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Monomorphic and polymorphic carbohydrate antigens on pig tissues: implications for organ xenotransplantation in the pig-to-human model.

The existence of the alpha Gal epitope in 137 pigs belonging to 23 different breeds suggests that this antigen is either monomorphic or occurs at a high incidence in the porcine species. Its histological location at the surface of pig vascular endothelial cells makes it a target for human natural anti-alpha Gal antibodies and complement, which may be responsible for the hyperacute vascular rejection of transplanted pig organs. The precursor carbohydrate chain (N-acetyllactosamine) and NeuAc-substituted epitopes are also exposed at the surface of pig vascular endothelium and were found in all pigs in this study. However, humans also have these two epitopes on vascular endothelium and, consequently, have not made natural antibodies against these carbohydrate antigens. Therefore, these two pig epitopes cannot be the main target of the hyperacute vascular rejection process. Three pig phenotypes-A+ (51%), A:H+ (38%), and A-H- I+ (11%) were identified among 37 Large-white pigs by the presence of polymorphic A, H, and I carbohydrate antigens on the brush border of the surface epithelium of small intestine. These antigens were also present in other exocrine secretions but were not detected on vascular endothelium of the same pigs, suggesting that they are not involved in the hyperacute vascular rejection, although the pig A tissue antigen can induce an immune response in 0 or B blood group recipients. Once the problem of the initial hyperacute vascular rejection directed against the alpha Gal epitope is overcome, typing donor pigs for A, H, and I, as well as for the protein swine leukocyte antigens (SLA) and other pig antigens, may help in elucidating antigens involved in acute or chronic xenograft rejection.

ABO Blood-Group System↗

Cell composition and lymphocyte subsets in the bronchoalveolar lavage of normal pigs of different ages in comparison with germfree and pneumonic pigs.

For studies of lymphocyte kinetics in the different compartments of the lung, basic data on the cellular composition and lymphocyte subsets of porcine bronchoalveolar lavage (BAL) are essential. Therefore BAL was performed in pigs, and cytologic findings were studied using cytologic staining, enzyme cytochemistry, and immunologic labeling techniques. Four groups of normal pigs at different ages and of different breeds, 1 group with bacterial pneumonia, and 1 group of germfree animals were used. The total cell recovery was 10 x 10(6) in germfree pigs, in normal pigs it was 1.2-4 times higher, and in pneumonic pigs 23 times higher. The main cell type was macrophages, approximately 80% in normal and germfree and 50% in pneumonic pigs. The BAL contained 20% lymphocytes. Granulocytes were absent in normal BAL, but formed 30% of pneumonic BAL. Here the total number of macrophages was 3-9 times higher than normal, the total number of lymphocytes was 11-23 times higher. The proportion of surface immunoglobulin positive (sIg+) cells was 4.5% in the young and 8.5% in the older pigs, and of T cells it was 6% in the young and 14.9% in the older pigs. The BAL of germfree pigs contained few sIg+ cells and half of the normal number of T cells. In pneumonic animals the BAL contained twice the number of sIg+ and a normal percentage of T cells. The percentage of T-helper lymphocytes was approximately 2% without marked differences between the groups, whereas the percentage of T-suppressor lymphocytes varied significantly from 0.8% in germfree to 4% in young normal and 8% in the older pigs. In this respect the pneumonic pigs did not differ from the normal groups. The study shows that the pig is a suitable animal model for further investigation of the cellular immune system of the lung.

Age Factors↗

Digestion in the pig between 7 and 3 d of age. 3. The digestion of nitrogen in pigs given milk and soya-bean proteins.

