Richard A. Hesse

A field evaluation of mortality rate and growth performance in pigs vaccinated against porcine circovirus type 2

OBJECTIVE To evaluate, under field conditions, the effects of a commercial porcine circovirus type 2 (PCV2) vaccine on mortality rate and growth performance in a herd infected with PCV2 that had a history of porcine circovirus disease. DESIGN Randomized controlled clinical trial. ANIMALS 485 commercial, cross-bred, growing pigs. PROCEDURES Prior to weaning, pigs were randomly assigned within litter to a vaccination or unvaccinated control group. Pigs in the vaccination group were given a commercial PCV2 vaccine at weaning and 3 weeks later. Mortality rate was recorded, and pigs were weighed prior to vaccination, when moved from the nursery, and prior to marketing. Infection status was assessed by serologic testing and detection of viral DNA in serum. RESULTS Compared with control pigs, pigs vaccinated against PCV2 had a significantly lower mortality rate during the finishing phase, significantly higher average daily gain during the finishing phase, and significantly lower likelihood of being lightweight at the time of marketing. For vaccinated pigs, overall mortality rate was reduced by 50% and average daily gain during the finishing period was increased by 9.3%. At the time of marketing, vaccinated pigs weighed an average of 8.8 kg (19.4 lb) more than control pigs, without any difference in days to marketing. Serum PCV2 antibody titers increased in control pigs, and PCV2 DNA was detected, indicating active PCV2 infection. CONCLUSIONS AND CLINICAL RELEVANCE Results suggested that vaccination against PCV2 was effective at reducing mortality rate and improving growth performance among pigs in a herd infected with PCV2.

Emergence of novel reassortant H3N2 swine influenza viruses with the 2009 pandemic H1N1 genes in the United States

Reassortant H1 swine influenza viruses (SIVs) carrying 2009 pandemic H1N1 virus (pH1N1) genes have been isolated from pigs worldwide. Seven novel reassortant H3N2 SIVs were identified from diseased pigs in the USA from winter 2010 to spring 2011. These novel viruses contain three or five internal genes from pH1N1 and continue to circulate in swine herds. The emergence of novel reassortant H3N2 SIVs demonstrates reassortment between pH1N1 and endemic SIVs in pigs and justifies continuous surveillance.

Restricted Enzooticity of Hepatitis E Virus Genotypes 1 to 4 in the United States

ABSTRACT Hepatitis E is recognized as a zoonosis, and swine are known reservoirs, but how broadly enzootic its causative agent, hepatitis E virus (HEV), is remains controversial. To determine the prevalence of HEV infection in animals, a serological assay with capability to detect anti-HEV-antibody across a wide variety of animal species was devised. Recombinant antigens comprising truncated capsid proteins generated from HEV-subgenomic constructs that represent all four viral genotypes were used to capture anti-HEV in the test sample and as an analyte reporter. To facilitate development and validation of the assay, serum samples were assembled from blood donors (n = 372), acute hepatitis E patients (n = 94), five laboratory animals (rhesus monkey, pig, New Zealand rabbit, Wistar rat, and BALB/c mouse) immunized with HEV antigens, and four pigs experimentally infected with HEV. The assay was then applied to 4,936 sera collected from 35 genera of animals that were wild, feral, domesticated, or otherwise held captive in the United States. Test positivity was determined in 457 samples (9.3%). These originated from: bison (3/65, 4.6%), cattle (174/1,156, 15%), dogs (2/212, 0.9%), Norway rats (2/318, 0.6%), farmed swine (267/648, 41.2%), and feral swine (9/306, 2.9%). Only the porcine samples yielded the highest reactivities. HEV RNA was amplified from one farmed pig and two feral pigs and characterized by nucleotide sequencing to belong to genotype 3. HEV infected farmed swine primarily, and the role of other animals as reservoirs of its zoonotic spread appears to be limited.

