In Canada and the USA alike, Senecavirus A is a challenge for producers and veterinarians because of its clinical similarity to Food and Mouth Disease (FMD). Indeed, Senecavirus A, is a causative agent of swine vesicular disease with lesions developing on the snout, around the mouth and on the coronary band of the feet. Therefore, being able to differentiate Senecavirus A infections from FMD rapidly is of utmost importance to be able to take the appropriate measures.
In the past months, several diagnostic tests have been developed at the University of Minnesota to detect antibodies against Senecavirus A. The difference between those tests and the in situ hybridization (ISH) described here is that ISH targets the genetic material included in the viral particle and marks it as a red spot as can be seen on the figure below. This advantage of this method is to be able to locate the virus and gives additional information to researcher wanting to study the behavior of Senecavirus A in the body of the pig.
Seneca Valley virus (SVV) is the causative agent of an emerging vesicular disease in swine, which is clinically indistinguishable from other vesicular diseases such as foot-and-mouth disease. In addition, SVV has been associated with neonatal mortality in piglets. While a commercial SVV qRT-PCR is available, commercial antibodies are lacking to diagnose SVV infections by immunohistochemistry (IHC). Thus, a novel in situ hybridization technique—RNAscope (ISH) was developed to detect SVVRNA in infected tissues. From a total of 78 samples evaluated, 30 were positive by qRT-PCR and ISH-RNA, including vesicular lesions of affected sows, ulcerative lesions in the tongue of piglets and various other tissues with no evidence of histological lesions. Nineteen samples were negative for SVV by qRT-PCR and ISH-RNA. The Ct values of the qRT-PCR from ISH-RNA positive tissues varied from 12.0 to 32.6 (5.12 x 106 to 5.31 RNA copies/g, respectively). The ISH-RNA technique is an important tool in diagnosing and investigating the pathogenesis of SVV and other emerging pathogens.
Molecular characterization tools such as p146 sequencing for Mycoplasma hyopneumoniae (M. hyopneumoniae) can provide insight towards investigating elimination failures or new introductions within swine herds.
Porcine Epidemic Diarrhea virus is highly contagious.
The 2013 Porcine Epidemic Diarrhea virus’ (PEDV) outbreak in the USA taught the swine industry that the virus is highly contagious. This event forced producers and veterinarians to review and upgrade their biosecurity procedures.
Drs. Torremorell, Cheeran, and Goyal from the University of Minnesota evaluated some of these measures and how they can prevent PEDV transmission.
Changing Personal Protective Equipment (PPE) and showering before entering a new room prevented contamination.
Among the measures included in this study were the use and change of PPE as well as showering in and out of a facility. In the low biosecurity setting, personnel went from a room with PEDV positive pig straight to a room with naive pigs, contaminating them after the very first movement. In the medium biosecurity setting, personnel washed their hands and face and change their PPE before being in contact with the naive pigs. In this situation, pigs stayed negative for PEDV but two personnel hair/face swabs came back positive for viral genetic material. On the contrary, personnel showered before getting in contact with the high biosecurity group. Those pigs as well as all personnel tests remained negative for PEDV during the study.
Rotaviruses are responsible for increased mortality in neonatal swine populations. They are different genetically and more studies are needed to characterize their diversity. This is the objective of this study coordinated by Dr. Marthaler’s lab focusing on rotaviruses strains found in Canada.
Viral proteins 7 and 4 are used for rotavirus A classification.
Rotaviruses are classified based on two viral proteins (VP) found on their outer capsid called respectively VP7 and VP4. Those two proteins are also essential to induce an efficient immune response against the virus. This project characterized VP7 and VP4 sequences in 136 Canadian samples and compared them with the strains used in a rotavirus commercial vaccine.
The VP7 (n=32) and partial VP4 (n=25) were analyzed, identifying the G3P, G5P, G5P[x], G9P, G9P, G9P, and G9P[x] genotypes.
Minimal differences in the antigenic epitopes for the G5, G9, and P strains were identified.
Major differences in the antigenic epitopes of the G3, P, and P may question the effectiveness of the ProSystems RCE RVA.
Surveillance of Rotavirus A (RVA) infections in North America swine populations are limited and not performed over a significant time period to properly assess the diversity of RVA strains in swine. The VP7 (G) and VP4 (P) genes of 32 Canadian RVA strains, circulating between 2009 and 2015 were sequenced, identifying the G3P, G5P, G9P, G9, and G9 genotype combinations. The Canadian RVA strains were compared to the RVA strains present in the swine ProSystems RCE rotavirus vaccine. The comparison revealed multiple amino acid differences in the G and P antigenic epitopes, regardless of the G and P genotypes but specifically in the Canadian G3, P and P genotypes. Our study further contributes to the characterization of RVA’s evolution and disease mitigation among swine, which may optimize target vaccine design, thereby minimizing RVA disease in this economically important animal population.
Mycoplasma hyopneumoniae is the causative agent enzootic pneumonia, an economically significant disease in pigs. In this study published by Drs. Pieters and Rovira from the University of Minnesota, pigs experimentally inoculated with M.hyopneumoniae were sampled 0, 2, 5, 9, 14, 21, and 28 post-inoculation.
Different sample types were compared:
Using different diagnostic tests:
ELISA IgG anti M.hyopneumoniae
ELISA Ig M anti M.hyopneumoniae
ELISA C-reactive protein
Laryngeal swab samples tested by PCR were highly sensitive for detection of Mycoplasma hyopneumoniae in live pigs.
Various commercial ELISA kits for detection of Mycoplasma hyopneumoniae antibodies showed similar sensitivity.
Oral fluids showed a low sensitivity for detection of Mycoplasma hyopneumoniae in experimentally infected pigs.
Detection of Mycoplasma hyopneumoniae in live pigs during the early stages of infection is critical for timely implementation of control measures, but is technically challenging. This study compared the sensitivity of various sample types and diagnostic methods for detection of M. hyopneumoniae during the first 28 days after experimental exposure. Twenty-one 8-week old pigs were intra-tracheally inoculated on day 0 with M. hyopneumoniae strain 232. Two age matched pigs were mock inoculated and maintained as negative controls. On post-inoculation days 0, 2, 5, 9, 14, 21 and 28, nasal swabs, laryngeal swabs, tracheobronchial lavage fluid, and blood samples were obtained from each pig and oral fluid samples were obtained from each room in which pigs were housed. Serum samples were assayed by ELISA for IgM and IgG M. hyopneumoniae antibodies and C-reactive protein. All other samples were tested for M. hyopneumoniae DNA by species-specific real-time PCR. Serum antibodies (IgG) to M. hyopneumoniae were detected in challenge-inoculated pigs on days 21 and 28. M. hyopneumoniae DNA was detected in samples from experimentally inoculated pigs beginning at 5 days post-inoculation. Laryngeal swabs at all samplings beginning on day 5 showed the highest sensitivity for M. hyopneumoniae DNA Detection, while oral fluids showed the lowest sensitivity. Although laryngeal swabs are not considered the typical M. hyopneumoniae diagnostic sample, under the conditions of this study laryngeal swabs tested by PCR proved to be a practical and reliable diagnostic sample for M. hyopneumoniae detection in vivo during early-stage infection.