1. In two separate experiments, forty-four pigs weaned at 4--5 d of age were fed on diets containing milk or soya-bean protein until slaughtered at 14, 28 or 35 d of age. 2. Daily amounts of nitrogen flowing through the stomach and proximal small intestine of pigs were similar with both sources of protein fed. 3. Endogenous N flows in the duodenum, jejunum and ileum of pigs given an N-free diet were 22 . 9, 7 . 4 and 4 . 6 g N/kg dry matter (DM) intake. Endogenous N in faeces was 0 . 970 g N/kg DM intake. 4. The apparent and true digestibility of N to the ileum of 28-d-old pigs was 0 . 86 and 0 . 92 for the pigs given the milk-protein diets, 0 . 80 and 0 . 86 for the pigs given the isolated soya-bean protein (ISP; Promine D) diets and 0 . 51 and 0 . 62 for pigs given the soya-bean meal (SBM)-protein diet. These values over the entire gastro-intestinal tract were 0 . 98 and 1 . 00 for milk, 0 . 92 and 0 . 95 for ISP and 0 . 82 and 0 . 85 for SBM. 5. The apparent and true digestibility of N to the ileum of 14-d-old pigs given a liquid milk diet was 0 . 92 and 0 . 94, while the values for the pigs given a pelleted milk diet were 0 . 86 and 0 . 87. At 35 d of age there were no differences in the apparent digestibility of the N in the liquid an pelleted milk diets. 6. The apparent digestibility of N to the ileum and over the entire gastro-intestinal tract of pigs given ISP (Supro 610) increased with increasing age of pigs from 0 . 83 at 14 d of age to 0 . 88 at 35 d of age. 7. Of the N in the stomach 14--32% could not be precipitated by trichloroacetic acid (TCA; 30 milligram). 8. In the jejunal and ileal contents an average of 46 and 24% of the N was precipitated by TCA in pigs given soya-bean proteins and milk proteins respectively.

Aging↗

Preterm birth affects the intestinal response to parenteral and enteral nutrition in newborn pigs.

Maturation of gastrointestinal (GI) function in neonates is stimulated by enteral nutrition, whereas parenteral nutrition induces GI atrophy and malfunction. We investigated whether preterm birth alters the GI responses to parenteral and enteral nutrition. Pigs were delivered either preterm (107 d gestation) or at term (115 d gestation) and fed total parenteral nutrition (TPN) or enteral sow's milk (ENT) for 6 d after birth. Immaturity of the preterm pigs was documented by reduced blood pH, oxygen saturation and neutrophil granulocyte function, impaired intestinal immunoglobulin G uptake from colostrum, and altered relative weights of visceral organs (small intestine, liver, spleen, pancreas, and adrenals). For both ages at delivery, increases occurred in pancreatic weight (30-75%) and amylase activity (0.5- to 13-fold) after birth, but much more in ENT than in TPN pigs (P < 0.05). Six days of TPN feeding was associated with reduced intestinal weight for both delivery groups (60% of values in ENT, P < 0.001), but only in term TPN pigs was the weight lower than at birth (-20%, P < 0.05). Likewise, it was only in term TPN pigs that intestinal maltase activity increased, compared with ENT, and the absorption of glucose and proline decreased. Only in preterm pigs did TPN feeding increase lactase activity (+50% compared with ENT, P < 0.05). For both delivery ages, the mRNA of lactase-phloridzin hydrolase and sodium-coupled glucose transporter 1 were increased in TPN, compared with ENT. In conclusion, the trophic effect of enteral vs. parenteral nutrition on the GI tract is also present after preterm birth, but the postnatal maturation of many GI functions is modified, compared with term birth. The effects of nutritional regimen on the maturation of the gut epithelium in neonates depend on gestational age at birth.

Animals↗

Removal of natural human xenoantibodies to pig vascular endothelium by perfusion of blood through pig kidneys and livers.

We have examined the nature of the binding of human xenoantibodies to pig liver and kidney vascular endothelium. Our results demonstrate that human serum contains IgM and IgG xenoantibodies that bind to pig vascular endothelium, and that the pattern of antibody binding is similar for both livers and kidneys. Immunohistochemical analysis of pig kidneys after perfusion with human blood demonstrated the binding of both IgM and IgG xenoantibodies, complement (C3), and fibrinogen to the vascular and glomerular endothelium. An ELISA assay of the perfusate after perfusion of 500 ml of human blood through a single pig kidney for 60 min demonstrated a significant reduction in the amount of human IgM (67%) and IgG (55%) binding to pig aortic endothelium. Similar perfusion experiments conducted with pig livers were associated with minimal immunohistochemical evidence of the binding of human xenoantibodies to liver vascular endothelium. Immunofluorescence staining for IgM, IgA, C3, and C1q was negative or minimally positive in the liver vascular endothelium. Sinusoidal endothelium were weakly positive for IgG and fibrinogen. The perfusion of the pig liver with human blood was, however, associated with a significant reduction in the subsequent binding of IgM and IgG to pig kidney vascular endothelium. Pig liver perfusion was also responsible for the removal of both IgM and IgG xenoantibodies capable of reacting with pig aortic endothelium, as measured by an ELISA assay of the perfusate. These results suggest that both pig kidney and livers are capable of absorbing the xenoantibodies that may be responsible for mediating a hyperacute rejection of pig xenografts and that the distribution of the target antigens for these antibodies is similar in the two organs.

