Canine Infectious Respiratory Disease Research Articles

Molecular identification and occurrence of bordetellosis among dogs in northern Kerala

Bordetella bronchiseptica is a gram–negative bacterium with a broad host range that is most commonly seen in animals kept in close quarters. It is considered to be one of the main causes of the canine infectious respiratory disease complex (CIRDC). In this study, 45 dogs with clinical signs suggestive of CIRDC were selected and nasal/ oropharyngeal swabs were collected aseptically. Total DNA was extracted from the swabs and subjected to polymerase chain reaction (PCR) targeting alc gene of Bordetella spp. for confirmation. Nine animals (20 per cent) tested positive for the presence of Bordetella antigen. All the dogs tested positive for Bordetellosis were less than one year of age. Most common clinical signs in positive dogs were cough and nasal discharge, and the majority of the bordetellosis positive dogs were from multiple dog houses.

Development of a Recombinase Polymerase Amplification Fluorescence Assay for the Detection of Canine Adenovirus 2.

Canine adenovirus type 2 (CAdV-2) is often found in co-infections with other pathogens causing canine infectious respiratory disease (CIRD). Rapid, efficient, and convenient pathogen detection is the best approach for early confirmatory diagnosis. In this study, we developed and evaluated a rapid real-time recombinase polymerase amplification (RPA) assay for detection of canine adenovirus 2 (CAV), which can detect CAV within 15 min at 39°C. The detection limit that assay was 214 copies/μl DNA molecules per reaction. The specificity was indicated by a lack of cross-reaction with canine distemper virus (CDV), canine coronavirus (CCV), and canine parvovirus (CPV). Field and clinical applicability of this assay were evaluated using 86 field samples. The coincidence rate of the detection results for clinical samples between CAV-RPA and qPCR was 97.7%. In summary, the real-time CAV-RPA analysis provides an efficient, rapid and sensitive detection method for CAV.

An emerging orthopneumovirus detected from dogs with canine infectious respiratory disease in China.

Canine infectious respiratory disease (CIRD) is a major cause of morbidity in dogs and is associated with several viral pathogens. The viral diversity associated with CIRD was investigated by analyzing the viral communities from nine CIRD-affected dogs using metagenomics. The results identified 10 mammalian viruses, including canine parvovirus, canid alphaherpesvirus 1, canine kobuvirus, Felis catus papillomavirus 3, canine respiratory coronavirus, canine adenovirus 2, Canis familiaris polyomavirus (DogPyV), canine coronavirus, human papillomavirus and canine pneumovirus (CPnV). Interestingly, CPnV and DogPyV were first discovered in China. Further investigation in 107 samples in China using specific PCR found only two CPnV positive strains in 51 CIRD samples and none in 56 healthy samples. Furthermore, a complete nucleotide sequence of CPnV strain SMU-2020-CB19 and a partial nucleotide sequence of strain SMU-2020-CB14 were obtained. Sequence comparisons and phylogenetic analysis showed that both novel CPnV strains were a close match to the detected swine orthopneumovirus strain in USA, but distantly related to other CPnV strains. Here, the first discovery and characterization of orthopneumovirus in dogs with CIRD in China were reported, highlighting the need for further research on pathogenicity and transmission in China.

Asymptomatic carriage of canine infectious respiratory disease complex pathogens among healthy dogs.

