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At least 1,297 records · Page 72Linked to original sources
PubMed2006

An MHC class II disparity raises the threshold for tolerance induction in pulmonary allografts in miniature swine.

OBJECTIVES: The mechanisms and treatment of chronic rejection in pulmonary allotransplantation remain elusive. We have induced robust tolerance to class I-disparate lung allografts in miniature swine using an intensive 12-day course of tacrolimus. Here, we tested whether a tolerant state can be induced in swine receiving fully mismatched lung allografts. METHODS: Orthotopic left lung allografts were performed using MHC class I-disparate (group 1: n = 3) or fully disparate (group 2: n = 6) donors. The recipients received a 12-day postoperative course of tacrolimus (continuous intravenous infusion; target level = 35-50 ng/mL) as their only immunosuppression. RESULTS: All swine in group 1 maintained their grafts long term without developing any lesions of chronic rejection (>497, >432, >451 days). These recipients exhibited donor-specific hyporesponsiveness in cell-mediated lymphocytotoxity (CML) and mixed lymphocyte reaction (MLR) assays. In group 2, five of the six recipients maintained their grafts long term (sacrificed on postoperative days 515, 389, 429, 481, and 438 with viable grafts). Isolated lesions of obliterative bronchiolitis were occasionally seen on biopsy, and donor-specific hyporesponsiveness on assays was consistently observed. The remaining recipient rejected its graft on day 103 with histologic findings of obliterative bronchiolitis. CONCLUSIONS: We report long-term graft acceptance without chronic immunosuppression in five of six recipients across a full MHC disparity, albeit with some evidence of obliterative bronchiolitis. These data suggest that the class II disparity inherent in a fully mismatched transplant increases the requirement for tolerance induction.

Animals↗
PubMed2004

Recombinant vesicular stomatitis (Indiana) virus expressing New Jersey and Indiana glycoproteins induces neutralizing antibodies to each serotype in swine, a natural host.

Vesicular stomatitis virus (VSV) is the most common cause of vesicular disease outbreaks in livestock throughout the Western Hemisphere. Two major serotypes, Indiana and New Jersey, cause epidemic disease in pigs, cattle, and horses. We generated recombinant viruses derived from the Indiana serotype genome that were engineered to contain and express: (1) a single copy of the glycoprotein gene from the Indiana serotype (VSIV-GI); (2) a single copy of the glycoprotein gene from the New Jersey serotype (VSIV-GNJ); or (3) two copies of the glycoprotein gene, one from each of the two major VSV serotypes (VSIV-GNJGI) [Martinez I, Rodriguez LL, Jimenez C, Pauszek SJ, Wertz GW. Vesicular stomatitis virus glycoprotein is a determinant of pathogenesis in swine, a natural host. J Virol 2003;77(14):8039-47]. These recombinant viruses and a highly pathogenic New Jersey field isolate (VSNJV), from which the GNJ gene was derived, were inoculated into swine, a natural host, and the induction of neutralizing antibodies to both serotypes was analyzed. The neutralizing antibody response induced by VSIV-GI, VSIV-GNJ and VSNJV was serotype-specific, according to the glycoprotein expressed. VSIV-GNJGI expressed both glycoproteins stably through multiple rounds of replication in swine and induced neutralizing antibodies against both VSV serotypes, with a dominance of the Indiana serotype in the serological response. Pigs immunized with VSIV-GI or VSIV-GNJ were protected against homologous high dose virus challenge. Pigs inoculated with VSIV-GNJGI were protected against challenge with VSIV-GI but three of four animals developed lesions after challenge with the highly pathogenic New Jersey field isolate.

Animals↗
PubMed2006

Reconstruction of a swine SLA-I protein complex and determination of binding nonameric peptides derived from the foot-and-mouth disease virus.