Animals↗

Carbohydrate antigens of pig tissues reacting with human natural antibodies as potential targets for hyperacute vascular rejection in pig-to-man organ xenotransplantation.

Pig tissues were screened by immunofluorescence with lectins, mAb, and human natural antibodies for the presence of carbohydrate antigens, which may be potential targets for hyperacute vascular rejection in pig to man xenotransplantation. The unfucosylated monomorph linear B-antigen was found at the surface of all porcine vascular endothelial cells. This pig linear-B antigen reacts strongly with the anti-alpha Gal isolectin B4 from Griffonia simplicifolia 1 and with human natural anti-alpha Gal antibodies specifically purified by affinity chromatography on synthetic oligosaccharides containing the terminal nonreducing alpha Gal1-->3 beta Gal-R disaccharide. This antigenic activity is destroyed by treatment of pig tissues with alpha-galactosidase. The localization of this linear-B epitope on vascular endothelium and its reactivity with natural human anti-alpha Gal antibodies suggest that it may play a major role in the hyperacute vascular rejection of pig to man organ xenografts. The lectin from Maackia amurensis reacting with alpha NeuAc2-->3 beta Gal1-->4GlcNAc/Glc was also positive on pig vascular endothelium, but we do not know yet whether there are human natural antibodies reacting with the carbohydrate recognized by this lectin. Epithelial cells of pig renal proximal convoluted tubules, respiratory epithelium, pancreatic ducts, and epidermis express the linear-B antigen, but they are less likely to trigger a hyperacute vascular rejection because they are not directly exposed to the blood. The genetically defined pig A+/A- system controls the expression of A and H antigens in pig epithelial cells from renal distal and collecting tubules, biliary ducts, pancreatic ducts, large bronchi, and digestive mucosa. The pig A antigen may trigger an immune response in human O or B recipients if they are transplanted with organs from A+ pigs, but the pig A antigen is probably not involved in the hyperacute vascular rejection of a xenograft because it is not expressed on vascular endothelium.

Acute Disease↗

Identification of glucose and nucleoside transport proteins in neonatal pig erythrocytes using monoclonal antibodies against band 4.5 polypeptides of adult human and pig erythrocytes.

Cytochalasin B and nitrobenzylthioinosine (NBMPR), which inhibit membrane transport of glucose and nucleosides, respectively, have served as photoaffinity ligands that become covalently linked at inhibitor binding sites on transporter-associated proteins. Thus, when membranes from erythrocytes of neonatal pigs with site-bound [3H]cytochalasin B or [3H]NBMPR were irradiated with uv light, two labeled membrane polypeptides (peak Mr values: 55,000 and 64,000, respectively) were identified. Treatment of the photolabeled membranes with endoglycosidase F increased the mobility of [3H]cytochalasin B- and [3H]NBMPR-labeled material (peak Mr values: 44,000 and 57,000, respectively) and limited digestion with trypsin yielded different polypeptide fragments (Mr values: 18,000-23,000 and 43,000, respectively). Identification of the photolabeled polypeptides as transporter components was established using monoclonal antibodies (MAbs) raised against partially purified preparations of band 4.5 from erythrocytes of adult pigs and humans. MAbs 65D4 and 64C7 (anti-human band 4.5), raised in this study, reacted with [3H]cytochalasin B-labeled material from membranes of human erythrocytes and bound to permeabilized erythrocytes but not to intact cells. MAb 65D4 also bound to erythrocytes of mice and neonatal pigs and to a variety of cultured cells (mouse, human, rat), including AE1 mouse lymphoma cells, which lack an NBMPR-sensitive nucleoside transporter. Also employed was MAb 11C4 (anti-pig band 4.5), which recognizes the NBMPR-binding protein of erythrocyte membranes from adult pigs. When membrane proteins from neonatal and adult pigs were subjected to electrophoretic analysis and blots were probed with different MAbs, MAb 65D4 (anti-human band 4.5) bound to material that comigrated with [3H]cytochalasin B-labeled polypeptides (band 4.5) from neonatal, but not adult, pig erythrocytes, whereas MAb 11C4 (anti-pig band 4.5) bound to material that comigrated with [3H]NBMPR-labeled band 4.5 polypeptides of erythrocytes from both neonatal and adult pigs. These results, which indicate structural differences in the cytochalasin B- and NBMPR-binding proteins of pig erythrocytes, establish the presence of both proteins in erythrocytes of neonatal pigs and suggest that only the NBMPR-binding protein is present in erythrocytes of adult pigs.

