Atypical case of subcutaneous filariosis in a cat caused by Dirofilaria immitis.

Subcutaneous dirofilariosis is a well-known disease caused mainly by Dirofilaria repens and described in several mammalian species including humans, dogs, and cats. Additionally, early developing stages of the heartworm Dirofilaria immitis are rarely reported in subcutaneous localization from humans and dogs. To our knowledge, confirmed clinical evidence of this condition has not been described in the cats yet, even if the feline hosts can be affected either by the classic adult-related heartworm form or heartworm-associated respiratory disease (HARD) caused by immature stages. A 2 year old, spayed male cat was presented for three subcutaneous nodules on the head and trunk. The cat lived in Northern Italy and was regularly vaccinated and treated monthly with an antiparasitic spot on formulation containing selamectin. One of the three nodules was surgically excised and examined. Histology showed the presence of a nodular lesion in the subcutis characterized by a severe inflammatory infiltrate composed of macrophages, small lymphocytes, with fewer eosinophils, and mast cells, supported by a proliferation of mature fibroblasts (fibrosis). Inflammatory cells were multifocally surrounding sections of parasites identified as adult nematodes. Microscopic features were compatible with D. immitis, which has been molecularly confirmed (98.2% identity to D. immitis isolate OP107739). The cat tested negative for D. immitis antigenemia and the two remaining nodules disappeared spontaneously in a few months. In region where heartworm is prevalent, aberrant localization of D. immitis should be considered in the differential diagnoses of subcutaneous filarial worms in cats and dogs.

Evaluation of a commercial coproantigen immunoassay for the detection of Toxocara cati and Ancylostoma tubaeforme in cats and Uncinaria stenocephala in dogs.

Coproantigen immunoassays (IDEXX Fecal Dx® antigen tests) were evaluated for their ability to identify Toxocara cati and Ancylostoma tubaeforme infections in cats and Uncinaria stenocephala infection in dogs. Five cats were experimentally infected with 500 embryonated eggs of T. cati, eight cats with 500 third-stage larvae (L3) of A. tubaeforme and seven dogs with 500 L3 of U. stenocephala. In addition to the three coproantigen tests, the course of infection was monitored by a combined sedimentation-flotation method with ZnSO4 as flotation medium (specific gravity: 1.28-1.30) and a modified McMaster method in case of copromicroscopically positive samples. Eggs of T. cati were first observed between 28 and 54 days post infection (dpi) in four of the five infected cats. In these four cats, positive roundworm coproantigen signals were obtained between 16 and 44 dpi. Positive coproantigen signal always preceded egg observations, but the interval varied between 6 and 30 days. Hookworm-specific positive coproantigen signals were detected in seven of the eight A. tubaeforme infected cats between 10 and 52 dpi, while consecutive egg excretion was observed in three cats between day 26 and 54 pi. Of these three, coproantigen signal preceded egg observation by 12 to 24 days. Four cats had positive coproantigen results in the absence of egg excretion, and one cat never achieved a positive result for egg or coproantigen. In six of seven U. stenocephala infected dogs, infection was confirmed by copromicroscopy between 16 and 24 dpi as well as for hookworm coproantigen between 10 and 14 dpi. Coproantigen signal was detected prior to egg observation by 2 to 14 days. No cross-reactions between the roundworm, hookworm und whipworm tests occurred in study animals. The results of this study demonstrate the ability of the coproantigen tests to detect the common roundworm and hookworm infections in cats and U. stenocephala infections in dogs as well as the ability to detect the prepatent stage of infection.

Estimated specific antibody-based true sero-prevalences of canine filariosis in dogs in Central Europe and the UK.

