Is there any change in the prevalence of intestinal or cardiopulmonary parasite infections in companion animals (dogs and cats) in Germany between 2004-2006 and 2015-2017? An assessment of the impact of the first ESCCAP guidelines.

Main objective of the present nationwide study was to assess the impact of the ESCCAP guideline for the control of worm infections in dogs and cats 8-10 years after its first publication in Germany. A secondary aim was to determine the prevalence of canine and feline cardiopulmonary nematodes and intestinal protozoa. Faecal samples of 53,693 dogs and 26,491 cats in 2004-2006 as well as of 129,578 dogs and 45,709 cats in 2015-2017 routinely submitted by veterinarians to a private veterinary laboratory were examined using appropriate parasitological methods. In dogs, the prevalence of Toxocara and taeniid egg shedding was significantly lower in 2015-2017 (3.8 % and 0.16 %, respectively) than in 2004-2006 (4.6 % and 0.27 %, respectively). The prevalence of hookworm and Capillaria eggs was higher in the second study period (2.3 % and 0.77 %, respectively) than in the first (1.3 % and 0.6 %, respectively). For Toxascaris leonina (0.55-0.6 %) and Trichuris (0.8-0.9 %), the difference was not significant between the study periods. Dogs shed more often Angiostrongylus vasorum larvae in the second study (3.1 %) than in the first (1.0 %), whereas the prevalence of Crenosoma vulpis did not change significantly (2.2-2.6 %). Cystoisospora canis and C. ohioensis-like infections were less detected in the second study period (1.0 % and 2.1 %, respectively) than in the first (1.8 % and 2.7 %, respectively). Neospora-like oocysts and Sarcocystis sporocysts were more prevalent in the second study period (0.19 % and 0.13 %, respectively) than in the first (0.13 % and 0.06 %, respectively). The percentage of Giardia or Cryptosporidium coproantigen-positive samples was lower in the second study period (18.9 % and 6.7 %, respectively) than in the first (22.8 % and 10.0 %, respectively). In cats, the prevalence of egg shedding of T. cati, Capillaria and taeniids was significantly lower in 2015-2017 (3.5 %, 0.25 % and 0.1 %, respectively) than in 2004-2006 (4.8 %, 0.54 % and 0.22 %, respectively). No difference was recorded for hookworms (0.12-0.13 %) and Ts. leonina (0.04-0.05 %). Aelurostrongylus-like larvae were detected more often in the second study period (6.5 %) than in the first (2.6 %). Infections with Cystoisospora felis, C. rivolta, Toxoplasma-like coccids and Sarcocystis were less prevalent in the second study period (1.9 %, 0.7 %, 0.24 % and 0.02 %, respectively) than in the first (2.7 %, 1.1 %, 0.36 % and 0.1 %, respectively). The percentage of Giardia or Cryptosporidium coproantigen-positive samples was significantly lower in the second study period (10.6 % and 4.8 %, respectively) than in the first (15.4 % and 8.3 %, respectively). Although these results indicate a decline of the occurrence of most canine and feline intestinal parasites in Germany over the years, a transmission risk of zoonotic parasites remains. Therefore, the control of helminth infections in domestic dogs and cats continues to be a challenge for veterinarians and pet owners.

GIS-supported epidemiological analysis on canine Angiostrongylus vasorum and Crenosoma vulpis infections in Germany.