Affinity Labels↗

Infectivity of a strain of Cryptosporidium found in the guinea-pig (Cavia porcellus) for guinea-pigs, mice and lambs.

Cryptosporidiosis was diagnosed in guinea-pigs bred by a commercial laboratory supplier on histological examination of the intestine. Oral transmission to laboratory guinea-pigs aged up to 16 weeks and to infant mice, with gut contents containing oocysts, was successful, but the organism failed to infect adult mice. From day 5 post-inoculation (pi) in guinea-pigs, infection of the ileum was associated with villous stunting and fusion, and with infiltrates of macrophages and other mononuclear cells, and eosinophils. Some guinea-pigs died; others were depressed and anorectic, with diarrhoea or watery caecal contents. Mouse infections were subclinical and caused no significant pathological changes. By contrast, a bovine Cryptosporidium isolate infected infant mice but failed to infect young guinea-pigs. Guinea-pigs and infant mice excreted oocysts in faeces after a prepatent period of 3 to 4 days. Some guinea-pigs excreted oocysts for up to 2 weeks, but excretion in mice lasted only about 4 days. Infection of guinea-pigs by contact with a contaminated environment occurred, with excretion of oocysts between 17 and 27 days after exposure. An indirect fluorescent antibody test (IFA) showed that antibody was present by day 17 pi with infected bowel contents, but none was detected in the guinea-pigs exposed to the contaminated environment. The IFA test demonstrated a serological relationship between the guinea-pig isolate and a bovine strain used to infect gnotobiotic lambs. Transmission electron microscopy of intestine from infected guinea-pigs and mice showed that more than one schizont generation occurred. The first consisted of 8 merozoite packets attached to enterocytes, but many packets of 2 or 4 merozoites of the second or subsequent generations were apparently released into the gut lumen. Fixed microgametocytes contained lipid vacuoles and had microneme-like structures in their cytoplasm. Oocysts and sporocysts were also identified, with sporulation occurring within the parasitiphorous vacuole. A sparse infection was established in 1 of 2 12-day-old specific pathogen-free lambs by day 3 pi, but no oocysts were detected in its caecal contents or those of a second lamb killed 4 days later.

Animals↗

Intestinal absorption and retention of cadmium in neonatal pigs compared to rats and guinea pigs.

The purpose of this study was to measure intestinal absorption and retention of cadmium in the newborn pig and to compare data from the pig, rat and guinea pig, three species that differ greatly in their ability to absorb macromolecules at birth. Newborn pigs were administered a single oral dose of 50 muCi of 115mCd (0.20 mCi/mg Cd) 24 hours after birth and killed at intervals between 1 and 14 days after dosing. Cd absorption and gastrointestinal retention were then determined; these data were compared with similar data from the rat and guinea pig. The cumulative Cd absorption was about 4.0, 5.0 amd 23% of the dose in the day-old pig, rat and guinea pig, respectively. The residence time of Cd in the intestinal tract was longer in the rat than in the pig or guinea pig, the 50% excretrion time being 15, 8, and 5 days in the rat, pig and guinea pig, respectively. Cd absorption in the neonate appears to be a two-step process: mucosal uptake of Cd from the lumen, probably by pinocytosis, followed by transfer of a portion of this Cd into the body. This transfer process is similar, but does not entirely coincide with changes associated with protein absorption in the neonate.

Age Factors↗

Association of insulin resistance with hyperglycemia in streptozotocin-diabetic pigs: effects of metformin at isoenergetic feeding in a type 2-like diabetic pig model.