No experimental system to date is available to identify viral T-cell epitopes in swine. In order to reconstruct the system for identification of short antigenic peptides, the swine SLA-2 gene was linked to the beta(2)m gene via (G4S)3, a linker encoding a 15-amino acid glycine-rich sequence (G4S)3, using splicing overlap extension-PCR (SOE-PCR). The maltose binding protein (MBP)-SLA-2-(G4S)3-beta(2)m fusion protein was expressed and purified in a pMAL-p2X/Escherichia coli TB1 system. The purified MBP-SLA-2-(G4S)3-beta(2)m protein was cleaved by factor Xa protease, and further purified by DEAE-Sepharose chromatography. The conformation of the SLA-2-(G4S)3-beta(2)m protein was determined by circular dichroism (CD) spectrum. In addition, the refolded SLA-2-(G4S)3-beta(2)m protein was used to bind three nonameric peptides derived from the foot-and-mouth disease virus (FMDV) O subtype VP1. The SLA-2-(G4S)3-beta(2)m-associated peptides were detected by mass spectrometry. The molecular weights and amino acid sequences of the peptides were confirmed by primary and secondary spectra, respectively. The results indicate that the SLA-2-(G4S)3-beta(2)m was 41.6kDa, and its alpha-helix, beta-sheet, turn, and random coil by CD estimation were 78 aa, 149 aa, 67 aa, and 93 aa, respectively. SLA-2-(G4S)3-beta(2)m protein was able to bind the nonameric peptides derived from the FMDV VP1 region: 26-34 (RRQHTDVSF) and 157-165 (RTLPTSFNY). The experimental system demonstrated that the reconstructed SLA-2-(G4S)3-beta(2)m protein complex can be used to identify nonameric peptides, including T-cell epitopes in swine.

Animals↗
PubMed2004

Efficacy of vaccines against bacterial diseases in swine: what can we expect?

This paper discusses what can be expected with regard to efficacy of antibacterial vaccines used in swine, based on the present knowledge of pathogen-host interactions. First, vaccination against bacteria that mainly cause disease by production of exotoxins is considered. Vaccines containing the inactivated toxin or a non-toxic but antigenic recombinant protein derived from the exotoxin can be expected to provide protection against disease. The degree of protection induced by such vaccines varies, however, depending amongst other things on the pathogenesis of the disease. Vaccination against clostridial infections, Actinobacillus pleuropneumoniae infections, progressive atrophic rhinitis and enterotoxigenic Escherichia coli, is considered. The second part of the article deals with vaccination against extracellular bacteria. Protection against these bacteria is generally mediated by antibodies against their surface antigens and certain secreted antigens, but cellular immunity may also play a role. Efficacy of vaccines against swine erysipelas, Streptococcus suis infections, Mycoplasma hyopneumoniae infections and swine dysentery is discussed. Finally, vaccination against facultatively intracellular bacteria is considered. For protection against these bacteria cell-mediated immunity plays an important role, but antibodies may also be involved. It is generally accepted that live-attenuated vaccines are more suitable for induction of cell-mediated immunity than inactivated vaccines, although this also depends on the adjuvant used in the vaccine. As an example, vaccination against Salmonella enterica serotype Typhimurium is discussed.

Animals↗
PubMed2004

Evaluation of the German cockroach (Blattella germanica) as a vector for verotoxigenic Escherichia coli F18 in confined swine production.

German cockroaches are common pests of confined swine production in North Carolina and other southeastern states. Vector competence of German cockroaches for one of the most important porcine bacterial pathogens, verotoxigenic Escherichia coli F18, was evaluated in laboratory bioassays using a culturing approach followed by multiplex PCR. In addition, the populations of fecal coliforms from the feces of piglets and cockroaches collected from a swine nursery were assessed. Viable and virulent cells of E. coli F18 were detected in cockroach feces for up to 8 days after the initial exposure. The population of fecal coliforms in cockroach feces was high (4.4 x 10(5) CFU g(-1)) and comparable to that of piglet feces (1.9 x 10(6) CFU g(-1)). This study demonstrates that cockroaches may serve as important mechanical vectors of pathogenic E. coli. Integrated management of cockroach populations should be incorporated into the disease prevention and control programs in the swine industry.