The objectives of this study were to determine the prevalence of canine infectious respiratory disease pathogens among asymptomatic client-owned dogs, and to compare the risks of asymptomatic pathogen carriage between client-owned dogs and dogs in an animal shelter. Pooled tonsillar, conjunctival and nasal cavity swabs from asymptomatic client-owned dogs (n=133) were tested using a real-time polymerase chain reaction canine respiratory panel. Identical samples from asymptomatic dogs in an animal shelter (n=295) were similarly tested for selected pathogens. Risk differences were calculated between client-owned dogs and shelter dogs for each of the respiratory pathogens included in the analyses. A total of 15 of 133 (11.3%) asymptomatic client-owned dogs were positive for at least one pathogen in the complex. Seven dogs (6.1%) were positive for M. cynos, six (5.2%) were positive for B. bronchiseptica, two (1.7%) were positive for canine herpesvirus type 1 and two (1.7%) were positive for canine respiratory coronavirus. For all eight pathogens tested in both groups, the proportion of positive cases was higher among shelter dogs than among client-owned dogs. Shelter dogs had a higher risk for M. cynos (0.18, 95% confidence interval: 0.12 to 0.25), canine respiratory coronavirus (0.15, 95% confidence interval: 0.10 to 0.19), canine distemper virus (0.06, 95% confidence interval: 0.03 to 0.09), and canine pneumovirus (0.05, 95% confidence interval: 0.03 to 0.08) than client-owned dogs. Odds ratios for M. cynos (0.31, 95% confidence interval: 0.08 to 0.92) and canine respiratory coronavirus (0.05, 95% confidence interval: 0.01 to 0.18) were significantly different between client-owned and shelter dogs. In all cases except for canine herpesvirus type 1, dogs within the shelter population were observed to be at higher risk of exhibiting asymptomatic carriage of a respiratory pathogen as compared to client-owned dogs. The strength of this association was strongest for M. cynos and canine respiratory coronavirus. The risk of canine infectious respiratory disease pathogen exposure posed by asymptomatic client-owned dogs is poorly defined. This study also corroborates previous reports of high canine infectious respiratory disease prevalence among clinically healthy shelter dogs, and further determined that the overall prevalence of canine infectious respiratory disease pathogen carriage among clinically healthy client-owned dogs is low but is highest for the traditional pathogen B. bronchiseptica and the emerging pathogen M. cynos.

A molecular survey of canine respiratory viruses in New Zealand

ABSTRACT Aims The aim of this study was to identify viruses associated with canine infectious respiratory disease syndrome (CIRDS) among a population of New Zealand dogs. Methods Convenience samples of oropharyngeal swabs were collected from 116 dogs, including 56 CIRDS-affected and 60 healthy dogs from various locations in New Zealand between March 2014 and February 2016. Pooled samples from CIRDS-affected (n = 50) and from healthy (n = 50) dogs were tested for the presence of canine respiratory viruses using next generation sequencing (NGS). Individual samples (n = 116) were then tested by quantitative PCR (qPCR) and reverse transcriptase qPCR (RT-qPCR) for specific viruses. Groups were compared using Fisher’s exact or χ2 tests. The effect of explanatory variables (age, sex, type of household, presence of viral infection) on the response variable (CIRDS-affected or not) was tested using RR. Results Canine pneumovirus (CnPnV), canine respiratory coronavirus (CRCoV), canine herpesvirus-1 (CHV-1), canine picornavirus and influenza C virus sequences were identified by NGS in the pooled sample from CIRDS-affected but not healthy dogs. At least one virus was detected by qPCR/RT-qPCR in 20/56 (36%) samples from CIRDS dogs and in 23/60 (38%) samples from healthy dogs (p = 0.84). CIRDS-affected dogs were most commonly positive for CnPnV (14/56, 25%) followed by canine adenovirus-2 (CAdV-2, 5/56, 9%), canine parainfluenza virus (CpiV) and CHV-1 (2/56, 4% each), and CRCoV (1/56, 2%). Only CnPnV (17/60, 28%) and CAdV-2 (14/60, 23%) were identified in samples from healthy dogs, and CAdV-2 was more likely to be detected healthy than diseased dogs (RR 0.38; 95% CI = 0.15–0.99; p = 0.045) Conclusions The frequency of detection of viruses traditionally linked to CIRDS (CAdV-2 and CPiV) among diseased dogs was low. This suggests that other pathogens are likely to have contributed to development of CIRDS among sampled dogs. Our data represent the first detection of CnPnV in New Zealand, but the role of this virus in CIRDS remains unclear. On-going monitoring of canine respiratory pathogens by NGS would be beneficial, as it allows rapid detection of novel viruses that may be introduced to the New Zealand canine population in the future. Such monitoring could be done using pooled samples to minimise costs. Clinical relevance Testing for novel respiratory viruses such as CnPnV and CRCoV should be considered in all routine laboratory investigations of CIRDS cases, particularly in dogs vaccinated with currently available kennel cough vaccines.