Insulin-mediated glucose metabolism was investigated in streptozotocin (STZ)-treated diabetic pigs to explore if the STZ-diabetic pig can be a suitable model for insulin-resistant, type 2 diabetes mellitus. Pigs (approximately 40 kg) were meal-fed with a low-fat (5%) diet. Hyperinsulinemic (1, 2, and 8 mU kg(-1) min(-1)) clamps and/or 6,6-(2)H-glucose infusion studies were performed in 36 pigs. Diabetic (slow, 30-minute infusion of 130 mg STZ/kg) vs normal pigs were nonketotic, showed fasting hyperglycemia (21.7 +/- 1.1 vs 5.3 +/- 0.2 mmol/L), comparable plasma insulin (9 +/- 7 vs 5 +/- 1 mU/L), and elevated triglyceride concentrations (1.0 +/- 0.3 vs 0.2 +/- 0.1 mmol/L). After a standard meal, plasma triglycerides, cholesterol, and nonesterified fatty acid concentrations were significantly higher in diabetic vs normal pigs (1.2 +/- 0.3 vs 0.3 +/- 0.1, 2.3 +/- 0.2 vs 1.7 +/- 0.1, and 1.5 +/- 0.5 vs 0.2 +/- 0.1 mmol/L, respectively, P < .05). Fasting whole-body glucose uptake, hepatic glucose production, and urinary glucose excretion were increased (P < .01) in diabetic vs normal pigs (9.1 +/- 0.6 vs 4.8 +/- 0.4, 11.4 +/- 0.6 vs 4.8 +/- 0.4, and 2.3 +/- 0.2 vs 0.0 +/- 0.0 mg kg(-1) min(-1)). During hyperinsulinemic euglycemia (approximately 6 mmol/L), whole-body glucose uptake was severely reduced (P < .01) and hepatic glucose production was moderately increased (P < .05) in diabetic vs normal pigs (6.7 +/- 1.3 vs 21.1 +/- 2.2 and 1.7 +/- 0.5 vs 0.8 +/- 0.3 mg kg(-1) min(-1)) despite plasma insulin concentrations of 45 +/- 5 vs 24 +/- 5 mU/L, respectively. Metformin vs placebo treatment of diabetic pigs (twice 1.5 g/d) for 2 weeks during isoenergetic feeding (1045 kJ/kg body weight(0.75)) resulted in a reduction in both fasting and postprandial hyperglycemia (14.7 +/- 1.5 vs 19.4 +/- 0.6 and 24.9 +/- 2.2 vs 35.5 +/- 4.9 mmol/L), a reduction in daily urinary glucose excretion (approximately 250 vs approximately 350 g/kg food), and an increase in insulin-stimulated glucose disposal (9.4 +/- 2.2 vs 5.8 +/- 1.7 mg kg(-1) min(-1); P < .05), respectively. In conclusion, a slow infusion of STZ (130 mg/kg) in pigs on a low-fat diet induces the characteristic metabolic abnormalities of type 2 diabetes mellitus and its sensitivity to oral metformin therapy. It is therefore a suitable humanoid animal model for studying different aspects of metabolic changes in type 2 diabetes mellitus. Insulin resistance in STZ-diabetic pigs is most likely secondary to hyperglycemia and/or hyperlipidemia and therefore of metabolic origin.

Animals↗

Salmonella in slaughter pigs: the effect of logistic slaughter procedures of pigs on the prevalence of Salmonella in pork.

