Parasitological and molecular investigation of consequences of raw meat feeding (BARF) in dogs and cats: implications for other pets living nearby.

Considering the original nature of carnivorism among dogs and cats, nowadays these pets are increasingly fed BARF (Biologically Appropriate Raw Food/Bone And Raw Food) diet to improve their health conditions. However, this diet may also carry health risks, such as vitamin or mineral deficiencies, and infection with pathogens including parasites. In our study, fecal samples of 89 pet animals kept on BARF diet were subjected to coprological examination followed by molecular analyses. Six of them shed eggs of Dicrocoelium dendriticum. This result was confirmed with PCR and sequencing, and in one case, the DNA of Fasciola hepatica was also demonstrated. In addition, oocysts of Cystoisospora canis, a Cystoisospora ohioensis-like sp. and Eimeria stiedai, as well as sporocysts of a Sarcocystis sp. were also detected. All samples were negative for Neospora caninum and Toxoplasma gondii. In conclusion, no evidence was found for the infection of BARF-fed dogs and cats with parasites that are usually associated with this diet and considered as clinico-pathological risk factors for these pets themselves (e.g., N. caninum, T. gondii). However, fluke eggs (probably originating from ruminants) and oocysts of E. stiedai (from rabbit liver in the food) were demonstrated as pseudoparasites. These species are usually not considered among parasite-associated risks of BARF-feeding, implying that other animals living near BARF-fed pets are neglected in this context. However, where intermediate hosts of D. dendriticum occur in urban areas, BARF-feeding may indirectly affect later other dogs and cats. It was also shown here that BARF-feeding may contribute to the contamination of the environment with E. stiedai oocysts, thus increasing the risks of biliary coccidiosis in nearby living pet rabbits that would otherwise not have access to oocysts of E. stiedai.

Establishment of a reference interval for urinary albumin-to-creatinine ratio in dogs.

BACKGROUND: Albuminuria is an important marker of renal damage and can precede proteinuria; thus, it can be a useful analyte in the early diagnosis of kidney diseases. Albuminuria has also been found in dogs with hypertension, inflammatory, infectious, and neoplastic diseases. OBJECTIVES: The aim of this study was to establish a reference interval (RI) for albuminuria in dogs. METHODS: One hundred sixty-four clinically healthy dogs were enrolled in the study. Urinary albumin was determined by the immunoturbidimetric method, and albumin excretion was expressed as the urinary albumin-to-creatinine (UAC) ratio. The RI for UAC was established. RESULTS: After exclusions, 124 dogs from 32 breeds remained. The median UAC of the study population was 3.0 mg/g (range: 0-48). The RI was defined as 0-19 mg/g (with a 90% CI for the upper limit of 13-28 mg/g). No significant difference was found between male and female dogs or between different age and body weight groups. The results of Sighthounds (n = 30) and Beagle dogs (n = 23) did not differ from the other breeds. CONCLUSION: The canine RI of UAC is similar but somewhat narrower than the human RI.

Dogs are final hosts of Sarcocystis morae (Apicomplexa: Sarcocystidae): First report of this species in Hungary and its region - Short communication.

In this study, faecal samples of four American Staffordshire terrier dogs (used for illegal fighting) were analysed by DNA extraction, molecular-phylogenetic and parasitological methods, in order to examine the occurrence of protozoan, apicomplexan parasites. In one sample, the DNA of Sarcocystis morae was shown to be present. This species was identified based on 100% identity with already reported sequences of S. morae from cervids in Lithuania and Spain. The result was also confirmed by phylogenetic analysis. The sporocysts of the canine S. morae isolate measured 14.95 × 9.75 µm on average. This is the first molecular evidence in support of the final host role of domestic dogs in the life cycle of S. morae. The most likely source of the infection was raw meat given to the examined dog to increase its physical achievement. In conclusion, under similar circumstances dogs may participate in the life cycle of S. morae in a 'natural way', shedding sporocysts/oocysts when used for hunting or taken to walks in forested areas.

Babesia gibsoni emerging with high prevalence and co-infections in "fighting dogs" in Hungary.

