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PubMed1998

Characterization of metabolic responses to low-flow ischemia in intact pig hearts and isolated blood-perfused neonatal pig hearts.

UNLABELLED: There are different well established experimental models of low-flow ischemia. We examined metabolic variables during reduced coronary blood flow (CBF) in intact pig hearts and isolated neonatal pig hearts, producing similar degrees of postischemic dysfunction without infarction. The isolated hearts were perfused with red blood cell enriched buffer. In eight open-chest pigs mid-LAD flow was reduced to 70% for 60 min, followed by 120 min reperfusion. Myocardial segment lengths were recorded and regional coronary venous blood was sampled. In isolated piglet hearts CBF was reduced to 50% (n = 4), 25% (n = 4), and 10% (n = 17). Only when flow was reduced to 10% did hearts show signs of anaerobic metabolism. Mechanical function was recorded by a balloon in the left ventricle and coronary venous blood was sampled. Intact pig hearts showed release of protons, CO2, and lactate which peaked after 5-10 min of ischemia and thereafter stabilized at reduced levels. In contrast, in isolated neonatal hearts exposed to 10% CBF releases of protons, CO2, and lactate were stable during ischemia with no adaptational changes over time. In a separate group (n = 4), repetitive biopsies revealed no adaptational changes over time for adenosine triphosphate and creatine phosphate during 10% CBF. Contractile function was stably reduced during ischemia in both models. CONCLUSION: During reduced CBF "metabolic adaptation" occurs in intact pig hearts. In contrast, this feature is not present in isolated blood-perfused piglet hearts. The mechanisms responsible for these differences are uncertain. However, differences in metabolism between adult and neonatal hearts and different loading conditions during ischemia might contribute.

Adaptation, Physiological↗

PubMed1994

A comparison of the performance of pig hearts perfused with pig or human blood using an ex-vivo working heart model.

The pathophysiology of xenograft hyperacute rejection is still poorly understood although it is believed to involve complement fixation to vascular endothelium, probably as a result of the presence of naturally occurring anti-species antibodies. Hyperacute rejection of pig hearts by human blood was studied in an ex-vivo working heart model. Cardiac performance and immunological reactions occurring in the perfusing blood were studied. Stroke work performed by pig hearts perfused with human blood and their survival (median 47 min: n = 10) was significantly reduced compared to survival (median 158 min: n = 10) and stroke work performed by pig hearts perfused with pig blood. Decomplementation of human blood resulted in improved performance and duration of the action (median survival > 240 min: n = 10) of hearts. Quantitative differences were seen in complement fixation between the groups. Our data demonstrate the central role of complement in the destruction of pig-to-man xenografts.

Animals↗

PubMed2005

Mass spectral analysis of pig (Sus scrofa) apo HDL: Identification of pig apoA-II, a dimeric apolipoprotein.

Comparative studies of mammalian high density lipoproteins have clearly indicated that the major apolipoprotein is apoA-I and in some mammals apoA-II is the second major apolipoprotein. However, in pigs, apoA-II has been considered to be either present in trace amounts or absent. Recently, cDNA sequences for pigs A-II have been entered into the database. Translation of these sequences revealed that pig A-II consisted of 77 amino acids and that a cysteine residue was at residue 6. The A-II of three other mammals, chimpanzees, horses and humans, also has a cysteine residue at this position. As a result of a disulfide bond formed between monomers, the A-II in each of these cases circulates as a homodimer. Using electrospray-ionization mass spectrometry (ESI-MS), we obtained molecular mass data demonstrating that dimeric apoA-II is also present in pig plasma. In addition to being the first to report on the presence of apoA-II in pig plasma, we also obtained values for the molecular masses of apoA-I, apoC-III, apoD and serum amyloid A protein.

Amino Acid Sequence↗

PubMed2004

Quantitation of porcine circovirus type 2 isolated from serum/plasma and tissue samples of healthy pigs and pigs with postweaning multisystemic wasting syndrome using a TaqMan-based real-time PCR.

