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Swine interleukin 2 activity produced by mesenteric lymph node cells.

Swine interleukin 2 (IL-2) activity was assayed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) colorimetric assay using murine IL-2 dependent cell line (CTLL-2). The culture supernatant of mesenteric lymph node (MLN) cells stimulated with phytohemagglutinin-P (PHA) induced a generation of MTT formazan in a dose-dependent manner, suggesting dose-dependent proliferation of CTLL-2. The maximal swine IL-2 activity was observed in the culture of MLN cells at 1 to 2 x 10(7) cells/ml when stimulated with 20 to 40 micrograms/ml PHA for 48 hr. Based on these findings, a large culture of MLN cells to prepare swine IL-2 were performed under the following condition; cell concentration of 1 x 10(7) cells/ml, PHA concentration of 20 micrograms/ml, a culture scale of 200 to 400 ml, and a stirring speed of 30 rpm. Swine IL-2 activity was detected from 4 hr after PHA stimulation, and rapidly increased until 16 hr. Almost maximal IL-2 activity in stirring culture was observed at the incubation of 20 hr. Swine IL-2 was partially purified by Sephacryl S-200 gel filtration and the estimated molecular weight was about 32 kD based on the peak of IL-2 activity. The pI value of swine IL-2 was estimated to be approximately pH 5.3. Swine IL-2 was sensitive to acid (pH 3.2) or alkaline (pH 10.5), 4 or 8 M urea, trypsin, and the heating at 70 degrees C. These physico-chemical properties of swine IL-2 was similar to those of human, murine or feline IL-2.

Animals↗

Purification of carbonic anhydrase isozyme VI (CA-VI) from swine saliva.

Salivary or secreted carbonic anhydrase (CA), which constitutes a new class of CA, designated CA-VI, was isolated. Swine CA-VI purified from swine saliva by inhibitor-affinity chromatography and ion exchange chromatography had a specific activity of 5,468 units/mg. The molecular weight was 250,000, as determined by gel filtration under non-denaturing conditions, and the subunit molecular weight was found to be 37,000 by sodium dodecyl sulfate polyacrylamide gel electrophoresis, indicating that swine CA-VI consists of 7 subunits. The treatment of the enzyme with endo-N-acetylglucosaminidase F reduced its subunit molecular weight from 37,000 to 35,000 and 32,000. We raised a rabbit antibody against purfied swine CA-VI. Double immunodiffusion showed that anti-swine CA-VI serum reacted with swine CA-VI and swine saliva, but not with hemolysate (containing CA-I and CA-Il) or muscle extracts (containing CA-III). The concentration of CA-VI in swine saliva, measured using single radial immunodiffusion, was 0.027 +/- 0.017 mg/mg total protein.

Animals↗

Respiratory health of swine producers. Focus on young workers.

In this report, we compare the respiratory health of swine producers, grain farmers, and nonfarming control subjects, separately in all age groups and in young subjects. We examined 249 swine producers (age 37.7 years), 251 grain farmers (age 44.7 years), and 263 nonfarming subjects (age 40.7 years). Swine producers had significantly more symptoms of chronic bronchitis (15.3 percent) than did grain farmers (7.2 percent) or nonfarming men (5.7 percent). After controlling for age, height, and smoking, the functional indices of airflow (FEV1, FEV1/FVC, FEF25-75, Vmax50, and Vmax25) were slightly but significantly lower in swine producers than in grain farmers. In comparison with nonfarming subjects, swine producers also had significantly lower FEV1/FVC, FEF25-75, and Vmax50. Respiratory symptoms were associated with the number of hours of work per day. This indirect index of exposure was also inversely associated with FVC (p < 0.01) and FEV1 (p = 0.06), after adjustment for age, height, smoking, and dust mask usage. A relative excess of respiratory symptoms and lower lung function variables were found in swine producers aged 26 to 35 years. Also in this age group, a multivariate analysis revealed statistically significant effects of daily duration of work on FVC and FEV1. The results confirm that working in swine confinement units is a risk factor for chronic respiratory symptoms and minor lung function changes. An increased risk in young workers may reflect more intense occupational exposure in this subgroup of swine producers.

Adult↗

Diabetes-induced accelerated atherosclerosis in swine.

