Comparative Evaluation of 2 In-Clinic Assays for Vector-Borne Disease Testing in Dogs

Vector-borne agents comprise medically important infections affecting dogs throughout much of the world. Sensitive detection of antibodies directed at tick-borne disease-causing organisms in dogs is diagnostically important for veterinarians, pets and their owners, and epidemiologically important for public health surveillance. The SNAP 4Dx Plus Test (IDEXX Laboratories, Inc., Westbrook, ME) identifies antibodies to or infection with multiple tick-borne pathogens and canine heartworm antigen in a single assay. Recently, VetScan FLEX4 Rapid Test (Abaxis, Inc., Union City, CA) was launched as a new assay to detect tick-borne pathogen antibodies and heartworm antigen. In the present study, we evaluated the comparative performance of SNAP 4Dx Plus (SNAP) and FLEX4 Rapid Test (FLEX4) using samples selected based on geographic distributions for canine vector borne diseases, including Borrelia burgdorferi (n = 105), Anaplasma phagocytophilum (160), Anaplasma platys (115), Ehrlichia canis (154), Ehrlichia ewingii (163), Ehrlichia chaffeensis (151) and Dirofilaria immitis (105). Canine vector borne diseases infection status was established for each sample by a combination of reference methods that included necropsy (D. immitis, heartworm disease), Western immunoblotting (B. burgdorferi), immunofluorescence assays (A. phagocytophilum and E. canis) and species-specific ELISAs (A. platys, E. canis, E. ewingii and E. chaffeensis). For comparisons among the 2 assays, samples were evaluated per the manufacturers’ instructions for each test kit.By testing each same sample set compared to the defined reference results, sensitivities differed substantially between SNAP and FLEX4, at 95.5 vs. 40.9%, respectively for B. burgdorferi, 97.1% vs. 61.4% for E. canis, 98.2% vs. 59.3% for E. ewingii, 64.3% vs. 35.7% for E. chaffeensis, 84.5% vs. 12.7% for A. phagocytophilum, 83.3% vs. 33.3% for A. platys, and 94.1% vs. 88.2% for D. immitis. Specificities for both rapid assay tests ranged from 98% to 100%.Based upon the comparative results derived from this study, the SNAP test was more sensitive than the FLEX4 test for detection of antibodies to all tick-borne pathogens and heartworm disease (Dirofilaria immitis) antigen in dogs.     

Canine lymphoma and vector‐borne diseases: Molecular and serological evaluation of a possible complicity

Lymphoma is the most common haematological malignancy in dogs and its aetiology is largely unknown. The presence of canine vector‐borne agents (CVBD) in lymphoma tissues has been described and its causative effects questioned. We intended to evaluate the presence and extent of Leishmania infantum, Ehrlichia canis, Anaplasma phagocytophilum and Bartonella henselae infection in dogs with lymphoma. Sixty‐one dogs, living in the Lisbon metropolitan area, with a diagnosis of lymphoma were enrolled. Immunofluorescence assays were used to detect serum IgG's. The presence of DNA from CVBD agents in tumour tissue was assessed by PCR. All dogs tested negative for B. henselae, A. phagocytophilum and E. canis by both serology and PCR. Regarding L. infantum, 8.2% (n = 5) of the dogs had a positive serologic result. L. infantum DNA was detected in two samples of diffuse large B‐cell lymphoma (DLBCL). These results show an increased, but not significant, seropositivity (8.2% vs 7.9%) and molecular detection (3.3% vs 1.2%) for L. infantum in dogs with lymphoma, when compared to the reported canine population in the same geographical area. We could not identify an association between lymphoma and E. canis, A. phagocytophilum, B. henselae or Leishmania infantum infection in the studied population. Nevertheless, further studies, following dogs trough their CVBD disease evolution, are worthwhile and may help clarify a possible role of CVBD agents in lymphomagenesis.     