Dirofilariosis is a vector-borne disease mainly caused by Dirofilaria immitis and Dirofilaria repens. In contrast to the known endemicity of dirofilariosis in southern and south-eastern Europe, information on the distribution of D. repens in Central-Europe is fragmentary. We tested 8877 serum samples from dogs from Austria, Denmark, Germany, Italy, Lithuania, Poland, Switzerland and the UK using an ELISA detecting filarial-specific antibodies, hypothesising higher occurrence of D. repens. Based on two overlapping frequency distributions, presumed negative samples had a mean optical density (OD) value of 0.097, representing 97.45% of all samples. Presumed positive samples, representing 2.55% of all sera, had a mean OD value of 0.287. Test prevalence based on the calculated cut-off was 3.51% for all sera (4.36% for Austria, 1.94% for Denmark, 1.39% for Germany, 3.37% for Italy, 6.90% for Lithuania, 6.99% for Poland, 0.77% for Switzerland and 0.0% for the UK, respectively). The bimodal distribution, representing overlapping distributions of OD values from positive and negative dogs, enabled the assignment of a probability of true infection status to each dog. Mean probabilities of true infection status across groups, based on the postal codes of origin, allowed us to estimate and map true prevalences. For all countries, except the UK, the true prevalence was lower than the test prevalence. The large number of serum samples and the use of a non-gold standard analytical method allowed us to create a more realistic picture of the distribution of D. repens in Central Europe and the UK.

HEALTH ASSESSMENT OF CAPTIVE AND FREE-LIVING EUROPEAN POND TURTLES (EMYS ORBICULARIS) IN SWITZERLAND.

The highly endangered European pond turtle (Emys orbicularis) was reintroduced in Switzerland in 2010. Up until 2019, no routine medical examinations have been carried out prior to its release or during recapture events. The aim of this study was to assess the health status of captive and free-living Emys orbicularis populations in Switzerland, taking into account the most important and frequently occurring health threats to freshwater turtles. A total of 141 European pond turtles, including captive (n = 89) and free-living (n = 52) individuals, underwent clinical examination (n = 136), choanal and cloacal swab collection for microbiology investigation (n = 140), blood sampling (n = 121), fecal examination for parasitology (n = 92), radiography (n = 84), and ultrasound (n = 46). Microbiology investigation included conventional PCR for herpesvirus, ranavirus, and Mycoplasma spp. Blood was used for the establishment of reference values for hematocrit, leukocyte count, and differential blood count as well as for biochemistry parameters tested with the VetScan VS2. An emydid Mycoplasma was detected in 40% (n = 56/140; 95%CI: 31.82-48.61%) of the turtles, including one individual with upper respiratory signs. Four animals positive for Mycoplasma arrived dead or were euthanized during the study period. Their necropsies revealed no evidence of respiratory disease. No ranavirus or herpesvirus was detected in any of the tested turtles. Two presumptively fatal infections with spirorchiid trematodes were reported during the study period. Endoparasites were detected in only 7.94% of the samples examined. This study provides comprehensive data on the current health status of the largest sample size of captive and free-living populations of Emys orbicularis ever assessed to date and serves as a baseline for future research investigations and management recommendations in this species.

Co-infection with Babesia canis and Babesia gibsoni in a dog.

A four-year-old intact male Boxer, that had a history of travelling to Serbia, was referred for lethargy and anaemia. Shortly before the dog was referred, it was diagnosed twice with an infection with Babesia canis and was treated with imidocarb both times. A blood smear evaluation was indicative of the presence of intraerythrocytic piroplasms. After receiving inconclusive results regarding the type of piroplasm, the dog was diagnosed with simultaneous infections with B. canis and Babesia gibsoni via real-time polymerase chain reaction (rt-PCR) testing. The dog was treated with imidocarb, atovaquone and azithromycin, and in a follow-up examination, the PCR results were negative for B. canis and B. gibsoni. Several weeks later, the dog was presented again, and a PCR was positive for B. gibsoni. After atovaquone and azithromycin failed to eliminate the parasites, a therapy attempt using metronidazole, clindamycin and doxycycline was initiated. Six months after diagnosis, the treatment appeared successful in eliminating B. gibsoni. This case report describes the clinical findings of the co-infection and the initiated diagnostic and therapeutic approaches.

