In vitro total-gas, CH4, H2, volatile fatty acid, and lactate kinetics studies on luminal contents from the small intestine, cecum, and colon of the pig

Two experiments were conducted to assess differences in fermentative activities of digesta obtained from various regions of the pig gastrointestinal tract. In experiment 1, the contents of small intestines, ceca, and colons of 110-kg pigs were collected, diluted twofold, and incubated for 2 h at 37 degrees C. In experiment 2, colonic samples from 16,100-kg pigs were similarly treated, except that the incubation period was 5 h. Total gas (gas pressure), CH4, H2, lactate, formate, acetate, propionate, butyrate, valerate, and isovalerate were measured in experiment 1. Only the gas variables were measured in experiment 2. Statistically significant differences (P greater than 0.05) were not observed among the gas production rate estimates across the small-intestinal, cecal, and colonic regions in experiment 1. Furthermore, all the small-intestinal samples and half the cecal samples assayed in experiment 1 were nonmethanogenic. The mean methanogenic and total-gas production rate estimates for the colonic samples in experiment 1 were 0.052 ml g of wet contents-1 h-1 and 1.7 ml of total gas g of wet contents-1 h-1, respectively. No differences in the methanogenic rate estimates were detected between the proximal, middle, and distal thirds of the pig colons (P greater than 0.05). The volatile fatty acid and lactate molar percentages measured in experiment 1 were consistent with previously published observations. Hydrogen accumulated to the greatest extent (7 microM on average) in the in vitro incubations of small-intestinal contents, whereas the H2 concentrations ranged from 0.5 to 1 microM for the incubated cecal and colonic samples in experiment 1.(ABSTRACT TRUNCATED AT 250 WORDS)     

Comparison of bacterial populations of the pig cecum and colon based upon enumeration with specific energy sources.

Concentrations of bacteria in the ceca and colons of pigs were measured by determinations of colony counts on rumen fluid-based media in anaerobic roll tubes. With our most complete medium (medium CCA), the mean colony count of cecal samples from 20 pigs was 2.37 X 10(10) +/- 1.0 X 10(10) (+/- standard deviation)/g (wet weight). The mean number of bacteria attached to or associated with cecal epithelial tissues from three pigs on medium CCA was 2.67 X 10(7) +/- 0.81 X 10(7)/cm2 of tissue. The proportions of gut bacterial populations able to use various energy substrates were estimated on the basis of relative colony counts. The following substrates are listed in descending order of their capacity to support growth of cecal bacteria: glucose, starch, cellobiose, xylose, Trypticase, gastric mucin from swine, mannitol, glycerol, and lactate. The effect of diet upon this distribution was not examined. The relative proportions of bacteria from a given population that were able to grow on various selective media were used as population profiles. Comparisons of populations in this way indicated that differences could be detected between (i) populations from the cecum of littermate pigs, (ii) populations from the cecum and colon of the same pig, and (iii) populations in the lumen of the cecum as compared with populations associated with cecal mucosa.     

Comparison of bacterial populations of the pig cecum and colon based upon enumeration with specific energy sources.

Concentrations of bacteria in the ceca and colons of pigs were measured by determinations of colony counts on rumen fluid-based media in anaerobic roll tubes. With our most complete medium (medium CCA), the mean colony count of cecal samples from 20 pigs was 2.37 X 10(10) +/- 1.0 X 10(10) (+/- standard deviation)/g (wet weight). The mean number of bacteria attached to or associated with cecal epithelial tissues from three pigs on medium CCA was 2.67 X 10(7) +/- 0.81 X 10(7)/cm2 of tissue. The proportions of gut bacterial populations able to use various energy substrates were estimated on the basis of relative colony counts. The following substrates are listed in descending order of their capacity to support growth of cecal bacteria: glucose, starch, cellobiose, xylose, Trypticase, gastric mucin from swine, mannitol, glycerol, and lactate. The effect of diet upon this distribution was not examined. The relative proportions of bacteria from a given population that were able to grow on various selective media were used as population profiles. Comparisons of populations in this way indicated that differences could be detected between (i) populations from the cecum of littermate pigs, (ii) populations from the cecum and colon of the same pig, and (iii) populations in the lumen of the cecum as compared with populations associated with cecal mucosa.     

