Ophthalmic Thelazia callipaeda infections: first feline and new canine imported cases in Germany.

The first case of feline ocular Thelazia callipaeda infection and two new canine imported infections in West Germany are here described. The three animals had a history of recent travel to/from other countries. The young adult cat imported from Spain presented an intermittent unilateral ocular discharge. During in-depth ophthalmic examination, a single alive nematode was removed from the conjunctival compartment of the affected eye. Referring to the canine cases, an adult female dog originated from Kenya presented epiphora and mucous whitish-grey discharge of the right eye. During flushing of the nasolacrimal duct two small, thin and long nematodes were removed. Furthermore, a male Borzoi racing dog with regular visit to racing tracks in different countries presented ocular mucous discharge. At ophthalmologic examination, two transparent-whitish vital nematodes were removed. All nematode specimens of the three cases were morphologically identified as adult T. callipaeda parasites. The animals were treated orally with milbemycin oxime (2.0 mg/kg; cat) or milbemycin oxime/praziquantel (0.5 mg/kg and 5.0 mg/kg; dogs) twice with 1-week interval resulting in complete resolution of symptoms. The repeated introduction of patent T. callipaeda-infected animals, especially from southern and eastern endemic countries, will ease the establishment of ophthalmic thelaziosis in Northern Europe. The male fruit fly, Phortica variegata, an intermediate host of T. callipaeda, is endemic within European countries. Considering the clinical and zoonotic relevance of ophthalmic thelaziosis, enhanced disease awareness of European medical and veterinarian doctors and in-depth eye examination for proper detection of T. callipaeda are crucial for appropriate anthelmintic treatments and to limit spreading of the infection.

Occurrence and Molecular Typing of Giardia psittaci in Parakeets in Germany-A Case Study.

A grey-hooded parakeet (Psilopsiagon aymara) and two budgerigars (Melopsittacus undulatus) from different owners presented with decreased activity, vomitus, and diarrhea. A microscopic examination of feces showed trophozoites of the protozoan flagellate Giardia. A commercial immunochromatographic dipstick test for Giardia sp. antigens confirmed the infection. These findings were assured by PCR of the small subunit ribosomal RNA (SSU rRNA) gene and coproantigen ELISA. Sequencing of PCR products of the SSU rRNA (292 bp) and ß-giardin genes (511 bp) identified Giardia psittaci as the species involved. Therefore, our results show that a GSA 65-based coproantigen ELISA, which was established for diagnosis of Giardia duodenalis is applicable for the detection of G. psittaci. A treatment with ronidazole was started. Additionally, fecal examination and dissection of the dead birds revealed coinfection with the fungal pathogen Macrorhabdus ornithogaster. One budgerigar survived and repeatedly tested negative after treatment with ronidazole. The described cases indicate that a single infection with G. psittaci has a good prognosis, whereas the prognosis is poor when coinfections occur, especially with M. ornithogaster.

Reporte de caso- Presentación y tipificación molecular de Giardia psittaci en periquitos en Alemania: Un estudio de caso. Un periquito catita aimará (Psilopsiagon aymara) y dos periquitos australianos (Melopsittacus undulatus) de diferentes propietarios presentaron actividad disminuida, vómito y diarrea. El examen microscópico de las heces mostró trofozoitos del protozoo flagelado Giardia. Una prueba de tira reactiva inmunocromatográfica comercial para antígenos de Giardia sp. confirmó la infección. Estos resultados fueron confirmados por PCR para el gene de ARN de la subunidad pequeña ribosomal (SSU rRNA) y por ELISA de coproantígeno. La secuenciación de los productos de PCR del ARNr de SSU (292 pb) y los genes de ß-giardina (511 pb) identificaron a Giardia psittaci como la especie involucrada. Por lo tanto, estos resultados muestran que el método de ELISA de coproantígeno basado en GSA 65, que se estableció para el diagnóstico de Giardia duodenalis, es aplicable para la detección de G. psittaci. Se inició un tratamiento con ronidazol. Además, el examen fecal y la disección de las aves muertas revelaron coinfección con el patógeno fúngico Macrorhabdus ornithogaster. Un periquito australiano sobrevivió y dio negativo repetidamente después del tratamiento con ronidazol. Los casos descritos indican que la infección única con G. psittaci tiene un buen pronóstico, mientras que el pronóstico es malo cuando ocurren coinfecciones, especialmente con M. ornithogaster. Abbreviations: GSA = Giardia-specific antigen; OD = optical density; rRNA = ribosomal ribonucleic acid; SSU = small subunit.

