Presumptive Acute Neural Toxoplasmosis in a Captive Red-Necked Wallaby (Macropus rufogriseus)

A red-necked male wallaby (Macropus rufogriseus) from a German zoo was presented for acute onset of severe neurological signs, including head tremor, lethargy, unresponsiveness, and weakness. Serum biochemical abnormalities included increased LDH- and AST-levels, hyperproteinaemia, and reduced ALT-, ALP-, and creatinine-levels. The wallaby was found serologically positive for Toxoplasma gondii by the indirect haemagglutination test. After initiation of therapy by subcutaneous injections of trimethoprim/sulfadoxin, amelioration of neurological signs was noted and after 10 days the affected wallaby recovered. T. gondii can be confirmed rapidly by serology, and immediate therapy may reduce clinical illness and fatality of the disease within captive macropods

Transmission of Armillifer armillatus Ova at Snake Farm, The Gambia, West Africa

Visceral pentastomiasis caused by Armillifer armillatus larvae was diagnosed in 2 dogs in The Gambia. Parasites were subjected to PCR; phylogenetic analysis confirmed relatedness with branchiurans/crustaceans. Our investigation highlights transmission of infective A. armillatus ova to dogs and, by serologic evidence, also to 1 human, demonstrating a public health concern

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

Symmetrical Dimethylarginine as a Diagnostic Parameter in Hermann's Tortoises (Testudo hermanni)

BackgroundDespite improvements in habitational conditions, kidney disease is relatively common in tortoises. ObjectivesPurpose of this study was the establishment of Symmetrical dimethylarginine (SDMA) reference values for clinically healthy Hermann's Tortoises. AnimalsClinically healthy Hermann's Tortoises (n = 131) were included in the period from October 2017 to September 2019. MethodsCreatinine and other biomarkers were tested at IDEXX Laboratories, Germany using residual blood samples from Hermann's tortoises. SDMA was measured with the IDEXX test and verified by liquid chromatography-mass spectrometry at IDEXX Laboratories, USA. ResultsSDMA values ranged from 1 to 21 mu g/dl (n = 131) for the IDEXX SDMA Test and SDMA values ranged from 1 to 17 mu g/dl (n = 82) for LC-MS. For the comparison of the two measuring systems, the following results were obtained R-2 = 0.75 (p < 0.001). Conclusion and Clinical ImportanceSDMA can be measured in Hermann's Tortoises and the reference values range in clinically healthy animals is comparable to that of dogs and cats

Detection of Leptospira DNA in urine and presence of specific antibodies in outdoor cats in Germany

Objectives Clinical manifestation of infection with Leptospira species in cats is rare. Nevertheless, cats can develop specific antibodies against the spirochetes after infection. In Canada, Taiwan and the USA it was recently demonstrated that naturally infected cats can also shed DNA from pathogenic Leptospira species in their urine, but the zoonotic potential of infected cats is still unclear. The objective of this study was to demonstrate if outdoor cats in Germany shed DNA from pathogenic Leptospira species in their urine. As a second aim, antibody prevalence was determined. Methods Two hundred and fifteen outdoor cats were prospectively recruited. Urine samples were tested by realtime PCR targeting the lipL32 gene of pathogenic Leptospira species. Antibody titres against eight serovars (Australis, Autumnalis, Bratislava, Canicola, Copenhageni, Grippotyphosa, Pomona, Saxkoebing) belonging to seven serogroups (Australis, Autumnalis, Canicola, Grippotyphosa, Icterohaemorrhagiae, Pomona, Sejroe) were determined by microscopic agglutination test. Results Urine samples from 7/215 cats (3.3%;95% confidence interval [CI] 0.9-5.7) were PCR-positive. Specific antibodies were detected in 35/195 cats (17.9%;95% CI: 12.5-23.3) with titres ranging from 1:100 to 1:6400. Australis, Bratislava and Grippotyphosa were the most common serovars. Conclusions and relevance Outdoor cats in Germany can shed DNA from pathogenic Leptospira species. Therefore, outdoor cats should be considered as a possible source of infection for dogs or humans. Further studies are needed to determine the role of Leptospira species as a cause of disease in cats

Isolation of canine Anaplasma phagocytophilum strains from clinical blood samples using the Ixodes ricinus cell line IRE/CTVM20

