Spatial heterogeneity and stability of bacterial community in the gastrointestinal tracts of broiler chickens.

Bacterial communities in the different regions of gastrointestinal tract (GIT) of broiler chickens were analyzed by pyrosequencing approach to understand microbial composition and diversity. The DNA samples extracted from 7 different regions along the GIT were subjected to bacterial-community analysis by pyrosequencing of the V1-V3 region of 16S rRNA gene. Major bacterial phyla in the chicken-gut microbiota included Firmicutes, Proteobacteria, Bacteroidetes, Actinobacteria, and Acidobacteria, but Firmicutes were mostly dominant (67.3 ± 16.1% of the total sequence reads identified). Among Firmicutes, Lactobacillales, including the genera Lactobacillus and Enterococcus, were the most dominant (51.8 ± 34.5% of the total sequence reads identified) from the crop to ileum. In contrast, in the cecum and large intestine, those genera were rarely detected, and Clostridiales were dominant (55.9 ± 31.4%). Fast UniFrac analysis showed that microbial communities from the crop to jejunum of the same individual chicken were grouped together, and those from ileum, cecum, and large intestine were clustered in a more GIT-specific manner. The numbers of shared operational taxonomic units between the neighboring segments of GIT were low, ranging from 2.9 to 20.3%. However, the abundance of shared operational taxonomic units in each segment was relatively high, ranging from 61.7 to 85.0%, suggesting that substantial proportions of microbial communities were shared between each segment and its neighboring segments, comprising a core microbiota. Our results suggested that the microbial communities of 7 main segments in the chicken GIT were distinctive according to both individuals and the different segments of GIT, but their stability was maintained along the GIT.

Lytic transglycosylase contributes to the survival of lipooligosaccharide-deficient, colistin-dependent Acinetobacter baumannii.

OBJECTIVES: The phenomenon of colistin dependence in Acinetobacter baumannii has been described in a situation in which colistin is now considered as the last resort for the treatment of infections caused by multidrug-resistant Gram-negative bacteria. In this study, we aimed to reveal a gene associated with colistin dependence in A. baumannii. METHODS: The colistin-dependent A. baumannii H08-391D strain was isolated from a patient, and target gene-inactivation mutants were constructed. We investigated the effects of target gene on colistin dependence with quantitative real-time PCR and endotoxin assay. Also, we observed the change of cell morphology by electron microscopy. RESULTS: The expression of ACICU_02898, encoding a soluble lytic transglycosylase associated with cell-wall degradation and recycling, was increased by eight-to 42-fold in colistin-dependent mutants, and deletion of ACICU_02898 in a colistin-dependent strain led to colistin susceptibility (MIC = 8 mg/L). Endotoxin activity was significantly low in a colistin-dependent derivative ACICU_02898-inactivated mutant and a complemented mutant. In addition, the ACICU_02898-inactivated mutant showed a highly reduced growth rate. The colistin-dependent derivative and ACICU_02898-inactivated mutant showed clearly distinguished absorption profiles in the red/green fluorescence dot blot with regard to their membrane potential. Electron microscopy revealed that the deletion mutant cells were elongated compared to the colistin-susceptible wild-type strain and colistin-dependent strain. CONCLUSIONS: A colistin-dependent A. baumannii strain exhibited a deficiency in its outer membrane integrity and high expression of lytic transglycosylase was required for survival. This study reveals why the colistin-dependent mutant can tolerate high antibiotic concentrations.

Influence of dietary avilamycin on ileal and cecal microbiota in broiler chickens.

The mechanisms by which antibiotic growth promoters (AGP) enhance growth rates, feed efficiencies, and disease resistance in poultry need to be understood for designing safer and alternative strategies to replace AGP. Avilamycin has been widely used as an AGP in poultry, but its impact on the structure and function of the gut microbiome of broiler chickens has not been fully elucidated. In this study, we investigated the bacterial communities of the ileum and cecum in broiler chickens fed with an avilamycin-supplemented diet, by high-throughput sequencing of bacterial 16S rRNA genes. Alpha diversity metrics indicated that the ileal bacterial diversity was higher in avilamycin-fed chickens than in the control group, whereas the opposite was true for the cecum. Multivariate analyses revealed that the ileal microbiota of the avilamycin-fed group were clearly distinguished from those of the control group, whereas the cecal bacterial communities were apparently not influenced by feeding diets containing avilamycin. In the ilea, 2 operational taxonomic units (OTU) that matched Lactobacillus reuteri and Clostridium were enriched (P = 0.016 and P = 0.007, respectively) in the avilamycin-fed group, and an OTU belonging to Lactobacillus crispatus was decreased (P = 0.016). In the cecal microbiota showing much higher diversity with 1,286 non-singleton OTU, 12 OTU were decreased, and 3 were increased in response to avilamycin treatment (P = 0.005-0.047). Functional profiling of bacterial communities based on PICRUSt analysis revealed that 10 functional categories were enriched by avilamycin treatments, and 4 functional categories were decreased. In conclusion, our results demonstrated that the influence of avilamycin supplementation on the diversity, taxonomic composition, and functional profiles of the microbiota was evidently different in the ileum and cecum. These results further our understanding of the impact of AGP on the composition and activity of commensal bacteria in the chicken gastrointestinal tract to develop novel feeding strategies for improving animal health and performance.