Central nervous signs, blindness and cerebral vermicosis in free-ranging peregrine falcons (Falco peregrinus) associated with aberrant larval migrations.

Four free-ranging peregrine falcons (Falco peregrinus) were submitted with a history of unilateral or bilateral blindness and central nervous signs to a veterinary clinic in Germany. There were no indications of trauma or ocular disease. Likewise, other differential diagnoses for CNS signs were ruled out within the diagnostic process. The clinical diagnostic panel in live falcons included general examination, radiography, endoscopy, hematology, ophthalmoscopy and parasitological examination of the feces, blood gas analysis and blood chemistry as well as computed tomography, and magnetic resonance imaging (MRI). A complete pathological and histopathological examination was performed post-mortem. The only common finding in all birds was an infection with the nematode parasite Serratospiculum tendo. The parasite was confirmed morphologically and via PCR. In two falcons intracerebral vermicoses was suspected in MRI and confirmed in subsequent histopathology, but molecular biological identification of the parasite species failed from brain tissue. Until today, S. tendo had been reported to affect the respiratory system, the liver and different parts of the gastrointestinal tract and to cause cachexia, inappetence, regurgitation, dyspnea and general signs of illness such as lethargy, poor plumage, and reduced reproduction. Our findings indicate that aberrant migration should be considered as cause for CNS signs in falcons. As S. tendo might be a possible cause for this, CNS signs might be included in the list of clinical signs of serratospiculiasis in falcons.

Survey of Baylisascaris spp. in captive striped skunks (Mephitis mephitis) in some European areas.

Skunks are popular carnivore species kept both in zoological institutions and in households where they are hand raised as exotic pets. These small carnivores are considered the main definitive hosts of the roundworm Baylisascaris columnaris. The purpose of this survey was to investigate the occurrence of Baylisascaris spp. in striped skunks kept as pets or in private zoo collections in some European areas. Copromicroscopic data from two laboratories, one in Italy and one in Germany, were used. A total of 60 animals were selected. Samples came from Germany (n = 30), Italy (n = 23), United Kingdom (n = 5), Austria (n = 1), and the Netherlands (n = 1). Twenty-eight animals were certainly kept as pets in private households in Italy and the UK. Fifteen out of 60 animals (25%) were positive for Baylisascaris spp. Molecular identification of adult parasites was performed in ten of those animals, revealing B. columnaris in all cases. To the authors' knowledge, this is the first survey of Baylisascaris spp. in captive skunks in Europe.

Seasonality in the proportions of domestic cats shedding Toxoplasma gondii or Hammondia hammondi oocysts is associated with climatic factors.

A previous study on domestic cats in Germany and neighbouring countries suggested seasonality in shedding Toxoplasma gondii oocysts. The aim of the present study was to elucidate whether this seasonality in shedding could be explained by climatic effects and whether differences between years in the proportions of cats shedding oocysts could also be explained by climatic factors. To this end, a long-term study over a period of 55 months on domestic cats for T. gondii and Hammondia hammondi oocysts was performed and the results compared with climatic data. Using species-specific PCR, T. gondii oocysts were identified in 0.14% (84/61,224) and H. hammondi in 0.10% (61/61,224) of the samples. Toxoplasma gondii oocysts were predominantly observed from summer to autumn, while H. hammondi oocysts were mainly found during autumn and winter. In statistical analyses using climatic data, even differences in parasitological findings between years could be partially modelled using monthly temperature, North Atlantic Oscillation indices and precipitation. Of the three climatic variables analysed, precipitation as an explanatory variable had the lowest impact in the statistical models while those taking only temperature and North Atlantic Oscillation indices into account were sufficiently predictive. Interestingly, time lags between the climatic event and the parasitological findings had to be implemented in all models. For T. gondii, North Atlantic Oscillation indices with a time lag of 7 months and temperature with a time lag of 2 months had the best predictive value. In contrast, temperature (with a time lag of 6 months) and the interaction of precipitation (with a time lag of 5 months) and North Atlantic Oscillation indices (with a time lag of 11 months) were optimal for predicting the seasonality of H. hammondi. These results suggest prominent differences in the life cycles of the two closely related parasites. Previous findings showed that H. hammondi lack avian hosts, in contrast to T. gondii, and the coincidence in the periods of high abundance of birds and high proportions of cats shedding T. gondii suggest that birds may play an important role in the epidemiology of this infection. The result that North Atlantic Oscillation index is an important variable in modelling variations in the proportion of cats shedding T. gondii and H. hammondi over the year is an indication that global warming may also influence the infection risk of animals and humans with T. gondii and H. hammondi. The findings have important implications for planning epidemiological studies and for estimating the risk of human infection.