Animals↗
PubMed2006

Full genome sequence and analysis of Indian swine hepatitis E virus isolate of genotype 4.

The full-length genomic sequence of an Indian swine hepatitis E virus (HEV) isolate (IND-SW-00-01) recovered from feces of a pig experimentally infected with swine HEV pool from western India was determined. The genome consisted of 7,240 nucleotides, excluding the poly (A) tail of at least 22 residues and contained three open reading frames (ORFs), ORF-1 encoding 1,707 amino acids, ORF-2 encoding 674 amino acids and ORF-3 encoding 114 amino acids. Comparative full-length genome sequence and phylogenetic analyses suggested that the Indian swine HEV represents a distinct variant among the genotype 4 isolates with a divergence of 15-16.6%. Analyses based on ORF-1, 2 and 3 as well as partial ORF-2 (227 nucleotides) yielded similar results. As compared to type 4 HEV isolates, 26 unique amino acid substitutions were recorded, 16 in ORF-1, 8 in ORF-2 and 2 in ORF-3. IND-SW-00-01 showed insertion of 'C' at 5159 position while all other type 4 isolates have insertion of 'U' at the same position. Whether these changes contribute towards observed absence of type 4 HEV infections in Indian patients needs to be determined.

3' Untranslated Regions↗
PubMed2006

Analytical verification of a multiplex PCR for identification of Bordetella bronchiseptica and Pasteurella multocida from swine.

Bordetella bronchiseptica and Pasteurella multocida are etiologic agents of progressive atrophic rhinitis (PAR) and bronchopneumonia in swine. Only dermonecrotic toxin-producing strains of P. multocida play a role in atrophic rhinitis while both toxigenic and nontoxigenic strains have been associated with pneumonia. Monitoring and investigation of outbreaks involving these bacteria require sensitive and accurate identification and reliable determination of the toxigenic status of P. multocida isolates. In the present study, we report the development, optimization, and performance characteristics of a multiplex PCR assay for simultaneous amplification of up to three different targets, one common to all P. multocida strains, one found only in toxigenic P. multocida strains, and one common to B. bronchiseptica strains. Based on analysis of 94 P. multocida isolates (31 toxigenic) and 126 B. bronchiseptica isolates assay sensitivity is 100% for all amplicons. Evaluation of 22 isolates of other bacterial genera and species commonly found in the swine respiratory tract demonstrated a specificity of 100% for all gene targets. The limit of detection for simultaneous amplification of all targets is 1-10pg of DNA per target, corresponding to a few hundred genomes or less. Amplicon mobility in agarose gels and sequence analysis indicate the amplicons are highly stable. The data presented establish this multiplex PCR as a reliable method for identification of B. bronchiseptica and both toxigenic and nontoxigenic P. multocida that may greatly simplify investigations of swine PAR and bronchopneumonia.

Animals↗
PubMed2006

World association for the advancement of veterinary parasitology (WAAVP): second edition of guidelines for evaluating the efficacy of anthelmintics in swine.

Guidelines are provided for evaluating the efficacy of anthelmintics in swine which, in conjunction with other sets of guidance such as those of the International Cooperation on Harmonization of Technical Requirements for Registration of Veterinary Medicinal Products (VICH GL7 and VICH GL16), should encourage the adoption of uniform registration requirements globally. Testing of efficacy should be carried out according to the principles of "Good Clinical Practice" (VICH GL9, 2000). Data obtained according to these guidelines should be internationally acceptable for the registration of anthelmintics for swine. Further, the use of the guidelines should expedite development, government review, and approval of anthelmintics for swine, as well as contribute towards reducing costs and the number of experimental animals used for drug testing.