Characterization of predominant bacteria from the colons of normal and dysenteric pigs.

Bacterial populations adherent to the mucosa of the proximal colons of weaned, healthy pigs were compared with populations from pigs with dysentery induced by inoculation with a culture of Treponema hyodysenteriae. Isolates (136) representative of the predominant flora adherent to colonic epithelia of normal pigs and isolates (162) from pigs with dysentery were cultured anaerobically on a rumen fluid-based medium and characterized. Most (71%) of the isolates from colonic epithelia of normal pigs were gram positive, whereas 88% of the epithelia-associated isolates from pigs with dysentery were gram negative. The geometric mean of colony counts was 5.7 X 10(7)/cm2 of colonic tissue from three normal pigs and 7.7 X 10(8)/cm2 from four pigs with dysentery. A number of isolates obtained from contents of the lumens of normal pigs with dysentery were also characterized. Comparison of isolates from epithelial tissue and from contents of the lumens of the same pig indicated that these populations were different. Our results indicate that physiological changes that occur in the colons of pigs with dysentery are accompanied by marked changes in the microbial populations in the colons. The factors which regulate the population changes are not yet understood.     

Characterization of predominant bacteria from the colons of normal and dysenteric pigs

Bacterial populations adherent to the mucosa of the proximal colons of weaned, healthy pigs were compared with populations from pigs with dysentery induced by inoculation with a culture of Treponema hyodysenteriae. Isolates (136) representative of the predominant flora adherent to colonic epithelia of normal pigs and isolates (162) from pigs with dysentery were cultured anaerobically on a rumen fluid-based medium and characterized. Most (71%) of the isolates from colonic epithelia of normal pigs were gram positive, whereas 88% of the epithelia-associated isolates from pigs with dysentery were gram negative. The geometric mean of colony counts was 5.7 X 10(7)/cm2 of colonic tissue from three normal pigs and 7.7 X 10(8)/cm2 from four pigs with dysentery. A number of isolates obtained from contents of the lumens of normal pigs with dysentery were also characterized. Comparison of isolates from epithelial tissue and from contents of the lumens of the same pig indicated that these populations were different. Our results indicate that physiological changes that occur in the colons of pigs with dysentery are accompanied by marked changes in the microbial populations in the colons. The factors which regulate the population changes are not yet understood.     

Enumeration of selected anaerobic bacterial groups in cecal and colonic contents of growing-finishing pigs

Selected anaerobic bacterial groups in cecal and colonic contents of clinically healthy pigs fed a corn-soybean meal production diet were determined at sacrifice after 4, 8, and 11 weeks on feed, corresponding to intervals within the growing-finishing growth period. By using ruminal fluid-based media, the densities of the culturable anaerobic population; the cellulolytic, pectin-fermenting, pectin-hydrolyzing, xylan-fermenting; and the xylan-hydrolyzing, sulfate-reducing, and methanogenic bacterial populations were estimated. An analysis of variance was performed on these bacterial group variables to examine the effects of phase (weeks on feed), site (cecum or colon), or the interaction of phase with site. The population of total anaerobic bacteria was twice as dense in the colon as it was in the cecum (2 x 10(10) versus 1 x 10(10)/g [wet weight]; P = 0.001). The proportion of cellulolytic bacteria was lower at 4 weeks on feed than at 8 or 11 weeks (23 versus 32%; P = 0.026), while the proportion of pectin-fermenting bacteria depended on the interaction of phase with site (P = 0.021). The numbers of sulfate-reducing bacteria were significantly higher in the colon than in the cecum (6 x 10(7) versus 3 x 10(7); P = 0.014), as were methanogenic bacteria (19 x 10(7) versus 0.6 x 10(7); P = 0.0002). The remaining bacterial groups were stable with respect to phase and site. The results suggest that except for density differences, the microbial communities of the pig cecum and colon are similar in composition throughout the growing-finishing phase.     

Intestinal colonization of laboratory rats with Oxalobacter formigenes.