Bartonella Seroepidemiology in Dogs from North America, 2008–2014

Background

Improved understanding of Bartonella species seroepidemiology in dogs may aid clinical decision making and enhance current understanding of naturally occurring arthropod vector transmission of this pathogen.

Objectives

To identify demographic groups in which Bartonella exposure may be more likely, describe spatiotemporal variations in Bartonella seroreactivity, and examine co‐exposures to other canine vector‐borne diseases (CVBD).

Animals

A total of 15,451 serology specimens from dogs in North America were submitted to the North Carolina State University, College of Veterinary Medicine Vector Borne Disease Diagnostic Laboratory between January 1, 2008, and December 31, 2014.

Methods

Bartonella henselae, Bartonella koehlerae, and Bartonella vinsonii subspecies berkhoffii indirect fluorescent antibody (IFA) serology results, as well as results from a commercial assay kit screening for Dirofilaria immitis antigen and Ehrlichia species, Anaplasma phagocytophilum, and Borrelia burgdorferi antibodies, and Ehrlichia canis, Babesia canis, Babesia gibsoni, and Rickettsia species IFA results were reviewed retrospectively.

Results

Overall, 3.26% of dogs were Bartonella spp. seroreactive; B. henselae (2.13%) and B. koehlerae (2.39%) were detected more frequently than B. vinsonii subsp. berkhoffii (1.42%, P < 0.0001). Intact males had higher seroreactivity (5.04%) than neutered males (2.87%, P < 0.0001) or intact or spayed females (3.22%, P = 0.0003). Mixed breed dogs had higher seroreactivity (4.45%) than purebred dogs (3.02%, P = 0.0002). There was no trend in seasonal seroreactivity; geographic patterns supported broad distribution of exposure, and co‐exposure with other CVBD was common.

Conclusions and Clinical Importance

Bartonella spp. exposure was documented throughout North America and at any time of year. Male intact dogs, mixed breed dogs, and dogs exposed to other CVBD have higher seroreactivity to multiple Bartonella species.     

Seroprevalence against Borrelia burgdorferi sensu lato and occurence of antibody co-expression with Anaplasma phagocytophilum in dogs in Latvia

Background

Lyme disease is commonly diagnosed in humans in Latvia, but up to date no studies have been performed to investigate its prevalence in dogs. The aim of this study was to evaluate if seroprevalence against B. burgdorferi sensu lato (B. burgdorferi s.l.) and co-expression of antibodies against B.burgdorferi s.l. and A. phagocytophilum is higher in dogs with clinical suspicion of tick-borne diseases compared to healthy dogs.

Findings

Venous blood was taken from healthy dogs (n=441) and dogs suspected to have borreliosis and/ or canine granulocytic anaplasmosis (n=29). The presence of antibodies was detected with SNAP 4Dx test (IDEXX, Westbrook, Maine, USA). The seroprevalence against B. burgdorferi s.l. in healthy dogs was 2.49% (11/441) and 36% (4/11) of seropositive dogs had antibodies against both of investigated bacteria. None of the dogs in sick dog group had detectable antibodies against B. burgdorferi s.l.

Conclusions

We conclude that seroprevalence to B. burgdorferi s.l. in dogs in Latvia is low and that dogs with suspicion of tick-borne disease do not have higher B. burgdorferi s.l. seroprevalence than healthy dogs. Dogs that express antibodies against B. burgdorferi s.l. frequently co-express antibodies against A. phagocytophilum.     