Canine Dracunculus Nematode Infection, Toledo, Spain.

A fragment of a Dracunculus-like worm was extracted from the hind limb of a 2-year-old dog from Toledo, Spain. Cytochrome oxidase I and rRNA sequences confirmed an autochthonous mammalian Dracunculus worm infection in Europe. Sequence analyses suggest close relation to a parasite obtained from a North American opossum.

Leptospira infection and shedding in dogs in Thailand.

BACKGROUND: Leptospirosis is a widespread zoonosis and has been recognized as a re-emerging infectious disease in humans and dogs, but prevalence of Leptospira shedding in dogs in Thailand is unknown. The aim of this study was to determine urinary shedding of Leptospira in dogs in Thailand, to evaluate antibody prevalence by microscopic agglutination test (MAT) and enzyme-linked immunosorbent assay (ELISA), and to assess risk factors for Leptospira infection. In Northern, Northeastern, and Central Thailand, 273 stray (n = 119) or client-owned (n = 154) dogs from rural (n = 139) or urban (n = 134) areas were randomly included. Dogs that had received antibiotics within 4 weeks prior to sampling were excluded. No dog had received vaccination against Leptospira. Urine was evaluated by real-time polymerase chain reaction (PCR) specific for lipL32 gene of pathogenic Leptospira. Additionally, urine was cultured for 6 months in Ellinghausen-McCullough-Johnson-Harris (EMJH) medium. Antibodies were measured by ELISA and MAT against 24 serovars belonging to 15 serogroups and 1 undesignated serogroup. Risk factor analysis was performed with backwards stepwise selection based on Wald. RESULTS: Twelve of 273 (4.4%; 95% confidence interval (CI): 2.0-6.8%) urine samples were PCR-positive. In 1/273 dogs (0.4%; 95% CI: 0.01-1.1%) Leptospira could be cultured from urine. MAT detected antibodies in 33/273 dogs (12.1%; 95% CI: 8.2-16.0%) against 19 different serovars (Anhoa, Australis, Ballum, Bataviae, Bratislava, Broomi, Canicola, Copenhageni, Coxi, Grippotyphosa, Haemolytica, Icterohaemorrhagiae, Khorat, Paidjan, Patoc, Pyrogenes, Rachmati, Saxkoebing, Sejroe). In 111/252 dogs (44.0%; 95% CI: 37.9-50.2%) immunoglobulin M (IgM) and/or immunoglobulin G (IgG) antibodies were found by ELISA. Female dogs had a significantly higher risk for Leptospira infection (p = 0.023). CONCLUSIONS: Leptospira shedding occurs in randomly sampled dogs in Thailand, with infection rates comparable to those of Europe and the USA. Therefore, the potential zoonotic risk should not be underestimated and use of Leptospira vaccines are recommended.

First report of Cytauxzoon sp. infection in Germany: organism description and molecular confirmation in a domestic cat.

Cytauxzoonosis is described as an emerging tick-borne disease of domestic and wild felids caused by protozoans of the genus Cytauxzoon. While in the Americas the condition is described as a fatal disease, in Europe, reports on the clinical expression of the infection are scarce. This study describes the first case of Cytauxzoon sp. infection in Germany, in a domestic cat. A 6-year-old male domestic cat living in Saarlouis (Saarland) was presented with anorexia, lethargy and weight loss. The cat had an outdoor lifestyle and had not travelled abroad. Serum clinical chemistry analysis revealed azotaemia with markedly increased symmetric dimethylarginine, hypercreatinemia, hyperphosphatemia and hypoalbuminemia. Moreover, a mild non-regenerative anaemia was present. Approximately 1 year prior to these findings, the domestic cat was diagnosed with a feline immunodeficiency virus (FIV) infection. These results pointed toward a decreased glomerular filtration rate, presumably as a result of kidney dysfunction. Round to oval signet ring-shaped intraerythrocytic organisms, morphologically suggestive for a piroplasm, were revealed during blood smear evaluation with a degree of parasitaemia of 33.0%. PCR analyses and sequencing of a region of the 18S rRNA gene confirmed the presence of a Cytauxzoon sp. infection, with 99-100% nucleotide sequence identity with previously published Cytauxzoon sp. isolates. As this is the first molecularly confirmed Cytauxzoon sp. infection in a domestic cat in Germany, these findings suggest that cytauxzoonosis should be considered as a differential diagnosis in cases of anaemia in outdoor domestic cats, particularly in areas where wild felid populations are present.