Six strains of Oxalobacter formigenes (anaerobic oxalate-degrading bacteria) were examined for their ability to colonize the gastrointestinal tracts of adult laboratory rats. These rats did not harbor O. formigenes. Strain OxCR6, isolated from the cecal contents of a laboratory rat that was naturally colonized by oxalate-degrading bacteria, colonized the ceca and colons of adult rats fed a diet that contained 4.5% sodium oxalate. Five days after rats were inoculated intragastrically with 10(9) viable cells of strain OxCR6, oxalate degradation rates in cecal and colonic contents increased by 19 and 40 times, respectively. Viable counts of strain OxCR6 from these rats averaged 10(8)/g (dry weight) of cecal contents. Strain OxCR6 was not detected in the cecal contents of inoculated rats fed diets that contained less than 3.0% sodium oxalate. Strains of O. formigenes isolated from the cecal contents of swine, guinea pigs, and wild rats and from human feces also colonized the ceca of laboratory rats; a ruminal strain failed to colonize the rat cecum.     

Vasoconstrictor-mediated release of lactate from the perfused rat hindlimb.

The effects of different vasomodulators on lactate release by the constant-flow-perfused rat hindlimb were examined and compared with that by perfused mesenteric artery, incubated preparations of aortas, soleus and epitrochlearis muscles, and perifused soleus muscles. Infusion of vasopressin (0.5 nM), angiotensin II (5 nM), norepinephrine (50 nM), and methoxamine (10 microM) into the hindlimbs of 180- to 200-g rats increased the perfusion pressure by 112-167% from 30.4 +/- 0.8 mmHg, O2 consumption by 26-68% from 6.4 +/- 0.2 mumol.g-1 x h-1, and lactate efflux by 148-380% from 5.41 +/- 0.25 mumol.g-1 x h-1. Hindlimbs of 100- to 120-g rats responded similarly to angiotensin II. Isoproterenol (1 microM) had no effect on O2 uptake or perfusion pressure but increased lactate release by 118%. Nitroprusside (0.5 mM) markedly inhibited the vasoconstrictor-mediated increases in lactate release, perfusion pressure, and O2 consumption by the hindlimb but had no effect on isoproterenol-mediated lactate efflux. Serotonin (6.7 microM) increased lactate release from the perfused mesenteric artery by 120% from 5.48 mol.g-1 x h-1. Lactate release by incubated aorta was increased by angiotensin II (50 nM), isoproterenol (1 microM), and mechanical stretch. The increase mediated by angiotensin II was blocked by glycerol trinitrate (2.2 microM), which had no effect on lactate release by isoproterenol. Neither angiotensin II (5 nM) nor vasopressin (0.5 nM) increased lactate release from incubated soleus and epitrochlearis muscles; however, lactate release was increased by isoproterenol, and this increase was unaffected by glycerol trinitrate (2.2 microM).(ABSTRACT TRUNCATED AT 250 WORDS)     

Enumeration of selected anaerobic bacterial groups in cecal and colonic contents of growing-finishing pigs.

Selected anaerobic bacterial groups in cecal and colonic contents of clinically healthy pigs fed a corn-soybean meal production diet were determined at sacrifice after 4, 8, and 11 weeks on feed, corresponding to intervals within the growing-finishing growth period. By using ruminal fluid-based media, the densities of the culturable anaerobic population; the cellulolytic, pectin-fermenting, pectin-hydrolyzing, xylan-fermenting; and the xylan-hydrolyzing, sulfate-reducing, and methanogenic bacterial populations were estimated. An analysis of variance was performed on these bacterial group variables to examine the effects of phase (weeks on feed), site (cecum or colon), or the interaction of phase with site. The population of total anaerobic bacteria was twice as dense in the colon as it was in the cecum (2 x 10(10) versus 1 x 10(10)/g [wet weight]; P = 0.001). The proportion of cellulolytic bacteria was lower at 4 weeks on feed than at 8 or 11 weeks (23 versus 32%; P = 0.026), while the proportion of pectin-fermenting bacteria depended on the interaction of phase with site (P = 0.021). The numbers of sulfate-reducing bacteria were significantly higher in the colon than in the cecum (6 x 10(7) versus 3 x 10(7); P = 0.014), as were methanogenic bacteria (19 x 10(7) versus 0.6 x 10(7); P = 0.0002). The remaining bacterial groups were stable with respect to phase and site. The results suggest that except for density differences, the microbial communities of the pig cecum and colon are similar in composition throughout the growing-finishing phase.     

Natural transmission of Salmonella choleraesuis in swine.