Porcine circovirus type 2 (PCV2) has been linked to several disease syndromes during the last decade. In this context, postweaning multisystemic wasting syndrome (PMWS) has emerged as a significant disease. As most pig herds are infected with PCV2, the determination of viral load in animals may be useful in discriminating between healthy and PMWS pigs. A TaqMan-based real-time PCR for quantitation of PCV2 in serum/plasma and tissue samples was established. A standard curve was created from serial dilutions of a plasmid encoding ORF2 (cap gene) of PCV2 and used to estimate the number of viral DNA copies in the analyzed samples. Comparison of viral load in mesenteric lymph nodes and serum/plasma from healthy animals and PMWS animals showed statistical significant difference between the two groups (p < 0.01). No healthy pigs had viral load greater than 10(6) PCV2 genomes per ml serum or 500 ng tissue sample, while all clinically sick PMWS pigs had PCV2 loads above 10(7) in both serum/plasma and in tissue samples. Furthermore, the estimated viral load in tissue samples from PMWS pigs was related to the immunohistochemical findings, with especially lymph nodes, ileum, and tonsil giving both high viral load, and a high degree of staining by immunohistochemistry.

Animals↗

PubMed2005

Jalpha-gene segment usage and the CDR3 diversity of porcine TCRalpha-chain cDNA clones from the PBL of a five-month-old pig and the thymus of a one-month-old pig.

Porcine T-cell receptor alpha (TCRalpha)-chain cDNA clones were isolated from libraries made from two different sources, the thymus of a 1-month-old LW strain pig and the peripheral blood lymphocytes (PBL) of a 5-month-old Clawn strain pig. Among 109 cDNA clones with the Jalpha-gene segment, 44 different Jalpha-gene segments were found out of the 61 Jalpha-gene segments previously identified in the porcine germline sequence. Among the 103 complete TCRalpha-chain cDNA clones with the rearranged Valpha- and Jalpha-gene segments, 33 different Valpha-gene segments were identified, which randomly rearranged to Jalpha-gene segments indicating lack of any specific combinations between Valpha- and Jalpha-gene segments with only one exception of the same set of Jalpha-gene segments in duplicate clones. Among the cDNA clones from PBL of an individual 5-month-old Clawn strain pig, a broad distribution of the Jalpha-gene segment usage was observed over the entire Jalpha-gene cluster. The Jalpha-gene segment usage in an individual 1-month-old thymus from a LW strain pig also gave a pattern consistent with the 5-month-old pig. These distributions of the Jalpha-gene segment usage were similar to the previously reported patterns for human T-cells and those of adult murine T-cells. Among the porcine cDNA clones isolated, TCRalpha-chain CDR3 length ranged from 4 to 14 amino acids with the average being 9.35 amino acids. Present report provides groundwork for further studies on porcine TCRalpha-chain expression.

Amino Acid Sequence↗

PubMed2006

Prevalence of hepatitis E virus antibodies in pigs: implications for human infections in village-based subsistence pig farming in the Lao PDR.

We report a high seroprevalence of hepatitis E virus (HEV) in pigs in the Lao PDR. HEV seroprevalence was 51.2% (300/586) amongst abattoir pigs and 15.3% (46/301) amongst village pigs. The age distribution suggested previous in-village HEV pig infections. These findings suggest a zoonotic risk associated with village-based smallholder pig farming.

Age Distribution↗

PubMed1998

No evidence of pig DNA or retroviral infection in patients with short-term extracorporeal connection to pig kidneys.

BACKGROUND: The xenotransplantation of organs and tissues, in particular those from pigs, is viewed as a means to alleviate the shortage of human donor organs and cells available for transplantation and also as a therapy for other diseases. The potential microbiological hazards of xenotransplantation have recently attracted much attention. One concern is over pig endogenous retroviruses (PERV). Until the possible consequences of infection by PERV are better understood it is unlikely that a significant number of porcine xenotransplants will proceed. However, a small number of patients have already been treated with or exposed to living porcine cells or tissue, and investigation of these patients may provide valuable information. METHODS: We took serial blood samples from two renal dialysis patients whose circulation had been linked extracorporeally to pig kidneys and tested them for pig DNA and PERV DNA by nested PCR. The patients' plasma was also tested for neutralising antibodies to two anthropotropic PERV strains. FINDINGS: Having established that the nested PCRs could detect single molecules of target sequence, we analysed DNA isolated from patients' peripheral blood mononuclear cells. We found no evidence of pig or PERV DNA in either patient, even in samples taken as early as 6 h after the perfusion. Furthermore, we found no evidence of seroconversion for PERV-specific antibodies. INTERPRETATION: The absence of porcine cells in the circulation of both patients, even in the samples taken soon after the perfusion experiment, suggests that any porcine cells dislodged from the kidney became rapidly sequestered from the circulation. Since cell-to-cell contact increases the efficiency of infection of PERV this removal of porcine cells may increase the risk of transmission of PERV to the xenograft recipient. We did not, however, detect indications of infection by PERV by PCR or neutralisation assay. The genetic and serological methods described here will be useful for detection of possible PERV infection in other patients.