Seroprevalence of Anaplasma phagocytophilum, Ehrlichia spp. and Borrelia burgdorferi infections in horses: first report from Northern Bulgaria - Short communication.

Lyme borreliosis, granulocytic anaplasmosis and monocytic ehrlichiosis are well studied in humans and dogs. In horses, these diseases are not widely investigated and limited information is available about their occurrence. The purpose of this study was to present the first ELISA-based report on the seroprevalence of Anaplasma phagocytophilum, Ehrlichia spp. and Borrelia burgdorferi in horses from Northern Bulgaria. A total of 192 horses were investigated from three regions in Northern Bulgaria (Northwestern, North-Central and Northeastern Bulgaria). All equine sera were tested for A. phagocytophilum, Ehrlichia spp. and B. burgdorferi antibodies by a commercial rapid ELISA test. Antibodies against A. phagocytophilum were found in all the three regions at a mean frequency of 12% (23/192), ranging from 9.38 to 15.63% by region. Antibodies against Ehrlichia spp. were found in horses from one region (Northeastern) at a rate of 0.5% (1/192). Anti-B. burgdorferi antibodies were detected in all the three regions with a mean frequency of 15.1% (29/192), ranging from 14.06 to 17.19% by region. A co-exposure to A. phagocytophilum and B. burgdorferi was observed in 6.3% of the cases (12/192). This is the first report on the natural exposure of horses to these bacteria (A. phagocytophilum, Ehrlichia spp. and B. burgdorferi) in Northern Bulgaria.

Canine Leishmania infantum infection: an imported case in UK after staying in the Canary Islands.

Leishmaniosis is reported in the Iberian Peninsula and the Balearic Islands, but the Canary Islands are deemed free. In the present communication, we report a clinical leishmaniosis due to Leishmania infantum in a dog that was presumptively infected during its stay on Tenerife. The result of Leishmania serology (whole-cell based ELISA with L. infantum antigen) was high positive (test score of 82.2 at a cut-off value of 12.0). This result was further confirmed with quantitative real-time polymerase chain reaction (qPCR) for Leishmania spp. on a blood sample. A medium load of parasites was detected (48 parasites/ml blood). L. infantum was identified by RFLP analysis of the ITS-1 PCR product. Confirmation that leishmaniosis is endemic to the Canary Islands would further require study on local dogs with no travel history as well as reassessment on frequency and distribution of Phlebotomus spp. as well as Leishmania spp. detection in the sand fly vector. However, this case strongly suggests that L. infantum is present on the Canary Islands. Although transmission seems to be still exceptional, preventive measures in dogs travelling to the Canaries should be considered.

Sensitivity and specificity of a real-time reverse transcriptase polymerase chain reaction detecting feline coronavirus mutations in effusion and serum/plasma of cats to diagnose feline infectious peritonitis.