Animals↗
PubMed2006

Comparison of detection methods for Toxoplasma gondii in naturally and experimentally infected swine.

Results from recent serological surveys and epidemiological studies show that pigs raised in a variety of management systems can be carriers of the tissue cyst stage of Toxoplasma gondi. This parasite can be transmitted to humans through the consumption of improperly prepared pork, making detection and removal of infected swine carcasses from the food chain an important food safety issue. Several methods are available for detection of T. gondii infected swine, including serological assays, polymerase chain reaction, and animal bioassays. The aim of the present study was to compare the detection sensitivities of six of these commonly used methods for detection of T. gondii infection in tissues from naturally and experimentally infected pigs. The results indicate that a serum-based ELISA is the most sensitive method, of those tested, for detection of T. gondii infected swine.

Animals↗
PubMed2006

Zoonotic implications of the swine-transmitted protozoal infections.

Pig production is an important part of the economy in many countries. Domestic and wild pigs (Sus scrofa) are susceptible to a wide range of infectious and parasitic diseases. Some of these diseases are specifically limited to pigs while some of the other diseases are shared with other species of wildlife and domestic livestock. As the numbers and geographic distribution of wild and domestic swines continue to increase, it is certain that the number of contacts between these swines and domestic livestock will also increase, as will the probability of human exposure to the parasites of swine directly or indirectly. Here, we will discuss the protozoal infections of pigs, which have the potential to infect humans and provide reasonable risk assessment for zoonotic transmission.

Animals↗
PubMed2006

Identification of novel foot-and-mouth disease virus specific T-cell epitopes in c/c and d/d haplotype miniature swine.

To identify foot-and-mouth disease virus (FMDV) specific T-cell epitopes within the non-structural protein 3D in swine, pentadecapeptides were tested in proliferation and Interferon-gamma ELISPOT assays using lymphocytes from two strains of inbred miniature pigs (c/c and d/d haplotype) experimentally infected with FMDV. Lymphocytes of c/c pigs recognized peptides from three different regions in 3D, d/d lymphocytes recognized peptides from two regions, one of them being adjacent to an epitope of c/c pigs and comprising amino acid residues 346-370. Analyses of the response of d/d lymphocytes against peptides representing the structural protein 1A revealed another novel T-cell epitope. Investigation of the phenotype of responding lymphocytes showed a response of CD4(+)CD8(+)MHC-class-II(+) cells, identifying them as activated T-helper cells. This is the first report on FMDV specific T-cell epitopes recognized by swine leukocyte antigen (SLA) inbred swine and provides information useful for the design of novel vaccines against FMDV.

Amino Acid Sequence↗
PubMed1996

Cycloheximide-induced apoptosis in melanoma cells derived from regressing cutaneous tumours of Sinclair swine.

The role of apoptosis in the spontaneous regression of Sinclair swine melanoma was investigated in vitro with swine melanoma cell lines. Growth characteristics and sensitivity to cycloheximide-induced apoptosis were determined in melanoma cell lines derived from tumours that were progressing or undergoing regression in vivo. In contrast to cell lines derived from progressing tumours, those derived from regressing tumours showed induction of apoptosis; this phenomenon was dependent on dose but independent of cell growth stage in culture. Chromatin condensation, cell shrinkage, and fragmentation into apoptotic bodies, as well as DNA fragmentation into large kilobase fragments, occurred in cell lines from regressing tumours but not from progressing tumours. These findings suggest that swine melanoma cells are inherently more sensitive to cell death during tumour regression. The apoptosis-sensitive and resistant cell lines will be important for further studies of the roles of cell signalling pathways and gene expression in tumour regression.

Animals↗
PubMed2001

Adenovirus-mediated expression of interleukin-1 receptor antagonist in swine cells in vitro and in vivo.

