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Khurana S.,U.S. Food and Drug Administration | Loving C.L.,U.S. Department of Agriculture | Manischewitz J.,U.S. Food and Drug Administration | King L.R.,U.S. Food and Drug Administration | And 3 more authors.
Science Translational Medicine | Year: 2013

Vaccine-induced disease enhancement has been described in connection with several viral vaccines in animal models and in humans. We investigated a swinemodel to evaluatemismatched influenza vaccine-associated enhanced respiratory disease (VAERD) after pH1N1 infection. Vaccinating pigs with whole inactivated H1N2 (human-like) virus vaccine (WIV-H1N2) resulted in enhanced pneumonia and disease after pH1N1 infection. WIV-H1N2 immune sera contained high titers of cross-reactive anti-pH1N1 hemagglutinin (HA) antibodies that bound exclusively to the HA2 domain but not to the HA1 globular head. No hemagglutination inhibition titers against pH1N1 (challenge virus) were measured. Epitope mapping using phage display library identified the immunodominant epitope recognized by WIV-H1N2 immune sera as amino acids 32 to 77 of pH1N1-HA2 domain, close to the fusion peptide. These crossreactive anti-HA2 antibodies enhanced pH1N1 infection of Madin-Darby canine kidney cells by promoting virus membrane fusion activity. The enhanced fusion activity correlated with lung pathology in pigs. This study suggests a role for fusion-enhancing anti-HA2 antibodies in VAERD, in the absence of receptor-blocking virus-neutralizing antibodies. These findings should be considered during the evaluation of universal influenza vaccines designed to elicit HA2 stem-targeting antibodies. Copyright © 2013 by the American Association for the Advancement of Science.


Henningson J.N.,Kansas State Veterinary Diagnostic Laboratory | Prickett J.,Carthage Veterinary Service | Gauger P.C.,Iowa State University | Hause B.M.,Kansas State Veterinary Diagnostic Laboratory
Journal of General Virology | Year: 2016

Porcine parainfluenza virus 1 (PPIV1) was first identified in 2013 in slaughterhouse pigs in Hong Kong, China. Here, two near-complete genomes were assembled from swine exhibiting acute respiratory disease that were 90.0-95.3 % identical to Chinese PPIV1. Analysis of the HN gene from ten additional PPIV1-positive samples found 85.0-95.5 % identity, suggesting genetic diversity between strains. Molecular analysis identified 17 out of 279 (6.1 %) positive samples from pigs with respiratory disease. Eleven nursery pigs from a naturally infected herd were asymptomatic; however, nasal swabs from six pigs and the lungs of a single pig were quantitative reverse transcriptase (qRT)-PCR positive. Histopathology identified PPIV1 RNA in the nasal respiratory epithelium and trachea. Two serological assays demonstrated seroconversion of infected pigs and further analysis of 59 swine serum samples found 52.5 % and 66.1 % seropositivity, respectively. Taken together, the results confirm the widespread presence of PPIV1 in the US swine herd. © 2016 The Authors.


News Article | December 2, 2015
Site: phys.org

The researchers identified the virus as a member of the aptly name pestivirus family. A sample submitted to the lab by a veterinarian in North Carolina came from a swine herd where uncontrollable shaking, or intention tremors, was observed and resulted in the death of nearly 700 pigs. Virus symptoms included tremors, said Benjamin Hause, a clinical assistant professor in the university's College of Veterinary Medicine. "The veterinarian described the tremors as similar to those seen with Parkinson's disease in humans—but more severe," Hause said. "A recent report described pigs born with congenital tremors caused by a novel pestivirus similar to one we identified this past summer, but this current situation involved older pigs with disease onset from 5 to 14 weeks of age. This has a significant economic impact, especially in a situation like this where 700 animals had died." Historically, the pestivirus family has been associated with important livestock diseases such as bovine viral diarrhea in cattle, border disease in sheep and classical swine fever virus. Pestivirus infections of pigs have caused a wide range of clinical symptoms, including neurological disease Hause's research sheds new light on the pathology associated with this newly described virus. Earlier this year, Hause and colleagues at the Kansas State Veterinary Diagnostic Laboratory identified and characterized a proposed new species of pestivirus, named atypical porcine pestivirus or APPV. It was identified through metagenomic sequencing of swine serum samples submitted to the diagnostic lab. This work, "Discovery of a novel putative atypical porcine pestivirus in pigs in the USA," was published in the Journal of General Virology in October. "While we identified a novel, highly divergent pestivirus in swine samples and went on to show that the virus was widely distributed in the U.S. swine herd, we had no idea what, if any, disease was caused by this virus," Hause said. The lab tested samples submitted from two separate outbreaks of grower pigs with uncontrolled tremors in North Carolina. "While we do not know how often this virus causes clinical disease, I have spoken to other veterinarians who have reported seeing similar symptoms in herds they care for," Hause said. "We're hoping the diagnostic tests that we've developed to detect this virus lay the groundwork to improving our understanding of disease caused by APPV." The Kansas State Veterinary Diagnostic Laboratory has developed quantitative reverse transcript PCR and immunohistochemistry assays to detect APPV in swine samples. Explore further: Researchers hope new tests will prevent enteric disease in pork industry More information: Lalitha Peddireddi et al. Discovery of a novel putative atypical porcine pestivirus in pigs in the USA, Journal of General Virology (2015). DOI: 10.1099/jgv.0.000251