BACKGROUND: Feline coronavirus (FCoV) exists as two pathotypes, and FCoV spike gene mutations are considered responsible for the pathotypic switch in feline infectious peritonitis (FIP) pathogenesis. The aim of this study was to evaluate sensitivity and specificity of a real-time reverse transcriptase polymerase chain reaction (RT-PCR) specifically designed to detect FCoV spike gene mutations at two nucleotide positions. It was hypothesized that this test would correctly discriminate feline infectious peritonitis virus (FIPV) and feline enteric coronavirus (FECV). METHODS: The study included 63 cats with signs consistent with FIP. FIP was confirmed in 38 cats. Twenty-five control cats were definitively diagnosed with a disease other than FIP. Effusion and/or serum/plasma samples were examined by real-time RT-PCR targeting the two FCoV spike gene fusion peptide mutations M1058 L and S1060A using an allelic discrimination approach. Sensitivity, specificity, negative and positive predictive values including 95% confidence intervals (95% CI) were calculated. RESULTS: FIPV was detected in the effusion of 25/59 cats, one of them being a control cat with chronic kidney disease. A mixed population of FIPV/FECV was detected in the effusion of 2/59 cats; all of them had FIP. RT-PCR was negative or the pathotype could not be determined in 34/59 effusion samples. In effusion, sensitivity was 68.6% (95% CI 50.7-83.2), specificity was 95.8% (95% CI 78.9-99.9). No serum/plasma samples were positive for FIPV. CONCLUSIONS: Although specificity of the test in effusions was high, one false positive result occurred. The use of serum/plasma cannot be recommended due to a low viral load in blood.

Characterization of an IgG monoclonal antibody targeted to both tissue cyst and sporocyst walls of Toxoplasma gondii.

Toxoplasma gondii infects animals habiting terrestrial and aquatic environments. Its oocysts and tissue cysts are important for the horizontal transmission of this parasite. The oocyst and tissue cyst walls are crucial for the ability of the parasite to persist in the environment or in animal tissues, respectively. However, the composition of these walls is not well understood. We report the generation of monoclonal antibodies directed against wall components using mice immunized with oocyst antigens of T. gondii. One monoclonal antibody (mAb) G1/19 reacted solely with T. gondii sporozoites. The respective antigen had a relative molecular weight (Mr) of 30 kDa. MAb G1/19 failed to react with sporozoites of any other coccidian parasite species tested (Hammondia hammondi, Hammondia heydorni, Cystoisospora felis, Eimeria bovis, Sarcocystis sp.). Another mAb, designated K8/15-15, recognized antigens in sporocyst walls of the parasite and in the walls of in vivo or in vitro produced tissue cysts, as demonstrated by immunofluorescence and immunoblot assays. Antigens of 80 to a high molecular weight protein of about 350 kDa Mr were recognized by this antibody using antigen extracts from sporocysts, and from in vitro or in vivo generated tissue cysts of the parasite. Tissue cyst and sporocyst walls of H. hammondi and H. heydorni, and tissue cysts of Neospora caninum were also recognized by mAb K8/15-15. Sporocyst walls of C. felis also reacted to this mAb. The cyst walls of Sarcocystis sp. and Besnoitia besnoiti were not recognized by mAb K8/15-15. Reactivity by a single mAb against T. gondii antigens in tissue cysts and sporocysts had not been reported previously. MAb K8/15-15 may be a practical tool for the identification of both cysts and sporocysts of the parasite, and may also be potentially employed in proteomic studies on the identification of new components of the cyst and sporocyst walls of T. gondii.

Emergence of cutaneous neosporosis in a dog receiving immunosuppressive therapy: molecular identification and management.

BACKGROUND: Neosporosis is a multisystemic disease caused by the intracellular protozoan Neospora caninum. In dogs the disease primarily affects the central nervous system. Canine cutaneous neosporosis is a rare condition often associated with old age or concurrent immunosuppressive treatments for different underlying conditions. ANIMALS: A 10-year-old female spayed golden retriever dog affected by primary immune-mediated myelofibrosis and treated with immunosuppressive therapies for 6 weeks that developed severe cutaneous lesions. METHODS: Definitive diagnosis was based on several investigation techniques including serology (immunoblotting), immunohistochemistry (IHC), species-specific conventional and real-time PCR, and DNA sequencing. RESULTS: Remission of cutaneous neosporosis was obtained with the administration of clindamycin while the concurrent immunosuppressive therapy was maintained to manage the underlying primary condition. CONCLUSIONS AND CLINICAL IMPORTANCE: To the best of the authors' knowledge this is the first report of species-specific PCR and DNA sequencing used as diagnostic methods for canine cutaneous neosporosis emerging in a dog receiving immunosuppressive therapy.