A substantial part of the finishing pigs in the Netherlands is infected with Salmonella. Infection of pigs with Salmonella can occur already on the farm. Pigs can also get infected or contaminated during transport, lairage or slaughter. The aim of this study was to evaluate the effect of separating pigs from Salmonella-infected farms from pigs from Salmonella-free farms during transport, lairage and slaughter on the prevalence of Salmonella on pork after slaughter. Two experiments were carried out. In the first experiment, farms were selected to participate, based on serology of the pigs (Dutch Salmonella ELISA). The pigs were slaughtered at the beginning of the day: firstly, sero-negative herds, secondly, sero-positive herds and thirdly, again sero-negative herds. The latter were slaughtered to investigate the effect of a contaminated slaughterline due to a previously slaughtered positive herd. In the second experiment, farms were selected to participate, based on both serology and bacteriology of the pigs on the farm. Two hundred pigs from Salmonella-free farms were slaughtered after 200 pigs from Salmonella-infected farms. Results showed that the prevalence of Salmonella in pork samples of sero-negative herds was lower than in samples of sero-positive herds. Results also showed that Salmonella contamination of carcasses after slaughter was partially caused by Salmonella-infected herds that were slaughtered before, and partially by residential flora of the slaughterhouse. It is concluded that separate slaughter of sero-negative pig herds can be useful to decrease the prevalence of Salmonella-contaminated pork after slaughter. To avoid cross-contamination by residential flora from trucks, lairage and slaughterline, cleaning and disinfection have to be improved.

Abattoirs↗

Feeding NOD mice with pig splenocytes induces transferable mechanisms that modulate cellular and humoral xenogeneic reactions against pig spleen or islet cells.

We have reported previously that oral administration of pig cells to NOD mice modified xenogeneic cellular response against pig islet cells (PICs), and hypothesized that it may have induced active suppression. This preliminary report evaluated only the effect of feeding pig cells by 'primary' proliferation, i.e. when splenocytes from fed mice are confronted with pig cells in vitro. The present study also considered 'secondary' proliferation and cytokine production after feeding and subsequent in vivo graft of pig cells. Additionally, serum IgM and IgG isotypes were quantified by ELISA using pig target cells. Induction of active mechanism by feeding was hypothetical, which led us here to transfer splenocytes from mice fed pig spleen cells (PSC) and evaluate 'primary' (after transfer) and 'secondary' (after transfer and subsequent graft of pig cells) proliferations and cytokine secretions in recipient mice. We also determined whether the effects of feeding pig cells persisted after depression of suppressor mechanisms by cyclophosphamide. Mice fed with PSC displayed increased 'primary' splenocyte proliferation to PSC or PIC (P < 0.0001), while 'secondary' responses were decreased (P < 0.03) in those fed PSC and subsequently grafted with PSC. The increased 'primary' and decreased 'secondary' proliferations were reduced (P < 0.04) by pretreatment with cyclophosphamide. The IL-10/ and IL-4/IFNgamma ratios produced in response to PSC increased (P < 0.04) in mice fed and grafted with PSC compared to those grafted only with PSC. IgM and IgG levels against pig cells were, respectively, increased (P < 0.04) and decreased (P < 0.04) in mice fed and grafted with PSC. IgG2a and IgG2b, but not IgG1, levels were lower (P < 0.01). These effects of feeding PSC on 'secondary' proliferation, cytokine and antibody productions, were not detected when mice were fed PSC only after graft with PSC. Transfer with splenocytes from mice fed PSC increased 'primary' proliferation of splenocytes from recipient mice in response to PSC (P < 0.02) or PIC (P < 0.05). After transfer with splenocytes from PSC-fed mice and graft with PSC, 'secondary' proliferation to pig cells were reduced (P < 0.04), and the IL-10/IFNgamma ratio produced in response to PSC was increased fourfold. Thus, oral administration of PSC induces active transferable mechanisms, characterized by a biphasic pattern with early increased 'primary' xenogeneic cellular reactions to both PSC and PIC, followed by decreased 'secondary' responsiveness and a concomitant shift of the Th1/Th2 balance towards greater Th2 influence. Decreased responsiveness may be due to active suppression, even though induction of anergy or deletion cannot be excluded.

Administration, Oral↗

Herd factors associated with the seroprevalences of four major respiratory pathogens in slaughter pigs from farrow-to-finish pig herds.