Adult↗

PubMed2003

Mitochondrial DNA sequence variation and phylogenetic relationships among Iberian pigs and other domestic and wild pig populations.

Nucleotide sequences of mitochondrial DNA (mtDNA) cytochrome B gene (1140 bp) and control region (707 bp) were used to determine the phylogenetic relationships among 51 pig samples representing ancient and current varieties of Iberian pigs (26), Spanish wild boars (seven) and other domestic pigs (18) of cosmopolitan (Duroc, Large White, Landrace, Pietrain and Meishan) and local (Spotted Black Jabugo, Basque and Mangalitza) breeds. A neighbour-joining tree constructed from pairwise distances provide evidence of the European origin of both Iberian pigs and Spanish wild boars. The introgression of Asian mtDNA haplotypes in the genetic pool of the Iberian breed seems unlikely. Four estimates of sequence divergence between European and Asian clades were calculated from the two main domains of the D-loop region and the synonymous and nonsynonymous nucleotide substitutions in the cytochrome B gene. The time since the divergence of pig ancestors was estimated at about 600,000 years before present.

Animals↗

PubMed2003

Salmonella on pig carcasses: positive pigs and cross contamination in the slaughterhouse.

AIMS: The purpose of this study was to investigate the prevalence of Salmonella in pigs at the moment of slaughter and in the slaughterhouse environment. METHODS AND RESULTS: In total, five different commercial slaughterhouses were sampled during eight slaughterhouse visits. Carcass swabs, colon content and mesenteric lymph nodes were taken to reflect the animal status and from the slaughterhouse environmental samples were taken. Salmonella was isolated from 37% of the carcass samples as a mean value. High variations were noticed between different slaughterhouses (between 0 and 70%) and sampling days in the same abattoir (between 3 and 52%). A correlation was found between the carcass contamination and the status of the delivered animals (P=0.01675). Cross contamination was estimated to account for 29% of the positive carcasses. The slaughterhouse environment was highly contaminated; before starting the slaughtering activities 25% of the samples were positive on average. The most prevalent serotypes isolated at the slaughterhouse environment and from the colon content were S. Typhimurium, S. Livingstone and S. Derby. On carcasses S. Typhimurium was predominately isolated (71%). The biggest variability of serotypes was found in the mesenteric lymph nodes. Serologically 56.3% of the pigs were found positive for Salmonella using a cut-off level of the optical density percentage higher than 10 (O.D.% > or = 10). While on individual pig level the correlation between the bacteriological and serological data was poor, because of recent Salmonella infections, a better correlation was found at the herd level on the moment of slaughtering. CONCLUSION: A high degree of carcass contamination is noticed after slaughtering. This contamination resulted from the delivery of Salmonella-positive pigs and cross-contamination from the slaughterhouse environment. SIGNIFICANCE AND IMPACT OF THE STUDY: In pigs, Salmonella carriage is high, but it is obvious that slaughterhouse hygiene is a determinative factor for managing carcass contamination.

Abattoirs↗

PubMed1993

Phosphorus studies in pigs. 2. Assessing phosphorus availability for pigs and rats.