Study on ticks and tick-borne zoonoses in public parks in Italy

A survey on tick density and on tick‐borne zoonoses was carried out in four public parks in the outskirts of Imola (northern Italy) from June to October 2006. All stages of Ixodes ricinus and only larvae of Riphicephalus sanguineus were recovered by dragging, performed on 100‐m transects. Almost all ticks (99%) were harvested in one park. I. ricinus density (nymphs/100 m²) ranged from 0 in park L to 6.3 in park F. Nymphs and adults of I. ricinus were subjected to PCR for Anaplasma phagocytophilum, Bartonella spp., Borrelia burgdorferi s. l. and Rickettsia spp. The observed prevalences were 38.3% for Bartonella henselae, 5.2% for Bartonella clarridgeiae, 10.4% for B. burgdorferi s. l., 2.6% for Rickettsia helvetica and 13% for Rickettsia monacensis, respectively. No DNA of A. phagocytophilum was found. Acarological risks (AR) were calculated as probabilities of collecting at least one infected nymph per transect. The AR values calculated for the various zoonotic agents were 11.4% for R. helvetica, 27.7% for B. clarridgeiae, 49.7% for B. burgdorferi s. l., 57.2% for R. monacensis and 90.4% for B. henselae, respectively. In this study, B. clarridgeiae was for the first time identified in I. ricinus ticks.     

Serum acute phase proteins as clinical phase indicators and outcome predictors in naturally occurring canine monocytic ehrlichiosis

Background: Canine monocytic ehrlichiosis (CME), caused by Ehrlichia canis, is an important tick‐borne disease of global importance. Currently, limited information is available on the diagnostic and prognostic value of acute phase proteins (APPs) in dogs naturally infected with E. canis. Hypothesis: APPs may be useful indicators of the clinical phase of CME and predictive of the clinical outcome (death or survival). Animals: Fifty‐six dogs naturally infected with E. canis and 7 clinically healthy control dogs. Methods: C‐reactive protein (CRP), serum amyloid A (SAA), haptoglobin (Hp), and albumin concentrations determined on admission were retrospectively compared among 27 dogs with nonmyelosuppressive CME, 29 dogs with myelosuppressive CME and 7 healthy dogs. Diagnosis of CME was based on clinical and clinicopathological findings, seropositivity to E. canis, polymerase chain reaction amplification of E. canis‐specific 16S rDNA, microscopic observation of Ehrlichia sp. morulae in blood monocytes or some combination of these. Results: Mean concentrations of CRP, SAA, and Hp were significantly higher in the myelosuppressed dogs compared with the other groups, but no significant differences were found in the concentration of albumin. Survival analysis of the affected animals indicated that APP concentrations were not associated with clinical outcome; the latter was strongly associated with pancytopenia (odds ratio for death 22.7) and neutropenia (odds ratio for death 7.7). Conclusions and Clinical Importance: CRP, SAA, and Hp serum concentrations on admission are useful indicators of the clinical phase of CME, but are not useful predictors of clinical outcome.     

Seroprevalence,spatial distribution and risk factors of Borrelia burgdorferi sensu lato in Jordan

Lyme borreliosis has not been studied in Jordan or in much of the Middle East. However, limited research indicates that the tick vector, Ixodes ricinus, exists in the region. This study examined the seroprevalence of B. burgdorferi s.l. in Jordan and potential demographic and zoonotic risk factors for seropositivity. Serum samples of 824 apparently healthy participants from 11 governorates in Jordan were tested for B. burgdorferi s.l. using Enzygnost Lyme link VlsE/IgG enzyme-linked immunosorbent assay. A validated questionnaire was used to collect demographic and animal exposure data. Univariate and multivariate logistic regression were used to identify factors associated with seropositivity. The results showed that 11.7 % (95 % CI, 9.3–14.0 %) of the participants were seropositive for B. burgdorferi s.l.. There was a bimodal age distribution of seroprevalence with higher seroprevalence among individuals <20 and>60 years old. After controlling for governorate of residence, females had 2.77 (95 % CI 1.53–5.00) times greater odds of seropositivity compared to males. Individuals living in the southeastern part of Jordan (Ma’an) had 2.32 (95 % CI, 1.02−5.31) greater odds of seropositivity compared to those living in Amman, the Capital of Jordan, while those living in the northeast had significantly lower odds of seropositivity. This study presents the first evidence of B. burgdorferi s.l. seropositivity in Jordan and suggests several risk factors which were reported in studies conducted elsewhere. This study suggests that Lyme borreliosis should be considered in the differential diagnosis for patients presenting with skin lesions in Jordan.     