Serum 25-hydroxyvitamin D concentration and infectious respiratory disease complex in shelter dogs.

BackgroundHypovitaminosis D is a risk factor for the development of respiratory infections in humans and repletion can be protective.ObjectivesDetermine if serum 25‐hydroxyvitamin (OH)D concentrations are lower in shelter dogs and if 25(OH)D concentrations are associated with clinical signs of canine infectious respiratory disease complex (CIRDC) or with time in the shelter.AnimalsOne hundred forty‐six shelter dogs (clinically ill n = 36, apparently healthy n = 110) and 23 nonshelter control dogs.MethodsProspective cohort study. Shelter dogs were grouped as clinically ill or apparently healthy based on the presence or absence, respectively, of clinical signs associated with CIRDC. Serum 25(OH)D concentrations were measured with a competitive chemiluminesence immunoassay. Nucleic acids of agents associated with the CIRDC were amplified by polymerase chain reaction assays.ResultsThe concentration of 25(OH)D was 7.3 ng/mL (4.5‐9.9, 95% confidence interval [CI]) lower in dogs with signs of CIRDC than apparently healthy shelter dogs (t(142) = 2.0, P = .04). Dogs positive for DNA of canine herpesvirus (CHV)‐1 had serum 25(OH)D concentrations 14.9 ng/mL (−3.7 to 29.6, 95% CI) lower than dogs that were negative (t(137) = 2.0, P = .04). Serum 25(OH)D concentrations in shelter dogs were not different from control dogs (t(45) = −1.4, P = .17). Serum 25(OH)D concentration was not associated with duration of time in the shelter (F(1, 140) = 1.7, P = .2, R 2 = 0.01).Conclusion and Clinical ImportanceVitamin D could have a role in acute respiratory tract infections in shelter dogs.

Canine Respiratory Coronavirus: A Naturally Occurring Model of COVID-19?

Discovered in 2003 at the Royal Veterinary College, London, canine respiratory coronavirus (CRCoV) is a betacoronavirus of dogs and major cause of canine infectious respiratory disease complex. Generally causing mild clinical signs of persistent cough and nasal discharge, the virus is highly infectious and is most prevalent in rehoming shelters worldwide where dogs are often closely housed and infections endemic. As the world grapples with the current COVID-19 pandemic, the scientific community is searching for a greater understanding of a novel virus infecting humans. Similar to other betacoronaviruses, SARS-CoV-2 appears to have crossed the species barrier, most likely from bats, clearly reinforcing the One Health concept. Veterinary pathologists are familiar with coronavirus infections in animals, and now more than ever this knowledge and understanding, based on many years of veterinary research, could provide valuable answers for our medical colleagues. Here I review the early research on CRCoV where seroprevalence, early immune response, and pathogenesis are some of the same key questions being asked by scientists globally during the current SARS-CoV-2 pandemic.

Analysis of the lung microbiota in dogs with Bordetella bronchiseptica infection and correlation with culture and quantitative polymerase chain reaction

Infection with Bordetella bronchiseptica (Bb), a pathogen involved in canine infectious respiratory disease complex, can be confirmed using culture or qPCR. Studies about the canine lung microbiota (LM) are recent, sparse, and only one paper has been published in canine lung infection. In this study, we aimed to compare the LM between Bb infected and healthy dogs, and to correlate sequencing with culture and qPCR results. Twenty Bb infected dogs diagnosed either by qPCR and/or culture and 4 healthy dogs were included. qPCR for Mycoplasma cynos (Mc) were also available in 18 diseased and all healthy dogs. Sequencing results, obtained from bronchoalveolar lavage fluid after DNA extraction, PCR targeting the V1–V3 region of the 16S rDNA and sequencing, showed the presence of Bb in all diseased dogs, about half being co-infected with Mc. In diseased compared with healthy dogs, the β-diversity changed (P = 0.0024); bacterial richness and α-diversity were lower (P = 0.012 and 0.0061), and bacterial load higher (P = 0.004). Bb qPCR classes and culture results correlated with the abundance of Bb (r = 0.71, P < 0.001 and r = 0.70, P = 0.0022). Mc qPCR classes also correlated with the abundance of Mc (r = 0.73, P < 0.001). Bb infection induced lung dysbiosis, characterized by high bacterial load, low richness and diversity and increased abundance of Bb, compared with healthy dogs. Sequencing results highly correlate with qPCR and culture results showing that sequencing can be reliable to identify microorganisms involved in lung infectious diseases.