The objective of this study was to investigate sero-epidemiological aspects of Mycoplasma hyopneumoniae (Mh), influenza H1N1 and H3N2 viruses and Aujeszky disease virus (ADV) in fattening pigs from 150 randomly selected farrow-to-finish pig herds. Different herd factors were examined as potential risk indicators for the percentage of pigs with antibodies against the 4 pathogens. The median within-herd seroprevalences of the pathogens were: Mh 76%, H1N1 100%, H3N2 40% and ADV 53%. There was a positive association between the seroprevalences of both influenza viruses, and a negative association between the seroprevalences of ADV and H1N1. The percentage of pigs seropositive for Mh increased with the purchase of gilts and with the season (slaughter date in March-April). The within-herd seroprevalences of both influenza viruses were higher in the case of a higher density of pig herds in the municipality. A higher number of fattening pigs per pen additionally increased the risk of being seropositive for H3N2. The percentage of pigs with anti-gE-antibodies against the wild type ADV increased with higher airspace stocking density in the finishing unit, increasing herd size, increasing number of pig herds in the municipality and slaughter date in March-April. Increased seroprevalences for these 4 respiratory pathogens were mostly associated with pig density in the herd and its vicinity, the winter period, and with the purchase of gilts. Purchase of gilts, number of fattening pigs per pen and airspace stocking density are risk factors that can be managed directly by farmers striving to attain a high respiratory health status of pigs.

Animal Husbandry↗

PIG-S and PIG-T, essential for GPI anchor attachment to proteins, form a complex with GAA1 and GPI8.

Many eukaryotic cell surface proteins are anchored to the plasma membrane via glycosylphosphatidylinositol (GPI). The GPI transamidase mediates GPI anchoring in the endoplasmic reticulum, by replacing a protein's C-terminal GPI attachment signal peptide with a pre-assembled GPI. During this transamidation reaction, the GPI transamidase forms a carbonyl intermediate with a substrate protein. It was known that the GPI transamidase is a complex containing GAA1 and GPI8. Here, we report two new components of this enzyme: PIG-S and PIG-T. To determine roles for PIG-S and PIG-T, we disrupted these genes in mouse F9 cells by homologous recombination. PIG-S and PIG-T knockout cells were defective in transfer of GPI to proteins, particularly in formation of the carbonyl intermediates. We also demonstrate that PIG-S and PIG-T form a protein complex with GAA1 and GPI8, and that PIG-T maintains the complex by stabilizing the expression of GAA1 and GPI8. Saccharomyces cerevisiae Gpi16p (YHR188C) and Gpi17p (YDR434W) are orthologues of PIG-T and PIG-S, respectively.

Acyltransferases↗

The induction of specific pig skin graft tolerance by grafting with neonatal pig thymus in thymectomized mice.

BACKGROUND: Xenogeneic donor-specific tolerance can be induced by transplanting fetal pig thymus and liver tissue (FP THY/LIV) to thymectomized (ATX), T/NK cell-depleted mice. By using neonatal pig tissue, we hoped to overcome two obstacles that arise with the use of fetal pig tissue: (1) the inability to keep fetal pigs alive after harvesting their thymic tissue, resulting in unavailability of their skin or other organs for grafting; and (2) the limited fetal thymic tissue yield, making application to large animals and humans more difficult. METHODS: Neonatal pig thymus tissue (NP THY) was grafted into ATX, T/NK cell-depleted, 3Gy whole body-irradiated, originally immunocompetent B6 mice to evaluate the ability of NP THY to reconstitute mouse CD4+ T cells and to induce xenogeneic tolerance to donor pig skin grafts. RESULTS: Repopulation of mouse CD4+ T cells in the peripheral tissues was observed in T/NK cell-depleted, ATX B6 mice that received NP THY with or without neonatal pig spleen (NP SPL), but not in those receiving NP SPL alone, indicating that pig thymus grafting was necessary and sufficient for mouse T cell recovery. Seven of nine NP THY/SPL-grafted ATX mice and two of six NP THY-grafted ATX mice that reconstituted >5% CD4+ cells in PBL accepted donor pig skin long-term without lymphocyte infiltration, whereas they rejected allogeneic BALB/c skin and third party pig skin grafts as rapidly as euthymic mice. CONCLUSIONS: NP THY can support the development of mouse CD4+ T cells that are functional and specifically tolerant to donor pig antigens in ATX, T/NK cell-depleted, 3 Gy whole body-irradiated, originally immunocompetent B6 mice. Additional grafting of NP SPL with NP THY improves the efficiency of tolerance induction in this model.

Animals↗

The effect of farrowing crate heat lamp location on sow and pig patterns of lying and pig survival.