This experiment was designed to study the natural transmission of Salmonella choleraesuis in swine. Forty pigs were divided into three groups. Group 1 (n = 12) was challenged with 10(8) CFU of S. choleraesuis per ml by intranasal inoculation. One day postinoculation (p.i.), group 2 (n = 24) was commingled with group 1. Group 3 (n = 4) served as uninoculated controls. Serum samples were collected weekly. Blastogenesis assays and necropsies were performed at 1, 2, 4, 6, 9, and 12 weeks p.i., and 16 tissue samples per pig were collected and cultured. Environmental (pooled feces from the pen floor) levels of S. choleraesuis were 2.61 log10 CFU/g of feces at 24 h p.i. (immediately prior to commingling). Severe clinical signs were observed in groups 1 and 2. The results indicated that at least 16% of group 2 pigs were shedding S. choleraesuis within 24 h of commingling. At 1 week p.i., 32 of 32 group 1 and 39 of 62 group 2 tissue samples were positive for S. choleraesuis. Only 3 of 12 group 2 pigs were positive at 6, 9, and 12 weeks (1 pig for each week), indicating that only a small proportion of infected swine become long-term carriers. At 12 weeks p.i., only the colon and colonic lymph node samples of one pig from group 2 were positive. Humoral, mucosal, and cellular immune responses were similar between groups 1 and 2. These data demonstrate that a few pigs shedding low levels of Salmonella organisms before slaughter can result in rapid transmission and subsequent shedding by many swine.     

Characterization of eosinophil cell activation by peptides. Differential effects of substance P, melittin, and FMET-Leu-Phe.

The ability of three different peptides, substance P (SP), FMLP and melittin, to activate eosinophils purified from the peritoneal cavity of human serum-treated guinea pigs was investigated. Degranulation (eosinophil peroxidase, EPO), oxidative burst (O2-), [Ca2+]i mobilization, and arachidonic acid metabolism (thromboxane B2, TXB2) were used as indices of eosinophil activation. SP (100 nM to 100 microM), FMLP (1 to 100 microM) and melittin (10 nM to 100 microM) induced EPO release but only FMLP (1 to 100 microM) led to an elevation of [Ca2+]i. The melittin- and SP-induced EPO secretion occurred at both cytotoxic and noncytotoxic concentrations as assessed by lactate dehydrogenase release. In addition, the effect of SP was not inhibited by the SP analogue (D-Pro4, D-Trp7,9,10)SP(4-11) and SP failed to promote the generation and subsequent release of TXA2. In contrast, FMLP (10 to 100 microM) stimulated the release of TXB2 from guinea pig eosinophils that was selectively inhibited by pretreatment of the cells with BOC-FMLP. On an equimolar basis (1 microM), melittin was approximately fivefold more active at promoting TXB2 release than FMLP. The results indicate that eosinophils respond to the three peptides, SP, melittin, and FMLP in differential fashion. We conclude that activation of guinea pig eosinophils by FMLP is likely to be receptor-mediated whereas the actions of SP and melittin may act through nonspecific peptide-membrane phospholipid interactions.     

Increase in colonic methanogens and total anaerobes in aging rats

Methanogens are present in the colons of our local Wistar rat colony. We studied the changes in concentrations of their fecal methanogenic and nonmethanogenic bacteria with age as a model of the development of these communities in humans. We found that the predominant methanogen in the rats is a Methanobrevibacter species. The log of the concentration of total anaerobes increased from 9.8/g (dry weight) at 3.0 weeks of age (shortly after weaning) to 10.7/g (dry weight) at 96 weeks (shortly before the end of the life span). In contrast, the log concentration of methanogens increased from 5.5 to 9/g (dry weight) during the same time period. Therefore, methanogens increased as a percentage of the total anaerobes from 0.005% at 3.0 weeks to 2.0% at 96 weeks. About 12 doublings of the methanogenic population and 3.3 doublings of the nonmethanogenic population took place from weaning until death. The slow increase in the ratio of methanogens to total anaerobes with age followed the same pattern in cecal contents as found in feces. There were no relationships between animal weights or fecal outputs and the increase in total anaerobe and methanogen concentrations in feces. A possible explanation for the slow increase in the Methanobrevibacter species in Wistar rats with age is a gradual shifting of the use of electrons from the reduction of CO2 to acetate by acetogens to the reduction of CO2 to CH4. The results provide the first evidence for an age-related change in the nonmethanogenic bacteria of the colon and supporting microbiological evidence for physiological studies that have shown age-related increases in colonic methane production in humans.     