In vitro cultivation of Hammondia heydorni: Generation of tachyzoites, stage conversion into bradyzoites, and evaluation of serologic cross-reaction with Neospora caninum.

Hammondia heydorni was in vitro isolated from oocysts shed by three dogs using a finite cell line from embryonal bovine heart (KH-R). The oocysts were purified and suspended in 2% potassium dichromate or 2% sulphuric acid for sporulation for 2-5 days at room temperature. The parasites were confirmed as H. heydorni by PCR using specific primers (JS4/JS5) and by negative reaction for Neospora caninum employing the primers Np6+/Np21+. H. heydorni sporulated oocysts (1 × 10(6)) from each dog were initially treated with sodium hypochlorite. For excystation of sporozoites, oocysts from one dog were lysed by ultrasound followed by incubation with 0.75% taurocholate. Excystation of sporozoites from the other two dogs was achieved by oocyst fragmentation with glass beads with no further chemical treatment. Tachyzoites were clearly seen in the cultures at three days post inoculation (dpi). Bradyzoite conversion and cyst formation were evaluated at different time points by using a polyclonal rabbit serum against a bradyzoite-specific antigen (anti-BAG1), and a rat monoclonal antibody (mAbCC2) against a cyst wall protein. Bradyzoites were firstly detected at 7 dpi. Between 18 and 21 dpi most of cultured parasites consisted of encysted bradyzoites. The H. heydorni cysts increased in size during cultivation and reached a length of up to 135 µm. The parasite was maintained in the bovine heart cells up to 4.5months. Sera from mice and sheep experimentally infected with H. heydorni oocysts reacted with H. heydorni by IFAT, but did not cross-react with N. caninum antigens using IFAT or immunoblot. These findings suggest that serological cross-reactivity between H. heydorni and N. caninum seems to be of minor importance.

Pyogranulomatous panniculitis in ferrets (Mustela putorius furo) with intralesional demonstration of Pseudomonas luteola.

One ferret (Mustela putorius furo) from Finland and two ferrets from Austria, aged 1-4.5 years and of both genders, were presented with pyogranulomatous subcutaneous inflammation affecting the inguinal, preputial and femoral regions, respectively. Histologically, microorganisms were detected within the lesions. The organisms had a capsule that stained positively by the periodic acid-Schiff reaction. Pseudomonas spp. were cultured from the lesions in two cases. In the third case, electron microscopy revealed a prokaryotic organism surrounded by an electron lucent matrix. 16S rRNA gene sequencing showed highest sequence homology to Pseudomonas luteola in all three cases. In combination with recent reports of pleuropneumonia in ferrets due to P. luteola infection, these cases might indicate a predisposition of ferrets for infection by these bacteria.

Occurrence and molecular typing of Giardia isolates in pet rabbits, chinchillas, guinea pigs and ferrets collected in Europe during 2006-2012.

A total of 1180 faecal samples (528 from rabbits, 531 from chinchillas and 121 from guinea pigs) collected during 2006-2012 by veterinarians in Germany and in other European countries were submitted to a diagnostic laboratory for Giardia testing by means of coproantigen ELISA. Of these samples, 40 rabbits (7.6 per cent), 326 chinchillas (61.4 per cent) and five guinea pigs (4.1 per cent ) were found to be positive. To gain insights into the genetic identity of Giardia in small mammals, ELISA-positive samples from 23 chinchillas, five ferrets, a rabbit, and a Desmarest's hutia were investigated by PCR and sequencing of fragments of the small subunit ribosomal DNA (ssu), the triose phosphate isomerase (tpi) and the ß-giardin (bg) genes. At the ssu locus, assemblage B was identified in 28 of 30 isolates, whereas assemblage A and D were each detected in one sample. The majority of isolates from chinchillas and those from ferrets had Giardia duodenalis sequences identical to sub-assemblages AI or BIV, based on either a single locus (tpi or bg) or multiple loci (tpi and bg). As sub-assemblages AI or BIV are associated with human infection, these results indicate that small mammals can act as reservoirs of cysts potentially infectious to humans.

