overall, 20.1% (30/149) of dogs were positive for L. infantum DNA. All samples were seronegative for anti-Leishmania antibodies. When adopting dogs from this region of Romania, owners should be aware of possible infection with especially L. infantum. The travel of infected dogs may introduce the infection to areas where leishmaniasis is not present.

Leishmania infantum and Dirofilaria immitis infections in Italy, 2009-2019: changing distribution patterns.

BACKGROUND: For long time, canine leishmaniosis (CanL) was considered endemic in the southern, central, and insular regions of Italy, whereas heartworm disease (HW) caused by Dirofilaria immitis was considered endemic in the northern region and in the swampy Po Valley. Following the reports of new foci of both diseases, in this study we update the distribution patterns and occurrence of new foci of CanL and HW discussing the main drivers for the changes in the epidemiology of these two important zoonotic canine vector-borne diseases. METHODS: Based on the statistical analyses of serological assays (n = 90,633) on L. infantum exposure and D. immitis infection performed by two reference diagnostic centres in Italy over a ten-year period (2009-2019) irrespective of the anamnesis of dogs. The distribution patterns of both parasites are herein presented along with the occurrence of new foci. RESULTS: Results highlighted the changing distribution patterns of L. infantum vs D. immitis infection in Italy. CanL is endemic in some areas of northern regions and HW has endemic foci in central and southern regions and islands. Significant differences in L. infantum exposure and HW infection prevalence among the study macroareas were detected. The overall results of the positive tested samples were 28.2% in southern Italy and islands, 29.6% in central Italy and 21.6% in northern Italy for L. infantum and 2.83% in northern Italy, 7.75% in central Italy and 4.97% in southern Italy and islands for HW. HW positivity significantly varied over years (χ2 = 108.401, df = 10, P < 0.0001), gradually increasing from 0.77% in 2009 to 8.47% in 2016-2017. CONCLUSIONS: New potential epidemiological scenarios are discussed according to a range of factors (e.g. environmental modifications, occurrence of competent insect vectors, transportation of infected animals to non-endemic areas, chemoprophylaxis or vector preventative measures), which may affect the current distribution. Overall, the results advocate for epidemiological surveillance programmes, more focussed preventative and control measures even in areas where few or no cases of both diseases have been diagnosed.

ZnO Nanolower-Based NanoPCR as an Efficient Diagnostic Tool for Quick Diagnosis of Canine Vector-Borne Pathogens.

: Polymerase chain reaction (PCR) is a unique technique in molecular biology and biotechnology for amplifying target DNA strands, and is also considered as a gold standard for the diagnosis of many canine diseases as well as many other infectious diseases. However, PCR still faces many challenges and issues related to its sensitivity, specificity, efficiency, and turnaround time. To address these issues, we described the use of unique ZnO nanoflowers in PCR reaction and an efficient ZnO nanoflower-based PCR (nanoPCR) for the molecular diagnosis of canine vector-borne diseases (CVBDs). A total of 1 mM of an aqueous solution of ZnO nanoflowers incorporated in PCR showed a significant enhancement of the PCR assay with respect to its sensitivity and specificity for the diagnosis of two important CVBDs, Babesia canis vogeli and Hepatozoon canis. Interestingly, it drastically reduced the turnaround time of the PCR assay without compromising the yield of the amplified DNA, which can be of benefit for veterinary practitioners for the improved management of diseases. This can be attributed to the favorable adsorption of ZnO nanoflowers to the DNA and thermal conductivity of ZnO nanoflowers. The unique ZnO nanoflower-assisted nanoPCR greatly improved the yield, purity, and quality of the amplified products, but the mechanism behind these properties and the effects and changes due to the different concentrations of ZnO nanoflowers in the PCR system needs to be further studied.

Canine vector-borne pathogens from dogs and ticks from Tamil Nadu, India.