Adenovirus-mediated gene delivery of anticytokines is a powerful tool for modulating the cytokine environment under conditions of respiratory disease. In order to determine the feasibility of cytokine modulation in the context of respiratory disease in swine, nonreplicating E1- and E3-deficient adenovirus constructs expressing a model protein, beta-galactosidase, and an anticytokine, the IL-1 receptor antagonist (IL-1Ra), were evaluated for in vitro expression in porcine PK15 cells, and in vivo following endotracheal instillation into the lungs. beta-Galactosidase and IL-1Ra were readily expressed in vitro in swine cells. Endotracheal administration of lacZ-containing adenovirus demonstrated that endothelial and epithelial cells in the alveolar spaces and bronchi of the middle and lower lobes were the principal sites of infection and expression, whereas beta-galactosidase staining was not observed in the upper lobe. Endotracheal administration of IL-1Ra recombinant adenovirus resulted in sustained expression of IL-1Ra into the alveolar spaces, where it was recovered in a concentration of 660 pg/ml in 500 ml of lavage fluid, equivalent to 330 ng IL-1Ra, in the lungs 7 days after treatment. Moreover, in vivo instillation of nonreplicating adenovirus did not induce an inflammatory response in the 1-week time frame of the study period. Lung weight as a percent of body weight, serum zinc, serum amyloid A, leukocyte differentials, neutrophil activity, and TNF levels all were the same between untreated pigs and pigs treated with either recombinant adenovirus. The results indicate that the delivery of IL-1Ra to swine lungs via nonreplicating, recombinant adenovirus may be an effective method for in vivo modulation of IL-1 activity and investigation of cytokine involvement in respiratory disease pathogenesis.

Actinobacillus Infections↗
PubMed2003

Development of a TaqMan PCR assay with internal amplification control for the detection of African swine fever virus.

A closed-tube polymerase chain reaction (PCR) was developed to allow the rapid detection of African swine fever virus (ASFV) DNA. This assay targets the VP72 gene of ASFV and uses the 5'-nuclease assay (TaqMan) system to detect PCR amplicons, avoiding tube opening and potential cross-contamination of post-PCR products. An artificial mimic was engineered with the TaqMan probe site replaced by a larger irrelevant DNA fragment allowing discrimination from ASFV by using two-colour TaqMan probe reporters. When added to the samples, successful amplification of this mimic demonstrated the absence of substances inhibitory to PCR, thereby validating negative results. Assay sensitivity was confirmed by obtaining positive signals with a representative selection of ASFV isolates. Many of the clinical and post-mortem features of ASF resemble those of classical swine fever (CSF) and porcine dermatitis and nephropathy syndrome (PDNS). Therefore, fast and reliable detection of ASFV is essential not only for the implementation of control measures to prevent the spread of ASF, but also in the differential diagnosis from CSF and PDNS. This assay should prove to be a valuable tool in the laboratory diagnosis of ASF and will complement existing molecular methods to provide rapid differential diagnosis in cases of suspected swine fever.

African Swine Fever Virus↗
PubMed1998

Risk factors for high sow mortality in French swine herds.

Episodes of high sow mortality rates affect profitability of swine farms. However, relevant control actions are difficult to implement. The objective of this study was to identify the risk factors for high levels of sow mortality rate (HM) in French swine herds. A case-control study was carried out in 102 swine herds located in Brittany (western France). Level of sow mortality of a herd was quantified by the annual mortality rate using sow-days as denominator. Fifty-five (53.9%) herds which experienced a sow mortality rate over 5% were classified as HM herds. Logistic regression was used to assess associations of managerial practices and disease prevalence with the odds of HM. High prevalence of urinary tract infections, metritis or lameness were significantly associated with a HM herd status (P < 0.10, OR ranging from 3.4 to 5.2). Multiplying herds were herds at higher risk for sow mortality than commercial farrow-to-finish herds. Providing three meals per day instead of two to dry sows decreased the odds of HM. Feeding plans where the maximum daily amount of feed provided to lactating sows was lower than 8 kg and was reached before 15 d in lactation were related to lower odds of HM (P < 0.10). Average age at weaning of 28 d or more and/or small average litter size at birth (12 piglets or less) were associated with higher odds of experiencing HM.