Massilamany C.,University of Nebraska - Lincoln | Mohammed A.,University of Nebraska Medical Center | Loy J.D.,University of Nebraska - Lincoln | Purvis T.,Kansas State Veterinary Diagnostic Laboratory | And 8 more authors.
BMC Genomics | Year: 2016

Background: We recently reported the identification of Bacillus sp. NRRL B-14911 that induces heart autoimmunity by generating cardiac-reactive T cells through molecular mimicry. This marine bacterium was originally isolated from the Gulf of Mexico, but no associations with human diseases were reported. Therefore, to characterize its biological and medical significance, we sought to determine and analyze the complete genome sequence of Bacillus sp. NRRL B-14911. Results: Based on the phylogenetic analysis of 16S ribosomal RNA (rRNA) genes, sequence analysis of the 16S-23S rDNA intergenic transcribed spacers, phenotypic microarray, and matrix-assisted laser desorption ionization time-of-flight mass spectrometry, we propose that this organism belongs to the species Bacillus infantis, previously shown to be associated with sepsis in a newborn child. Analysis of the complete genome of Bacillus sp. NRRL B-14911 revealed several virulence factors including adhesins, invasins, colonization factors, siderophores and transporters. Likewise, the bacterial genome encodes a wide range of methyl transferases, transporters, enzymatic and biochemical pathways, and insertion sequence elements that are distinct from other closely related bacilli. Conclusions: The complete genome sequence of Bacillus sp. NRRL B-14911 provided in this study may facilitate genetic manipulations to assess gene functions associated with bacterial survival and virulence. Additionally, this bacterium may serve as a useful tool to establish a disease model that permits systematic analysis of autoimmune events in various susceptible rodent strains. © 2016 The Author(s).


Palinski R.M.,800 Denison Avenue | Mitra N.,Kansas State Veterinary Diagnostic Laboratory | Hause B.M.,800 Denison Avenue | Hause B.M.,Kansas State Veterinary Diagnostic Laboratory
Virus Genes | Year: 2016

Parvoviruses are a diverse group of viruses containing some of the smallest known species that are capable of infecting a wide range of animals. Metagenomic sequencing of pooled rectal swabs from adult pigs identified a 4103-bp contig consisting of two major open reading frames encoding proteins of 672 and 469 amino acids (aa) in length. BLASTP analysis of the 672-aa protein found 42.4 % identity to fruit bat (Eidolon helvum) parvovirus 2 (EhPV2) and 37.9 % to turkey parvovirus (TuPV) TP1-2012/HUN NS1 proteins. The 469-aa protein had no significant similarity to known proteins. Genetic and phylogenetic analyses suggest that PPV7, EhPV2, and TuPV represent a novel genus in the family Parvoviridae. Quantitative PCR screening of 182 porcine diagnostic samples found a total of 16 positives (8.6 %). Together, these data suggest that PPV7 is a highly divergent novel parvovirus prevalent within the US swine. © 2016 Springer Science+Business Media New York