BACKGROUND: Angiostrongylus vasorum infections are the cause of severe cardiopulmonary diseases in dogs. In the past, canine angiostrongylosis has largely been neglected in Europe, although some recent studies indicated an expansion of historically known endemic areas, a phenomenon that might also apply to Crenosoma vulpis. The aim of the present study was to analyse temporal and spatial trends of canine A. vasorum and C. vulpis infections and to perform GIS-supported risk factor analysis to evaluate the role of landscape, age and seasonality in the life-cycle of these nematodes. METHODS: A total of 12,682 faecal samples from German dogs (collected in 2003-2015) with clinical suspicion for lungworm infection were examined for the presence of A. vasorum and C. vulpis larvae by the Baermann funnel technique and respective epidemiological data (location and age of the sampled dogs, date of sampling) were subjected to GIS-supported risk factor analysis. RESULTS: Overall, A. vasorum and C. vulpis larvae were detected in 288 (2.3%) and 285 (2.2%) faecal samples, respectively. In general, both lungworm infections were found to be widely spread in Germany. GIS-supported analyses demonstrate spatial differences in the occurrence of canine A. vasorum and C. vulpis infections in Germany. also, risk factor analyses revealed an overlap but also diverging risk and protective factors for A. vasorum and C. vulpis infections. The current data also indicate a significant increase of A. vasorum and C. vulpis prevalences from 2003 to 2015 and from 2008 until 2015, respectively, and a potential spread of A. vasorum endemic areas to the northeastern part of Germany. CONCLUSIONS: The results of the present study show an insight into the epidemiological situation of lungworm infections (A. vasorum and C. vulpis) of the past 13 years in Germany. The data clearly demonstrate an increase of diagnosed A. vasorum prevalence in the tested dog population between 2003 and 2015 as well as spatial differences in the occurrence of diagnosed A. vasorum and C. vulpis infections of dogs in Germany. Risk factor analyses suggest possible differences in the biology of these parasites, presumably at the intermediate host level.

Seropositivity of Borrelia burgdorferi in a cohort of symptomatic cats from Europe based on a C6-peptide assay with discussion of implications in disease aetiology.

There are only few reports on Lyme borreliosis (LB) in cats. The reasons might be a different tick infestation in cats compared to dogs, a low susceptibility for tick-borne infections or a low awareness of veterinarians for tick-borne diseases in feline patients. The aim of this study was to determine the proportion of antibodies against Borrelia burgdorferi sensu lato (Bbsl) in feline sera, to compare the significance of feline versus canine LB, as well as to evaluate possible implications on disease occurrence. Specific antibodies against the C6-peptide of Bbsl in cats were detected by a rapid test based on enzyme immunoassay technique. The serum samples were sent to a diagnostic laboratory by veterinarians from Germany and other European countries with request for Borrelia serology in the years 2009-2011. Veterinarians were asked for information regarding the cats' location, age, gender, clinical signs, treatment and follow-up. In six of 271 (2.2%; 95% CI: 0.8-4.8%) cat sera, antibodies against the C6-peptide of Bbsl were detected. Proportion of Borrelia antibody-positive cat sera was significantly lower than the one determined for dogs during the same time period. All positive cats lived in countries endemic for LB (Germany, Sweden and Belgium), and all C6-antibody positive cats with the exception of one cat showed clinical signs. Possible implications on disease occurrence are discussed. Data presented here demonstrate a lower prevalence of Borrelia specific C6-antibodies in European cats when compared to dogs residing in the same regions. The absence of antibodies against Bbsl in 97.8% (95% CI: 95.2-99.2%) of the submitted samples indicate that diagnosis "feline LB"is rare in cats. Nevertheless, LB should be considered in cats with compatible clinical signs (e.g. shifting leg lameness, to less extent neurological signs) when other differential diagnoses are ruled out.

Use of Ronidazole and Limited Culling To Eliminate Tritrichomonas muris from Laboratory Mice.

Tritrichomonas muris is occasionally identified during routine fecal screening of laboratory mice. Frequently, entire racks are affected, and because no effective treatment is available, culling of affected mice and rederivation by embryo transfer have been suggested. The current study evaluated whether treatment with ronidazole, a nitroimidazole efficacious against T. fetus infections in cats, combined with limited culling was effective against T. muris in laboratory mice (Mus musculus). A subset (n = 39) of mice were treated with ronidazole (400 mg/L in drinking water) for 15 d, after which 6 of the mice still shed T. muris. Consequently all mice in the affected rack received ronidazole (500 mg /L in drinking water) for 25 d. All mice were retested by using pooled samples, and those positive for T. muris (except for a valuable breeding pair) were culled. The remaining mice continued to receive ronidazole for another 17 d. At the end of the treatment period, all mice were tested (days 60 and 81) and were shown to be negative for T. muris. Over the following year, sentinel mice from the rack were tested every 3 mo and remained negative for tritrichomonads by fecal smear. Thus, a combination of limited culling and treatment with ronidazole in the drinking water successfully cleared research mice of infection with T. muris.

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 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.

Diagnosis of gastrointestinal parasites in reptiles: comparison of two coprological methods.