Animal Husbandry↗
PubMed1999

Salmonella level of Danish swine herds based on serological examination of meat-juice samples and Salmonella occurrence measured by bacteriological follow-up.

Analyses of data collected in the Danish Salmonella Control Program in swine herds were conducted to assess the association between bacteriological and serological findings (including the degree to which changing serological status over time modulated the relationship) and to assess the degree to which type of swine operation and age group affected the relationship between bacteriological and serological findings. Although serological status was significantly associated with bacteriological prevalence of salmonella enterica in swine herds, the association was clarified by adding information on the change in serological status over time and the age group being tested. Salmonella infection assessed by serological tests at a single time-point and data on age group tested would allow better targeting and managing control programs.

Animals↗
PubMed1999

Development of DNA vaccines for foot-and-mouth disease, evaluation of vaccines encoding replicating and non-replicating nucleic acids in swine.

We have developed naked DNA vaccine candidates for foot-and-mouth disease (FMD), an important disease of domestic animals. The virus that causes this disease, FMDV, is a member of the picornavirus family, which includes many important human pathogens, such as poliovirus, hepatitis A virus, and rhinovirus. Picornaviruses are characterized by a small (7-9000 nucleotide) RNA genome that encodes capsid proteins, processing proteinases, and enzymes required for RNA replication. We have developed two different types of DNA vaccines for FMD. The first DNA vaccine, pP12X3C, encodes the viral capsid gene (P1) and the processing proteinase (3C). Cells transfected with this DNA produce processed viral antigen, and animals inoculated with this DNA using a gene gun produced detectable antiviral immune responses. Mouse inoculations with this plasmid, and with a derivative containing a mutation in the 3C proteinase, indicated that capsid assembly was essential for induction of neutralizing antibody responses. The second DNA vaccine candidate, pWRMHX, encodes the entire FMDV genome, including the RNA-dependent RNA polymerase, permitting the plasmid-encoded viral genomes to undergo amplification in susceptible cells. pWRMHX encodes a mutation at the cell binding site, preventing the replicated genomes from causing disease. Swine inoculated with this vaccine candidate produce viral particles lacking the cell binding site, and neutralizing antibodies that recognize the virus. Comparison of the immune responses elicited by pP12X3C and pWRMHX in swine indicate that the plasmid encoding the replicating genome stimulated a stronger immune response, and swine inoculated with pWRMHX by the intramuscular, intradermal, or gene gun routes were partially protected from a highly virulent FMD challenge.

Animals↗
PubMed1994

Delivery of pseudorabies virus envelope antigens enclosed in immunostimulating complexes (ISCOMs); elicitation of neutralizing antibody and lymphoproliferative responses in swine and protection in mice.

An experimental subunit vaccine that consisted of pseudorabies virus (PRV) envelope glycoproteins enclosed into immunostimulating complexes (PRVenv/ISCOM) was constructed, and evaluated in DBA/2 mice and inbred swine of the SLA haplotype c/c. Two to three weeks after the first vaccine dose, specific anti-PRV antibodies could be demonstrated by ELISA or virus neutralization (VN) assays. Booster PRVenv/ISCOM vaccinations resulted in rapid and significant increases in antibody titres in both mice and swine. In addition, a week after receiving the third PRVenv/ISCOM vaccine dose swine peripheral blood mononuclear cells exhibited significant proliferation in response to stimulation with PRV virion antigen. Moreover, two doses of vaccine sufficed to protect mice fully against lethal virus challenge. Therefore, the data presented here support the ISCOM as a viable antigen delivery system for subunit PRV vaccines.

Animals↗