Deoxynivalenol (DON) naturally contaminated feed impairs the immune response induced by porcine reproductive and respiratory syndrome virus (PRRSV) live attenuated vaccine

Cereal commodities are frequently contaminated with mycotoxins produced by the secondary metabolism of fungal infection. Among these contaminants, deoxynivalenol (DON), also known as vomitoxin, is the most prevalent type B trichothecene mycotoxin worldwide. Pigs are very sensitive to the toxic effects of DON and are frequently exposed to naturally contaminated feed. Recently, DON naturally contaminated feed has been shown to decrease porcine reproductive and respiratory syndrome virus (PRRSV) specific antibody responses following experimental infection. The objective of this study was to determine the impact of DON naturally contaminated feed on the immune response generated following vaccination with PRRSV live attenuated vaccine. Eighteen pigs were randomly divided into three experimental groups of 6 animals based on DON content of the diets (0, 2.5 and 3.5 mg DON/kg). They were fed these rations one week prior to the vaccination and for all the duration of the immune response evaluation. All pigs were vaccinated intra-muscularly with one dose of Ingelvac® PRRSV modified live vaccine (MLV). Blood samples were collected at day −1, 6, 13, 20, 27 and 35 post vaccination (pv) and tested for PRRSV RNA by RT-qPCR and for virus specific antibodies by ELISA. Results showed that ingestion of DON-contaminated diets significantly decreased PRRSV viremia. All pigs fed control diet were viremic while only 1 (17%) and 3 (50%) out of 6 pigs were viremic in the groups receiving 3.5 and 2.5 mg of DON/kg, respectively. Subsequently, all pigs fed control diet developed PRRSV specific antibodies while only viremic pigs that were fed contaminated diets have developed PRRSV specific antibodies. These results suggest that feeding pigs with DON-contaminated diet could inhibit vaccination efficiency of PRRSV MLV by severely impairing viral replication

Postvaccinal reovirus infection with high mortality in breeder chicks

A broiler breeder flock was subcutaneously vaccinated at the hatchery with a live avian orthoreovirus (ARV) vaccine against viral arthritis. Chicks began to die at 3 days of age and postmortem examination revealed massive subcutaneous hemorrhages and edema on the dorsal aspect of the neck at the site of vaccination, a severe necrotic hepatitis, and pulmonary edema. Microscopically, the main lesion was a multifocal vacuolar degeneration and necrosis of randomly distributed small groups of hepatocytes with presence of apoptotic and multinucleated syncytial cells. Necrotic foci were also found in the lungs as well as a hemorrhagic, granulomatous, and heterophilic cellulitis and myositis of the neck and a generalized depletion and lymphocytolysis of lymphoid organs. At 8 days of age, birds also began to show hock swelling histologically characterized by a fibrinoleucocytic inflammation of the articulation and tendon sheaths, with hyperplasia of the synovial membrane, and lymphoplasmocytic infiltration. PCR and viral culture of livers were positive for ARV. Partial sequencing of the S1 gene from the virus isolate showed 99.2% to 99.8% homology with three vaccinal strains (ARV S1133, 1733, and 2408). Viral particles compatible with reovirus virions were observed at transmission electron microscopy. Investigation at the hatchery revealed that chicks were inadvertently administered an S1133 reovirus vaccine labeled for water administration in 10- to 17-week-old chickens. This human error is most likely the reason for this unusually severe viremic reovirus infection that affected this flock at such an early age

Coding-complete genome sequence of a Falcon aviadenovirus A strain associated with necrotizing hepatitis in an American Kestrel (Falco sparverius)

A necropsy was performed on an American kestrel (Falco sparverius) with necrotizing hepatitis associated with inclusion bodies, suggesting an adenovirus infection. A next-generation sequencing assay was conducted on the liver, and the coding-complete genome sequence of a Falcon aviadenovirus A strain was revealed

Porcine reproductive and respiratory syndrome virus whole-genome sequencing efficacy with field clinical samples using a poly(A)-tail viral genome purification method

