Immunization for Influenza A Virus by Intranasal Administration of Alphavirus Replicon Particles

Improved vaccines are necessary to prevent swine influenza, especially in young growing pigs. The objective of this study is to determine whether intranasal vaccination with Alphavirus replicon particle (RP) vector vaccine prevents influenza A virus (IAV) in pigs. RP vaccine was prepared with the hemagglutinin (HA) gene of pandemic H1N1 influenza virus (A/California/04/2009, pH1N1). The efficacy of intranasal (IN) administration with pH1N1 HA RP was evaluated in two pig experiments. In the first experiment, prime/boost RP vaccination was administered IN/IN to pigs. In the second experiment, pigs were administered a one dose intramuscular (IM) or IN HA RP vaccine, or with a combination of IN/IM routes with an interval of three weeks. Results showed that two doses IN administration of HA RP did not protect pigs against IAV; one dose IM and combination IN/IM routes vaccination with HA RP reduced pneumonia significantly and partially inhibited virus shedding following homologous challenge

Effects of Label Dose Permethrin Use in Yearling Angus Beef Bulls on Reproductive Function and Testicular Histopathology

To eliminate the potential for insect borne diseases and improve productivity in cow-calf operations, many producers use pour-on pyrethroids. However, popular press literature has identified potential links between use of pyrethroids and reproductive function failures in bulls. While literature in mice and rats have reported potential endocrine disruption of sex steroids resulting from pyrethroid exposure, effects of pyrethroid use on bull fertility is still questionable. The objective of this study was to measure reproductive parameters in peripubertal Angus bulls using a commonly used pyrethroid pour-on. We hypothesized that use of a pyrethroid pour-on at labeled dose would have limited effects on semen and testicular characteristics. Results from this study revealed pyrethroid-treated bulls had greater spermatozoa head and midpiece abnormalities compared to controls, resulting in higher primary morphological abnormalities. Although some morphological semen parameters appear to be negatively affected by use of pyrethroid administration in bulls, biological relevance of this result needs to be further elucidated as the ability to pass a breeding soundness exam was not affected. Moreover, further research should be conducted to determine the effects of pyrethroid pour-ons on fertility when used with other pyrethroids such fly tags or perimeter sprays. Based on these results, pour-on pyrethroid use in yearling bulls is still recommended for label dose administration

Infectivity of porcine circovirus type 2 DNA in semen from experimentally-infected boars

Porcine circovirus type 2 (PCV2) is an economically important pathogen. It has been demonstrated that PCV2 DNA can be detected in boar semen by PCR; however, the biological relevance of this is unknown. The objectives of this study were to determine if semen positive for PCV2 DNA is infectious (1) in a swine bioassay, or (2) when used for artificial insemination. For the first objective, 4-week-old pigs were inoculated intraperitoneally with PCV2 DNA-negative (bioassay-control; n = 3), PCV2a DNA-positive (bioassay-PCV2a; n = 3), or PCV2b DNA-positive (bioassay-PCV2b; n = 3) raw semen, or PCV2 live virus (bioassay-positive; n = 3), respectively. Pigs inoculated with PCV2 DNA-positive semen and PCV2 live virus became viremic and developed anti-PCV2 antibodies indicating that the PCV2 DNA present in semen was infectious. For the second objective, three Landrace gilts were inseminated with PCV2 DNA-negative semen (gilts-controls) from experimentally-infected boars, and six gilts were artificially inseminated with semen positive for PCV2a DNA (gilts-PCV2a; n = 3) or PCV2b DNA (gilts-PCV2b; n = 3). Serum samples collected from the gilts in all groups remained negative for anti-PCV2 antibodies for the duration of the experiment. In addition, fetal serum samples from all 105-day-gestation fetuses were negative for anti-PCV2 antibodies or PCV2 DNA. Under the conditions of this study, PCV2 DNA-positive semen was not infectious when used to artificially inseminate gilts; however, it was demonstrated to be infectious in a swine bioassay model and therefore is a potential means of PCV2 transmission amongst swine herds

Difference in severity of porcine circovirus type two-induced pathological lesions between Landrace and Pietrain pigs