Differential expression and activity of the porcine type I interferon family

Type I interferons (IFNs) are central to innate and adaptive immunity, and many have unique developmental and physiological functions. However, in most species, only two subtypes, IFN-alpha and IFN-beta, have been well studied. Because of the increasing importance of zoonotic viral diseases and the use of pigs to address human research questions, it is important to know the complete repertoire and activity of porcine type I IFNs. Here we show that porcine type I IFNs comprise at least 39 functional genes distributed along draft genomic sequences of chromosomes 1 and 10. These functional IFN genes are classified into 17 IFN-alpha subtypes, 11 IFN-delta subtypes, 7 IFN-omega subtypes, and single-subtype subclasses of IFN-alphaomega, IFN-beta, IFN-epsilon, and IFN-kappa. We found that porcine type I IFNs have diverse expression profiles and antiviral activities against porcine reproductive and respiratory syndrome virus (PRRSV) and vesicular stomatitis virus (VSV), with activity ranging from 0 to >10(5) U.ng(-1).ml(-1). Whereas most IFN-alpha subtypes retained the greatest antiviral activity against both PRRSV and VSV in porcine and MARC-145 cells, some IFN-delta and IFN-omega subtypes, IFN-beta, and IFN-alphaomega differed in their antiviral activity based on target cells and viruses. Several IFNs, including IFN-alpha7/11, IFN-delta2/7, and IFN-omega4, exhibited minimal or no antiviral activity in the tested target cell-virus systems. Thus comparative studies showed that antiviral activity of porcine type I IFNs is virus- and cell-dependent, and IFN-alphas are positively correlated with induction of MxA, an IFN-stimulated gene. Collectively, these data provide fundamental genomic information for porcine type I IFNs, information that is necessary for understanding porcine physiological and antiviral responses.

Discovery of a novel putative atypical porcine pestivirus in pigs in the USA.

Pestiviruses are some of the most significant pathogens affecting ruminants and swine. Here, we assembled a 11 276 bp contig encoding a predicted 3635 aa polyprotein from porcine serum with 68 % pairwise identity to that of a recently partially characterized Rhinolophus affinis pestivirus (RaPV) and approximately 25-28 % pairwise identity to those of other pestiviruses. The virus was provisionally named atypical porcine pestivirus (APPV). Metagenomic sequencing of 182 serum samples identified four additional APPV-positive samples. Positive samples originated from five states and ELISAs using recombinant APPV Erns found cross-reactive antibodies in 94 % of a collection of porcine serum samples, suggesting widespread distribution of APPV in the US swine herd. The molecular and serological results suggest that APPV is a novel, highly divergent porcine pestivirus widely distributed in US pigs.

Antibody Recognition of Porcine Circovirus Type 2 Capsid Protein Epitopes after Vaccination, Infection, and Disease

ABSTRACT Open reading frame 2 (ORF2) of porcine circovirus type 2 (PCV2) codes for the 233-amino-acid capsid protein (CP). Baculovirus-based vaccines that express only ORF2 are protective against clinical disease following experimental challenge or natural infection. The goal of this study was to identify regions in CP preferentially recognized by sera from experimentally infected and vaccinated pigs and to compare these responses to those of pigs diagnosed with porcine circovirus-associated disease (PCVAD), including porcine multisystemic wasting syndrome (PMWS) and porcine dermatitis and nephropathy syndrome (PDNS). The approach was to react porcine sera with CP polypeptide fragments followed by finer mapping studies using overlapping oligopeptides that covered amino acids 141 to 200. The results showed that vaccinated pigs preferentially recognized only the largest polypeptide fragment, CP(43-233). A subset of experimentally infected pigs and pigs with PDNS showed strong reactivity against a CP oligopeptide, 169-STIDYFQPNNKR-180. Alanine scanning identified Y-173, F-174, Q-175, and K-179 as important for antibody recognition. The results from this study support the notion of PCV2 modulation of immunity, including antibody responses that may represent a precursor for disease. The recognition of CP(169-180) and other polypeptides provides opportunities to devise diagnostic tests for monitoring the immunological effectiveness of vaccination.

Comparison of Host Immune Responses to Homologous and Heterologous Type II Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) Challenge in Vaccinated and Unvaccinated Pigs

Porcine reproductive and respiratory syndrome (PRRS) is a high-consequence animal disease with current vaccines providing limited protection from infection due to the high degree of genetic variation of field PRRS virus. Therefore, understanding host immune responses elicited by different PRRSV strains will facilitate the development of more effective vaccines. Using IngelVac modified live PRRSV vaccine (MLV), its parental strain VR-2332, and the heterologous KS-06-72109 strain (a Kansas isolate of PRRSV), we compared immune responses induced by vaccination and/or PRRSV infection. Our results showed that MLV can provide complete protection from homologous virus (VR-2332) and partial protection from heterologous (KS-06) challenge. The protection was associated with the levels of PRRSV neutralizing antibodies at the time of challenge, with vaccinated pigs having higher titers to VR-2332 compared to KS-06 strain. Challenge strain did not alter the cytokine expression profiles in the serum of vaccinated pigs or subpopulations of T cells. However, higher frequencies of IFN-γ-secreting PBMCs were generated from pigs challenged with heterologous PRRSV in a recall response when PBMCs were re-stimulated with PRRSV. Thus, this study indicates that serum neutralizing antibody titers are associated with PRRSV vaccination-induced protection against homologous and heterologous challenge.