Experiments were conducted with pigs and rats to determine the availability of P in feeds. Initially, the availability of P in a soya-bean meal and field peas (Pisum sativum cultivar Early Dun) was assessed using a slope-ratio assay for grower pigs. Three different levels of either monosodium phosphate (MSP), soya-bean meal or field peas were added to a basal sucrose-soya-bean meal diet (2.5 g/kg P) to give three levels of P (3.0, 3.5 and 4 g/kg) for each source. The diets were offered for 35 d at three times maintenance energy requirements to female pigs initially weighing 20 kg live weight. Several bone variables and the ash and P concentration and retention levels in the empty body were used as criteria of availability. The responses to MSP were linear for all variables. However, responses to P in the test proteins, particularly soya-bean meal, were mostly non-linear, except for ash and P concentrations and retentions in the empty body. The estimates of P availability in the soya-bean meal and in the field peas were dependent on the criteria used to assess availability. Using bone variables as the criterion, the availabilities of P in soya-bean meal and field peas were approximately 0.17 and 0.38 respectively. Using ash or P concentrations or retentions in the empty body as the response criterion, the availabilities of P in soya-bean meal and field peas were 0.61 and 0.38 respectively. The mean retention values for P from MSP, soya-bean meal and field peas were 0.74, 0.33 and 0.21 respectively. Experiments were then conducted to define the conditions necessary for a slope-ratio assay for P availability with growing rats to determine if they could be used as a model to assess P availability for pigs. These experiments examined level of P in the diet, the effect of Ca:P ratio and criteria for assessing response (growth rate, feed intake, feed conversion ratio, bone ash and bone bending moment). The results indicated that a suitable range of dietary P for a slope-ratio assay in rats was 1.8-3.5 g/kg, with Ca:P ratio of between 1.3 and 6.2, and that ash content or bone bending moment of the femur bone were suitable criteria of response. However, using bone bending moment as the criterion of response, the availability of P in soya-bean meal was 0.81, which was considerably higher than the estimate with pigs.(ABSTRACT TRUNCATED AT 400 WORDS)

Animal Feed↗

PubMed1989

[The energy preservation requirements of growing pigs of different sexes fed normal and high protein diets. 5. Comparison with the results obtained from castrated pigs, sows and boars].

In one experiment each with castrated pigs, sows and boars (hybrid pigs of line 150), two groups of six animals received rations containing 17 and 45% crude protein resp. over the complete test period from 35 to 130 kg (castrated pigs), 150 kg (sows), 170 kg live weight (boars) and passed alternatively through a total of 60 metabolism periods on growth and 48 on maintenance level. There were significant differences between the 3 categories of animals with regard to energy maintenance requirement in the relation of 100:105:110 for castrated pigs less than sows less than boars. In contrast to the estimated values derived from previous studies for energetic maintenance requirement of 650 kJ/kg LW0.62.d the measured values for all 3 animal categories are by approximately 50% higher. The protein content did not have an influence on the maintenance requirement of metabolizable energy of the animals. The utilization of metabolizable energy of the two rations for body energy retention corresponds to the expectations with regard to its dependence on nutrients. The requirement of metabolizable energy for protein and fat retention is 1.7:1.0. The energy content of the weight gain of boars was, on average, 85% of that of castrated pigs and sows. The studies of blood parameters did not show any deviations in the parameters tested.

Animal Feed↗

PubMed2005

Progeny of somatic cell nuclear transfer (SCNT) pig clones are phenotypically similar to non-cloned pigs.

Systematic studies of cloned animals generated from adult somatic cell nuclei are critical in assessing the utility of somatic cell cloning in various applications, including the safety of food products from cloned animals and their offspring. Previously, we compared somatic cell derived cloned pigs with naturally bred control pigs on a series of physiological and genetic parameters. We have extended our studies to the F1 progeny of these clones to see whether these phenotypic differences are transmitted to the next generation. There were no differences in the average litter size between litters from cloned gilts and naturally bred controls (7.78 +/- 2.6 and 7.40 +/- 3.0, respectively; mean +/- SD) or in the degree of litter size variation (coefficients of variation of 33.4% and 40.5% for litters of clones and controls, respectively). Similarly there were no statistical differences between sex ratios of cloned litters (51-49%, M:F) and control litters (59-41%, M:F). Blood profiles between cloned pigs, control pigs, and their progeny were compared at two time points (i.e., 15 and 27 weeks) to quantify the effect of cloning on various blood parameters and their transmission to the next generation. Although the range of values for all traits overlapped between different classes, the variability between all the traits in F1 progeny of clones and the control pigs was similar at 15 and 27 weeks, with one exception. Combined, our data and previous results in mice strongly support the hypothesis that offspring of clones are similar to offspring of naturally bred animals, and as such there should not be any increased risks associated with consumption of products from these animals.