Patients with diabetes are at higher risk for atherosclerotic disease than nondiabetic individuals with other comparable risk factors. Studies examining mechanisms underlying diabetes-accelerated atherosclerosis have been limited by the lack of suitable humanoid animal models. In this study, diabetes was superimposed on a well-characterized swine model of atherosclerosis by injection of the beta-cell cytotoxin streptozotocin (STZ), resulting in a >80% reduction in beta-cells and an increase in plasma glucose to diabetic levels. Animals were maintained without exogenous insulin for up to 48 weeks. Plasma glucose and cholesterol levels and lesion extent and severity were quantified in swine with diabetes and hyperlipemia alone and in combination compared with controls. Diabetes had no effect on plasma cholesterol levels, but diabetic/hyperlipemic (D-HL) swine developed hypertriglyceridemia and showed a doubling in aortic sudanophilia over nondiabetic/hyperlipemic (N-HL) swine as early as 12 weeks (47.25 +/- 4.5 vs. 24.0 +/- 4.6%). At 20 weeks, coronary artery stenosis was significantly greater in D-HL than in N-HL animals (86 +/- 10 vs. 46 +/- 8%). Coronary lesions predominantly arose in the first 2-3 cm of the vessels and displayed humanoid morphology. Aortic lesions in D-HL swine had double the cholesterol content of those in N-HL swine, and incorporation of oleate into cholesteryl ester was significantly greater in grossly normal aortic areas of D-HL swine compared with N-HL and was attributed to similar elevated incorporation in monocytes. This large study demonstrates that a model of diabetes with humanoid characteristics, including hypertriglyceridemia and severe, accelerated atherosclerosis can be reproducibly induced and maintained in swine. This model should potentially be of great value in elucidating mechanisms underlying the accelerated atherosclerosis seen in human diabetic individuals.

Acetates↗

Sustainability issues of U.S. swine production.

The incorporation of livestock into agricultural systems ensures a more sustainable agriculture. Sustainable swine production systems are defined as those that combine production and management techniques to enhance profit and improve the ecological and socioeconomic surroundings. Over the long term, the systems should maintain or enhance the environment and resource base, the quality of life for the producers and society as a whole, the profit level of producers, and the quality of pork produced. Swine production in the United States is rapidly changing to fewer and larger production units. At the interface of sustainable agriculture and swine production are several levels of issues. Four levels of issues are the farm, the rural community, the society or consuming public, and the ecosystem or environment. By examining each level, long-term sustainability issues for U.S. swine production emerge. Some of these issues include swine care and husbandry, producer health, management and production systems, access to markets, information, technology and genetics, producer entrance requirements, specialization, manure utilization, industry structure, pork quality and leanness, and the impact of modern intensive systems on the environment, farms, and rural communities. The challenge beyond identifying issues is to incorporate sustainable concepts into profitable, ecologically based swine production systems. Several major groups of issues are discussed, including swine industry structure change issues, access and entrance issues, manure, by-product and nuisance issues, and pork quality issues. Many of these issues can be addressed. The solutions often rely on increased education, management, or technology applications. These processes provide broad opportunities for animal scientists. Examples of current solutions are quality assurance programs, segregated age rearing techniques, educational and promotional commodity programs, manure management programs, phase feeding, diet manipulation to alter manure nutrient concentrations, young producer assistance programs, producer networking, and environmental awareness activities. There are many more opportunities for improving the sustainability of U.S. swine production, when a long-term, issue-oriented viewpoint is maintained.

Animal Husbandry↗

Considerations of specific pathogen-free swine (SPF) in xenotransplantation.