Canine vector-borne diseases (CVBDs) pose a major health problem in dogs globally, with the potential to cause zoonoses, in particular in developing countries where scientific knowledge on the topic is minimal. Blood samples and ticks were collected from stray dogs in Tamil Nadu, South India to assess the prevalence of CVBD-causing pathogens (Anaplasma spp., Babesia spp., Ehrlichia spp., Hepatozoon spp., filarioids and Leishmania spp.). Of the 230 dogs examined, 229 (99.6%) were infested by ticks (mean intensity, 5.65) with Rhipicephalus sanguineus sensu lato and Rhipicephalus haemaphysaloides being morphologically identified in the 98.3% and 1.7% of the infested dogs, respectively. Overall, the 67.8% (n = 156) of dogs was positive for at least one pathogen with Hepatozoon canis being the most prevalent (37.8%) followed by Anaplasma platys (22.6%), Ehrlichia canis (16.1%) Babesia vogeli (10%), Anaplasma phagocytophilum (0.4%) and Babesia gibsoni (0.4%). Two filarioids (Dirofilaria sp. "hongkongensis" and Brugia malayi, 0.4%) were diagnosed in sampled animals. Co-infection with H. canis and A. platys was the most prevalent (8.3%, P = 0.00001), whilst all animals scored negative for Leishmania spp.. Out of 295 ticks analysed, 215 R. sanguineus s.l. (75.4%) and 8 R. haemaphysaloides (88.9%) were positive for at least one pathogen with H. canis as the predominant species (42.5%), followed by A. platys (33.8%), E. canis (16.9%), B. vogeli (3.8%) and A. phagocytophilum (0.3%). Fifty-six dogs (35.9%) harboured the same pathogen as the respective tick specimens, while 29 dogs (18.6%) had a different pathogen. Thirteen sequence types (STs) were identified for H. canis, with ST2 (49.4%) as the most representative in dogs and ST1 (73.5%) in ticks. Similarly, seven STs were found for Anaplasma spp. (i.e., five for A. platys, one for A. phagocytophilum and one for Anaplasma sp.), with ST2 as the most representative in dogs (70.6%) and ST3 (52.5%) in ticks for A. platys. Only one ST was identified for B. vogeli, B. gibsoni, E. canis, D. sp. "hongkongensis" and B. malayi. Regular surveillance and adoption of adequate treatment and control measures are needed to reduce the risk of disease-causing pathogens in stray dogs and of pathogens with zoonotic potential.

Simultaneous Detection of Parasitic Vector Borne Diseases: A Robust Cross-Sectional Survey in Hunting, Stray and Sheep Dogs in a Mediterranean Area.

Canine vector-borne diseases (CVBDs) are a spectrum of diseases caused by different pathogens transmitted by blood-feeding arthropoda. The aim of this study was to investigate leishmaniosis, babesiosis, and filarial infections in dogs with three different lifestyles (hunting, stray, and sheep dogs) in Molise, the smallest region of southern Italy, where data available about these parasitic infections are very scant. A cross-sectional survey was conducted on 318 hunting, 180 stray, and 218 sheep dogs. Immunofluorescence antibody test, blood smear, molecular techniques and Knott's test were performed to detect Leishmania infantum, Babesia spp. and filarial nematodes. Association between positivity to CVBDs, age, sex, and living conditions was evaluated. An overall prevalence of 12.3% of CVBDs caused by L. infantum (10.2%), B. canis canis (0.3%) and filarial nematodes (2.1%) was detected. Three dogs showed co-infections of L. infantum and B. c. canis (0.1%) or Acanthocheilonema reconditum (0.3%). A significantly association was found only for filarial infection in hunting dogs. These parasites were reported also in dogs without clinical signs. It is very important to plan effective control programs for CVBDs to guarantee not only the health and welfare of pets, but also the public safety, because some of mentioned parasites are of zoonotic importance.

Causative agent of canine heartworm (Dirofilaria immitis) detected in wild lemurs.

The lemurs of Madagascar are threatened by human activities. We present the first molecular detection of canine heartworm (Dirofilaria immitis) in a wild non-human primate, the mouse lemur (Microcebus rufus). Zoonotic D. immitis infection has been associated with clinical pathology that includes serious and often fatal cardiac and pulmonary reactions. With human encroachment and associated increases in free-roaming dog populations in Madagascar, we examined lemurs for zoonotic canid pathogens. D. immitis presents a new potential conservation threat to lemurs. We highlight the need for wide-ranging and effective interventions, particularly near protected areas, to address this growing conservation issue.

Six-month field efficacy and safety of the combined treatment of dogs with Frontline Tri-Act® and NexGard Spectra®.