Animals↗

PubMed1990

Tissue penetration of cefotaxime in normal pigs and pigs with haemorrhagic pancreatitis.

This study was designed to evaluate the tissue penetration of cefotaxime in normal pigs and pigs with haemorrhagic pancreatitis. Serum, peritoneal fluid, bile, gallbladder wall and pancreatic tissue concentrations of cefotaxime in these groups of pigs exceeded the MIC90 for susceptible species of Gram-negative aerobic bacteria. Cefotaxime penetration into pancreatic tissue and peritoneal fluid was increased from 2% to 2.6% and 73% to 89%, respectively, in pigs with pancreatitis in comparison with normal pigs. These increases however, were not statistically significant.

Animals↗

PubMed1996

Antibiotic resistance among Escherichia coli isolated from faecal samples of pig farmers and pigs.

The prevalence and degree of antibiotic resistance of Escherichia coli found in 266 faecal samples of pig farmers was significantly lower than that observed in 285 samples obtained from their pigs. Moreover, porcine E. coli were significantly more resistant to chloramphenicol, nitrofurantoin, oxytetracycline, streptomycin and sulphamethoxazole than were farmer isolates. Resistant pig isolates showed resistance patterns mainly to oxytetracycline and streptomycin and oxytetracycline and streptomycin and sulphamethoxazole whereas the corresponding farmer isolates were mainly resistant to a single agent such as amoxycillin or sulphamethoxazole although 41 were resistant to both streptomycin and sulphamethoxazole. The resistance patterns of only 4% of farmer E. coli and those of pigs from the same farm (n = 259) showed resistance to the same antibiotics. These results suggest that the resistance of the faecal E. coli of farmers and their pigs is distinctly different.

Agricultural Workers' Diseases↗

PubMed1999

Characterization of human xenoreactive antibodies in liver failure patients exposed to pig hepatocytes after bioartificial liver treatment: an ex vivo model of pig to human xenotransplantation.

BACKGROUND: There are limited experimental data on the nature of the humoral response elicited in humans against pig antigens. In this study, we have examined the xenoantibody (XAb) response in eight patients with acute liver failure exposed to pig hepatocytes after treatment with the bioartificial liver (BAL). METHODS: Patients' plasma samples obtained before and after BAL treatment were tested for IgM and IgG XAbs, IgG subclasses, and XAb cytotoxicity, using enzyme-linked immunosorbent assay and flow-cytometric assays. The characterization of pig aortic endothelial cell (PAEC) surface xenoantigens was analyzed by immunoprecipitation. RESULTS: We observed by day 10, a strong anti-pig IgG and IgM XAb response in patients undergoing two or more BAL treatments, with a significant increase in all the IgG subclasses; in contrast, XAb titers did not change if the patients received only one BAL treatment. The majority of the XAbs produced to porcine antigens were primarily specific for the alphaGal epitope. Both IgG and IgM XAbs were cytotoxic to PAECs, and the cytotoxic activity of IgG was associated with high levels of IgG1 and IgG3 subclasses, known to be efficient on complement activation. The characterization of porcine surface antigens demonstrated that IgM human XAbs, before and after BAL exposure, recognized xenoantigens on PAECs with similar molecular weights, suggesting that the same population of XAbs were present in the patients before and after exposure to pig antigens. CONCLUSIONS: Repetitive exposure of humans to porcine antigens after BAL treatment, results in a strong IgG and IgM XAb responses that are primarily directed against the alphaGal epitope. These XAbs are cytotoxic to PAECs and the IgG toxicity correlates with high IgG1 and IgG3 levels. Our data also suggest that no new XAb specificity emerges after porcine exposure.

Animals↗

PubMed1999

Neonatal pig islets induce a lower T-cell response than adult pig islets in IDDM patients.