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

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

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

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

Canine vector-borne disease in travelled dogs in Germany--a retrospective evaluation of laboratory data from the years 2004-2008

When importing dogs from various Mediterranean countries into Western Europe canine vector-borne infections are often considered as a major issue. Several diseases including babesiosis, leishmaniosis, hepatozoonosis, canine heartworm disease or ehrlichiosis can potentially be endemic in this region and pose a potential health risk for travelling dogs. Information on such infections in travelled dogs is scarce and therefore this study has been undertaken to examine the frequency of vector-borne infections in travelled dogs from the years 2004-2008. A total of 997 samples were screened by direct and/or indirect methods. Total seroprevalence was 7.5% with individual seroprevalence for the 3 species Leishmania spp., Ehrlichia canis and Babesia canis spp. ranging from 3.1 to 4.9%. Total detection rate for pathogens by direct methods was 3.5%. Ninteen Giemsa-stained blood smears were positive for large Babesia. None of the samples screened for microfilariae by Knott's test or for Dirofilaria immitis antigen by DiroChek^® were positive. Using PCR methods Leishmania-DNA was detected in 1/42 samples but none of 59 animals screened for E. canis-DNA was positive. The prevalence values as established by indirect and direct pathogen detection are considered as rather low.     

Ehrlichia canis in dogs of Mexico: Prevalence,incidence, co–infection and factors associated

Rickettsial infections in dogs of Mexico were investigated. A total of 246 dogs were blood sampled and initially screened to detect Ehrlichia canis, E. chaffeensis, E. ewingii, Anaplasma phagocytophilum and Rickettsia rickettsii by a quantitative real–time PCR (qPCR) assay. Sixty–five dogs were monitored and sampled twice 7–8 months apart. Using the qPCR, 72 positive dogs to E. canis were detected (prevalence of 29.26%). These dogs were also tested by nested PCR to detect the same pathogens. None of the studied dogs were positive to E. chaffeensis, E. ewingii, R. rickettsii nor A. phagocytophilum by both PCR assays. The cumulative incidence of E. canis infection was 38.46%. Sequencing analysis of the nested PCR products revealed 100% and 98.1% identity of E. canis and R. parkeri, respectively. We found a dog co–infected with E. canis and R. parkeri.     

Evidence for Unapparent Brucella canis Infections among Adults with Occupational Exposure to Dogs

Human serological assays designed to detect brucellosis will miss infections caused by Brucella canis, and low levels of periodic bacteremia limit diagnosis by blood culture. Recent B. canis outbreaks in dogs and concomitant illnesses in caretakers suggest that unapparent human infections may be occurring. With more than a quarter of a million persons in occupations involving dogs, and nearly 80 million dog owners in the United States, this pathogen is an under‐recognized human health threat. To investigate occupational exposure to B. canis, we adapted a commercial canine serological assay and present the first controlled seroepidemiological study of human B. canis infections in recent years. 306 adults with occupational exposure to dogs and 101 non‐matched, non‐canine‐exposed subjects were enrolled. Antibodies were detected using the canine D‐Tec^® CB rapid slide agglutination test (RSAT) kit with a secondary 2‐mercaptoethanol (ME)‐RSAT. Results were validated on a blinded subset of sera with an additional RSAT and indirect enzyme‐linked immunoassay at the National Administration of Laboratories and Health Institutes (ANLIS) in Argentina. Seroprevalence ranged from 10.8% (RSAT) to 3.6% (ME‐RSAT) among canine‐exposed subjects. Kennel employees were more likely to test RSAT seropositive compared with other canine exposures (OR = 2.7; 95% CI, 1.3–5.8); however, low seroprevalence limited meaningful occupational risk factor analyses. Two seropositive participants reported experiencing symptoms consistent with brucellosis and having exposure to B. canis‐infected dogs; however, temporality of symptom onset with reported exposure could not be determined. D‐Tec^® CB results had substantial agreement with ANLIS assays (Cohen's kappa = 0.60–0.68). These data add to a growing body of literature suggesting that people occupationally exposed to dogs may be at risk of unapparent B. canis infection. It seems prudent to consider B. canis as an occupational public health concern and encourage the development of serological assays to detect human B. canis infections.     