BACKGROUND: Safety and efficacy of the combined monthly use of spot-on fipronil 6.76% w/v / permethrin 50.48% w/v (Frontline Tri-Act®) and chewable tablets of afoxolaner 1.9% w/w / milbemycin oxime 0.4% w/w (NexGard Spectra®) in dogs was evaluated in a field study over a period of 6 months. METHODS: Forty-one healthy dogs living in highly endemic area for canine leishmaniosis and other canine vector borne diseases (VBD) were included in the study at the beginning of the Leishmania transmission season. Sixteen dogs were pet dogs living each in a single household; twenty-five dogs were hunting dogs living in three kennels. At inclusion, the dogs were ELISA (rapid test) negative for antibodies to Anaplasma, Borrelia, Ehrlichia, and for antigens of Dirofilaria. The dogs were also negative for blood microfilariae at the Knott's test, and no clinical or haematological abnormalities were observed. Of the included dogs, six hunting, apparently healthy, dogs were ELISA (rapid test) positive to Leishmania, and some were naturally infected by gastrointestinal nematodes (58.5%) and/or infested by fleas (58.5%) and ticks (9.8%). All the included dogs were treated at Days 0, 28, 56, 84, 112 and 140, and followed-up for efficacy until the study end (Day 168). RESULTS: No adverse events related to the two products, nor skin reactions, general signs, or changes in the haematological profile, were observed during the study. At Day 14, anthelminthic efficacy was 100% for Toxocara canis, Toxascaris leonina and Capillaria aerophila, while few hunting dogs were still shedding eggs of Trichuris vulpis (1/25 hunting dog) and Ancylostomatidae (9/25 hunting dogs). All pet dogs were nematode free at the end of the study. Hunting dogs were free of roundworms and whipworms. Twenty-four hours after the first treatment, 95.8% of the ectoparasite infested dogs were free from fleas and ticks. Ectoparasites were significantly controlled during the 6-month study period, with 100% efficacy on both fleas and ticks from Day 56 to Day 168. Blood and serum samples collected on Day 168 were tested for vector-borne pathogens using same methods of the inclusion and no new seroconversions or circulating blood microfilariae were observed. CONCLUSIONS: Concomitant use of Frontline Tri-Act® and NexGard Spectra® in dogs for six months was well tolerated. The combination was effective in controlling fleas, ticks, gastro-intestinal nematodes, and neither new seroconversion to the tested vector-borne pathogens nor blood microfilariae were detected in treated dogs at the end of the study.

Vector-borne parasitic infections in dogs in the Baltic and Nordic countries: A questionnaire study to veterinarians on canine babesiosis and infections with Dirofilaria immitis and Dirofilaria repens.

Canine vector-borne diseases have been spreading northwards in Europe, and canine babesiosis and infections with Dirofilaria immitis (heartworm) and Dirofilaria repens have been diagnosed also in the Baltic and the Nordic countries. We used an online questionnaire to survey how large a proportion of veterinarians in the Baltic (Estonia, Latvia, and Lithuania) and the Nordic countries (Denmark, Finland, Iceland, Norway, and Sweden) saw canine babesiosis cases and dogs infected with D. immitis and D. repens in 2016. In addition, questions regarding transmission, zoonotic potential, clinical signs, and treatment of the infections were asked. The questionnaire was completed by 122 veterinarians. In 2016, 23% of them had seen at least one case of canine babesiosis, 15% at least one dog with D. immitis infection, and 9% at least one dog with D. repens infection. A veterinarian working in the Baltic countries had 12.2 times higher odds to have seen a canine babesiosis case and 9.3 times higher odds to have seen a dog with D. repens infection than a veterinarian working in the Nordic countries did. While 48% of the veterinarians knew that canine babesiosis is not considered a zoonosis, 26% knew that D. immitis is zoonotic and 34% knew that D. repens is zoonotic. The results suggested that autochthonous cases of the three vector-borne parasitic infections were seen by veterinarians in the Baltic countries, whereas most cases seen by veterinarians in the Nordic countries appeared to be imported. A substantial proportion of the veterinarians did not know whether the parasites are zoonotic.

Serological and molecular prevalence of canine vector-borne diseases (CVBDs) in Korea.

BACKGROUND: Previous surveys in dogs from Korea indicated that dogs are exposed to a variety of vector- borne pathogens, but perception for a nation-wide canine vector-borne disease (CVBD) occurrence has been missing. We report here results of both serological and molecular prevalence studies for major CVBDs of dogs from all over the South Korean Peninsula except for Jeju Island. RESULTS: Serological survey of 532 outdoor dogs revealed the highest prevalence for Dirofilaria immitis (25.2%), followed by Anaplasma phagocytophilum (15.6%), Ehrlichia canis (4.7%) whereas Borrelia burgdorferi showed the lowest prevalence (1.1%). The number of serologically positive dogs for any of the four pathogens was 216 (40.6%). Concurrent real-time PCR assay of 440 dogs in the study indicated that DNA of "Candidatus M. haematoparvum", Mycoplasma haemocanis, Babesia gibsoni, A. phagocytophilum, and Hepatozoon canis was identified in 190 (43.2%), 168 (38.2%), 23 (5.2%), 10 (2.3%) and 1 (0.2%) dogs, respectively. DNA of Bartonella spp., Ehrlichia spp., Leishmania spp., Rickettsia spp. and Neorickettsia risticii was not identified. Analysis of questionnaires collected from owners of 440 dogs showed that the number of dogs with heartworm preventive medication was 348 (79.1%) among which dogs still positive to D. immitis infection were 60 (17.2%), probably due to the mean months of heartworm preventive medication being only 6.5. The high prevalence rates of both "Ca. M. haematoparvum" and Mycoplasma haemocanis in dogs from Korea indicate that these organisms may be transmitted by vectors other than Rhipicephalus sanguineus because this tick species has rarely been found in Korea. This is the first nationwide survey for canine haemotropic mycoplasma infections in Korea. CONCLUSIONS: This study showed that the risk of exposure to major vector-borne diseases in dogs is quite high throughout all areas of South Korean Peninsula. Since achieving full elimination of many pathogens causing CVBDs from infected animals is often impossible even when they are clinically cured, dogs once exposed to CVBDs can remain as lifetime reservoirs of disease for both other animals and humans in the close vicinity, and should therefore be treated with preventative medications to minimise the risk of pathogen transmission by the competent vectors.

