Automated highly multiplex detection system for respiratory pathogens in canines

In this investigation a population of dogs at a rehoming center was monitored over a period of 2 years. Despite regular vaccination of incoming dogs against distemper, canine adenovirus type 2 (CAV-2), and canine parainfluenza virus (CPIV), respiratory disease was endemic. Tissue samples from the respiratory tract as well as paired serum samples were collected for analysis. The development of PCR assays for the detection of CPIV, canine adenovirus types 1 and 2, and canine herpesvirus (CHV) is described. Surprisingly, canine adenovirus was not detected in samples from this population, whereas 19.4% of tracheal and 10.4% of lung samples were positive for CPIV and 12.8% of tracheal and 9.6% of lung samples were positive for CHV. As reported previously, a novel canine respiratory coronavirus (CRCoV) was detected in this population (K. Erles, C. Toomey, H. W. Brooks, and J. Brownlie, Virology 310:216-223, 2003). Infections with CRCoV occurred mostly during the first week of a dog's stay at the kennel, whereas CPIV and CHV were detected at later time points. Furthermore, the evaluation of an enzyme-linked immunosorbent assay for detection of antibodies to CPIV and an immunofluorescence assay for detection of antibodies to CHV is described. This study shows that CPIV is present at kennels despite vaccination. In addition, other agents such as CHV and CRCoV may play a role in the pathogenesis of canine respiratory disease, whereas CAV-2 and canine distemper virus were not present in this population, indicating that their prevalence in the United Kingdom is low due to widespread vaccination of dogs.

Infectious tracheobronchitis (ITB), also known as the kennel cough, is a respiratory syndrome of dogs and usually appears to be contagious among dogs housed in groups. Etiologic agent of ITB is multiple and sometimes complex. In the present study, 68 household dogs showing clinical signs of respiratory infection were examined, and 20 dogs (29.4%) were found to be positive for either of following agents. Bordetella bronchiseptica (B.b.) was most frequently detected from nasal and oropharynx sites of 7 dogs (10.3%). Among the viruses examined, canine parainfluenza virus (CPIV) was detected with the highest frequency (7.4%). Other pathogens included in the order of frequency group 1 canine coronavirus (4.4%), canine adenovirus type 2 (2.9%), group 2 canine respiratory coronavirus (1.5%), and canine distemper virus (1.5%). Only 2 cases showed mixed infections. Neither influenza A virus nor canine bocavirus (minute virus of canines) was found in any dogs examined. These results indicate that both B.b. and CPIV are likely to be the principal etiologic agents of canine ITB in Japan, and they may be considered as the target for prophylaxis by vaccination.

Eight puppies (group 1) were vaccinated once with a bivalent modified-live vaccine against infectious tracheobronchitis by the intranasal route and at the same time with an injectable trivalent vaccine against...