Complement C3 in Bernese Mountain dogs

Background: Previous research suggests that low serum concentrations of the third component of complement (C3) are associated with both the susceptibility to infectious agents such as Borrelia burgdorferi and the development of glomerular disease. We hypothesized that low levels of C3 are associated with the coincident occurrence of B. burgdorferi infection and glomerulonephritis in Bernese Mountain dogs. Objectives: The aims of this study were to evaluate the serum concentration of C3 in Bernese Mountain dogs with and without antibodies against B. burgdorferi and to compare this concentration with that of healthy control dogs. Methods: Eighty‐three clinically healthy Bernese Mountain dogs and 46 control dogs were included. Antibodies against B. burgdorferi were determined using an ELISA with a whole cell sonicate as antigen. Results were confirmed using Western blot. C3 was measured using a single radial immunodiffusion test. Results were reported as the percentage concentration of C3 compared with that in pooled preserved canine serum (100% C3 concentration). Results: Median C3 concentration was 128.5% in Bernese Mountain dogs with antibodies against B. burgdorferi, 133.5% in B. burgdorferi‐negative Bernese Mountain dogs, 87.8% in positive control dogs, and 102.2% in negative control dogs. Within Bernese Mountain and control groups, C3 was lower in dogs with antibodies against B. burgdorferi compared with those without. Percentage concentration of C3 was higher in healthy Bernese Mountain dogs compared with control dogs. Conclusion: Low C3 concentration is not an explanation for the high prevalence of B. burgdorferi infections and glomerular disease in Bernese Mountain dogs.     

THE PREVALENCE OF DIROFILARIA IMMITIS AND OTHER PARASITES IN URBAN POUND DOGS IN NORTH-EASTERN VICTORIA

SUMMARY The results of a survey investigating the prevalence of dirofilaria immitis and other helminth and protozoan infections in urban dogs in north-eastern Victoria are presented. D. immitis was detected in 2.7% of the general population (8.3% of dogs older than 2 years). Microfilariae of D. immitis were not detected in the peripheral circulation of 30% of dogs infected with the adult parasites. The most prevalent gastro-intestinal parasites were Dipylidium caninum 57%; Trichuris vulpis 41%; Toxocara canis 38%; and Uncinaria stenocephala 26%. The prevalence of T. canis in dogs less than one year of age was 73% and this decreased with increasing age. No Echinococcus granulosus adults were detected. Sarcocystis spp and Isospora ohioensis were the most commonly seen coccidians.     

The incidence of canine haematozoa in peninsular Malaysia

In 3 urban areas in Selangor, Peninsular Malaysia between 1973 and 1981, blood from 4080 dogs was examined for haematozoa. The following frequencies were found: Babesia gibsoni 17.7%; microfilariae of Dirofilaria immitis 9.6%; Hepatozoon canis 1.2%; B. canis 1.1%; Ehrlichia canis 0.2%; Trypanosoma evansi 0.1%. A detailed examination of B. gibsoni infections and microfilariasis due to D. immitis with regards to monthly distribution, breed frequency, sex and age, revealed that pedigree and non-pedigree dogs were equally susceptible to Babesia and microfilariae infections.     