Current Surveys of the Seroprevalence of Borrelia burgdorferi, Ehrlichia canis, Anaplasma phagocytophilum, Leishmania infantum, Babesia canis, Angiostrongylus vasorum and Dirofilaria immitis in Dogs in Bulgaria.

Canine vector-borne diseases (CVBDs) have increasingly become a focus of interest in recent years. Some of the CVBDs are zoonotic and may therefore also represent a risk for the human population. Different factors are in discussion to explain the expansion of vectors and pathogens into formerly unaffected areas. Knowledge of the prevalence and distribution of CVBDs in Bulgaria is scant overall and most data rely on single case descriptions. The aim of the present study was to determine the seroprevalence of important CVBDs in 167 dogs from central-southern Bulgaria (Stara Zagora), with special emphasis on hitherto uninvestigated babesiosis and angiostrongylosis, on poorly investigated Lyme borreliosis and canine granulocytic anaplasmosis, and on the potentially zoonotic dirofilariosis and leishmaniosis. Relatively high prevalence rates were documented for anti-Babesia canis antibodies, Dirofilaria immitis antigen (16.2 %; 27/167 each), anti-Ehrlichia canis (21 %; 35/167) and anti-Anaplasma phagocytophilum antibodies (30.5 - 46.1 %; 51 - 77/167), while Borrelia burgdorferi seroprevalence was low (2.4 %; 4/167). All samples were negative for Leishmania infantum antibodies and Angiostrongylus vasorum antigen and antibodies. In total, 64.7 % (108/167) of the samples indicated infection or exposure to at least one agent and a high proportion of dual infections (39.8 %; 43/108) was demonstrated. Multiple infections with up to four different organisms were also detected. Our data underline the importance of CVBDs and especially of co-infections which could influence the clinical outcome in dogs.

Tick-borne Diseases (Borreliosis, Anaplasmosis, Babesiosis) in German and Austrian Dogs: Status quo and Review of Distribution, Transmission, Clinical Findings, Diagnostics and Prophylaxis.

Tick-borne diseases (TBD) in dogs have gained in significance in German and Austrian veterinary practices. The widespread European tick species Ixodes ricinus represents an important vector for spirochaetes of the Borrelia burgdorferi sensu lato group and Rickettsiales such as Anaplasma phagocytophilum. The meadow or ornate dog tick (Dermacentor reticulatus) is an important vector for Babesia canis, as is the brown dog tick (Rhipicephalus sanguineus) for Babesia vogeli in the Mediterranean region. The present work covers pathogen transmission by tick vectors, including the mechanisms and the minimum intervals required, in conjunction with possible non-vector-borne transmission routes. It also addresses the incubation periods, pathogenicity and clinical findings associated with each pathogen and genospecies and presents case examples. Current data on prevalence, annual fluctuations and distribution in various pre-selected dog populations (symptomatic versus asymptomatic) in both countries are depicted in maps. Reasons for changes in prevalence (especially of Borrelia) are discussed. Criteria and algorithms for clinical diagnosis and monitoring in dogs, including case history, direct detection (blood smears, molecular detection by species-specific PCR and sequencing) and indirect methods (whole-cell and peptide-based antibody tests), are presented, together with laboratory abnormalities (haematology, clinical chemistry, urine). The role of anti-C6 antibody concentration (ACAC) and its correlation with proteinuria and Lyme nephritis are assessed on the basis of new data. Consideration is also given to the importance of blood smears, PCR and serology in the case of anaplasmosis and babesiosis, and the diagnostic value of combining these methods. The relevance of molecular differentiation of Anaplasma species (A. phagocytophilum versus A. platys) and Babesia spp. (large versus small forms) in cases of serological cross-reaction is emphasized. A summary is given of methods for prophylaxis using acaricide products (collars, spot-on solutions and oral treatments in both countries), vaccination (Borrelia and Babesia vaccines) and imidocarb-based chemoprophylaxis for large Babesia.