Characterization of a New Disease Syndrome Associated with Porcine Circovirus Type 2 in Previously Vaccinated Herds

ABSTRACT Porcine circovirus-associated disease (PCVAD) encompasses a group of wasting syndromes linked to porcine circovirus type 2 (PCV2). This paper describes a new PCV2 disease syndrome, called acute pulmonary edema (APE), which, unlike other PCVAD syndromes, has a peracute onset and is associated with herds vaccinated for PCV2.

Bovine rhinitis viruses are common in U.S. cattle with bovine respiratory disease.

Bovine rhinitis viruses (BRV) are established etiological agents of bovine respiratory disease complex however little research into their epidemiology and ecology has been published for several decades. In the U.S., only bovine rhinitis A virus 1 (BRAV1) has been identified while bovine rhinitis A virus 2 (BRAV2) and bovine rhinitis B virus (BRBV) were previously only identified in England and Japan, respectively. Metagenomic sequencing of a nasal swab from a bovine respiratory disease (BRD) diagnostic submission from Kansas identified contigs with approximately 90% nucleotide similarity to BRAV2 and BRBV. A combination of de novo and templated assemblies using reference genomes yielded near complete BRAV2 and BRBV genomes. The near complete genome of bovine rhinitis A virus 1 (BRAV1) was also determined from a historical isolate to enable further molecular epidemiological studies. A 5’-nuclease reverse transcription PCR assay targeting the 3D polymerase gene was designed and used to screen 204 archived BRD clinical specimens. Thirteen (6.4%) were positive. Metagenomic sequencing of six positive samples identified mixed BRAV1/BRAV2, BRAV1/BRBV and BRAV2/BRBV infections for five samples. One sample showed infection only with BRAV1. Seroprevalence studies using a cell culture adapted BRBV found immunofluorescence assay-reactive antibodies were common in the herds analyzed. Altogether, these results demonstrate that BRV infections are common in cattle with respiratory disease and that BRAV1, BRAV2 and BRBV co-circulate in U.S. cattle and have high similarity to viruses isolated more than 30 years ago from diverse locations.

Renal Lesions Associated with Experimental Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) Infection

Viral upper respiratory disease is considered to be the single most important infectious feline respiratory disease because of the contagious nature of the agents and because of the chronic carrier state. FCV and FHV-1 are the 2 viral agents most commonly associated with feline upper respiratory infections (URI). It has been estimated that FHV-1 and FCV cause 80-90% of all cases of infectious feline URI. Chlamydia psittaci is the other member of the triad of infectious agent most commonly incriminated as a cause of URI. Clinical signs are similar for caliciviral and herpesviral infections and include sneezing, mucopurulent oculonasal discharge, coughing, open-mouth breathing, ptyalism, conjunctivitis, rhinitis, sinusitis, gingivitis, and stomatitis. FCV has also been isolated from cats with nonrespiratory signs, including footpad and interdigital ulcers, arthritis, myalgia, and enteritis. Clinical manifestations of viral upper respiratory disease are seen primarily in cats < 6 months of age. Although morbidity is high for both FHV-1 and FCV infections, mortality is seen commonly only in kittens < 10 weeks of age. Highly virulent pneumotropic FCV isolates have been associated with a high mortality rate such as that observed in this case. Both calicivirus and herpesvirus infections may progress to a chronic carrier state, and carrier animals are an important source of infection for kittens. In 1 study, FCV was isolated from 18.5% of the asymptomatic adult cats tested. During the 7 months following the removal of the nonclinical adult carriers from the premises, the colony has experienced no morbidity or mortality associated with upper or lower respiratory disease, suggesting that these cats may have been a significant source of infection. A killed vaccine containing FCV had been administered to all of the queens and to all of the kittens ≥ 8 weeks of age in this colony. Although vaccination induces prompt seroconversion, it does not prevent infection and the manifestation of mild clinical signs . A recent study also demonstrated that vaccine strains derived from FCV-F9, the original calicivirus vaccine strain, may not be as broadly crossprotective as the parent strain. It was also found that vaccination does not prevent the chronic carrier state and may contribute to both acute and chronic infection and clinical disease. In 1 study, 43% (417) of all of the cats in the test group (963) were fully vaccinated. Bordetella bronchiseptica was isolated only from the kitten with gross evidence of lower respiratory infection. This bacterium is most commonly associated with canine and porcine respiratory disease and not usually thought of as a feline pathogen. A recent report described B. bronchiseptica infections in 11 cats from 10 different households or catteries. The infection was fatal in 7 of the 11 affected cats, suggesting that B. bronchiseptica may be a significant respiratory pathogen in cats.