The purpose of this manuscript was to introduce the complexities involved in defining both the appropriate health status and the management programs for SPF swine as they may relate to xenotransplantation. One factor that must always be considered in using swine is that they are considered agricultural animals regardless of their purpose and are regulated as such by municipalities, states, and countries. There are nonflexible regulations related to the interstate transportation of swine and even greater hurdles to international transportation. For example, I tried unsuccessfully for three years to transport a single breeding animal with a congenital heart defect to collaborators in the United Kingdom. Bureaucratic factors on both sides of the Atlantic proved insurmountable because the regulations were formulated for economic and isolation purposes and no exceptions could be granted for research animals. Another factor to be considered is the designation of SPF swine, which in this species has a specific proprietary connotation. If the finalized program for production of swine for xenotransplantation proves to be different from the already established SPF program, then a different designation may have to be used. For example, a term like "xenografic defined flora (XDF)" swine could be used to avoid confusion with the already established registry. If xenotransplantation using swine becomes a reality, then it is important that the parameters used to select such animals be defined in advance. It is unlikely that conventionally reared animals of uncertain health status will be widely accepted because of legal and ethical considerations. Likewise, I do not believe that the use of swine for xenotransplantation will be able to avoid the regulatory measures already in place for laboratory animals. The multidisciplinary approach to the problem initiated by the Institute of Medicine workshop is the most appropriate mechanism for resolving the issues.

Animals↗

[Origin of internal genes of two strains of swine influenza A (H1N1) virus].

BACKGROUND: Through the analysis of internal genes of viruses to understand whether the internal genes of two strains of swine influenza A(H1N1) virus contain the gene segment deriving from avian influenza A viruses, and whether the reassortment of the internal genes occurred between swine H1N1 and swine H9N2 viruses. METHODS: Viruses were passaged in embryonated hen eggs and virion RNA was extracted from allantoic fluids and reverse transcribed to synthesize cDNA. cDNA was amplified by PCR and the PCR product was purified with a purification kit. Afterwards, RNA sequence analysis was performed by dideoxynucleotide chain termination and a cloning method. Finally, phylogenetic analysis of the sequencing data was performed with MegAlign (version 1.03). RESULTS: All the six, but PB2 gene segment internal gene segments were the same between A/swine/Beijing/1/2002(H1N1) and A/Swine/Beijing/3/2002(H1N1) viruses. Whereas all the six internal gene segments in two trains of swine (H1N1) virus were similar to those of swine H1N1 viruses, but different from those of classical strain of swine (H1N1) virus. CONCLUSION: Two strains of Beijing swine H1N1 influenza A virus were not reassortant, All the six internal gene segments were closely related to swine influenza A (H1N1) viruses.

Animals↗

[Clinical and microbiological investigations of atrophic rhinitis of Upper Austrian swine herds].

The aim of this study was to determine the distribution of the progressive and non-progressive atrophic rhinitis of Upper Austrian swine herds. Further on the resistance pattern of the pathogens involved to chemotherapeutics was tested. In the period of May 1993 to June 1996 a total of 56 Upper Austrian swine herds were examined and on the occasion of the animal herd health management 997 nasal swab-samples of young pigs taken. The area of this investigation included 14 Upper Austrian districts and the herds examined were divided into 3 types. Type 1 were swine herds of the swine breeding association (SZV), type 2 piglet producing farms (FP) and type 3 closed swine herds (GB). Sucking- and weaning piglets aged from 4 to 10 weeks were selected for these examinations. On the average 10 nasal swab-samples (2 swabs per animal) per herd were taken, microbiologically examined and the toxin by means of ELISA-tests determined. In terms of resistance testing (antibiogram) 20 different chemotherapeutics, which consisted of 11 different groups of drugs, were used. As a result of 997 nasal swab-samples examined, 304 (30.5%) P. m., 111 (11.1%) tox. P. m. and 35 (3.5%) B. b. were isolated. 50% of the Upper Austrian swine herds showed PAR by means of microbiological examination and ELISA-tests as well. The resistance pattern of P. m. and B. b. exhibited significant differences. Penicillin and lincospectin were highly resistant concerning isolates of B. b., but were highly sensitive for P. m. Enrofloxacin turned out as the most effective drug to the P. m.-toxin-negative- and B. b.-strains tested, because no resistance was observed. Finally efficient PAR control programmes of swine herds belonging to the Upper Austrian swine herd health service are described.

Agricultural Workers' Diseases↗

Immunogenetic analysis of Trichinella spiralis infections in swine.