Respiratory infections represent an important and frequent cause of disease in cats and dogs worldwide. Many pathogens have been associated with feline (FIRD) and canine infectious respiratory disease (CIRD) including feline calicivirus (FCV) and herpesvirus (FeHV-1) in cats and canine distemper virus (CDV), parainfluenza virus (CPiV), adenovirus type 2 (CAV-2), herpesvirus (CHV) and Bordetella bronchiseptica in dogs. All these pathogens are currently included in either core or non-core vaccines, leading to a reduction in overall disease burden. Despite widespread use of vaccines, disease is still present at significant levels. In dogs, several new pathogens, potentially associated with respiratory disease, have recently emerged including canine respiratory coronavirus (CRCoV) and canine pneumovirus (CnPnV). In cats, it has been theorised that the high rates of evolution of viruses such as FCV may lead to a reduction of vaccine efficacy. Finally, even though respiratory disease of suspected infectious origin is frequent, the culprit often remains unidentified. This thesis aims to build understanding of canine and feline infectious respiratory disease epidemiology, re-evaluate the roles of known and suspected pathogens in this syndrome and to aid in informing the development of new disease prevention strategies. During a cross-sectional study across 6 European countries, samples were collected from 1521 veterinary practice attending cats. The prevalence of FCV in this population was 9.2%. Phylogenetic analysis of these field isolates showed high viral variability with a radial phylogeny. In vitro viral neutralisation suggested that antibodies raised to the FCV-F9 vaccine strain (which has been widely in use for decades) are still broadly cross-reactive to contemporary field isolates. A retrospective serosurvey of 200 canine and 179 feline samples screened for influenza A and B viruses suggested that UK dogs and cats have been rarely exposed to influenza viruses in recent years with only 1.5% of canine samples being seropositive for equine H3N8 and 0.56% of feline samples being seropositive to human pandemic 2009 H1N1. Two case control studies were conducted in British veterinary practices in order to re-appraise the role of known and potential upper respiratory pathogens in pet dogs and cats. In the feline study, FCV was detected in 21.1% of cases and 4.7% controls. FeHV-1 was detected in 10.5% cases and 1.6% controls. Bordetella bronchiseptica was identified in 5.3% cases and 9.4% controls. Finally, 65.8% cases and 48.8% controls were positive for M. felis. In the canine study, CRCoV was identified in 6.1% of cases, CnPnV in 4.4% cases and 1.2% controls, M.cynos in 4.4% cases and 2.3% controls, and B.bronchiseptica in 6.7% cases and 20.7% controls. No samples tested positive for CAV-2, CPiV, CHV, CDV, influenza A, Streptococcus equi subsp. zooepidemicus or C.felis. A cross-sectional sampling of dogs and cats was conducted in four British shelters with one of these also taking part in a 10-week longitudinal study. FCV, FeHV-1 and M. felis were detected in 7.4%, 4.6% and 16.8% feline samples. Cross-sectional dog samples tested negative for all pathogens included in the study. In the longitudinal study, M. felis was the most frequently encountered microorganism followed by FCV in cats. In dogs, CnPnV was identified during two weeks that coincided with reports of canine respiratory disease by members of the shelter staff. Together, findings confirm the importance of CIRD and FIRD as multifactorial syndromes in both pet and unowned shelter populations where pathogen shedding is often observed without clinical signs. In FIRD, FCV is reaffirmed as a majorly important disease causing pathogen. Finally, where CIRD is concerned, findings suggest that recently emerged pathogens such as CRCoV are of increasing importance in this syndrome whereas classic pathogens seem to be less and less present, likely due to widespread vaccination.

Fifty-six red foxes (Vulpes vulpes), 18 gray foxes (Urocyon cinereoargenteus), and 13 coyotes (Canis latrans) obtained by the South Carolina Wildlife and Marine Resources Department during an investigation of suspected illegal wildlife translocation were examined for diseases and parasites. Red foxes and coyotes were confiscated from an animal dealer based in Ohio (USA), and gray foxes were purchased from an animal dealer in Indiana (USA). Emphasis was placed on detection of pathogens representing potential health risks to native wildlife, domestic animals, or humans. All animals were negative for rabies; however, 15 gray foxes were incubating canine distemper at necropsy. Serologic tests disclosed antibodies to canine parvovirus, canine distemper virus, canine adenovirus, canine coronavirus, canine herpesvirus, and canine parainfluenza virus in one or more host species. Twenty-three species of parasites (two protozoans, three trematodes, four cestodes, eleven nematodes, and three arthropods) were found, including species with substantial pathogenic capabilities. Echinococcus multilocularis, a recognized human pathogen not enzootic in the southeastern United States, was found in red foxes. Based on this information, we conclude that the increasingly common practice of wild canid translocation for stocking fox-chasing enclosures poses potential health risks to indigenous wildlife, domestic animals, and humans and, therefore, is biologically hazardous.