Increase in colonic methanogens and total anaerobes in aging rats.

Methanogens are present in the colons of our local Wistar rat colony. We studied the changes in concentrations of their fecal methanogenic and nonmethanogenic bacteria with age as a model of the development of these communities in humans. We found that the predominant methanogen in the rats is a Methanobrevibacter species. The log of the concentration of total anaerobes increased from 9.8/g (dry weight) at 3.0 weeks of age (shortly after weaning) to 10.7/g (dry weight) at 96 weeks (shortly before the end of the life span). In contrast, the log concentration of methanogens increased from 5.5 to 9/g (dry weight) during the same time period. Therefore, methanogens increased as a percentage of the total anaerobes from 0.005% at 3.0 weeks to 2.0% at 96 weeks. About 12 doublings of the methanogenic population and 3.3 doublings of the nonmethanogenic population took place from weaning until death. The slow increase in the ratio of methanogens to total anaerobes with age followed the same pattern in cecal contents as found in feces. There were no relationships between animal weights or fecal outputs and the increase in total anaerobe and methanogen concentrations in feces. A possible explanation for the slow increase in the Methanobrevibacter species in Wistar rats with age is a gradual shifting of the use of electrons from the reduction of CO2 to acetate by acetogens to the reduction of CO2 to CH4. The results provide the first evidence for an age-related change in the nonmethanogenic bacteria of the colon and supporting microbiological evidence for physiological studies that have shown age-related increases in colonic methane production in humans.     

Fatty acid effects on gluconeogenesis in goat, calf and guinea pig hepatocytes

1. Regulation of hepatic gluconeogenesis by fatty acid was studied in goat, calf and guinea pig hepatocytes. 2. Fatty acid effects on gluconeogenesis were dependent upon species; fatty acid and gluconeogenic substrate. 3. Oleate and octanoate inhibited gluconeogenesis from propionate in guinea pig hepatocytes and stimulated it in goat hepatocytes. 4. Oleate and octanoate markedly inhibited gluconeogenesis from lactate in guinea pig hepatocytes whereas octanoate, but not oleate, decreased glucose production from lactate in goat hepatocytes. 5. Effects of fatty acids on gluconeogenesis in calf hepatocytes were similar to goat hepatocytes suggesting control of gluconeogenesis is similar among ruminant species but differs from guinea pigs.     

Porcine colonic lymphoglandular complex: distribution, structure, and epithelium

Lymphoepithelium and cells specialized for uptake and transport of foreign matter are characteristic of antigen sampling organs, including lymphoglandular complexes (LGCs). Distribution, histologic structure, and epithelial ultrastructure of colonic lymphoglandular complexes were determined in 5- to 13-week-old pigs. LGCs averaged 1,231 in number per colon, displayed a characteristic distribution pattern, and were most evenly distributed in colons of older pigs. LGCs consisted of well-defined submucosal masses composed of lymphatic nodules and internodular lymphoid tissue penetrated by radially branching extensions of mucosal glands. Epithelial diverticula of each LGC entered the submucosa as a group through a circular collar derived from the muscularis mucosae. LGC epithelium contained goblet cells, cuboidal and columnar enterocytes, enteroendocrine cells, individual and clustered intraepithelial leukocytes, and cells morphologically compatible with follicle-associated epithelial cells/M cells. We regard the colonic LGC as a distinct mucosal lymphoid organ and suggest a significant role for it in local and systemic immune responses. The porcine colonic LGC may serve as a model for the human LGC.     