Molecular and serological detection of animal and human vector-borne pathogens in the blood of dogs from Côte d’Ivoire

In Côte d’Ivoire, limited information are available on vector-borne pathogens, their prevalence and distribution. Here, we assess the occurrence and diversity of canine vector-borne diseases (CVBDs) in Abidjan and Yamoussoukro cities. Blood from a total of 123 dogs were tested for Leishmania infantum and Ehrlichia canis antibodies and screened for Leishmania and Trypanosoma spp., Piroplasmida, Filariidae and Anaplasmataceae by PCR and sequencing. Among dogs, 39 % were positive for at least one pathogen. Seroprevalences were: 15.4 % and 12.2 % for L. infantum and E. canis, respectively. DNA of L. infantum and T. congolense (4.1 %), Baabesia vogeli (1.6 %), Filariidae (Dirofilaria immitis, D. repens and Acanthocheilonema reconditum) (10.6 %) has been detected. Anaplasmataceae were detected in (17.1 %) and E. canis was the only identified specie. Co-infections were observed in 13.8 % of dogs: E. canis-L. infantum co-infection was the most prevalent (4.9 %). Age, breed and sex of dogs do not seem to influence infections. Village dogs were more susceptible to CVBDs than kennel dogs (PV = 0.0000008). This study reports for the first time the presence of L. infantum, B. vogeli, A. reconditum, D. immitis and D. repens in dogs from Côte d’Ivoire and determines the prevalence and diversity of CVBD pathogens. The results indicate that human and animal pathogens are abundant in Ivoirian dogs which requires attention of veterinarians, physicians and authorities against these diseases, especially against major zoonosis such as visceral leishmaniasis (L. infantum).     

First record of autochthonous canine ehrlichiosis caused by Ehrlichia canis in Romania

This case study describes the first genetically confirmed and clinically manifested autochthonous Ehrlichia canis infection in a 9‐year‐old female mixed‐breed dog from Romania. Health screening of the dog included clinical examination, evaluation of stained peripheral blood smear and hematologic variables, as well as serologic testing and molecular analysis. Clinical signs included fever, apathy, dehydration, pale mucous membranes, and weakness. The microscopic examination of the blood smear and immunologic assays for Borrelia burgdorferi, Anaplasma phagocytophilum, and E canis antibodies, and for Dirofilaria immitis antigen yielded negative results. Hematologic abnormalities included moderate nonregenerative anemia, leucopenia with neutropenia, and moderate thrombocytopenia. The biochemical abnormalities identified were hypoalbuminemia, and mildly increased serum enzyme activities of AST and ALT. In addition, increased urea and creatinine levels associated with low urine specific gravity and proteinuria were also present. Nested PCR amplification of the partial E canis 16S rRNA gene demonstrated the presence of this rickettsial pathogen in the dog's blood, which subsequently was confirmed through sequencing based on the 100% homology with GenBank deposited E canis isolates. After specific treatment with doxycycline (10 mg/kg, orally, SID) for one month, the proteinuria, and hematologic and serum biochemical abnormalities with the exception of mild azotemia resolved. This report supports the geographical expansion of canine ehrlichiosis caused by E canis in nonendemic regions of Europe.     

Exposure of client-owned cats to zoonotic vector-borne pathogens: Clinic-pathological alterations and infection risk analysis

Zoonotic Vector-Borne Diseases (VBDs) represent a relevant health issue for pets and humans. Italy is a major epidemiological hub for feline VBDs, because of suitable conditions for vector biology and disease transmission patterns. The present study investigated the exposure to major zoonotic arthropod-borne pathogens of cats in Italy, along with the evaluation of clinic-pathological features and a risk factor analysis. Out of 167 examined cats, 52 (31.1%) were seropositive for at least one vector-borne pathogen, being positivity for Bartonella henselae the most recorded (18%). Also, various cats seroreacted for Rickettsia felis (10.8%) and Rickettisa typhi (4.2%), Leishmania infantum (3%), Anaplasma phagocytophilum (2.4%) and Ehrlichia canis (2.4%). Forty-six cats were tested also for antibodies against D. immitis and two (4.3%) scored positive. The statistical analysis showed a positive association between flea infestation and seropositivity to B. henselae, other than an association between the administration of monthly ectoparasiticide treatments and seronegativity for Rickettsia spp.; seropositive cats were older than negative animals and the lifestyle (i.e. indoor vs outdoor) was not correlated with exposure to vector-borne pathogens. The majority of seropositive cats appeared clinically healthy or showed aspecific clinical signs. Around 80% of seropositive cats had one or more biochemical and/or complete blood count abnormalities. The present data confirm the endemicity of zoonotic feline VBDs in Italy and indicate that awareness on arthropod infections and transmitted pathogens should be kept high and possible implemented, towards the protection of animal and human health with adequate surveillance plans.     