Anecdotal information from the field suggests that there are host genetic differences in susceptibility to porcine circovirus type 2 (PCV2) associated disease among Landrace and Pietrain breeds. The objective of this study was to determine if a difference exists in PCV2 susceptibility between Landrace and Pi-etrain pigs under experimental conditions. Thirty-nine Landrace pigs and 39 Pietrain pigs were blocked by breed, sire, dam, and litter and randomly divided into the following 4 groups: Landrace noninoculated negative control (Landrace-NEG; n = 13), Pietrain noninoculated negative control (Pietrain-NEG; n = 13), Landrace-PCV2 (n = 26; Landrace), and Pietrain-PCV2 (n = 26; Pietrain). After waning of passively acquired anti-PCV2 antibodies, Landrace-PCV2 and Pietrain-PCV2 groups were inoculated with PCV2 isolate ISU-40895. The Landrace-NEG and Pietrain-NEG groups were housed in a separate room, remained noninoculated, and served as negative controls. All pigs in all groups were necropsied at 21 d post PCV2-inoculation. Onset of seroconversion and concentrations of anti-PCV2-IgM, anti-PCV2-IgG, and anti-PCV2 neutralizing antibodies were similar in Landrace-PCV2 and Pietrain-PCV2 groups. Furthermore, the amount of PCV2 DNA and cytokine concentrations in serum and plasma samples were not different between the 2 PCV2-inoculated groups. The severity of PCV2-associated microscopic lesions was different between Landrace and Pietrain pigs; Landrace-PCV2 pigs had significantly (P \u3c 0.05) more severe lymphoid lesions than the Pietrain-PCV2 pigs. Although the pigs originated from the same farm where their dams were commingled, passively acquired anti-PCV2-antibodies waned in Pietrain pigs by approximately 12 wk of age, whereas the majority of the Landrace pigs remained PCV2 seropositive until 18 wk of age and beyond. The results from this study indicate that a genetic difference exists between these 2 breeds of pigs in susceptibility to PCV2-associated lesions

Two clinical isolates of \u3ci\u3eMycoplasma hyosynoviae\u3c/i\u3e showed differing pattern of lameness and pathogen detection in experimentally challenged pigs

Mycoplasma (M.) hyosynoviae is known to colonize and cause disease in growing-finishing pigs. In this study, two clinical isolates of M. hyosynoviae were compared by inoculating cesarean-derived colostrum-deprived and specific-pathogen-free growing pigs. After intranasal or intravenous inoculation, the proportion and distribution pattern of clinical cases was compared in addition to the severity of lameness. Tonsils were found to be the primary site of colonization, while bacteremia was rarely detected prior to the observation of clinical signs. Regardless of the clinical isolate, route of inoculation, or volume of inocula, histopathological alterations and tissue invasion were detected in multiple joints, indicating an apparent lack of specific joint tropism. Acute disease was primarily observed 7 to 10 days post-inoculation. The variability in the severity of synovial microscopic lesions and pathogen detection in joint cavities suggests that the duration of joint infection may influence the diagnostic accuracy. In summary, these findings demonstrate that diagnosis of M. hyosynoviae-associated arthritis can be influenced by the clinical isolate, and provides a study platform to investigate the colonization and virulence potential of field isolates. This approach can be particularly relevant to auxiliate in surveillance and testing of therapeutic and/or vaccine candidates

Investigation of the Impact of Increased Dietary Insoluble Fiber through the Feeding of Distillers Dried Grains with Solubles (DDGS) on the Incidence and Severity of Brachyspira-Associated Colitis in Pigs

Diet has been implicated as a major factor impacting clinical disease expression of swine dysentery and Brachyspira hyodysenteriae colonization. However, the impact of diet on novel pathogenic strongly beta-hemolytic Brachyspira spp. including “B. hampsonii” has yet to be investigated. In recent years, distillers dried grains with solubles (DDGS), a source of insoluble dietary fiber, has been increasingly included in diets of swine. A randomized complete block experiment was used to examine the effect of increased dietary fiber through the feeding of DDGS on the incidence of Brachyspira-associated colitis in pigs. One hundred 4-week-old pigs were divided into five groups based upon inocula (negative control, Brachyspira intermedia,Brachyspira pilosicoli, B. hyodysenteriae or “B. hampsonii”) and fed one of two diets containing no (diet 1) or 30% (diet 2) DDGS. The average days to first positive culture and days post inoculation to the onset of clinical dysentery in the B. hyodysenteriae groups was significantly shorter for diet 2 when compared to diet 1 (P = 0.04 and P = 0.0009, respectively). A similar difference in the average days to first positive culture and days post inoculation to the onset of clinical dysentery was found when comparing the “B. hampsonii” groups. In this study, pigs receiving 30% DDGS shed on average one day prior to and developed swine dysentery nearly twice as fast as pigs receiving 0% DDGS. Accordingly, these data suggest a reduction in insoluble fiber through reducing or eliminating DDGS in swine rations should be considered an integral part of any effective disease elimination strategy for swine dysentery

Evaluation of serological cross-reactivity and cross-neutralization between the United States porcine epidemic diarrhea virus prototype and S-INDEL-variant strains