Gluconeogenesis predominates in periportal regions of the liver lobule

Rates of gluconeogenesis from lactate were calculated in periportal and pericentral regions of the liver lobule in perfused rat livers from increases in O2 uptake due to lactate. When lactate (0.1-2.0 mM) was infused into livers from fasted rats perfused in either anterograde or the retrograde direction, a good correlation (r = 0.97) between rates of glucose production and extra O2 uptake by the liver was observed as expected. Rates of oxygen uptake were determined subsequently in periportal and pericentral regions of the liver lobule by placing miniature oxygen electrodes on the liver surface and measuring the local change in oxygen concentration when the flow was stopped. Basal rates of oxygen uptake of 142 +/- 11 and 60 +/- 4 mumol X g-1 X h-1 were calculated for periportal and pericentral regions, respectively. Infusion of 2 mM lactate increased oxygen uptake by 71 mumol X g-1 X h-1 in periportal regions and by 29 mumol X g-1 X h-1 in pericentral areas of the liver lobule. Since the stoichiometry between glucose production and extra oxygen uptake is well-established, rates of glucose production in periportal and pericentral regions of the liver lobule were calculated from local changes in rates of oxygen uptake for the first time. Maximal rates of glucose production from lactate (2 mM) were 60 +/- 7 and 25 +/- 4 mumol X g-1 X h-1 in periportal and pericentral zones of the liver lobule, respectively. The lactate concentrations required for half-maximal glucose synthesis were similar (0.4-0.5 mM) in both regions of the liver lobule in the presence or absence of epinephrine (0.1 microM). In the presence of epinephrine, maximal rates of glucose production from lactate were 79 +/- 5 and 59 +/- 3 mumol X g-1 X h-1 in periportal and pericentral regions, respectively. Thus, gluconeogenesis from lactate predominates in periportal areas of the liver lobule during perfusion in the anterograde direction; however, the stimulation by added epinephrine was greatest in pericentral areas. Differences in local rates of glucose synthesis may be due to ATP availability, as a good correlation between basal rates of O2 uptake and rates of gluconeogenesis were observed in both regions of the liver lobule in the presence and absence of epinephrine. In marked contrast, when livers were perfused in the retrograde direction, glucose production was 28 +/- 5 mumol X g-1 X h-1 in periportal areas and 74 +/- 6 mumol X g-1 X h-1 in pericentral regions.(ABSTRACT TRUNCATED AT 400 WORDS)     

PCO(2) in the large intestine of mice, rats, guinea pigs, and dogs and effects of the dietary substrate.

PCO(2) in the lumen and serosa of cecum and colon was measured in rats, guinea pigs, and dogs to examine the relationship between serosal PCO(2) and the incidence of intestinal necrotic lesions after administration of gas-carrier contrast agents in rodents. The effects of the dietary substrate were tested in a group of mice maintained on a diet based on glucose as the only carbohydrate source. The anesthetic used was a fentanyl-fluanison-midazolam mixture (rodents) and pentobarbital (dogs). PCO(2) was measured in vivo and postmortem, and the kinetics of the postmortem serosal PCO(2) [transmural CO(2) flux (J(CO(2)))] was calculated. PCO(2) in the cecal serosa and lumen, respectively, was 64 +/- 4 and 392 +/- 18 Torr in rats, 67 +/- 3 and 276 +/- 17 Torr in guinea pigs, and 73 +/- 6 and 137 +/- 7 Torr in mice on glucose-based diet. In the colon serosa and lumen of dogs, PCO(2) was 30 +/- 6 and 523 +/- 67 Torr, respectively. Serosal PCO(2) increased rapidly after death in rats and slower in guinea pigs and mice, and the slowest change was observed in dogs. Compared with dogs, serosal PCO(2) and J(CO(2)) of rats and guinea pigs were significantly higher. Serosal PCO(2) of guinea pigs was similar to that of rats, whereas the J(CO(2)) of guinea pigs was significantly lower. These data suggest a causal relationship between the ability of the cecal and colonic wall to act as a barrier to CO(2) diffusion and the presence of characteristic gas-carrier contrast agent-induced intestinal lesions in mice and rats and their absence in guinea pigs, dogs, and other species.     

Dietary supplementation with a nucleotide-rich yeast extract modulates gut immune response and microflora in weaned pigs in response to a sanitary challenge