Establishment of reverse transcription loop-mediated isothermal amplification for rapid detection and differentiation of canine distemper virus infected and vaccinated animals

ObjectivesTo assess the prevalence of canine parainfluenza virus, canine adenovirus type 2, canine distemper virus, canine respiratory coronavirus and influenza virus A infections in: (1) privately‐owned or, (2) kennelled dogs showing signs consistent with canine infectious respiratory disease and, (3) clinically healthy dogs in Vienna, Austria.Materials and MethodsProspectively, nasal and tonsillar swabs from 214 dogs affected with infectious respiratory disease, and 50 healthy control dogs were tested for nucleic acids specific to the various viral infections. Concurrent bronchoalveolar lavage fluid from 31 dogs with chronic respiratory disease was investigated for the same viral pathogens. Additionally, anti‐canine respiratory coronavirus antibody concentrations were measured in paired blood samples from 30 acutely diseased dogs.ResultsCanine respiratory coronavirus (7.5%) and canine parainfluenza virus (6.5%) were the most commonly detected viruses in samples from the upper airways of dogs with respiratory infections. Serological results showed a significant seroconversion in response to coronavirus in 50% of the examined cases. None of the samples was positive for influenza virus A‐specific nucleic acid. Canine coronavirus‐specific nucleic acid was detected in 4.0% of healthy dogs.Clinical SignificanceCanine coronavirus should be considered as a clinically relevant cause of infectious respiratory disease in crowded dog populations. For sample collection, the nasal mucosa can be recommended as the favoured site. Analysis of paired serum samples aids verification of canine coronavirus infection in respiratory disease.

Objectives To evaluate the safety and efficacy of a live canarypox virus recombinant-canine distemper virus (CDV) combination vaccine against virulent CDV challenge exposure, and to document lack of interference among the other modified-live virus (MLV) components. Animals 33 specific-pathogen-free (SPF) Beagle pups (7 to 10 weeks old). Procedure A canarypox virus recombinant-CDV combination vaccine was tested for safety and efficacy along with MLV components (canine adenovirus type 2, canine coronavirus, canine parainfluenza virus, and canine parvovirus) in 26 SPF Beagle pups. The combination vaccine was rehydrated with either Leptospira canicola-L icterohaemorrhagiae combination bacterin (vaccine 1) or sterile diluent (vaccine 2). An additional group of 7 seronegative SPF pups received the control MLV components devoid of the combination vaccine (vaccine 3). Two vaccinations were administered 21 days apart, either IM or SC. The dose of the combination vaccine used to inoculate these pups was 40 times lower than the recommended commercial dose. At 21 days after the booster vaccination, all pups were challenge exposed with a virulent CDV strain, then were observed for 21 days to record morbidity and mortality. Results Adverse local or generalized reactions were not induced by vaccinations. All vaccinates seroconverted to CDV. Serum antibody titers to MLV components were not different, with or without inclusion of the combination vaccine. After challenge exposure, morbidity and mortality in vaccinates were 0% (0/26); in control dogs, values were 100% morbidity and 86% mortality (6/7). Brain impression smear slides made from all dogs that did not survive challenge exposure were CDV positive by use of a direct fluorescein isothiocyanate method. Conclusions The canarypox virus-CDV combination vaccine, administered SC or IM, is a safe product that elicits CDV seroconversion, does not interfere with other vaccine components, and protects vaccinated pups against virulent CDV challenge exposure. (Am J Vet Res 1997;58:833–836)

Pathogen exposure in endangered island fox ( Urocyon littoralis) populations: Implications for conservation management

SUMMARY Serum samples collected from 100 dogs in eastern Washington were tested for serum antibody against several viruses having roles in canine respiratory tract infections. The viruses used in the serologic procedures were: canine distemper virus (cdv), canine adenovirus I (cav-1), canine adenovirus II (cav-2), canine parainfluenza virus (cpv), and canine herpesvirus (chv). Of the 100 serum samples, 76% had significant antibody titer against cdv, 42% against cav-1, 45% against cav-2, 19% against cpv, and 6% against chv.