BACKGROUND: Exotic reptiles have become increasingly common domestic pets worldwide and are well known to be carriers of different parasites including some with zoonotic potential. The need of accurate diagnosis of gastrointestinal endoparasite infections in domestic reptiles is therefore essential, not only for the well-being of captive reptiles but also for the owners. Here, two different approaches for the detection of parasite stages in reptile faeces were compared: a combination of native and iodine stained direct smears together with a flotation technique (CNF) versus the standard SAF-method. RESULTS: A total of 59 different reptile faeces (20 lizards, 22 snakes, 17 tortoises) were coprologically analyzed by the two methods for the presence of endoparasites. Analyzed reptile faecal samples contained a broad spectrum of parasites (total occurence 93.2%, n = 55) including different species of nematodes (55.9%, n = 33), trematodes (15.3%, n = 9), pentastomids (3.4%, n = 2) and protozoans (47.5%, n = 28). Associations between the performances of both methods to detect selected single parasite stages or groups of such were evaluated by Fisher's exact test and marginal homogeneity was tested by the McNemar test. In 88.1% of all examined samples (n = 52, 95% confidence interval [CI] = 77.1 - 95.1%) the two diagnostic methods rendered differing results, and the McNemar test for paired observations showed highly significant differences of the detection frequency (P < 0.0001). CONCLUSION: The combination of direct smears/flotation proved superior in the detection of flagellates trophozoites, coccidian oocysts and nematode eggs, especially those of oxyurids. SAF-technique was superior in detecting larval stages and trematode eggs, but this advantage failed to be statistically significant (P = 0.13). Therefore, CNF is the recommended method for routine faecal examination of captive reptiles while the SAF-technique is advisable as additional measure particularly for wild caught animals and individuals which are to be introduced into captive collections.

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.

Cryptosporidiosis outbreak in captive chelonians (Testudo hermanni) with identification of two Cryptosporidium genotypes.

An outbreak of diarrhea in an outdoor group of captive Hermann's tortoises (Testudo hermanni) was associated with fecal shedding of cryptosporidial oocysts, as determined by coproscopic and immunoassay examinations. With partial sequencing of the 18S ribosomal RNA gene, 2 different Cryptosporidium genotypes could be identified in the fecal samples. Cryptosporidium tortoise genotype has previously been found in tortoise and ophidian species, and Cryptosporidium ducismarci has been reported from a snake and a chameleon, and it has been linked to intestinal disease in tortoises. The Hermann's tortoises described were also infected with oxyurid nematodes. Treatment specific for reptilian cryptosporidiosis was administered. The clinical signs and fecal shedding ceased, but 9 months later, diarrhea and fecal shedding were seen in 3 animals again. Either the oocyst shedding was temporarily suppressed below detection limits, or the animals were reinfected by oocysts still present in the environment. At least 1 of the detected Cryptosporidium genotypes was presumed to contribute to the clinical symptoms.

Comparison of different commercial DNA extraction kits to detect Toxoplasma gondii oocysts in cat faeces.

The cat is the definitive host of Toxoplasma gondii and plays an important role in the transmission of this and other coccidian parasites, e.g. Hammondia hammondi, a protozoon closely related and morphologically similar to T. gondii. A number of techniques to detect T. gondii nucleic acids in feline faeces are described and several extraction kits for isolating pathogen DNA from faeces or soil are commercially available. To compare the performance of such kits with regard to isolating oocyst DNA, a feline sample that had tested negative for coccidian parasites including T. gondii and H. hammondi was spiked with 10(4), 10(3), 10(2), 50 and 10 H. hammondi oocysts. Several ready-to-use stool or soil kits and an in-house method were then used to extract parasite DNA from these spiked faecal samples. Of six kits tested, two were found suitable for the detection of H. hammondi oocysts DNA by the polymerase chain reaction (PCR) in faecal samples with a detection limit of 250 oocysts per 1 g of faecal sample. These two kits revealed a similar, even slightly lower detection limit (50 oocysts per 1 g of sample) when tested with faecal samples spiked with T gondii oocysts.

Aelurostrongylus abstrusus and Troglostrongylus sp. (Nematoda: Metastrongyloidea) infections in cats inhabiting Ibiza, Spain.