BACKGROUND: Pancreatic pig islets may provide a substitute in the future for difficult to obtain human islets for transplantation in insulin-dependent diabetes millitus (IDDM) patients. However, the immune response to xenografts may significantly hamper this approach. Because neonatal tissue is believed to be less immunogenic, we examined whether the T-cell response to neonatal pig islets differs from the response to adult islets. METHODS: The T-cell proliferative response to different concentrations of sonicated neonatal and adult pig islets, as well as to insulin and mitogens, was tested in 21 recent onset IDDM patients and 21 healthy controls. We determined the presence of various circulating islet autoantibodies and their association with the T-cell response in IDDM patients. RESULTS: In the IDDM patients, sonicated adult pig islets (at 1 microg protein/ml) induced a significantly higher frequency (12 of 21 vs. 1 of 21, p<0.001) and magnitude (2.58+/-0.44 vs. 1.38+/-0.13, p<0.02) of positive T-cell responses than neonatal islets at the same concentration. Similar results were obtained with a 10-fold higher concentration of islet sonicate. There was no significant association between the individual T-cell responses and the presence of circulating autoantibodies in IDDM patients. CONCLUSION: These results indicate that neonatal pig islets induce a lower T-cell reactivity than adult islets, suggesting that the neonatal tissue may be immunologically more suitable for future islet xenotransplantation.

Adolescent↗

PubMed1985

A comparison of the ecology of Escherichia coli in the intestine of healthy unweaned pigs and pigs after weaning.

A total of 51 clinically healthy pigs (14 unweaned and 37 weaned) from five litters, and aged 21 to 35 d, were studied. Escherichia coli isolates from the duodenum, jejunum, ileum, caecum and colon were differentiated on the basis of O-serogroup, biotype and resistance pattern. The complexity of the flora was influenced considerably by the presence or absence of the enterotoxigenic serotype 0149: K91, K88a,c (Abbotstown strain). When it was absent the E. coli flora of both weaned and unweaned pigs was complex with up to 25 strains being identified. The majority of these E. coli strains identified in each pig were isolated from only one of the five intestinal sites sampled. On the other hand, when the enterotoxigenic strain was present (14 pigs) it tended to dominate the E. coli flora at all levels of the intestine and this dominance was reflected in a corresponding fall in the total number of E. coli strains isolated per pig.

Animals↗

PubMed2004

Hyperacute rejection of hDAF-transgenic pig organ xenografts in cynomolgus monkeys: influence of pre-existing anti-pig antibodies and prevention by the alpha GAL glycoconjugate GAS914.

BACKGROUND: Our introductory pig-to-cynomolgus monkey heart or kidney transplantation using organs from pigs transgenic for human decay-accelerating factor (hDAF), showed a high incidence of hyperacute rejection (HAR), which was ascribed to extraordinary high levels of anti-pig antibodies. We evaluated the efficacy of GAS914, a Gal alpha 1-3Gal trisaccharide linked to a poly-l-lysine backbone, in inhibition of HAR. METHODS: hDAF transgenic heterotopic heart (n = 15) or life-supporting kidney (n = 8) transplantation included induction with cyclophosphamide or anti-thymocyte globulin, and maintenance with cyclosporine or tacrolimus, steroids and mycophenolate sodium/mofetil. Four doses of GAS914 were given before transplantation. Rejection was confirmed by graft histology, and anti-pig antibody levels were determined in various assays. RESULTS: Four of six heart transplants without GAS914 treatment showed HAR. Nine subsequent transplants with GAS914 pre-treatment, did not show HAR (chi-square, P < 0.05). Two of four kidney transplants without GAS914 treatment ended with HAR. Four subsequent transplants with GAS914 did not show HAR. Animals with HAR showed extremely high antibody levels. Samples just before transplantation showed significantly higher antibody levels in recipients presenting with HAR. In all assays antibody levels were significantly lowered by GAS914 pre-treatment. CONCLUSIONS: HAR of hDAF solid organs could be ascribed to high levels of anti-pig antibodies. It is hypothesized that the hDAF transgene shows a threshold in efficacy, above which an overwhelming attack by antibodies and complement activation cannot be modulated to prevent HAR. HAR does not occur when animals with lower levels are used, or when antibodies are effectively depleted from the circulation by GAS914 treatment.

Acute Disease↗