Encephalitozoonosis in two inland bearded dragons (Pogona vitticeps).

Microsporidiosis is reported rarely in reptiles. Sporadic multisystemic granulomatous disease of captive bearded dragons (Pogona vitticeps) has been associated with microsporidia showing Encephalitozoon-like morphology. Two such cases are described herein. Both animals displayed clinical signs suggestive of renal failure. Necropsy examination revealed granulomatous lesions in the liver and adrenal area in both animals, and in several other organs in one animal. The lesions were associated with intracellular protozoa consistent with microsporidia. Ultrastructural examination of the organisms revealed morphology similar to Encephalitozoon spp. Immunohistochemistry and chromogenic in-situ hybridization for Encephalitozoon cuniculi were positive in both animals. Nucleotide sequencing of the partial small subunit ribosomal RNA gene and the complete internal transcribed spacer (ITS) region revealed high similarity with published E. cuniculi sequences in both animals. However, the ITS region showed a GTTT-repeat pattern distinct from mammalian E. cuniculi strains. This may be a novel E. cuniculi strain associated with multisystemic granulomatous disease in bearded dragons.

Feasibility and safety of endoscopic ultrasound-guided fine needle aspiration of the pancreas in dogs.

BACKGROUND: Endoscopic ultrasound (EUS)-guided fine needle aspiration (EUS-FNA) has proven a useful and safe diagnostic tool for assessing pancreatic disease in human medicine. No information about pancreatic EUS-FNA is available in dogs. OBJECTIVES: To assess the feasibility and safety of pancreatic EUS-FNA in healthy dogs. ANIMALS: Thirteen beagles with a median body weight of 13.4 kg. METHODS: Experimental study. An ultrasound endoscope (insertion tube outer diameter 11.8 mm) was used, and FNA was carried out with 19 G needles. The optimal puncture site was chosen with the aid of Doppler imaging. Complete clinicopathologic assessments including pain scoring and pancreas-specific lipase measurements were obtained before EUS as well as on day 1 and day 2 after EUS-FNA. RESULTS: The pancreatic body was identified in all dogs, the left lobe was clearly identified in 9/13 and appeared indistinctly marginated in 4/13 dogs, and the distal third of the right lobe could not be identified in 7/13 dogs. EUS-FNA was carried out in 12/13 dogs. Cellularity of smears was adequate for evaluation in 8/12 cases, in which samples were obtained transgastrically (n = 4) or transduodenally (n = 4). All dogs recovered uneventfully and no clinical and laboratory abnormalities occurred during the 48 hour monitoring period after the procedure. CONCLUSION AND CLINICAL IMPORTANCE: Although the healthy canine pancreas is difficult to visualize in its entirety with EUS, pancreatic EUS-FNA with a 19 G needle is feasible in medium-sized dogs and can be considered a safe procedure. Its diagnostic usefulness should be evaluated in dogs with pancreatic disease.

Evaluation of fecal elastase and serum cholecystokinin in dogs with a false positive fecal elastase test.

BACKGROUND: An assay for the measurement of pancreatic elastase in dog feces has been introduced. HYPOTHESIS/OBJECTIVES: The goal of this study was to evaluate the rate of false-positive fecal-elastase test results in dogs with suspected exocrine pancreatic insufficiency (EPI) and to assess serum cholecystokinin (CCK) concentrations in dogs with a false positive fecal elastase test result. ANIMALS: Twenty-six fecal and serum samples from dogs suspected of EPI, for which samples had been submitted to a commercial laboratory (Vet Med Labor) for analysis. METHODS: Prospective study. Serum trypsin-like immunoreactivity (TLI) was measured in 26 dogs with a decreased fecal elastase concentration of <10 microg/g feces. Serum CCK concentrations were measured in 21 of these dogs. RESULTS: Of 26 dogs with a decreased fecal elastase concentration, 6 (23%) had serum TLI concentrations within or above the reference range. Serum CCK concentrations were significantly higher in dogs with a true positive fecal elastase test result (median: 1.1 pmol/L; range: 0.1-3.3 pmol/L) than in those with a false positive fecal elastase test result (median: 0.1 pmol/L; range: 0.1-0.9 pmol/L; P value = .0163). CONCLUSIONS AND CLINICAL IMPORTANCE: The rate of false positive fecal elastase test results was high in this group of dogs, suggesting that diagnosis of EPI must be confirmed by other means. The decreased CCK concentration in dogs with a false positive fecal elastase test result could suggest that false positive results are because of decreased stimulation of exocrine pancreatic function caused by other conditions.