Evolutionary genetics of canine respiratory coronavirus and recent introduction into Swedish dogs

Canine respiratory coronavirus (CRCoV) has been identified as a causative agent of canine infectious respiratory disease, an upper respiratory infection affecting dogs. The epidemiology is currently opaque, with an unclear understanding of global prevalence, pathology, and genetic characteristics. In this study, Swedish privately-owned dogs with characteristic signs of canine infectious respiratory disease (n = 88) were screened for CRCoV and 13 positive samples (14.7%, 8.4–23.7% [95% confidence interval (CI)]) were further sequenced. Sequenced Swedish CRCoV isolates were highly similar despite being detected in dogs living in geographically distant locations and sampled across 3 years (2013–2015). This is due to a single introduction into Swedish dogs in approximately 2010, as inferred by time structured phylogeny. Unlike other CRCoVs, there was no evidence of recombination in Swedish CRCoV viruses, further supporting a single introduction. Finally, there were low levels of polymorphisms, in the spike genes. Overall, we demonstrate that there is little diversity of CRCoV which is endemic in Swedish dogs.

Antimicrobial Prescribing Practices in Small Animal Emergency and Critical Care.

Background: Antimicrobial use contributes to emergence of antimicrobial resistance. It was hypothesized that antimicrobial prescribing behavior varies between the emergency (ER) and critical care (CC) services in a veterinary teaching hospital. This study aimed to: (i) describe antimicrobial prescribing patterns in the ER and CC services; (ii) assess adherence to stewardship principles; (iii) evaluate the prevalence of multidrug resistant (MDR) bacterial isolates.Methods: Institution electronic medical records were queried for all antimicrobial prescriptions from the ER and CC services between 1/1/2017 and 12/31/2017. Prescriptions were manually reviewed, and the following data recorded: drug, dosage, duration, diagnosis, outcome, hospitalization duration, culture submission, and susceptibility results.Results: There were 5,091 ER visits, of which 3,125 were not transferred to another service. Of these emergency visits, 516 (16.5%) resulted in 613 antimicrobial drug prescriptions. The most commonly prescribed drugs for the ER were amoxicillin/clavulanate (n = 243, 39.6%), metronidazole (n = 146, 23.8%), and ampicillin/sulbactam (n = 55, 9.0%). The most common reasons for antimicrobial prescriptions were skin disease (n = 227, 37.0%), gastrointestinal disease (n = 173, 28.2%), and respiratory disease (n = 50, 8.2%). For ER patients 18 cultures were submitted, equivalent to a 3.5% submission rate. The CC service managed 311 case visits for 822 patient days. Of these, 133 case visits (42.7%) resulted in 340 prescriptions. The most commonly prescribed drugs for the CC service were ampicillin/sulbactam (n = 103, 30.3%), enrofloxacin (n = 75, 22.1%), and metronidazole (n = 59, 17.4%). The most common reasons for antimicrobial prescriptions were gastrointestinal disease (n = 106, 31.2%), respiratory disease (n = 71, 20.9%), and sepsis (n = 61, 17.9%). On the CC service, 46 patients had ≥1 culture submitted, equivalent to a 34.6% submission rate. Of patients prescribed antimicrobials, 13/38 (34%) with urinary tract disease, 2/28 (7%) with pneumonia, 1/11 (9%) with canine infectious respiratory disease complex and 2/8 (25%) with feline upper respiratory infection were compliant with published guidelines.Conclusions: Antimicrobial prescription was common in both ER and CC services and followed similar patterns. Adherence to published guidelines for urinary and respiratory infections was poor.

Molecular detection of canine respiratory pathogens between 2017 and 2018 in Japan.