PubMed | Breeding and Animal Health Unit, University of Nebraska - Lincoln, University of Nebraska Medical Center and Kansas State Veterinary Diagnostic Laboratory
Type: | Journal: BMC genomics | Year: 2016

We recently reported the identification of Bacillus sp. NRRL B-14911 that induces heart autoimmunity by generating cardiac-reactive T cells through molecular mimicry. This marine bacterium was originally isolated from the Gulf of Mexico, but no associations with human diseases were reported. Therefore, to characterize its biological and medical significance, we sought to determine and analyze the complete genome sequence of Bacillus sp. NRRL B-14911.Based on the phylogenetic analysis of 16S ribosomal RNA (rRNA) genes, sequence analysis of the 16S-23S rDNA intergenic transcribed spacers, phenotypic microarray, and matrix-assisted laser desorption ionization time-of-flight mass spectrometry, we propose that this organism belongs to the species Bacillus infantis, previously shown to be associated with sepsis in a newborn child. Analysis of the complete genome of Bacillus sp. NRRL B-14911 revealed several virulence factors including adhesins, invasins, colonization factors, siderophores and transporters. Likewise, the bacterial genome encodes a wide range of methyl transferases, transporters, enzymatic and biochemical pathways, and insertion sequence elements that are distinct from other closely related bacilli.The complete genome sequence of Bacillus sp. NRRL B-14911 provided in this study may facilitate genetic manipulations to assess gene functions associated with bacterial survival and virulence. Additionally, this bacterium may serve as a useful tool to establish a disease model that permits systematic analysis of autoimmune events in various susceptible rodent strains.


PubMed | 800 Denison Avenue and Kansas State Veterinary Diagnostic Laboratory
Type: Journal Article | Journal: Virus genes | Year: 2016

Parvoviruses are a diverse group of viruses containing some of the smallest known species that are capable of infecting a wide range of animals. Metagenomic sequencing of pooled rectal swabs from adult pigs identified a 4103-bp contig consisting of two major open reading frames encoding proteins of 672 and 469 amino acids (aa) in length. BLASTP analysis of the 672-aa protein found 42.4% identity to fruit bat (Eidolon helvum) parvovirus 2 (EhPV2) and 37.9% to turkey parvovirus (TuPV) TP1-2012/HUN NS1 proteins. The 469-aa protein had no significant similarity to known proteins. Genetic and phylogenetic analyses suggest that PPV7, EhPV2, and TuPV represent a novel genus in the family Parvoviridae. Quantitative PCR screening of 182 porcine diagnostic samples found a total of 16 positives (8.6%). Together, these data suggest that PPV7 is a highly divergent novel parvovirus prevalent within the US swine.


Braucher D.R.,Virus and Prion Research Unit | Braucher D.R.,Boehringer Ingelheim | Henningson J.N.,Virus and Prion Research Unit | Henningson J.N.,Kansas State Veterinary Diagnostic Laboratory | And 6 more authors.
Clinical and Vaccine Immunology | Year: 2012

Influenza A virus (IAV) is widely circulating in the swine population and causes significant economic losses. To combat IAV infection, the swine industry utilizes adjuvanted whole inactivated virus (WIV) vaccines, using a prime-boost strategy. These vaccines can provide sterilizing immunity toward homologous virus but often have limited efficacy against a heterologous infection. There is a need for vaccine platforms that induce mucosal and cell-mediated immunity that is cross-reactive to heterologous viruses and can be produced in a short time frame. Nonreplicating adenovirus 5 vector (Ad5) vaccines are one option, as they can be produced rapidly and given intranasally to induce local immunity. Thus, we compared the immunogenicity and efficacy of a single intranasal dose of an Ad5-vectored hemagglutinin (Ad5-HA) vaccine to those of a traditional intramuscular administration of WIV vaccine. Ad5-HA vaccination induced a mucosal IgA response toward homologous IAV and primed an antigen-specific gamma interferon (IFN-γ) response against both challenge viruses. The Ad5-HA vaccine provided protective immunity to homologous challenge and partial protection against heterologous challenge, unlike the WIV vaccine. Nasal shedding was significantly reduced and virus was cleared from the lung by day 5 postinfection following heterologous challenge of Ad5-HA-vaccinated pigs. However, the WIV-vaccinated pigs displayed vaccine-associated enhanced respiratory disease (VAERD) following heterologous challenge, characterized by enhanced macroscopic lung lesions. This study demonstrates that a single intranasal vaccination with an Ad5-HA construct can provide complete protection from homologous challenge and partial protection from heterologous challenge, as opposed to VAERD, which can occur with adjuvanted WIV vaccines. Copyright © 2012, American Society for Microbiology. All Rights Reserved.