Maned wolves (Chrysocyon brachyurus) are neotropic mammals, listed as a CITES Appendix II species, with a distribution south of the Amazon forest from Bolivia, through northern Argentina and Paraguay and into eastern Brazil and northern Uruguay. Primary threats to the survival of free-ranging maned wolves include habitat loss, road kills, and shooting by farmers. An additional threat to the conservation of maned wolves is the risk of morbidity and mortality due to infectious and parasitic diseases. Captive maned wolves are susceptible to, and die from, common infectious diseases of domestic dogs (Canis familiaris) including canine distemper virus (CDV), canine parvovirus (CPV), rabies virus, and canine adenovirus (CAV). Results from this study show that free-ranging maned wolves in a remote area of Bolivia have been exposed to multiple infectious and parasitic agents of domestic carnivores, including CAV, CDV, CPV, canine coronavirus, rabies virus, Leptospira interrogans spp., Toxoplasma gondii, and Dirofilaria immitis, and may be at increased risk for disease due to these agents.

The WSAVA Vaccination Guidelines Group (VGG) was convened in order to develop guidelines for the vaccination of dogs and cats that have global application. The first version of these guidelines was published in 2007 and they were updated in 2010. The present document provides a further updated and expanded version of these international guidelines for the vaccination of small companion animals and indicates the scientific evidence base on which the recommendations are made. The VGG recognises that the keeping of pet small animals is subject to considerable variation in practice and associated economics throughout the world and that vaccination recommendations that might apply to a developed country may not be appropriate for a developing country. These guidelines are not a mandatory edict, but rather should be used by national associations and individual veterinary practices to develop vaccination schedules relevant locally. However, the VGG strongly recommends that wherever possible ALL dogs and cats receive the benefit of vaccination. This not only protects the individual animal, but provides optimum ‘herd immunity’ that minimises the likelihood of infectious disease outbreaks. With this background in mind, the VGG has defined core vaccines as those which ALL dogs and cats, regardless of circumstances or geographical location, should receive. Core vaccines protect animals from severe, life-threatening diseases that have global distribution. Core vaccines for dogs are those that protect against canine distemper virus (CDV), canine adenovirus (CAV) and the variants of canine parvovirus type 2 (CPV-2). Core vaccines for cats are those that protect against feline parvovirus (FPV), feline calicivirus (FCV) and feline herpesvirus-1 (FHV-1). In areas of the world where rabies virus infection is endemic, vaccination against this agent should be considered core for both species, even if there is no legal requirement for routine vaccination. The VGG recognizes that maternally-derived antibody (MDA) interferes with the efficacy of most current core vaccines administered to pups and kittens in early life. Because the level of MDA varies among litters, the VGG recommends administration of multiple core vaccine doses to pups and kittens, with the final dose given at 16 weeks or older and followed by a booster at 6- or 12-months of age. In cultural or financial situations where a pet animal may only be permitted the benefit of a single vaccination, that vaccination should be with core vaccines at 16 weeks of age or older. The VGG supports the use of simple in-practice tests for determination of seroconversion to the core vaccine components (CDV, CAV, CPV-2 and FPV) following vaccination, for determination of seroprotection in adult dogs and for management of infectious disease outbreaks in shelters. Vaccines should not be given needlessly. Core vaccines should not be given any more frequently than every three years after the 6- or 12-month booster injection following the puppy/kitten series, because the duration of immunity (DOI) is many years and may be up to the lifetime of the pet. The VGG has defined non-core vaccines as those that are required by only those animals whose geographical location, local environment or lifestyle places them at risk of contracting specific infections. The VGG has also classified some vaccines as not recommended (where there is insufficient scientific evidence to justify their use) and has not considered a number of minority products which have restricted geographical availability or application. The VGG strongly supports the concept of regular (usually annual) health checks which removes the emphasis from, and client expectation of, annual revaccination. The annual health check may still encompass administration of selected non-core vaccines which should be administered annually, as the DOI for these products is generally 1 year. The VGG has considered the use of vaccines in animal shelters, again recognising the particular circumstances of such establishments and the financial constraints under which they sometimes operate. The VGG minimum shelter guidelines are simple: that all dogs and cats entering such an establishment should be vaccinated before, or at the time of entry, with core vaccines. Where finances permit, repeated core vaccines should be administered as per the schedules defined in the guidelines and non-core vaccines against respiratory disease may be included. The VGG recognizes the importance of adverse reaction reporting schemes, but understands that these are variably developed in different countries. Wherever possible, veterinarians should be actively encouraged to report all possible adverse events to the manufacturer and/or regulatory authority to expand the knowledge base that drives development of improved vaccine safety. These fundamental concepts proposed by the VGG may be encapsulated in the following statement: We should aim to vaccinate every animal with core vaccines. Non-core vaccines should be given no more frequently than is deemed necessary.

