Outcome of parasitological examinations in dogs in Germany: a retrospective survey.

Dog faecal samples examined from January 2019 to December 2019 were retrospectively analysed for frequency of endoparasites. The examinations were performed with several different methods: 29,219 samples were examined by flotation method and sodium acetate-acetic acid-formalin concentration (SAFC) technique, 1,330 samples by Baermann-Wetzel migration technique, 12,221 samples using a Giardia coproantigen enzyme-linked-immunosorbent assay (ELISA), 1,180 samples using a Cryptosporidium coproantigen ELISA, 1,671 samples by polymerase chain reaction (PCR) testing for Giardia duodenalis and 447 samples by PCR testing for Cryptosporidium spp.. A total of 7.1% of the samples were positive for parasites in the microscopical examination using the flotation method and SAFC technique. The parasites found included Cystoisospora spp. (2.8%), Giardia duodenalis (2.3%), Ancylostomatidae (1.8%), Toxocara canis (1.6%), Trichuris vulpis (0.7%), Toxascaris leonina (0.5%), Capillaria spp. (0.2%), Angiostrongylus vasorum (0.2%), Crenosoma vulpis (0.1%), Taeniidae (0.1%), Sarcocystis spp. (0.03%), Dipylidium caninum (0.01%), Diphyllobothrium latum (< 0.01%), Spirurida (< 0.01%) and Opisthorchiidae (< 0.01%). Using the Baermann-Wetzel migration technique, Angiostrongylus vasorum was found in 0.75% and Crenosoma vulpis in 0.3% of the samples. ELISAs for Giardia duodenalis and Cryptosporidium spp. revealed 13.9% and 1.0% positive faecal samples, and Giardia duodenalis and Cryptosporidium spp. PCRs 19.4% and 2.0%, respectively. Dogs in the first year of life were more frequently infected with parasites than older animals. In the microscopic examination using flotation method and SAFC technique, the significantly highest detection rates were found in dogs up to six months of age (p < 0.001).

[Occurrence of Ehrlichia canis in dogs living in Germany and comparison of direct and indirect diagnostic methods]. / Vorkommen von Ehrlichia canis bei in Deutschland lebenden Hunden sowie Vergleich direkter und indirekter Diagnostikmethoden.

OBJECTIVE: To study the occurrence of Ehrlichia (E.) canis in dogs living in Germany and evaluate the possibilities and limits of direct and indirect diagnostic methods. MATERIAL AND METHODS: The first part of the study was a retrospective analysis of routine samples, which had been examined for E. canis antibodies using an indirect immunofluorescence antibody test (IFAT). The examination was part of a laboratory profile for the detection of travel-related diseases (travel disease profile) or was performed on an individual basis. In the second part, samples which were examined within a travel disease profile, including E. canis antibodies, were further evaluated for E. canis DNA by a polymerase chain reaction (PCR). Medical histories were obtained of E. canis-positive dogs (animals which were positive in IFAT and/or PCR) and for a part of the negative animals. RESULTS: For 2015, 11.8% of 12220 samples had antibodies for E. canis. Dogs, which were examined with a travel disease profile, displayed antibodies in 5.6% cases of 1172 animals (investigation period February to March 2016). Of the E. canis-positive dogs (n = 67), 91% were positive with only the IFAT, 7.5% with the IFAT and PCR and 1.5% were only positive by PCR. Anamnesis showed that particularly imported dogs without symptoms were controlled for travel diseases during their first year in Germany. CONCLUSION AND CLINICAL RELEVANCE: Although endemic E. canis infections play a minor role in Germany, infections with this pathogen are still of importance in this country, particularly because of the import of dogs. Therefore, a medical history helps in early ehrlichiosis diagnosis and to start an adequate treatment. Pathogen detection in imported E. canis-seropositive dogs, which often did not display clinical symptoms in this study, was frequently negative in blood samples by PCR. The diagnostic method should be chosen depending on the disease phase and the underlying question (symptoms or preventive screening).

Detection of Encephalitozoon cuniculi in the feline cataractous lens.