Current surveys on the prevalence and distribution of Dirofilaria spp. and Acanthocheilonema reconditum infections in dogs in Romania.

During the last decades, Dirofilaria spp. infection in European dogs has rapidly spread from historically endemic areas towards eastern and northeastern countries, but little or no information is available from these geographical regions. The present study provides a picture of filarial infections in dogs from Romania and compares two tests for the diagnosis of Dirofilaria immitis. From July 2010 to March 2011, blood samples were collected from 390 dogs from nine counties of Romania and serological SNAP tests were performed for the detection of D. immitis antigen. The remaining blood clots were subsequently used for DNA extraction followed by multiplex PCR for assessing filarioid species diversity (i.e. D. immitis, Dirofilaria repens and Acanthocheilonema reconditum). Based on molecular detection, an overall prevalence of 6.92 % (n = 27; 95 % confidence interval (CI) 4.70-10.03 %) for D. repens, 6.15 % (n = 24; 95 % CI 4.07-9.14 %) for D. immitis and 2.05 % (n = 8; 95 % CI 0.96-4.16 %) for A. reconditum was recorded, with significant variations according to sampling areas. Coinfections of D. immitis and D. repens were recorded in 23.91 % (n = 11) positive dogs. A slightly higher prevalence for D. immitis was detected at the SNAP test (n = 28, 7.17 %; 95 % CI 4.91-10.33 %), but this difference was not statistically significant (p = 0.66). However, only 53.57 % (n = 15) of antigen-positive dogs were confirmed by PCR, while other dogs (n = 9) PCR positive for D. immitis were negative at the serology. The present study shows that Dirofilaria species are endemic in the southern and southeastern areas of Romania, This article also provides, for the first time, an epidemiological picture of the distribution of A. reconditum in Romania.

Molecular identification of Sarcocystis spp. helped to define the origin of green pythons (Morelia viridis) confiscated in Germany.

Sarcocystis spp. represent apicomplexan parasites. They usually have a heteroxenous life cycle. Around 200 species have been described, affecting a wide range of animals worldwide, including reptiles. In recent years, large numbers of reptiles have been imported into Europe as pets and, as a consequence, animal welfare and species protection issues emerged. A sample of pooled feces from four confiscated green pythons (Morelia viridis) containing Sarcocystis spp. sporocysts was investigated. These snakes were imported for the pet trade and declared as being captive-bred. Full length 18S rRNA genes were amplified, cloned into plasmids and sequenced. Two different Sarcocystis spp. sequences were identified and registered as Sarcocystis sp. from M. viridis in GenBank. Both showed a 95-97% sequence identity with the 18S rRNA gene of Sarcocystis singaporensis. Phylogenetic analysis positioned these sequences together with other Sarcocystis spp. from snakes and rodents as definitive and intermediate hosts (IH), respectively. Sequence data and also the results of clinical and parasitological examinations suggest that the snakes were definitive hosts for Sarcocystis spp. that circulate in wild IH. Thus, it seems unlikely that the infected snakes had been legally bred. Our research shows that information on the infection of snakes with Sarcocystis spp. may be used to assess compliance with regulations on the trade with wildlife species.

Serological diagnosis of canine leishmaniosis: comparison of three commercially available tests.

