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Many puppies from commercial breeding kennels (CBKs) are transported by ground from their kennels of origin to a distributor. This experience may elicit fear and stress during a sensitive developmental period, which may in turn negatively impact the puppies' short- and long-term welfare. This study aimed to measure short-term effects of transportation on puppy welfare metrics. Eight-week-old puppies (n = 383) from 12 CBKs were tested at their kennels (pre-trans) and ~48 h after arriving at a distributor (post-trans). At each location, puppies underwent an isolation test, a stranger-approach test, and a physical health assessment. Behavioral responses to testing were scored from videos. Fecal glucocorticoid metabolites (FGM), fecal secretory immunoglobulin A (sIgA), and presence of intestinal parasites were also analyzed. Linear mixed-effects models identified decreased exploration (p < 0.001), and increased locomotion (p < 0.001) and escape attempts (p = 0.001) during the post-trans isolation test. Increased affiliative behavior (p < 0.001), FGM (p < 0.001) and sIgA (p = 0.014) were also observed post-trans. Findings support good physical health both pre- and post-trans, while behavioral and physiological changes suggest increased puppy distress post-trans. Higher post-transport affiliative behavior may indicate that puppies sought social support as a coping strategy after experiencing transport-related distress. Future studies should explore the efficacy of transportation-related interventions to mitigate puppy distress.

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BACKGROUND: Current guidelines recommend parvovirus revaccination of adult dogs no more frequently than every 3 years. The aim of this study was to determine the prevalence of dogs showing protective serum antibody titres against canine parvovirus 2 in breeding kennels in Northern Italy and to assess the effect of time from vaccination and the sex of the dog on antibody titres. The study was carried out on 370 animals of different breeds kept in 33 breeding kennels. Antibodies to canine parvovirus 2 in serum samples were measured with an indirect immunoenzymatic assay validated by the manufacturer in relation to the 'gold standard' haemagglutination inhibition test. The number of months that had elapsed since the last vaccination was calculated for each animal and categorized into the following classes: < 12 months; 13-24 months; 25-36 months; 37-48 months; and > 49 months. RESULTS: The prevalence of 'unprotected' dogs was 4.6%. A satisfactory solid herd immunity was present in the majority of breeding kennels, although some vaccination failures were detected. A significant negative correlation was found between antibody titre and months since last vaccination. Comparable antibody titres were found in the first 3 years after vaccination. Although the antibody titre over time was not affected by the sex of the dog, 'unprotected' females had been vaccinated more recently than males with analogous low titres. CONCLUSIONS: Parvovirus revaccination of adult dogs every 3 years, as currently recommended, is also the appropriate recommendation for breeding kennels. Serological tests could be a useful tool to assess the effectiveness of vaccination.

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Cryptosporidium is a common intestinal protozoan that can lead to diarrhea in humans and dogs. The predominant species of infection are C. hominis and C. parvum in humans, and C. canis in dogs. However, C. canis can infect immunocompromised humans. Considering the close contact with humans, dogs have the potential to be reservoirs for human cryptosporidiosis. Breeding kennels are the major supply source of puppies for pet shops. The present study is to determine the molecular prevalence and characteristics of Cryptosporidium spp. found in breeding kennel dogs. A total of 314 fecal samples were collected from young and adult dogs kept in 5 breeding kennels. A polymerase chain reaction targeting the small subunit rRNA gene was employed for the detection of Cryptosporidium spp. To determine the species, the DNA sequences were compared to GenBank data. Overall, 21.0% of the fecal samples were positive for Cryptosporidium spp. infection. Cryptosporidium spp. was detected in all 5 facilities. A sequencing analysis demonstrated that all isolates shared 99-100% similarity with C. canis. The results suggest that Cryptosporidium spp. infection is present at a high-level in breeding kennel dogs. However, because dominant species in this survey was C. canis, the importance of breeding kennel dogs as reservoirs for Cryptosporidium spp. transmission to humans is likely to be low in Japan.

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Strongyloides stercoralis is a worldwide-distributed intestinal nematode affecting mainly humans and dogs. Canine strongyloidosis is generally characterised by diarrhoea, malabsorption and bronchopneumonia, and may be fatal in cases of impaired immunity. In recent years, molecular and epidemiological studies suggested that host-adapted populations of S. stercoralis with different zoonotic potential may exist. Clinical and subclinical cases of S. stercoralis infection have been increasingly diagnosed in imported (France, Belgium, Bulgaria) and locally born dogs in Switzerland, showing that this parasite is currently circulating in Europe. Three of these clinical cases will be described here. All three dogs presented severe disease, characterised by harsh diarrhoea, dehydration, vomiting, respiratory and/or neurologic signs, and needed intensive care and hospitalisation. One of these dogs was related to a Swiss breeding kennel, in which the infection was subsequently diagnosed in several other dogs. Faeces were analysed by three coproscopical methods including (i) the Baermann technique, which consistently identified the typical S. stercoralis first-stage larvae in both clinical and subclinical infections, (ii) the sedimentation-zinc chloride flotation and (iii) sodium acetate-acetic acid-formalin concentration (SAFC) methods, which allowed the additional identification of parasitic females and/or eggs in two of the clinical cases. Interestingly, S. stercoralis isolated from all three independent clinical cases exhibited an identical genetic background on the nuclear 18S rDNA (fragment involving hypervariable regions I and IV) and the mitochondrial cytochrome oxidase subunit I (cox1) loci, similar to that of zoonotic isolates from other geographical regions, and not to that of dog-adapted variants. Due to the clinical relevance and zoonotic potential of this parasite, the awareness of both diagnosticians and clinicians is strongly required.

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Cryptosporidium is a common intestinal protozoan that can lead to diarrhea in humans and dogs. The predominant species of infection are C. hominis and C. parvum in humans, and C. canis in dogs. However, C. canis can infect immunocompromised humans. Considering the close contact with humans, dogs have the potential to be reservoirs for human cryptosporidiosis. Breeding kennels are the major supply source of puppies for pet shops. The present study is to determine the molecular prevalence and characteristics of Cryptosporidium spp. found in breeding kennel dogs. A total of 314 fecal samples were collected from young and adult dogs kept in 5 breeding kennels. A polymerase chain reaction targeting the small subunit rRNA gene was employed for the detection of Cryptosporidium spp. To determine the species, the DNA sequences were compared to GenBank data. Overall, 21.0% of the fecal samples were positive for Cryptosporidium spp. infection. Cryptosporidium spp. was detected in all 5 facilities. A sequencing analysis demonstrated that all isolates shared 99–100% similarity with C. canis. The results suggest that Cryptosporidium spp. infection is present at a high-level in breeding kennel dogs. However, because dominant species in this survey was C. canis, the importance of breeding kennel dogs as reservoirs for Cryptosporidium spp. transmission to humans is likely to be low in Japan.

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