Detection and molecular characterization of Hepatozoon canis, Babesia vogeli, Ehrlichia canis, and Anaplasma platys in dogs from Metro Manila, Philippines

The study of canine vector-borne diseases in the Philippines started in the 1970s but only gained interest in the past decade. Characterization of such diseases in the Philippines remains incomplete, thus, it is necessary to obtain additional information on the prevalence and diversity of canine tick-borne diseases in the country. In this study, blood samples were obtained at two veterinary clinics in Metro Manila, Philippines from 114 dogs suspected of having canine tick-borne pathogens. Polymerase chain reaction (PCR) was performed on whole blood DNA extracts followed by sequencing, and the following pathogens were detected: Hepatozoon (H.) canis (5.26%), Babesia (B.) vogeli (5.26%), Ehrlichia (E.) canis (4.39%), and Anaplasma platys (3.51%). Additionally, a set of multiplex PCR primers were developed to detect H. canis, Babesia spp. (B. canis and B. vogeli), and E. canis in canine blood. Multiplex and conventional single-reaction PCR results for the 114 dog blood samples were similar, except for one H. canis sample. Multiplex PCR is, therefore, a useful tool in screening infected dogs in veterinary clinics. This study's results, together with those of previous studies in the country, show that canine vector-borne pathogens are an emerging veterinary concern in the Philippines.

Countrywide serological evaluation of canine prevalence for Anaplasma spp., Borrelia burgdorferi (sensu lato), Dirofilaria immitis and Ehrlichia canis in Mexico.

BACKGROUND: Canine vector-borne diseases (CVBD) have become a major concern for canine and human public health. The aim of the study described here is to add epidemiological data regarding four pathogens responsible for CVBD, namely anaplasmosis, borreliosis, dirofilariosis and ehrlichiosis in a national survey conducted in Mexico. METHODS: Seventy-four veterinary centres located in 21 federal Mexican states were asked to test dogs with clinical signs suspect for CVBD and healthy dogs, for detection of Dirofilaria immitis antigen and antibodies against Anaplasma spp., Borrelia burgdorferi (sensu lato) and Ehrlichia canis using the SNAP® 4DX® from IDEXX® Laboratories. RESULTS: A total of 1706 dogs were tested, including 943 apparently healthy and 722 CVBD-suspect dogs. Infected dogs were 36.7 %. The highest percentages of infection with E. canis (51.0 %) and Anaplasma spp. (16.4 %) were obtained in the northwestern region, while D. immitis was most frequently found in the northeastern region of the country (8.9 %). Four dogs from the northwestern, northeastern, eastern and southeastern regions, respectively, were positive for B. burgdorferi (sensu lato). Northcentral regions showed lowest overall prevalence of infection (2.4 %). Co-infections were detected in 8.8 % of the dogs tested. Statistically significant lower positivity was found among dogs aged less than one year (23.2 %) and small-sized dogs (27.6 %), while higher prevalence of infection was found in dogs living outdoors (42.0 %), dogs with detectable tick infestation (43.3 %) and dogs that received treatment for tick-transmitted infections (58.8 %). Seropositivity was a risk factor for the presence of clinical signs as follows: Anaplasma spp. (OR = 2.63; 95 % CI: 1.88-3.67; P < 0.0001), D. immitis (OR = 2.52; 95 % CI: 1.61-3.95; P < 0.0001), E. canis (OR = 3.58; 95 % CI: 2.88-4.45; P < 0.0001), mixed infections (OR = 4.08; 95 % CI: 2.79-5.96; P < 0.0001), one or more agents (OR = 3.58; 95 % CI: 2.91-4.42; P < 0.0001). CONCLUSIONS: Canine serological evidence supports that dogs from Mexico are at risk of acquiring Anaplasma spp., D. immitis and/or E. canis, while B. burgdorferi (sensu lato) transmission is minimal in the country. Practitioners play a fundamental role in the detection and control of these diseases to protect dogs and humans.