PURPOSE: Identification of Encephalitozoon cuniculi (E. cuniculi) as a possible causative agent for cataracts and uveitis in cats. METHODS: Within a 12-month study period, cats that were presented with focal anterior cortical or mature cataract and secondary uveitis underwent a complete ophthalmic examination, complete blood count, serum biochemistry, serologic tests for E. cuniculi and tests for feline immunodeficiency virus (FIV), feline infectious peritonitis (FIP), feline leukemia virus (FeLV) and Toxoplasma gondii (T. gondii). PCR for DNA detection of E. cuniculi and T. gondii as well as cytologic examination of aqueous humor after paracentesis and phacoemulsified lens material were also performed. In addition histopathologic examination of the resected anterior lens capsule and attached lens epithelial cells was performed. Serologic testing for antibodies against E. cuniculi was also performed in 100 ophthalmologically healthy cats. RESULTS: Eleven (19 eyes) European shorthair cats with a median age of 3.5 years were included. Nine of 11 cats had bilateral cataracts, with 12/19 eyes having focal anterior cortical cataracts and 7/19 eyes having mature cataracts. In 14/19 eyes anterior uveitis was present. All cats had a positive antibody titer (1:80-1:10,000) for E. cuniculi. Encephalitozoon cuniculi DNA was detected by PCR and sequencing in 18/19 lenses and in 10/19 aqueous samples. Five tentative positive results were detected by cytologic examination. Spores were detected in 15/19 samples of lens material with histopathologic staining. Only 2/100 ophthalmologically healthy cats showed a positive antibody titer for E. cuniculi. CONCLUSION: Encephalitozoon cuniculi is a cause of focal anterior cortical cataract and anterior uveitis in cats.

Diagnostic markers for encephalitozoonosis in pet rabbits.

Encephalitozoonosis is a common disease in pet rabbits, routinely diagnosed in vivo by serological examination or post mortem by histopathology. Recently molecular techniques have become increasingly important as diagnostic tools. The application of different diagnostic markers for in vivo and post mortem determination of E. cuniculi in naturally infected pet rabbits were compared. The examined population was divided into 33 rabbits with symptoms of encephalitozoonosis (group I) and 38 animals without symptoms (group II) which were all tested by serological analysis (Indirect Immunofluorescence Test), histological examination including special spore staining (Ziehl-Neelsen, acid-fast trichrome) and conventional and nested PCR (organs, body fluids). Additionally, in group III lens material (n=10) of animals (n=9) with phacoclastic uveitis were examined by conventional PCR. Infections with E. cuniculi could be determined post mortem in 78.8% of the rabbits of group I and in 57.9% of group II by histological examination combined with spore staining. In group I 69.7% and in group II 50.0% showed seroconversion. Conventional PCR was only sufficiently sensitive in samples of eyes with phacoclastic uveitis (n=10; 100%). Therefore nested PCR was performed in tissue samples, urine and cerebrospinal fluid (CSF) with positive results in 63.6% of group I and 42.1% of group II. Positive results in serology, pathohistology (spore detection and histological changes in the brain and/or kidneys) and nested PCR were obtained in 52.1% of the rabbits (group I and II, n=71), whereas 31.0% showed negative results in all three diagnostic techniques. 5.6% of the rabbits were positive in two methods and 11.3% in one method. In 37 rabbits (group I and II) with positive results in nested PCR, E. cuniculi DNA could be detected more frequently in the brain (91.9%) than in the kidney (54.1%). Furthermore nested PCR of urine revealed positive results in 29.7% of the rabbits (n=37) with seroconversion and/or confirmed E. cuniculi infection by spore detection. All 25 samples of CSF tested negative in nested PCR. Conventional PCR of eyes with phacoclastic uveitis was an excellent diagnostic marker in living rabbits, while nested PCR of urine or CSF was not reliable. Histological examination combined with special staining was the most sensitive method in post mortem diagnostics. Nested PCR appears to be a good post mortem method to investigate organs, especially brains, of chronically infected animals.

Encephalitozoonosis in pet rabbits (Oryctolagus cuniculus): pathohistological findings in animals with latent infection versus clinical manifestation.

Encephalitozoon cuniculi is a common infectious agent of rabbits. The aim of this study was to determine the distribution and extent of histological lesions in the brain and in the kidney of naturally infected pet rabbits with or without clinical encephalitozoonosis. In 71 animals (33 with symptoms) which died or were euthanised, histopathological examination including staining of spores (Ziehl-Neelsen, acid-fast trichrome) was performed and changes were described quantitatively. The cerebrum was the most frequently affected brain region (97.5%), whilst the cerebellum (55%) and the vestibular cores (37.5%) were less commonly concerned. Granulomas were found in 77.5% of animals with encephalitis and in 12.5% of rabbits with interstitial nephritis. Although cerebral granulomas were found irrespective of the grade of histological changes, they were significantly correlated with changes at higher grades. There was no correlation between the severity of encephalitis and neurological symptoms. Since severe lesions were also found in clinically inconspicuous animals, histological findings of inflammatory lesions are not indicative of overt encephalitozoonosis as the causative agent for neurological signs. Other diseases causing neurological symptoms, such as suppurative encephalitis, otitis media as well as malignant lymphoma were also detected in the rabbit population that was examined in the present study.