A molecular survey was conducted to understand recent distribution of pathogens associated with canine infectious respiratory disease (CIRD) in Japan. Nasal and/or pharyngeal swabs were collected from asymptomatic dogs and those with CIRD, living in private house or in kennels. PCR-based examination was conducted for detecting nine pathogens. Among private household dogs, 50.8% with CIRD, 11.1% with respiratory disease other than CIRD, and 4.3% asymptomatic were positive for more than one pathogen, whereas in kennel-housed dogs, 42.9% with CIRD and 27.3% asymptomatic were positive. Bordetella bronchiseptica was most frequently detected, followed by canine herpesvirus 1, canine parainfluenza virus, canine pneumovirus, Mycoplasma cynos, and canine adenovirus type 2. In kennel environment, asymptomatic dogs might act as reservoirs carrying the respiratory pathogens.

Characterization of a novel Mycoplasma cynos real-time PCR assay.

Mycoplasma cynos is recognized as an emerging causative pathogen of canine infectious respiratory disease (CIRD) worldwide. We developed a new open-source real-time PCR (rtPCR) assay for M. cynos that performs well under standard rtPCR conditions. Primers and probes were designed to target the M. cynos tuf gene. Reaction efficiencies for the M. cynos tuf gene assay on 2 platforms were based on amplification of standard curves spanning 8 orders of magnitude: ABI 7500 platform, 94.3–97.9% (r2 ≥ 0.9935); QuantStudio OpenArray platform, 119.1–122.5% (r2 = 0.9784). The assay performed very well over a range of template input, from 109 copies to the lower limit of quantification at 4 copies of the M. cynos genome on the ABI 7500 platform. Diagnostic performance was estimated by comparison with an in-house legacy assay on clinical specimens as well as testing isolates that were characterized previously by intergenic spacer region (ISR) sequencing. Exclusivity was established by testing 12 other Mycoplasma species. To substantiate the high specificity of the M. cynos tuf gene assay, sequence confirmation was performed on ISR PCR amplicons obtained from clinical specimens. One ISR amplicon sequence revealed M. mucosicanis rather than M. cynos. The complete protocol of the newly developed M. cynos tuf assay is provided to facilitate assay harmonization.

Cross-sectional investigation and risk factor analysis of community-acquired and hospital-associated canine viral infectious respiratory disease complex

Canine infectious respiratory disease complex (CIRDC) is associated with multiple factors. The possible transmission source can be via community-acquired infection (CAI) or hospital-associated infection (HAI), but the variable factors within these two routes are not well described. This study aimed to (i) investigate a cross-sectional incidence of canine respiratory viruses, including influenza (CIV), parainfluenza, distemper (CDV), respiratory coronavirus (CRCoV), adenovirus-2, and herpesvirus, in respiratory-diseased dogs, and (ii) analyze the possibly related risk factors. In total 209 dogs with respiratory illness, consisting of 133 CAI and 76 HAI dogs, were studied. Both nasal and oropharyngeal swabs were sampled from each dog and subjected for CIRDC virus detection using multiplex PCRs. Common six viruses associated with CIRDC were detected in both groups with CIV and CRCoV being predominantly found. Only CDV was significantly more prevalent in CAI than HAI dogs. Multiple virus detections were found in 81.2% and 78.9% of CAI and HAI dogs, respectively. Co-detection of CIV and CRCoV was represented the highest proportion and most often found with other CIRD viruses. Moreover, the clinical severity level was notably related to the age of infected dogs, but not to the vaccination status, sex and transmission route. Since healthy or control dogs were not included in this study, the prevalence of the CIRD virus infections could not be assessed.

Detection of selected viral pathogens in dogs with canine infectious respiratory disease in Austria

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.

Canine infectious respiratory disease: New insights into the etiology and epidemiology of associated pathogens.