Babesia gibsoni is considered as an emerging protozoan parasite of dogs in North America and Europe. However, no data have been published on its prevalence, molecular-phylogenetic characteristics and associated co-infections in dogs used for illegal fighting (i.e. predisposed to acquiring this piroplasm via biting) in Europe. In this study, blood samples from 79 American Staffordshire Terrier dogs, confiscated for illegal dog fights, were molecularly analyzed for tick-borne pathogens. B abesia gibsoni was detected in 32 dogs, i.e. with a prevalence of 40.5%. In addition, Babesia vulpes was found in 8 samples (prevalence of 10.1%), for the first time in dogs in Hungary. Canine hemoplasmas were also identified in 49 samples (62%): only Mycoplasma haemocanis in 32 (40.5%) dogs, only "Candidatus Mycoplasma haematoparvum" in 9 (11.4%) dogs, and both hemoplasmas in 8 (10.1%) dogs. Thus, hemoplasma infections also showed a particularly high prevalence in this dog population. Based on a partial fragment of the 18S rRNA gene, B. gibsoni from Hungary exhibited complete sequence identity with conspecific strains reported from Europe and Asia. The cytochrome c oxidase subunit 1 (cox1) gene sequence of this isolate showed the closest identity with B. gibsoni reported from Japan but had a nonsynonymous mutation (M33I). Furthermore, the 11 B. gibsoni-positive samples analyzed for sequence variants of the cytochrome b (cytb) gene showed the presence of a common mutation (P310S). Most importantly, B. gibsoni had two further nonsynonymous mutations, M121I and F258L, in a dog with severe and relapsing anemia following atovaquone treatment. Phylogenetically, both cytb sequence variants clustered together, with a clear geographical pattern showing the closest relationship of both haplotypes identified in Hungary with those from China and Japan. To the best of our knowledge, this is the first cox1 and cytb characterization of B. gibsoni in Europe, as well as the first report on the emergence of this piroplasm and hemoplasmas with high prevalence among "fighting dogs" north of the Mediterranean Basin.

Serum hepcidin measurements in healthy dogs using liquid chromatography/tandem mass spectrometry.

BACKGROUND: Hepcidin is a key regulator of iron homeostasis. The measurement of this hormone is essential for the diagnosis of iron deficiency anemia and might be useful as a prognostic factor in many diseases. Serum hepcidin levels have been infrequently evaluated in dogs. OBJECTIVES: The aim of this study was to measure serum hepcidin in a population of healthy dogs using liquid chromatography/tandem mass spectrometry (LC-MS/MS), considered the gold standard assay for measuring hepcidin like in human medicine. METHODS: Blood samples from 86 healthy dogs of 25 different breeds were measured with the LC-MS/MS method. Synthetic canine hepcidin was used as the standard reagent. Reference values were calculated based on the results. RESULTS: The mean hepcidin concentration of the study population was 16.6 ± 7.7 ng/mL. There reference interval (RI) was defined as 5.3-36.4 ng/mL. No significant difference was found between male and female dogs, or between different age and body weight groups. Hepcidin concentrations did not correlate with red blood cell counts, hemoglobin concentrations, iron levels, iron-binding capacities, and C-reactive protein concentrations in this healthy population. A weak negative correlation was found between hepcidin and the mean corpuscular volume. CONCLUSION: LC-MS/MS proved to be a reliable and time-effective method for the detection of canine hepcidin. The RI was similar but narrower compared with that of human studies.

Pan-European Study on the Prevalence of the Feline Leukaemia Virus Infection - Reported by the European Advisory Board on Cat Diseases (ABCD Europe).

Feline leukaemia virus (FeLV) is a retrovirus associated with fatal disease in progressively infected cats. While testing/removal and vaccination led to a decreased prevalence of FeLV, recently, this decrease has reportedly stagnated in some countries. This study aimed to prospectively determine the prevalence of FeLV viraemia in cats taken to veterinary facilities in 32 European countries. FeLV viral RNA was semiquantitatively detected in saliva, using RT-qPCR as a measure of viraemia. Risk and protective factors were assessed using an online questionnaire to report geographic, demographic, husbandry, FeLV vaccination, and clinical data. The overall prevalence of FeLV viraemia in cats visiting a veterinary facility, of which 10.4% were shelter and rescue cats, was 2.3% (141/6005; 95% CI: 2.0%-2.8%) with the highest prevalences in Portugal, Hungary, and Italy/Malta (5.7%-8.8%). Using multivariate analysis, seven risk factors (Southern Europe, male intact, 1-6 years of age, indoor and outdoor or outdoor-only living, living in a group of ≥5 cats, illness), and three protective factors (Northern Europe, Western Europe, pedigree cats) were identified. Using classification and regression tree (CART) analysis, the origin of cats in Europe, pedigree, and access to outdoors were important predictors of FeLV status. FeLV-infected sick cats shed more viral RNA than FeLV-infected healthy cats, and they suffered more frequently from anaemia, anorexia, and gingivitis/stomatitis than uninfected sick cats. Most cats had never been FeLV-vaccinated; vaccination rates were indirectly associated with the gross domestic product (GDP) per capita. In conclusion, we identified countries where FeLV was undetectable, demonstrating that the infection can be eradicated and highlighting those regions where awareness and prevention should be increased.