No presente estudo, foi realizada uma pesquisa em busca de anticorpos neutralizantes contra os vírus da cinomose (CDV) e da parainfluenza (CPIV) caninos em amostras de soro de 173 cães recolhidos a canis municipais em Novo Hamburgo (n=82) e Porto Alegre (n=91), RS. A pesquisa de anticorpos neutralizantes foi realizada pela técnica de soroneutralização frente a duas amostras vacinais de CDV (Rockborn e Snyder Hill) e frente a uma amostra de CPIV (V660). Em relação ao CDV, 95,9% das amostras de soros foram negativas para anticorpos neutralizantes contra a amostra Snyder Hill e 90,7% soronegativas para a amostra Rockborn. Entre os soropositivos (n=20; 11,6%), somente três deles apresentaram anticorpos neutralizantes frente às duas amostras de CDV testadas, indicando pouca reatividade cruzada entre as mesmas. Quanto ao CPIV, a prevalência de anticorpos neutralizantes encontrada frente à amostra V660 foi de 51,4%. Esses achados indicam que a maioria dos cães examinados não teve contato prévio com o CDV, seja por infecção natural ou por imunização prévia. O CPIV, porém, parece estar amplamente difundido na população canina examinada, provavelmente por exposição natural ao vírus.

Infectious canine hepatitis: An “old” disease reemerging in Italy

Animal shelters provide an ideal environment for the spread of disease. Dogs are often housed in close quarters with others of unknown vaccine histories, and experience high levels of sustained stress. As a result, Canine Infection Respiratory Disease (CIRD) is often prevalent and difficult to control. The aims of this study were to (1) identify specific pathogens responsible for CIRD in a city shelter in West Texas, USA, and (2) determine whether intake vaccinations decrease proportion of dogs exhibiting signs of CIRD even during an outbreak. A laboratory analysis of conjunctival, pharyngeal, and nasal swabs (n = 15 dogs) and fecal samples (n = 6 kennels) showed prevalence of various CIRD pathogens (e.g., canine adenovirus-2, canine parainfluenza virus, canine distemper virus). All fifteen dogs tested positive for at least one pathogen, with the most prevalent pathogen being Canine Distemper Virus (CDV; n = 12). All of the kennels (n=6) tested positive for Canine Distemper Virus. Health data on dogs (n = 1,258) over the age of 6 weeks were assessed from May to August 2017. Beginning in July, both stray and owner-surrendered dogs were vaccinated with Nobivac® Canine 1-DAPPv 5 Way and Nobivac® Intra-Trac® 3 upon intake, which differed from the previous policy. For each day in the study, we calculated the proportion of dogs in each nasal discharge category, the proportion of dogs observed coughing, and the mean fecal score across all dogs. We conducted a linear regression between the proportion of the shelter vaccinated and the proportion of dogs coughing. At the beginning of the vaccination phase, ~25% of the dogs were coughing. However, as the proportion of the dogs vaccinated increased, the proportion of dogs coughing decreased. There was a significant decrease of 7% of the proportion of dogs coughing when vaccination was at least at 90% compared to when it was <90%. These data suggest that the shelter in this study was experiencing a CIRD outbreak, with CDV being primary pathogen, and that it is possible to substantially reduce illness by implementing a vaccination on intake protocol. The current study provides support for the importance of vaccination in animal shelter welfare.