Canine infectious respiratory disease (CIRD) is a syndrome where multiple viral and bacterial pathogens are involved sequentially or synergistically to cause illness. There is limited information regarding the prevalence of pathogens related to CIRD in the United States as well as the role of co-infections in the pathogenesis of the syndrome. We aimed to conduct a comprehensive etiologic and epidemiologic study of multiple CIRD agents in a diverse dog population using molecular methods and statistical modeling analyses. In addition, a novel probe-based multiplex real-time PCR was developed to simultaneously detect and differentiate two species of Mycoplasma (M. canis and M. cynos). Canine adenovirus, canine distemper virus, canine parainfluenza virus, coronavirus, influenza A virus (H3N2 and H3N8), Bordetella bronchiseptica, M. canis, M. cynos and Streptococcus equi subsp. zooepidemicus were investigated in specimens from clinically ill and asymptomatic dogs received at the Athens Veterinary Diagnostic Laboratory. Results showed low occurrence of classical CIRD agents such as B. bronchiseptica, canine adenovirus and distemper virus, while highlighting the potential role of emerging bacteria such as M. canis and M. cynos. Statistical modeling analyses of CIRD pathogens emphasized the impact of co-infections on the severity of clinical presentation, and showed that host factors, such as animal age, are the most important predictors of disease severity. This study provides new insights into the current understanding of the prevalence and role of co-infections with selected viruses and bacteria in the etiology of CIRD, while underscoring the importance of molecular diagnosis and vaccination against this disease.

FOXA2 depletion leads to mucus hypersecretion in canine airways with respiratory diseases.

Because of exposure to environmental pollutants, infectious agents, and genetic predisposition, companion animals develop respiratory illnesses similar to those in humans. Older dogs of smaller breeds develop canine infectious respiratory disease, chronic bronchitis, and chronic obstructive pulmonary disease, with chronic lung infection, airway goblet cell hyperplasia and metaplasia, and mucus hypersecretion. Excessive mucus clogs airways, reduces gas exchanges, disables the mucociliary clearance, and reduces drug penetration. The Forkhead box protein A2 (FOXA2) is a key transcriptional regulator that maintains airway mucus homeostasis. Prior studies have shown that FOXA2 expression is frequently depleted in diseased human airways. Unfortunately, FOXA2 depletion has not been examined in dogs. Our current study indicated that both single bacterial infection by Pseudomonas aeruginosa and Bordetella bronchiseptica and polymicrobial infection by viral/bacterial pathogens depleted FOXA2 in canine airways, resulting in goblet cell hyperplasia and metaplasia and excessive mucus production. Furthermore, P.aeruginosa virulence factor pyocyanin activated the antagonistic STAT6 and epidermal growth factor receptor signalling pathways to inhibit FOXA2. Unravelling the mechanism of FOXA2 inactivation will hasten the development of non-antibiotic therapeutics to improve mucociliary clearance of pathogens in canine airway.

European surveillance of emerging pathogens associated with canine infectious respiratory disease

Canine infectious respiratory disease (CIRD) is a major cause of morbidity in dogs worldwide, and is associated with a number of new and emerging pathogens. In a large multi-centre European study the prevalences of four key emerging CIRD pathogens; canine respiratory coronavirus (CRCoV), canine pneumovirus (CnPnV), influenza A, and Mycoplasma cynos (M. cynos); were estimated, and risk factors for exposure, infection and clinical disease were investigated.CIRD affected 66% (381/572) of the dogs studied, including both pet and kennelled dogs. Disease occurrence and severity were significantly reduced in dogs vaccinated against classic CIRD agents, canine distemper virus (CDV), canine adenovirus 2 (CAV-2) and canine parainfluenza virus (CPIV), but substantial proportions (65.7%; 201/306) of vaccinated dogs remained affected.CRCoV and CnPnV were highly prevalent across the different dog populations, with overall seropositivity and detection rates of 47% and 7.7% for CRCoV, and 41.7% and 23.4% for CnPnV, respectively, and their presence was associated with increased occurrence and severity of clinical disease. Antibodies to CRCoV had a protective effect against CRCoV infection and more severe clinical signs of CIRD but antibodies to CnPnV did not.Involvement of M. cynos and influenza A in CIRD was less apparent. Despite 45% of dogs being seropositive for M. cynos, only 0.9% were PCR positive for M. cynos. Only 2.7% of dogs were seropositive for Influenza A, and none were positive by PCR.