AbstractAnaplasma phagocytophilum is an intracellular tick-borne rickettsial pathogen, which causes granulocytic anaplasmosis in various species of livestock and companion animals and also in humans. Previously A. phagocytophilum has been isolated and propagated in cell lines derived from the tick Ixodes scapularis and in the human promyelocytic cell line HL60. In this study we used the Ixodes ricinus-derived cell line IRE/CTVM20 to isolate and propagate two new canine strains of A. phagocytophilum.Blood samples were collected by veterinarians from two dogs, one from Germany and the other from Austria. Suspicion of clinical canine granulocytic anaplasmosis was raised by the treating veterinarians and after confirmation of A. phagocytophilum infection by real-time PCR, buffy coat cells were isolated and co-cultivated with IRE/CTVM20 cells maintained at 28°C in L15/L15B medium.In the tick cells, rickettsial inclusions were first recognised after 86 days of incubation. Electron microscopic examination of tick cells infected with one of the isolates revealed cytoplasmic vacuoles containing pleomorphic organisms with individual bacteria enveloped by a bilayer membrane. Sequencing of 16S rRNA genes confirmed the isolation of A. phagocytophilum and showed the highest identity to the A. phagocytophilum human HZ strain. The two A. phagocytophilum isolates were passaged several times in IRE/CTVM20 cells and transferred to the I. scapularis cell line ISE6. This confirms for the first time the successful establishment and continuous cultivation of this pathogen in I. ricinus cells as well as infectivity of these canine strains for I. scapularis cells

First case of Anaplasma platys infection in a dog from Croatia

<p>Abstract</p> <p>Background</p> <p>It is known that <it>Anaplasma (A.) platys</it>, the causative agent of infectious canine cyclic thrombocytopenia, is endemic in countries of the Mediterranean basin. However, few reports are available from the Balkans. This case report describes a dog, which was imported from Croatia to Germany in May 2010. One month later the dog was presented to a local veterinarian in Germany due to intermittent/recurrent diarrhoea. Diagnostic tests were performed to identify infections caused by <it>Anaplasma </it>spp., <it>Ehrlichia </it>spp., <it>Hepatozoon canis, Babesia </it>spp., <it>Leishmania </it>spp., <it>Borrelia burgdorferi </it>and/or <it>Dirofilaria immitis</it>.</p> <p>Findings</p> <p>Haematological examination of a blood smear revealed basophilic inclusions in thrombocytes, which were confirmed as <it>A. platys </it>with a species-specific real-time PCR. Additionally, an infection with <it>Babesia (B.) vogeli </it>was also detected (PCR and serology). No specific antibodies against <it>Anaplasma </it>antigen were detectable. Although the dog showed no specific clinical signs, thrombocytopenia, anaemia and elevated C-reactive protein (CRP) were observed. Sequencing of a 1,348-bp partial ribosomal RNA gene revealed highest homology to <it>A. platys </it>sequences from Thailand, Japan and France.</p> <p>Conclusions</p> <p><it>A. platys </it>was detected for first time in a dog imported from Croatia. As the dog was also co-infected by <it>B. vogeli</it>, unique serological and haematological findings were recorded. Thrombocytopenia, anaemia and elevated values of C-reactive protein were the laboratory test abnormalities observed in this case. <it>A. platys </it>infections should be considered in dogs coming from Croatia and adjacent regions.</p

The Mitochondrial Genomes of the Zoonotic Canine Filarial Parasites Dirofilaria (Nochtiella) repens and Candidatus Dirofilaria (Nochtiella) Honkongensis Provide Evidence for Presence of Cryptic Species

Background Cutaneous dirofilariosis is a canine mosquito-borne zoonosis that can cause larva migrans disease in humans. Dirofilaria repens is considered an emerging pathogen occurring with high prevalence in Mediterranean areas and many parts of tropical Asia. In Hong Kong, a second species, Candidatus Dirofilaria hongkongensis, has been reported. The present study aimed to compare mitochondrial genomes from these parasites and to obtain population genetic information. Methods and Findings Complete mitochondrial genomes were obtained by PCR and Sanger sequencing or ILLU-MINA sequencing for four worms. Cytochrome oxidase subunit 1 sequences identified three as D. repens (all from Europe) and one as C. D. hongkongensis (from India). Mitochondrial genomes have the same organization as in other spirurid nematodes but a higher preference for thymine in the coding strand. Phylogenetic analysis was in contradiction to current taxonomy of the Onchocercidae but in agreement with a recent multi-locus p hylogenetic analysis using both mitochondrial and nuclear markers. D. repens and C. D. hongkongensis sequences clustered together and were the common sister group to Dirofilaria immitis. Analysis of a 2.5 kb mitochondrial genome fragment from macrofilaria or canine blood samples from Europe (42), Thailand (2), India (1) and Vietnam (1) revealed only small genetic differences in the D. repens samples including all European and the Vietnam sample. The Indian C. D. hongkongensis and the two Thai samples formed separate clusters and differences were comparatively large. Conclusion Genetic differences between Dirofilaria spp. causing cutaneous disease can be considerable whereas D. repens itself was genetically quite homogenous. C. D. hongkongensis was identified for the first time from the Indian subcontinent. The full mitochondrial genome sequence strengthens the hypothesis that it represents an independent species and the Thai samples might represent another cryptic species, Candidatus Dirofilaria sp. 'Thailand II', or a quite divergent population of C. D. hongkongensis