Occurrence and molecular characterization of Cryptosporidium spp. genotypes in European hedgehogs (Erinaceus europaeus L.) in Germany.

Juvenile hedgehogs having insufficient body weight are often brought for overwintering to hedgehog rehabilitation centres. Faecal samples of juvenile hedgehogs and overwintering hedgehogs (n=188) collected prior to releasing them back into the wilderness were examined for the presence of Cryptosporidium coproantigen and oocysts. Altogether 56 (29.8%) submitted samples were positive for coproantigen. Forty-five (39.5%, n=114) of the positive samples originated from newly rescued hedgehogs, while 11 (14.8%, n=74) positive samples were from animals that spent several months at the station. Fifteen samples subjected to PCR-RFLP analysis on the partial 18S rRNA locus suggested the presence of C. parvum. Multilocus sequence typing on partial 60 kDa glycoprotein gene, 18S rRNA, actin gene, 70 kDa heat shock protein gene sequences revealed 3 different subtype families: IIa, IIc and a new, proposed as VIIa subtype family. Cryptosporidium sp. genotype belonging to VIIa subtype family is closely related to C. parvum but is genetically distinct being probably a hedgehog-specific Cryptosporidium sp. genotype with unknown zoonotical potential. Hedgehogs excreting Cryptosporidium oocysts represent a potential source for human infections, but also an anthroponotic nature of the IIc subtype family should be reviewed.

Atypical Toxoplasma gondii genotypes identified in oocysts shed by cats in Germany.

A total of 18,259 feline faecal samples from cats in Germany were collected and analysed for the presence of Toxoplasma gondii oocysts between June 2007 and December 2008. The proportion of T. gondii-positive samples collected between January and June was significantly lower than between July and December. The age of cats shedding T. gondii oocysts was not significantly different from the age of negative control cats. Forty-six T. gondii-positive samples were genetically characterised using nine PCR-restriction fragment length polymorphism (RFLP) markers which included newSAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1 and Apico. In addition, 22 isolates that had already been partially characterised in a previous study were further typed using PCR-RFLP markers c22-8, c29-2, L358, PK1 and Apico. Genotyping of the 68 isolates revealed that the majority of T. gondii isolates (n=54) had Type II patterns at all loci but displayed a Type I pattern at the Apico locus. Three isolates displayed Type II patterns at all loci, including the Apico locus. In addition, we detected one isolate with clonal Type III patterns at all loci and three isolates with atypical and mixed genotypes. Seven isolates could not be fully genotyped. One of those isolates displayed alleles of both Types I and II at the Apico locus. To our knowledge this is the first description of the presence of T. gondii genotypes different from the clonal Types I, II and III in the faeces of naturally infected cats.

First isolation of Neospora caninum from the faeces of a dog from Portugal.

We report the in vitro isolation of Neospora caninum from the faeces of a naturally infected 8-year-old male stray boxer from Portugal. Vero cell cultures were infected using parasite stages obtained after oral inoculation of gamma-interferon knockout mice with 10(2) sporulated oocysts. The isolate was identified by microscopical examination, as well as histological, immunological and molecular methods including a DNA-microsatellite-based typing technique, and was subsequently named NC-P1. The DNA-microsatellite pattern observed in the NC-P1 isolate was not previously reported for any N. caninum isolate. To our knowledge, this is the first isolation of N. caninum from the faeces of a naturally infected dog from Portugal.

Molecular comparison of Neospora caninum oocyst isolates from naturally infected dogs with cell culture-derived tachyzoites of the same isolates using nested polymerase chain reaction to amplify microsatellite markers.