Viral molecular and pathological investigations of Canid herpesvirus 1 infection associated respiratory disease and acute death in dogs

Abstract Canid herpesvirus 1 (CaHV-1) is a member of the canine infectious respiratory disease complex (CIRDC). The outcome of CaHV-1 infection can be occasionally fatal. So far, no information on CaHV-1 circulation in Thailand has been reported resulting in a lack of preventive strategies. In this study, nasal (NS) and oropharyngeal (OS) swabs were collected from 100 live dogs with respiratory distress. Among them, 23 pleural effusions were aspirated. A panel of CIRDC-associated viruses was screened by (RT)-PCR, including CaHV-1, CIV, CPIV, CDV, CRCoV and CAdV-2, for all collected samples. The CaHV-1 was detected in 32 dogs. Additionally, CaHV-1 was consistently detected in six pleural effusions. Most CaHV-1 infected dogs were over 5 years of age (43.8%) and expressed a mild nasal discharge. Pathological results of four three-month-old puppies, naturally moribund from respiratory disease, revealed a severe multifocal necrotic-hemorrhagic disease in several organs without pathognomonic inclusion bodies. They were only found to be CaHV-1 positive by PCR. Phylogenetic analysis demonstrated concordant results of CaHV-1 circulation in Thailand. Although mostly found as a co-infection with other CIRDC viruses (68.8%) it also occurred alone. Therefore, rapid ante-mortem diagnosis might facilitate the investigation of unclassical CaHV-1 infection, which is fatal in neonates and causes illness in annually core-vaccinated adults.

Molecular surveillance of traditional and emerging pathogens associated with canine infectious respiratory disease

A molecular survey for traditional and emerging pathogens associated with canine infectious respiratory disease (CIRD) was conducted in Italy between 2011 and 2013 on a total of 138 dogs, including 78 early acute clinically ill CIRD animals, 22 non-clinical but exposed to clinically ill CIRD dogs and 38 CIRD convalescent dogs. The results showed that canine parainfluenza virus (CPIV) was the most commonly detected CIRD pathogen, followed by canine respiratory coronavirus (CRCoV), Bordetella bronchiseptica, Mycoplasma cynos, Mycoplasma canis and canine pneumovirus (CnPnV). Some classical CIRD agents, such as canine adenoviruses, canine distemper virus and canid herpesvirus 1, were not detected at all, as were not other emerging respiratory viruses (canine influenza virus, canine hepacivirus) and bacteria (Streptococcus equi subsp. zooepidemicus). Most severe forms of respiratory disease were observed in the presence of CPIV, CRCoV and M. cynos alone or in combination with other pathogens, whereas single CnPnV or M. canis infections were detected in dogs with no or very mild respiratory signs. Interestingly, only the association of M. cynos (alone or in combination with either CRCoV or M. canis) with severe clinical forms was statistically significant. The study, while confirming CPIV as the main responsible for CIRD occurrence, highlights the increasing role of recently discovered viruses, such as CRCoV and CnPnV, for which effective vaccines are not available in the market.

Development and application of multiplex PCR assays for detection of virus-induced respiratory disease complex in dogs.

Canine infectious respiratory disease complex (CIRDC) viruses have been detected in dogs with respiratory illness. Canine influenza virus (CIV), canine parainfluenza virus (CPIV), canine distemper virus (CDV), canine respiratory coronavirus (CRCoV), canine adenovirus type 2 (CAdV-2) and canine herpesvirus 1 (CaHV-1), are all associated with the CIRDC. To allow diagnosis, two conventional multiplex polymerase chain reactions (PCR) were developed to simultaneously identify four RNA and two DNA viruses associated with CIRDC. The two multiplex PCR assays were then validated on 102 respiratory samples collected from 51 dogs with respiratory illness by sensitivity and specificity determination in comparison to conventional simplex PCR and a rapid three-antigen test kit. All six viruses were detected in either individual or multiple infections. The developed multiplex PCR assays had a >87% sensitivity and 100% specificity compared to their simplex counterpart. Compared to the three-antigen test kit, the multiplex PCR assays yielded 100% sensitivity and more than 83% specificity for detection of CAdV-2 and CDV, but not for CIV. Therefore, the developed multiplex PCR modalities were able to simultaneously diagnose a panel of CIRDC viruses and facilitated specimen collection through being suitable for use of nasal or oral samples.