Jacob M.E.,Kansas State University | Almes K.M.,Kansas State Veterinary Diagnostic Laboratory | Shi X.,Kansas State University | Sargeant J.M.,University of Guelph | Nagaraja T.G.,Kansas State University
Journal of Food Protection | Year: 2011

Escherichia coli O157:H7 causes foodborne illness in humans; cattle are considered a primary reservoir for the organism, and transmission is often through contaminated food products or water. The objective of this study was to determine the genetic diversity of E. coli O157:H7 within a single individual bovine fecal sample based on pulsed-field gel electrophoresis (PFGE) typing. Fecal samples (n=01) were collected from dairy and beef cattle at three separate facilities, and E. coli O157:H7 was isolated by enrichment, immunomagnetic separation, and plating on selective medium. The prevalence of E. coli O157:H7 was 46 (7.7%) of 601. From each positive fecal sample, up to 10 putative colonies were tested, and isolates from samples with at least seven positive colonies were subtyped using PFGE and tested for six major virulence genes by multiplex PCR. A total of 254 E. coli O157:H7 isolates from 27 samples met these criteria and were included in PFGE analysis. Fifteen PFGE subtypes (lt;% Dice similarity) were detected among the 254 isolates, and there were no common subtypes between the three locations. Seven (26%) of 27 fecal samples had E. coli O157:H7 isolates with different PFGE subtypes (mean=.1) within the same sample. The virulence gene profiles of different isolates from the same sample were always identical, regardless of the number of PFGE types. The results of this study suggest that determining the PFGE pattern of a single isolate from a bovine sample may not be sufficient when comparing isolates from feces, hides, or carcasses, because multiple PFGE subtypes are present. © International Association for Food Protection.


Dargatz D.A.,United Health Centers | Bai J.,Kansas State Veterinary Diagnostic Laboratory | Lubbers B.V.,Kansas State Veterinary Diagnostic Laboratory | Kopral C.A.,United Health Centers | And 2 more authors.
Foodborne Pathogens and Disease | Year: 2013

While efforts to control foodborne illness associated with the Shiga toxin-producing Escherichia coli (E. coli) O157 through processes and procedures implemented at harvest facilities have been very successful, there is concern about the burden of illness associated with other Shiga toxin-producing E. coli. The U.S. Department of Agriculture Food Safety and Inspection Service announced plans to classify an additional six non-O157 Shiga toxin-producing E. coli as adulterants. Little is known about the prevalence and distribution of these E. coli in the animal production environment. An investigation of the prevalence of O157 and the six major non-O157 E. coli serogroups was conducted in 21 feedlots over the period July 2011 to October 2011. Individual fecal swabs were collected from cattle approximately 60 days after their arrival in the feedlot and were pooled for evaluation using a polymerase chain reaction assay to identify the presence of seven E. coli O-types (O157, O45, O103, O121, O145, O26, and O111) and four virulence genes (stx1, stx2, eaeA, and ehxA). Overall, 1145 fecal pools were evaluated, with 506 (44.2%) being positive for one or more of the E. coli O-serogroups. The pool prevalences for E. coli O157, O45, O26, O103, O121, O145, and O111 were 19.7%, 13.8%, 9.9%, 9.3%, 5.5%, 1.1%, and 0.5%, respectively. Nearly all pools were positive for ehxA (99.7%) or stx2 (98.6%). The pool level prevalence for stx1 and eae was 65.5% and 69.3%, respectively. Pools that were positive for one or more of the other E. coli O-serogroups were 1.37 times more likely to be positive for E. coli O157. Conversely, pools that were positive for E. coli O157 were 1.43 times more likely to be positive for at least one of the other E. coli O-serogroups evaluated. These data will be useful to understand the expected prevalence of potential Shiga toxin-producing E. coli in cattle feedlots. © 2013 Mary Ann Liebert, Inc.

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