The immune responses of outbred swine, inoculated with several different low doses of Trichinella spiralis muscle larvae (ML), was followed over 5-6 weeks of primary infection, in order to determine an inoculation dose which could be used to identify genetic controls on the response to this helminth parasite. Reproducible infections were established when swine were inoculated with 100-300 ML. Humoral antibody responses to different larval stages were evident at 4 weeks using enzyme-linked immunosorbent assay (ELISA) of antibody-binding to excretory-secretory (ES) products of ML, and flow cytometric (FCM) analysis of antibody-binding to newborn larvae. T-cell blastogenesis to T. spiralis ML antigens was predominantly in the CD4+, class II restricted, T-cell subset. Having established an appropriate inoculation dose, swine leukocyte antigen (SLA) inbred miniature swine were then inoculated with this low dose of T. spiralis ML, to determine whether major histocompatibility complex (MHC) genes regulate swine immune responses to T. spiralis, as has been found in rodent models. Preliminary evidence indicated that swine of the SLA c/c haplotype may exhibit a lower burden of T. spiralis larvae in the tongue and diaphragm. This lower muscle burden correlated with the earlier development of a humoral antibody response in these genetically-defined swine.

Animals↗

[Effect of passive immunity on subsequent vaccination against swine fever].

Studied was the effect of a serum paratyph and Aujeszky gammaglobulin injected 10, 15, and 20 days prior to the crystal-violet and the lapinized vaccine. If the animals were infected with a pathogenic swine fever virus 10 to 15 days after their treatment with serum at the rate of 0.5 cu. cm per kg, part of them died, and the remaining contracted the disease showing a protracted course. The control vaccination on the 20th day after the animals were injected with serum led to the death of all of them showing the signs of acute swine fever. Most of the pigs injected with 5 cu. cm gamma-globulin each and 10--15 days later being infected with a swine fever virus manifested the disease, however, none of them died of the disease. If, however, after treatment with gamma-globulin they were challenged on the 20th day all of them died of acute swine fever. In the immunization of pigs with a crystal-violet vaccine on the 14th day of their treatment with Aujeszky gamma-globulin, part of them developed an atypical form of the disease, while most of them did not show symptoms whatever of swine fever. Of the immunized with a lapinized vaccine some died, and the remaining part did not contract the disease at all. All this pointed to the fact that the residual antibodies against swine fever contained in the specific Aujeszky gamma-globulin could still be in a position to inhibit the vaccinal virus, and more specifically that of the lapinized K vaccine. The vaccination against swine fever should be performed 14 days after the injection of the serum paratyph and about 20 days after the injection of the Aujeszky gammaglobulin.

Animals↗

Evaluation of criteria for the postmortem diagnosis of mycoplasmal pneumonia of swine.

Ten swine from each of five herds believed to be affected with mycoplasmal pneumonia of swine and ten swine from each of five herds believed to be mycoplasmal pneumonia-free were selected for postmortem study. Lungs from the 100 swine were examined; grossly and microscopically for lesions typical of mycoplasmal pneumonia of swine and culturally and by an indirect immunofluorescent procedure for the presence of Mycoplasma hyopneumoniae. Nineteen of the lungs had both gross and microscopic lesions typical of mycoplasmal pneumonia of swine and 13 (68%) of these were infected, i.e. were culturally and/or indirect immunofluorescent positive. Absence of gross lesions did not prove freedom from mycoplasmal pneumonia, 14 of 73 (19%) grossly normal lungs were found to be infected with M. hyopneumoniae. Comparison of the indirect immunofluorescent and cultural examination, as methods of diagnosing mycoplasma pneumonia, revealed that neither procedure alone was reliable in the case of negative results. Ten lungs were indirect immunofluorescent negative and culturally positive and seven were culturally negative and indirect immunofluorescent positive (11 lungs were positive by both procedures). It was concluded that a definitive diagnosis of mycoplasmal pneumonia of swine requires that M. hyopneumoniae be visualized in indirect immunofluorescent stained lung sections or that it be recovered culturally.

Animals↗

Swine influenza virus infections. Transmission from ill pigs to humans at a Wisconsin agricultural fair and subsequent probable person-to-person transmission.