An experiment was carried out to evaluate the short-term effect of supplementing a nucleotide-rich yeast extract (NRYE) on growth performance, gut structure, immunity and microflora of piglets raised under sanitary and unsanitary conditions. A total of 84, 21-day old piglets were used in this study; 42 piglets were raised in a room designated as the clean room that was washed once per week, whereas the other 42 piglets were raised in a room designated as the unclean room in which 7 kg of manure from the sow herd was spread on each pen floor on day 1 and 7 and the room was not washed throughout the experiment. The pigs were fed a corn–soybean meal-based diet without or with 0.1% NRYE. Each treatment had 7 replicate pens in each room, and each pen housed 3 pigs. Feed disappearance and BW were recorded on day 1 and 14. On day 14, one pig per pen was euthanized to collect ileum, mesenteric lymph nodes and spleen tissues, and cecum and colon digesta. Overall, NRYE supplementation did not affect growth performance in both clean and unclean conditions, improved kidney weight in both clean (P=0.0002) and unclean room (P<0.0001) and tended to improve the villus height/crypt depth ratio in the clean room (P=0.073). Supplementing NRYE was associated with upregulation of Ileal programmed cell death gene-1 (P=0.0003), interleukin (IL)-1β (P<0.0001), IL-6 (P=0.0003), IL-10 (P<0.0001) and tumor necrosis factor-α (TNF-α) (P<0.0001) in pigs raised in the unclean room. Supplementing the NRYE in pigs raised in the clean room suppressed growth of cecal Enterobacteriacea (P<0.0001) members and colonic Enterococcus spp. (P<0.019), improved proliferation of cecal Lactobacillus spp. (P<0.002) and colonic Clostridium cluster IV (P<0.011) and XVIa members (P<0.0002). Supplementing the NRYE in the unclean room improved proliferation of cecal Clostridium cluster IV (P<0.026) and suppressed proliferation of colonic Enterococcus spp. (P<0.037). In conclusion, supplementing the NRYE to piglets under unsanitary conditions improved ileal immune response by upregulating inflammatory cytokines, and positively modulated proliferation of beneficial gut bacteria and suppression of harmful ones in both clean and unclean rooms.     

Effects of meal intake on materno-foetal exchanges of energetic substrates in the pig.

Catheters were implanted in 18 gilts at 99 days of pregnancy to study the effects of meal intake on uterine and umbilical uptake of energetic substrates in the conscious pig. Blood samples were withdrawn at 105 days of pregnancy from 10 min before and up to 90 min after feeding of a 2.5-kg meal. Plasma glucose was 2.2 to 2.5 times lower and blood lactate 2 to 3 times higher in the foetus than in the sow. Glucose and lactate increased after the meal. Their umbilical uptake amounted to 0.32 and 0.26 mmol x L(-1), respectively. Fructose was found in large amounts in foetal plasma (4.3 mmol x L(-1)), but it did not seem to be metabolised by the foetus. Meal intake decreased plasma levels of FFA and glycerol in the sows, whereas they increased in the foetuses. A small FFA and glycerol umbilical uptake was recorded (14 and 6 micromol L(-1), respectively). Most features of the materno-foetal exchanges in the porcine species resemble those of other species, especially ruminants.     

Rapidly growing rumen methanogenic organism that synthesizes coenzyme M and has a high affinity for formate

Methanogenic bacteria with a coccobacillus morphology similar to Methanobrevibacter ruminantium were isolated from the bovine rumen. One isolate, 10-16B, represented a previously undescribed rumen population that, unlike M. ruminantium, synthesized coenzyme M, grew rapidly (mu = 0.24 h-1) on H2-CO2 in a complex medium, had simple nutritional requirements, and metabolized formate at reported rumen concentrations. H2 was metabolized to partial pressures 10-fold lower than those reported for the rumen. After H2 starvation for 26 h, strain 10-16B rapidly resumed growth when H2 was made available. The minimum concentrations of acetate (6 mM) and ammonia (less than 7 mM) that were required for optimal growth were lower than the reported acetate and ammonia concentrations in the rumen. Isoleucine and leucine stimulated growth, but only at concentrations (greater than 50 microM) higher than those reported for the rumen. Another coccobacillary methanogenic organism that synthesized coenzyme M was isolated from a different animal as were organisms that required an exogenous supply of coenzyme M. In general, methanogenic bacteria that required an exogenous supply of coenzyme M had lower maximum growth rates and more complex nutritional requirements than organisms that synthesized the cofactor. The ability of all isolates to metabolize formate below the detection limit of 10 microM indicated that, in contrast to previous reports, methanogenic bacteria have the potential to directly metabolize formate in the rumen. This study demonstrated that there are physiologically diverse populations of coccobacillary methanogenic bacteria in the rumen that can interact competitively and cooperatively.