Quantitative serology is an important tool in canine leishmaniosis diagnostics from clinical and epidemiological points of view. Serologic diagnosis in laboratories is traditionally carried out by immunofluorescent antibody test (IFAT), but enzyme-linked immunosorbent assays (ELISA) are being increasingly employed. Two commercially available ELISAs (LEISHMANIA-ELISA DOG® [LED] and INGEZIM LEISHMANIA® [IL]) for the detection of Leishmania infantum infection in dogs were compared with the classical IFAT technique. Ninety-two canine serum samples covering a broad range of IFAT titers were chosen for evaluation. Titers ranged from negative (<1:50) to high (>1:3,200). Statistical analysis showed high correlation between all three assays for both negative and positive IFAT-tested samples as described by respective Spearman's rank correlation coefficient (r s), but results varied for samples with inconclusive IFAT titers (1:50-1:100) with IL stating samples predominantly negative. The highest accordance was found between LED and IFAT (percentage of identical results = 83.7%; r(s) = 0.90, p < 0.0001). IL showed higher analogy with LED (accordance = 81.5%; r(s )= 0.88, p < 0.0001) than with IFAT (73.9%; r(s) = 0.80, p < 0.0001). The distribution of the different ELISA scores is discussed and grouped according to correspondent IFAT titers to familiarize practitioners with the range of these tests since antibody levels play an important role in clinical management of canine patients with L. infantum infection.

Serological detection of Anaplasma phagocytophilum, Borrelia burgdorferi sensu lato and Ehrlichia canis antibodies and Dirofilaria immitis antigen in a countrywide survey in dogs in Poland.

Canine vector-borne diseases (CVBDs) have increasingly become a focus of attention in the past few years. Nevertheless, in many parts of Europe information on their occurrence is still scarce. In a large study in Poland 3,094 serum samples taken from dogs throughout all 16 Polish provinces were tested using a commercial kit for the detection of circulating antibodies against Anaplasma phagocytophilum, Borrelia burgdorferi sensu lato and Ehrlichia canis and of Dirofilaria immitis antigen. A total of 12.31% (381/3,094; 95% confidence interval [CI]: 11.18-13.52%) and 3.75% (116/3,094; 95% CI: 3.11-4.48%) of the dogs were positive for A. phagocytophilum and B. burgdorferi s.l. antibodies, respectively. Furthermore, 0.26% (8/3,094; 95% CI: 0.11-0.51%) were positive for E. canis antibodies and 0.16% (5/3,094; 95% CI: 0.05-0.38%) for D. immitis antigen. The highest percentages of A. phagocytophilum-positive dogs were noted in Lesser Poland, Silesia and Lódz Provinces. For B. burgdorferi s.l., the highest prevalence was recorded in Lódz Province. Co-infections with A. phagocytophilum and B. burgdorferi s.l. were recorded in 1.71% of all examined dogs (53/3,094; 95% CI: 1.29-2.23%). One dog even had a triple infection, testing positive for E. canis too. Both A. phagocytophilum and B. burgdorferi s.l. have previously been reported in Poland and were confirmed in the present study by positive samples from all 16 provinces. Concerning E. canis and D. immitis travel history or importation cannot be excluded as factors which may have determined the occurrence of these pathogens in the relevant animals. Practitioners in Poland should be aware of the above mentioned CVBDs and of prophylactic measures to protect dogs and their owners.

Anaplasma phagocytophilum infection associated with strong inflammatory response in 3 cats.

Anaplasmosis is a vector-borne disease caused by Anaplasma (A.) spp. which currently is still rarely diagnosed in cats. This article describes 3 independent cases of anaplasmosis in cats from different regions of Germany presented to veterinarians in 2021. All cats showed unspecific clinical signs, such as fever, reduced general condition, and decreased appetite. One cat additionally had generalized limb pain, another showed reluctance to move as well as vomiting. On complete blood cell count, only 1 of 3 cats showed mild thrombocytopenia. A. phagocytophilum was detected in blood samples of all 3 cats by polymerase chain reaction. Additionally, in 2 cats (in which blood smears were evaluated) morulae could be detected within neutrophilic granulocytes. Initially, all 3 cats had highly elevated serum amyloid A (SAA) concentrations. Treatment with doxycycline caused a rapid improvement of clinical signs, followed by a decrease of SAA concentrations to normal levels as well as negative PCR results after a treatment duration of at least 28 days. In cats with fever, otherwise unspecific clinical signs with only mild or no hematological changes, elevated SAA concentrations, and previous exposure to ticks, attending veterinarians should consider anaplasmosis as differential diagnosis.