In September 1988, a previously healthy 32-year-old pregnant woman was hospitalized for pneumonia and died 8 days later. The only pathogen detected was an influenza virus antigenically related to the swine influenza virus (SIV). Four days before illness onset, the patient visited a county fair swine exhibition where there was widespread influenzalike illness among the swine. To detect other persons who were possibly infected by contact with the ill swine, we measured serum SIV hemagglutination-inhibition antibody titer in 25 swine exhibitors who were 9 to 19 years old. Nineteen (76%) had SIV hemagglutination-inhibition titers of 20 or greater. Antibody was undetectable in serum samples from 25 swine exhibitors from a neighboring county. Additional studies suggest that one to three health care personnel who had contact with the patient developed influenzalike illnesses with laboratory evidence of SIV infection. An outbreak of apparent SIV infection in swine resulted in multiple human infections, and, although no recognized community outbreak resulted, there was evidence of virus transmission from the patient to health care personnel.

Adolescent↗

Flow cytometric quantification of cholesteryl ester-containing "foam" cells. I. Analysis of aortas from normolipidemic swine.

We have quantified using flow cytometry foam cells of aortas from normolipidemic swine varying in age from 6 months to 12 years. These swine were maintained throughout their lives on a low-fat, cholesterol-free diet. Intimal-medial tissues removed from the swine aortas were enzymatically dissociated to prepare Formalin-fixed cell suspensions. Foam cells were labeled by specific staining of their intracellular cholesteryl ester using the fluorescent dye filipin. This was carried out by first removing cellular unesterified cholesterol with ethanol, then enzymatically hydrolyzing cellular cholesteryl ester, and finally staining with filipin the unesterified cholesterol derived from hydrolysis of cholesteryl ester. Results of flow cytometric analysis indicated that thoracic foam cell densities of female swine became more variable and tended to increase with age. It appeared that there were two subgroups of female swine with either high or low levels of thoracic foam cells. A similar finding was not observed for abdominal foam cells in female swine nor for thoracic or abdominal foam cells in the smaller number of older male swine. Abdominal foam cell densities in younger males, however, also appeared to be comprised of low and high foam cell groups. Foam cell densities did not correlate with serum cholesterol or triglyceride levels. However, a genetic basis for some of the variability in foam cell densities among these animals was suggested by the observation that females with high thoracic foam cell densities had greater commonality among ancestors than did females with low thoracic foam cell densities.

Age Factors↗

Extracellular matrix maturation in the left ventricle of normal and diabetic swine.

The main objective of this study is to determine the transmural distribution of extracellular matrix (ECM) collagen and maturation in non-diabetic and diabetic hearts. The Yucatan miniature swine heart ECM was analyzed in eight streptozotocin (STZ) induced diabetic pigs (Diabetic-Swine) and age matched normal control pigs (Nondiabetic-Swine). After 12 weeks of STZ induced diabetes, transmural biopsies were obtained from the left ventricular free wall divided into subendocardial, mid- and subepicardial layers. Collagen concentration and maturation were measured by RP-HPLC determination of hydroxyproline (Hyp) and content of hydroxylysylpyridinoline (HP) cross-links, respectively. Results showed a significant elevation in arterial glucose (P<0.05) and reduction in arterial plasma insulin levels in the Diabetic-Swine. Hyp concentration was significantly greater (P<0.05) in the subendocardial layers in both the Diabetic and Nondiabetic animals. The HP cross-link content was significantly greater (17%) in the Diabetic-swine subendocardial layer compared to Nondiabetic-Swine (P<0.05), but not in other layers. In summary, the accumulation and/or increase in HP cross-link content in the Diabetic-Swine subendocardial layer suggests that myocardial fibrosis may be greater in this specific region.

Amino Acids↗

Immune responses and protection against foot-and-mouth disease virus (FMDV) challenge in swine vaccinated with adenovirus-FMDV constructs.