Three experiments were conducted to study sow and pig behavior during the 1st 3 d after birth and pig survival during the 1st 2 wk after farrowing. In Exp. 1, 23 sows were housed in conventional farrowing crates that were divided into five sections: a .5- x 1.5-m front creep section and the remaining area divided into four sections, .75 x 1.05 m each. Air temperature was maintained at 19 degrees C, and a 250-W heat lamp was placed at the right side of the front creep in Treatment 1 (T1), or in the creep at the right side of the sow for Treatment 2 (T2). The percentage of pigs within 8 cm of the sow's trunk was not affected by treatment, but it decreased (P < .001) from 61.8 +/- 3.4% on d 1 to 28.1 +/- 3.5% on d 3. As the percentage of pigs near the sow decreased, the percentage of pigs within the section containing the heat lamp increased (T1, P < .05; T2, P < .10). Experiment 2 involved 15 sows and litters housed as in Exp. 1, except that heat lamps were not provided, and average air temperature was 27.3 +/- .2 degrees C during behavioral observations. Even though the portion of the litter near the sow decreased (P < .001) from d 1 to d 3 (d 1, 57.0 +/- 3.4%; d 2, 42.9 +/- 3.3%; d 3, 31.7 +/- 3.3%), pigs did not concentrate in any specific section as they moved away from the sow. The average number of pigs within the front creep section (Section 1) for the 3-d period was less than (P < .01) the number in any other crate section. Experiment 3 involved 147 sows and tested the effect of solid creep floor covering on pig survival for each of the heat lamp locations used in Exp. 1. Neither heat lamp location nor floor covering affected pig survival. During the 1st 3 d of life, pigs tend to lie near the sow regardless of heat lamp location or air temperature. Heat lamp position and floor covering under the lamp do not affect pig survival.

Animals↗

[Effect of the Stellamune Mycoplasma vaccine on growth, energy conversion, death, and medication use in fattening pigs on a pig farm chronically infected with Mycoplasma hyopneumoniae].

The effect of Stellamune Mycoplasma vaccine, administered to piglets aged 2-15 days and then 13-15 days later, on daily weight gain, energy conversion, and use of medication was examined in fattening pigs on a chronically Mycoplasma hyopneumoniae infected pig farm. Half of the piglets were vaccinated and the other half acted as controls. In the study design, half of the pens in the fattening unit were allocated to vaccinated pigs; the other half to non-vaccinated pigs, pen was the experimental unit. In the fattening pens sows and castrated boars were separated. The study consisted of a total of 37 pens with vaccinated, and 37 pens with non-vaccinated pigs in 12 different compartments within the pig herd. In the finishing period, mean growth performance and mean energy conversion (EV/kg) of vaccinated animals was 65 grams/day higher and 0.07 EV/kg lower than in control pigs. Furthermore, the incidence of individual curative medication against respiratory problems was more than 4 times higher in control pigs than in vaccinated pigs. There was a tendency for a higher number of group medications against respiratory problems in control pigs than in vaccinated pigs. It is concluded that, in this herd, vaccination against M. hyopneumoniae was successful from an economic point of view.

Animals↗

Unidirectional fluxes of short-chain fatty acids across segments of the large intestine in pig, sheep and pony compared with guinea pig.

Unidirectional fluxes of short-chain fatty acids across pig, sheep and pony caecum, proximal and distal colon were studied under short-circuit current conditions in Ussing chambers. Findings are compared with results from guinea pig. Marked species differences are apparent; highest mucosal-to-serosal fluxes of acetate, propionate and butyrate were seen in guinea pig, lower values in pig and smallest fluxes in sheep and pony. Segmental differences between caecum, proximal and distal colon exist mainly in guinea pig and are less developed in pig, sheep and pony. Inhibition of Na+/H+ exchange by amiloride added to the mucosal solution decreased the mucosal-to-serosal fluxes of short-chain fatty acids clearly in guinea pig caecum and proximal colon, and very little in distal colon. This effect was somewhat less pronounced in pig caecum and distal colon, in caecum and distal colon of sheep and caecum of the pony. In pig, sheep and pony proximal colon and pony distal colon no significant inhibition was observed. Inhibition of the K(+)-H+ ATPase by addition of ouabain to the mucosal solution diminished mucosal-to-serosal fluxes of short-chain fatty acids in the guinea pig distal colon extensively. No comparable inhibition was seen in any of the other segments in the animals studied.

Amiloride↗