The genomic surveillance of porcine reproductive and respiratory syndrome virus (PRRSV) is based on sequencing of the ORF5 gene of the virus, which covers only 4% of the entire viral genome. It is expected that PRRSV whole-genome sequencing (WGS) will improve PRRSV genomic data and allow better understanding of clinical discrepancies observed in the field when using ORF5 sequencing. Our main objective was to implement an efficient method for WGS of PRRSV from clinical samples. The viral genome was purified using a poly(A)-tail viral genome purification method and sequenced using Illumina technology. We tested 149 PRRSV-positive samples: 80 sera, 33 lungs, 33 pools of tissues, 2 oral fluids, and 1 processing fluid (i.e., castration liquid). Overall, WGS of 67.1% of PRRSV-positive cases was successful. The viral load, in particular for tissues, had a major impact on the PRRSV WGS success rate. Serum was the most efficient type of sample to conduct PRRSV WGS poly(A)-tail assays, with a success rate of 76.3%, and this result can be explained by improved sequencing reads dispersion matching throughout the entire viral genome. WGS was unsuccessful for all pools of tissue and lung samples with Cq values > 26.5, whereas it could still be successful with sera at Cq ≤ 34.1. Evaluation of results of highly qualified personnel confirmed that laboratory skills could affect PRRSV WGS efficiency. Oral fluid samples seem very promising and merit further investigation because, with only 2 samples of low viral load (Cq = 28.8, 32.8), PRRSV WGS was successful

Whole genome sequencing of porcine reproductive and respiratory syndrome virus 2 (PRRSV) from field clinical samples improves the genomic surveillance of the virus

Porcine reproductive and respiratory syndrome virus (PRRSV) is a major economic concern worldwide. There are currently large data sets available about the ORF5 gene of the virus, with thousands of sequences available, but little data are currently available on the full-length genome of PRRSV. We hypothesized that whole-genome sequencing (WGS) of the PRRSV genome would allow better epidemiological monitoring than ORF5 gene sequencing. PRRSV PCR-positive serum, oral fluid, and tissue clinical samples submitted to the diagnostic laboratory for routine surveillance or diagnosis of PRRSV infection in Québec, Canada, swine herds were used. The PRRSV reverse transcription-quantitative PCR Cq values of the processed samples varied between 11.5 and 34.34. PRRSV strain genomes were isolated using a poly (A)-tail method and were sequenced with a MiSeq Illumina sequencer. Ninetytwo full-length PRRSV genomes were obtained from 88 clinical samples out of 132 tested samples, resulting in a PRRSV WGS success rate of 66.67%. Three important deletions in ORF1a were found in most wild-type (i.e., not vaccine-like) strains. The importance of these deletions remains undetermined. Two different full-length PRRSV genomes were found in four different samples (three serum samples and one pool of tissues), suggesting a 4.55% PRRSV strain coinfection prevalence in swine. Moreover, six PRRSV whole genomes (6.52% of PRRSV strains) were found to cluster differently than they did under the ORF5 classification method. Overall, WGS of PRRSV enables better strain classification and/or interpretation of results in 9.10% of clinical samples than ORF5 sequencing, as well as allowing interesting research avenues

Dual infections of CD163 expressing NPTr epithelial cells with influenza A virus and PRRSV

In the pig, respiratory co-infections involving various pathogens are far more frequent than single infections. Amongst respiratory viruses, swine influenza type A virus (swIAV) and porcine reproductive and respiratory syndrome virus (PRRSV) are frequently associated. Previously, we performed co-infections with swIAV and PRRSV in porcine alveolar macrophages (PAM) and precision cut lung slices (PCLS). With these two approaches it was practically impossible to have co-infections of the same cells as the main target cell of swIAV is the epithelial cell while the main target of PRRSV is the PAM. This constraint makes the study of interference between the two viruses difficult at the cellular level. In the current report, an epithelial cell line expressing, CD163, the main receptor of PRRSV was generated. This cell line receptive for both viruses was used to assess the interference between the two viruses. Results showed that swIAV as well as PRRSV, even if they interacted differently with the modified epithelial cells, were clearly interfering with each other regarding their replication when they infected a same cell with consequences within the cellular antiviral response. Our modified cell line, receptive to both viruses, can be used as a tool to assess interference between swIAV and PRRSV in a same cell as it probably happens in the porcine host

Exposure of feral swine (\u3ci\u3eSus scrofa\u3c/i\u3e) in the United States to selected pathogens