BACKGROUND: At least two genetically different porcine epidemic diarrhea virus (PEDV) strains have been identified in the United States (U.S. PEDV prototype and S-INDEL-variant strains). The current serological assays offered at veterinary diagnostic laboratories for detection of PEDV-specific antibody are based on the U.S. PEDV prototype strain. The objectives of this study were: 1) isolate the U.S. PEDV S-INDEL-variant strain in cell culture; 2) generate antisera against the U.S. PEDV prototype and S-INDEL-variant strains by experimentally infecting weaned pigs; 3) determine if the various PEDV serological assays could detect antibodies against the U.S. PEDV S-INDEL-variant strain and vice versa. RESULTS: A U.S. PEDV S-INDEL-variant strain was isolated in cell culture in this study. Three groups of PEDV-negative, 3-week-old pigs (five pigs per group) were inoculated orally with a U.S. PEDV prototype isolate (previously isolated in our lab), an S-INDEL-variant isolate or virus-negative culture medium. Serum samples collected at 0, 7, 14, 21 and 28 days post inoculation were evaluated by the following PEDV serological assays: 1) indirect fluorescent antibody (IFA) assays using the prototype and S-INDEL-variant strains as indicator viruses; 2) virus neutralization (VN) tests against the prototype and S-INDEL-variant viruses; 3) PEDV prototype strain whole virus based ELISA; 4) PEDV prototype strain S1-based ELISA; and 5) PEDV S-INDEL-variant strain S1-based ELISA. The positive antisera against the prototype strain reacted to and neutralized both prototype and S-INDEL-variant viruses, and the positive antisera against the S-INDEL-variant strain also reacted to and neutralized both prototype and S-INDEL-variant viruses, as examined by IFA antibody assays and VN tests. Antibodies against the two PEDV strains could be detected by all three ELISAs although detection rates varied to some degree. CONCLUSIONS: These data indicate that the antibodies against U.S. PEDV prototype and S-INDEL-variant strains cross-reacted and cross-neutralized both strains in vitro. The current serological assays based on U.S. PEDV prototype strain can detect antibodies against both U.S. PEDV strains

Porcine circovirus type 2 (PCV2)-infection and re-inoculation with homologous or heterologous strains: virological, serological, pathological and clinical effects in growing pigs.

Long-term PCV2 infection and/or concurrent infection with genotypes PCV2a and PCV2b may play a role in the development of clinical porcine circovirus-associated disease (PCVAD). To evaluate this premise, 24 11-week-old specific pathogen-free (SPF) pigs were randomly assigned to 1 of 4 treatments: negative controls, a single inoculation with PCV2a, single inoculation followed by re-inoculation with a homologous PCV2a strain, or repeated inoculations with heterologous strains (PCV2a, PCV2b). Pigs were evaluated for clinical signs daily through 140 days post inoculation (dpi). Serum samples were collected every other day from dpi 0 through 14 and weekly thereafter. PCV2-inoculated pigs were viremic by dpi 2 and 13 of 18 pigs remained viremic at 140 dpi. No statistical differences in the onset, level, or duration of PCV2 viremia were detected among treatment groups. Anti-PCV2 antibodies were detected between 14 and 28 dpi and were present through 140 dpi without statistical differences in antibody response among treatment groups. In the current study, pigs had extended viremia combined with detectable tissue PCV2 antigen levels despite the presence of high levels of anti-PCV2 antibody; however, no clinical disease was observed

First retrospective studies with etiological confirmation of porcine transmissible gastroenteritis virus infection in Argentina

Background: In 2014, a notification of porcine transmissible gastroenteritis virus (TGEV) was made by the National Services of Animal Health of Argentina (SENASA) to the World Organization of Animal Health (OIE). The notification was based on a serological diagnosis in a small farm with a morbidity rate of 2.3% without enteric clinical signs. In order to determine if TGEV was circulating before the official report, a retrospective study on cases of neonatal diarrhea was performed. The selection criteria was a sudden increase in mortality in 1- to 21-day-old piglets with watery diarrhea that did not respond to antibiotics. Based on these criteria, three clinical cases were identified during 2010–2015. Results: All animals that were evaluated presented histological lesions consistent with enteric viral infection. The feces and ultrathin sections of intestine that were evaluated by electron microscopy confirmed the presence of round particles of approximately 80 nm in size and characterized by finely granular electrodense nucleoids consistent with complete particles of coronavirus. The presence of the TGEV antigen was confirmed by monoclonal specific immunohistochemistry, and final confirmation of a metabolically-active virus was performed by in situ hybridization to detect a TGE mRNA encoding spike protein. All sections evaluated in this case were negative for PEDV and rotavirus A. Conclusions: This is the first case series describing neonatal mortality with etiological confirmation of TGEV in Argentina. The clinical diagnosis of TGEV infections in endemic regions is challenging due to the epidemiological distribution and coinfection with other enteric pathogens that mask the clinical presentation.Facultad de Ciencias Veterinaria