A multiplex quantitative real-time polymerase chain reaction panel for detecting neurologic pathogens in dogs with meningoencephalitis

Meningoencephalitis (ME) is a common inflammatory disorder of the central nervous system in dogs. Clinically, ME has both infectious and non-infectious causes. In the present study, a multiplex quantitative real-time polymerase chain reaction (mqPCR) panel was optimized for the detection of eight canine neurologic pathogens (Blastomyces dermatitidis, Cryptococcus spp., Neospora caninum, Borrelia burgdorferi, Bartonella spp., Toxoplasma gondii, Ehrlichia canis, and canine distemper virus [CDV]). The mqPCR panel was subsequently applied to 53 cerebrospinal fluid (CSF) samples collected from dogs with ME. The analytic sensitivity (i.e., limit of detection, expressed as molecules per 1 µL of recombinant vector) was 3.8 for CDV, 3.7 for Ehrlichia canis, 3.7 for Bartonella spp., 3.8 for Borrelia burgdorferi, 3.7 for Blastomyces dermatitidis, 3.7 for Cryptococcus spp., 38 for Neospora caninum, and 3.7 for Toxoplasma gondii. Among the tested CSF samples, seven (15%) were positive for the following pathogens in decreasing order of frequency: Cryptococcus spp. (3/7), Blastomyces dermatitidis (2/7), and Borrelia burgdorferi (2/7). In summary, use of an mqPCR panel with high analytic sensitivity as an initial screen for infectious agents in dogs with ME could facilitate the selection of early treatment strategies and improve outcomes.     

A survey of veterinarians’ knowledge,attitudes and practices regarding an emerging disease: Coccidioidomycosis in Washington State

Coccidioides immitis is an emerging fungal pathogen in Washington State (WA). While the geographical boundaries of C. immitis in WA have not been well characterized, human infections have resulted from exposure in the south‐central region of the state. Since 2010, only one dog has been identified as a probable locally acquired animal case, despite pilot canine serological surveys from south‐central WA revealing a 9.8% seroprevalence. On the suspicion that clinical animal cases may be underdiagnosed and/or underreported, we assessed WA veterinarians’ knowledge, attitudes and practices regarding coccidioidomycosis. All veterinarians with active licenses in WA were invited to complete a self‐administered, web‐based survey from November 2018 to January 2019. Four hundred and twenty‐five of 2,211 (19.2%) veterinarians returned the survey and were eligible for inclusion in the final analysis. Nearly, all respondents (98.8%) had heard of coccidioidomycosis, but only 31.5% and 25.6% knew that locally acquired animal and human cases, respectively, had been reported in WA. Only half (52.6%) of participants knew that the disease was reportable. Fewer than 20% of respondents reported confidence that their knowledge of coccidioidomycosis was up to date, and 76.9% “never” or “rarely” considered the risk of disease in their patients. A statistically significant predictor of a high knowledge score (≥70%), however, included practicing in endemic counties where the Washington State Department of Health had previously delivered outreach and education services. While the results suggest some success with these activities, it is clear that information on the emergence of coccidioidomycosis is not adequately reaching veterinary practitioners in WA. The data support that more frequent CE opportunities, and dissemination of public health communications through veterinary‐specific modalities, would help fill the knowledge gap. In turn, improved diagnosis and reporting of animal cases would assist in our overall understanding the epidemiology of coccidioidomycosis in WA.