Multiple species of metastrongylid lungworm (Nematoda: Metastrongyloidea) have been reported to infect members of the Felidae. This study describes two metastrongylid species infecting cats in Ibiza, Spain, including clinical features of infection and diagnosis via morphological and molecular characterisation of larval stages. Cats (n=7) presented with suspect lungworm infection, exhibiting coughing and other respiratory signs of infection. Faecal samples were collected from each cat and were subjected to the Baermann method for the detection of first stage larvae. In four cats, two different species of larvae were observed on the basis of morphology and were further molecularly characterised by PCR and sequencing of the 18S rRNA gene. Sequence data confirmed the presence of Aelurostrongylus abstrusus and an unknown species of Troglostrongylus. Molecular characterisation of Oslerus rostratus is also reported for the first time. Given the diversity of metastrongylid species capable of infecting cats, and morphological similarity of larval stages, an emphasis should be placed on the use of molecular characterisation for accurate diagnosis of infection.

Lungworm infections (Angiostrongylus vasorum, Crenosoma vulpis, Aelurostrongylus abstrusus) in dogs and cats in Germany and Denmark in 2003-2007.

Faecal samples of 4151 dogs from Denmark, 958 dogs from Germany and 231 cats from Germany with clinical signs were examined for lungworm larvae using the Baermann funnel technique between 2003 and 2007. In total, 3.6% of Danish and German dogs shed lungworm larvae. In Denmark, patent infections of dogs with Angiostrongylus vasorum were more prevalent (2.2%) than those with Crenosoma vulpis (1.4%). In Denmark, the majority of A. vasorum- (98%) and C. vulpis-infected (80%) dogs originated from Northern Zealand. The frequency of A. vasorum and C. vulpis infections in Danish dogs obviously decreased from 2003 to 2006. In Germany, canine faecal samples were found more frequently positive for C. vulpis than for A. vasorum larvae (2.4% and 1.2%, respectively). Lungworm-infected dogs originated mainly from southern and western Germany. Larvae of Aelurostrongylus abstrusus were detected in 5.6% of cats from Germany. Overall, a distinct seasonal pattern in the detection of infected dogs was apparent for A. vasorum in Denmark and C. vulpis in Germany. The relatively high number of lungworm-infected dogs and cats indicate that these parasitic diseases should be considered in differential diagnosis of cases of treatment-resistant respiratory/cardiopulmonary distress.

Echinococcus multilocularis infections in domestic dogs and cats from Germany and other European countries.

A cross-sectional survey was conducted to estimate the prevalence of Echinococcus multilocularis and E. granulosus infections in domestic dogs and cats from Germany and other European countries. Faecal samples of 21,588 dogs and 10,650 cats routinely submitted to a private veterinary laboratory between June 2004 and June 2005 were examined using the ZnSO(4)-NaCl flotation method. Taeniid eggs were detected in 54 (0.25%) and 37 (0.34%) of the canine and feline faecal samples, respectively. Taeniid eggs were separated and subjected to a DNA preparation and a modified two-step PCR for the detection of Echinococcus spp. based on mitochondrial 12S rRNA genes. PCR products from Echinococcus-negative but cestode-positive reactions were cloned and sequenced to determine the Taenia species. E. multilocularis DNA was specifically amplified in 43 (0.24%) and 25 (0.23%) of the samples from dogs and cats, respectively. E. granulosus DNA was not detected in any sample, while, E. multilocularis-positive samples were detected in dogs from Germany only, those of cats originated from Germany, Denmark and The Netherlands. The prevalence of E. multilocularis egg-positive canine samples was significantly higher in southern (0.35%) than in northern Germany (0.13%). In contrast, no significant regional difference was observed in cats from Germany. Taeniid eggs from Echinococcus-negative samples and from a few samples with macroscopically detected Taenia sp. proglottids were identified as eggs of T. crassiceps (n=8), T. martis, T. serialis, T. polyacantha, T. taeniaeformis and T. pisiformis in dogs (n=1 of each) and T. taeniaeformis (n=11) in cats. The spectrum of cestodes detected in domestic dogs and cats indicate the consumption of small rodents as infection source. The high proportion of E. multilocularis-positive samples, suggest domestic dogs and cats as a possible source of E. multilocularis infection for humans.