Feral swine (Sus scrofa) are widely distributed in the United States. In 2011 and 2012, serum samples and tonsils were recovered from 162 and 37 feral swine, respectively, in the US to evaluate exposure to important swine endemic pathogens. Antibodies against porcine reproductive and respiratory syndrome virus (PRRSV) and porcine circovirus type 2 (PCV2) were found in 2.5% and 25.3% of tested sera, respectively. Positive serological reactions against Mycoplasma hyopneumoniae and Actinobacillus pleuropneumoniae have been detected in 19.7% and 69.7% of animals. More than 15% of animals presented antibodies against these 2 pathogens simultaneously. Most animals were also seropositive for Lawsonia intracellularis. Feral swine can also be involved in transmission of zoonotic agents. Almost 50% of animals possessed antibodies against Salmonella. In addition, 94.4% of animals were carriers of Streptococcus suis in their tonsils. In conclusion, feral swine may be considered as a potential reservoir for different endemic diseases in domestic pigs, as well as for important zoonotic agents. Les porcs sauvages (Sus scrofa) sont largement distribués aux États-Unis. En 2011 et 2012, aux États-Unis des échantillons de sérum et d’amygdales furent obtenus de 162 et 37 porcs sauvages, respectivement, afin d’évaluer l’exposition à d’importants agents pathogènes porcins endémiques. Des anticorps contre le virus du syndrome reproducteur et respiratoire porcin (VSRRP) et le circovirus porcin de type 2 (CVP2) furent détectés chez 2,5 % et 25,3 % des sérums testés, respectivement. Des réactions sérologiques positives envers Mycoplasma hyopneumoniae et Actinobacillus pleuropneumoniae ont été détectées chez 19,7 % et 69,7 % des animaux. Plus de 15 % des animaux avaient des anticorps contre ces deux agents pathogènes simultanément. La plupart des animaux étaient également séropositifs pour Lawsonia intracellularis. Les porcs sauvages peuvent également être impliqués dans la transmission d’agents zoonotiques. Près de 50 % des animaux avaient des anticorps contre Salmonella. De plus, 94,4 % des animaux étaient porteurs de Streptococcus suis dans leurs amygdales. En conclusion, les porcs sauvages peuvent être considérés comme des réservoirs potentiels de différentes maladies endémiques des porcs domestiques, aussi bien que d’agents zoonotiques importants

Exposure of feral swine (Sus scrofa) in the United States to selected pathogens

Les porcs sauvages (Sus scrofa) sont largement distribués aux États-Unis. En 2011 et 2012, aux États-Unis des échantillons de sérum et d’amygdales furent obtenus de 162 et 37 porcs sauvages, respectivement, afin d’évaluer l’exposition à d’importants agents pathogènes porcins endémiques. Des anticorps contre le virus du syndrome reproducteur et respiratoire porcin (VSRRP) et le circovirus porcin de type 2 (CVP2) furent détectés chez 2,5 % et 25,3 % des sérums testés, respectivement. Des réactions sérologiques positives envers Mycoplasma hyopneumoniae et Actinobacillus pleuropneumoniae ont été détectées chez 19,7 % et 69,7 % des animaux. Plus de 15 % des animaux avaient des anticorps contre ces deux agents pathogènes simultanément. La plupart des animaux étaient également séropositifs pour Lawsonia intracellularis. Les porcs sauvages peuvent également être impliqués dans la transmission d’agents zoonotiques. Près de 50 % des animaux avaient des anticorps contre Salmonella. De plus, 94,4 % des animaux étaient porteurs de Streptococcus suis dans leurs amygdales. En conclusion, les porcs sauvages peuvent être considérés comme des réservoirs potentiels de différentes maladies endémiques des porcs domestiques, aussi bien que d’agents zoonotiques importants.Feral swine (Sus scrofa) are widely distributed in the United States. In 2011 and 2012, serum samples and tonsils were recovered from 162 and 37 feral swine, respectively, in the US to evaluate exposure to important swine endemic pathogens. Antibodies against porcine reproductive and respiratory syndrome virus (PRRSV) and porcine circovirus type 2 (PCV2) were found in 2.5% and 25.3% of tested sera, respectively. Positive serological reactions against Mycoplasma hyopneumoniae and Actinobacillus pleuropneumoniae have been detected in 19.7% and 69.7% of animals. More than 15% of animals presented antibodies against these 2 pathogens simultaneously. Most animals were also seropositive for Lawsonia intracellularis. Feral swine can also be involved in transmission of zoonotic agents. Almost 50% of animals possessed antibodies against Salmonella. In addition, 94.4% of animals were carriers of Streptococcus suis in their tonsils. In conclusion, feral swine may be considered as a potential reservoir for different endemic diseases in domestic pigs, as well as for important zoonotic agents