Neospora caninum infection is an important cause of bovine abortion. The infection can be transmitted transplacentally or by ingestion of oocysts shed by definitive hosts. There are few reports of dogs naturally shedding N. caninum oocysts and only some oocyst isolates were transferred into cell culture. The aim of the present study was to analyse N. caninum oocysts from the faeces of naturally infected dogs using a microsatellite-based typing technique and to compare them with cell culture-derived tachyzoites of the same isolates. To this end, N. caninum oocysts from six naturally infected dogs were inoculated into gamma-interferon knockout mice. After these mice had developed disease, tissue samples or peritoneal washings from necropsied mice were transferred into cell culture. Nested-PCR techniques were developed for the sensitive and specific amplification of N. caninum microsatellite-containing regions (MS1B, MS2, MS3, MS4, MS5 and MS10). DNA was extracted from oocysts and cell culture tachyzoites of each isolate, followed by amplification and sequence analysis of microsatellite-containing regions. Each parasite isolate examined yielded a unique microsatellite genotype, while no differences were revealed when data for N. caninum oocysts were compared with cultured tachyzoites of the same isolate. Our technique may allow the typing of clinical samples and different strains of N. caninum at the molecular level. This method may prove useful for the identification of infection sources in molecular epidemiological studies.

Characterization of a repetitive DNA fragment in Hammondia hammondi and its utility for the specific differentiation of H. hammondi from Toxoplasma gondii by PCR.

Hammondia hammondi and Toxoplasma gondii are closely related protozoan parasites. Both species use felids as definitive hosts and a broad spectrum of warm-blooded animals as intermediate hosts. Morphologically and serologically, the two parasites are difficult to differentiate. While T. gondii is an important pathogen of humans and a broad range of other vertebrates, disease has not yet been associated with H. hammondi infection. The aim of the present study was to identify and characterize a repetitive DNA fragment in H. hammondi and to evaluate its suitability for diagnostic purposes. With two primers considered to be specific for a 529 bp repetitive DNA fragment in T. gondii, weak products were amplified by polymerase chain reaction (PCR) from genomic DNA from H. hammondi oocysts. These amplicons (of approximately 150, 300 and 450 bp) were sequenced. The 292 bp consensus sequence of these three fragments revealed 84% identity with parts of the 529-bp repeat in T. gondii. Based on this sequence, a pair of primers was selected which amplified products of 98 and 630 bp from genomic DNA from H. hammondi oocysts but not from DNA from T. gondii. The 630-bp product was purified and cloned into a plasmid vector and the consensus sequence determined from seven randomly selected clones; comparison of this sequence with those available in current databases for T. gondii revealed an 84.0-88.1% identity over a length of 529 bp. The sequence data obtained was used for the development of a sensitive PCR which is entirely specific for H. hammondi and incorporates an internal control. The sequence data for the repetitive DNA element of H. hammondi provides a foundation for the design of primers specific to T. gondii, and the future optimisation of conventional and real-time PCR assays for the specific diagnosis of toxoplasmosis in definitive and intermediate hosts.

Cost-effective method of DNA extraction from taeniid eggs.

A new cost-effective method using silicon dioxide- and guanidine isothiocyanate-containing buffers, after previous alkaline lysis, was established for the DNA extraction from taeniid eggs isolated from canine faeces. The purified DNA can be used to amplify the species-specific 12S mitochondrial DNA of Echinococcus multilocularis in direct and nested polymerase chain reaction in order to differentiate between E. multilocularis and Taenia spp.

Occurrence of Toxoplasma gondii and Hammondia hammondi oocysts in the faeces of cats from Germany and other European countries.

Faecal samples of 24,106 cats from Germany and other European countries were examined microscopically in a veterinary laboratory in Germany between October 2004 and November 2006 to estimate the prevalence of animals shedding Toxoplasma gondii or Hammondia hammondi oocysts. Oocysts of 9-15 microm size with a morphology similar to that of H. hammondi and T. gondii were found in 74 samples (0.31%). A total of 54 samples were further characterised to achieve a species diagnosis and to determine the genotype of T. gondii isolates by PCR and PCR-RFLP. From these samples, 48 isolates were obtained: 26 (0.11%) were finally identified as T. gondii and 22 (0.09%) as H. hammondi. T. gondii-positive samples came from Germany, Austria, France and Switzerland while H. hammondi was detected in samples from Germany, Austria and Italy. In two samples (one T. gondii and one H. hammondi), PCR indicated the presence of Hammondia heydorni DNA. No Neospora caninum DNA was detected in any of the feline faecal samples. Twenty-two of the 26 T. gondii isolates could be genotyped. A PCR-RFLP analysis for the SAG2, SAG3, GRA6 and BTUB genes revealed T. gondii genotype II in all cases. Morphologically, H. hammondi oocysts exhibited a statistically significantly smaller Length-Width-Ratio than T. gondii oocysts.