A replication-defective adenovirus 5 encoding foot-and-mouth disease virus (FMDV) capsid and 3C proteinase coding regions (Ad5-FMDV3CWT) was used to vaccinate swine. A single inoculation utilizing 1 x 10(8) plaque forming units (pfu) or an inoculation of 1 x 10(8) followed by a boost of 5 x 10(8) pfu Ad5-FMDV3CWT were tested, along with an inoculation and boost using an adenovirus encoding the FMDV capsid coding region and an inactive form of the 3C proteinase (Ad5-FMDV3CMUT). Sera collected from these animals were examined for the presence of FMDV-specific antibodies using immunoprecipitation, neutralization, and ELISA assays specific for IgM, IgG1 and IgG2. Efficacy studies were performed by placing the vaccinated swine in contact with an FMDV-infected swine and monitoring for signs of disease and changes in serum antibody levels. Ad5-FMDV3CMUT, which is unable to produce FMDV capsid structures, did not elicit FMDV-neutralizing antibodies or protect against FMD. Single inoculation with Ad5-FMDV3CWT generated FMDV-specific neutralizing antibodies, and reduced clinical signs in challenged swine, but failed to completely protect the majority of swine from FMD. Swine which received a primary vaccination with Ad5-FMDV3CWT followed by the boost at 4 weeks generated high levels of FMDV-neutralizing antibodies resulting in complete protection of five of the six swine and limited disease in the remaining animal. Increased efficacy of the two-dose regimen was associated with heightened levels of FMDV-specific IgG1 and IgG2 antibodies.

Adenoviruses, Human↗

Prevalence of sparganosis by county of origin in Florida feral swine.

Sparganosis is a parasitic infection in amphibians, reptiles and mammals including feral swine and man. It is caused by migration of the metacestode (spargana) of Spirometra. The primary objective of this study was the determination of the prevalence of gross sparganosis in Florida county of origin in slaughtered feral swine. Tracebacks to county of origin were conducted for Florida feral swine with and without gross sparganosis. Feral swine trapped in Florida and presented for slaughter in a Texas slaughter establishment from May to December 1999 was the sample population. Overall prevalence of sparganosis in Florida feral swine was 6.9%. Because Highlands county had the same prevalence, other counties were compared to it. Sparganosis was detected in 17 Florida counties. Swine originating from Osceola or Hillsborough counties (4.3 and 1.8% prevalence, respectively) had lower prevalence of sparganosis than in Highlands, whereas those from Marion county (21.7% prevalence) had a higher prevalence. Transmission to humans may occur via consumption of infected feral swine, other species of secondary intermediate hosts or the primary intermediate hosts.

Abattoirs↗

Morphometric analysis of miniature swine hearts as potential human xenografts.

Miniature swine are considered to be potential donors for clinical cardiac transplantation. However, it is unclear how an appropriately sized porcine donor will be selected for a particular human recipient. To address this issue, we performed a morphometric study of the swine heart using transthoracic echocardiography (n = 26) to determine the diameters of the aortic annulus and root, pulmonary artery annulus, and mitral valve annulus. We also obtained direct ex vivo measurements of swine heart weight and linear dimensions (n = 71). Relationships between a swine's height, weight, length, chest circumference and these internal and external cardiac dimensions are described. The strongest correlations were found between a pig's body length and its aortic annulus and root diameters (r-values = 0.97). These relationships are accurately described by univariate linear regression models. By cross-relating our morphometric measurements of aortic annulus diameter in the miniature swine with normative human data, we were able to develop a nomogram, relating swine length and human height, which predicts which miniature swine would donate the best size-matched heart for a particular human recipient.

Animals↗

Cocaine-induced microvascular vasoconstriction but differential systemic haemodynamic responses in Yucatan versus Yorkshire varieties of swine.

1. Systemic and coronary haemodynamics were measured in 6 Yorkshire swine and 6 Yucatan miniature swine under isoflurane anaesthesia to investigate the influence of cocaine following its intravenous administration at 1, 3 and 7 mg kg-1. 2. Cocaine in Yorkshire swine decreased mean arterial pressure and rate pressure product (systolic pressure x heart rate), suggesting a cardiac depressant effect, whereas cocaine in Yucatan miniature swine increased these parameters, consistent with a hyperadrenergic state. 3. Cocaine in both Yorkshire swine and Yucatan miniature swine decreased coronary blood flow and coronary flow reserve, and increased coronary vascular resistance. 4. A modest generalized epicardial coronary artery constriction was observed by angiography, without evidence of focal spasm. 5. Our results confirm a marked vasoconstrictor effect of cocaine on the coronary arterial circulation, predominantly distal to the epicardial coronary arteries, but also indicate important differences in the systemic cardiovascular responses to the drug between two closely related strains of animals within the same species. Due to the similarities between the swine and human coronary arterial vasculature, we suggest that vasoconstriction in the coronary microcirculation may produce cardiac toxicity in man.

Angiocardiography↗