Jejunoileal bypass for morbid obesity: early results and body composition changes in forty-five patients.

Forty-five patients who underwent a 14 by 4 inch jejunoileal bypass for morbid obesity were studied before the operation and at periodic intervals after operation to determine the complications and changes in body composition resulting from this procedure. Body composition studies were determined using 3H2O and 42K. Rapid weight loss occurred in the first 3 months, with a mean loss of 30 percent of excess weight. This weight loss was accompanied by a decrease in exchangeable potassium (Ke) and total body water (TBW) during this interval by 14 and 10 percent, respectively. Although most patients continued to lose excess weight Ke and TBW stabilized at the end of the first year and returned to preoperative values in six patients at the end of 24 months. Analysis of the ratios of body cell mass and total body water to weight shows an improvement of body composition 12 months after operation. Body composition studies permit a quantitative assessment of the nutritional status in patients undergoing jejunoileal bypass. In spite of significant complications (23 percent), surgery for morbid obesity appears to satisfy the objective of allowing desirable loss of fat with relative sparing of muscle and other supporting tissues.

Maternal hyperglycemia in pregnant rats: its effect on growth and carbohydrate metabolism in the offspring.

Mild hyperglycemia during the last half of pregnancy was achieved by administration of streptozotocin to pregnant rats on the fifth day of gestation. Citrate buffer (vehicle for streptozotocin) was administered to control rats also on the fifth day of gestation. The pups born to the streptozotocin-treated mothers had higher birth weight, pancreatic insulin content, plasma insulin, and C-peptide concentrations compared with pups born to control mothers. The plasma glucose concentrations of the pups were similar between the two groups. The pups who were identified as macrosomic (birth weight greater than 1.7 SD than the mean of the control pups) maintained an accelerated postnatal growth through the first 10 weeks of age in female rats and in the first 3, and at 5 and 6 weeks of age for the male rats. The accelerated growth in the female rats was associated with higher perirenal-ovarian and salpingeal fat weight at 6 weeks. At 10 weeks of age, higher plasma insulin and glucose concentrations were observed following oral glucose challenge in both male and female macrosomic rats than in the control rats. At 12 weeks of age, only the female macrosomic rats showed abnormal glucose response due to peripheral insulin resistance. In the male rats at 12 weeks of age, a higher plasma insulin concentration in the macrosomic group was associated with a normal plasma glucose response to oral glucose challenge. We conclude that mild maternal hyperglycemia in rats resulted in fetal hyperinsulinemia and accelerated fetal growth.(ABSTRACT TRUNCATED AT 250 WORDS)

Postnatal growth and fatty acid synthesis in overgrown rat pups induced by fetal hyperinsulinemia.

Fetal hyperinsulinemia in the rat results in increased body weight, lipid content, and enhanced lipogenesis in liver and carcass. The purpose of our study was to determine whether the macrosomia and enhancement of fatty acid (FA) synthesis and/or content persisted postnatally in this animal model. Fetal hyperinsulinemia was produced in Sprague-Dawley rats by injecting fetuses with 2 units of insulin at 20.5 days of gestation. Alternate pups in the same litter were injected with saline. Pups were delivered surgically at 22.5 days of gestation, were weighed daily and sacrificed on day 15. FA content and synthesis rates of liver and skeletal muscle were measured. We found: (1) At birth, insulin-treated pups were 12% heavier than saline littermates, (5.88 +/- 0.14 g v 5.26 +/- 0.14 g, P less than .01); and (2) The enhanced growth associated with prenatal insulin treatment persisted during the suckling period, ie, compared with saline-treated controls, insulin pups were 15.7% heavier at 15 days of age (P less than .01); growth velocity of insulin pups, beginning on day 3, significantly exceeded that of control pups (P less than .05). FA contents of liver and muscle in insulin pups, (62.6 +/- 5.7 mumol/g and 62.7 +/- 13.2 mumol/g) were significantly greater (P less than .05) than in saline littermates (45.1 +/- 5.6 mumol/g and 30.2 +/- 4.7 mumol/g, respectively). We conclude that.(ABSTRACT TRUNCATED AT 250 WORDS)

Purification and characterization of a glutathione S-transferase from the fungus Cunninghamella elegans.

Cunninghamella elegans grown on Sabouraud dextrose broth had glutathione S-transferase (GST) activity. The enzyme was purified 172-fold from the cytosolic fraction (120000 x g) of the extract from a culture of C. elegans, using Q-Sepharose ion exchange chromatography and glutathione affinity chromatography. The GST showed activity against 1-chloro-2,4-dinitrobenzene, 1,2-dichloro-4-nitrobenzene, 4-nitrobenzyl chloride, and ethacrynic acid. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis gel filtration chromatography revealed that the native enzyme was homodimeric with a subunit of M(r) 27000. Comparison by Western blot analysis implied that this fungal GST had no relationship with mammalian alpha-, mu-, and pi-class GSTs, although it showed a small degree of cross-reactivity with a theta-class GST. The N-terminal amino acid sequence of the purified enzyme showed no significant homology with other known GSTs.

Homology of lubricin and superficial zone protein (SZP): products of megakaryocyte stimulating factor (MSF) gene expression by human synovial fibroblasts and articular chondrocytes localized to chromosome 1q25.

We have previously identified megakaryocyte stimulating factor (MSF) gene expression by synovial fibroblasts as the origin of lubricin in the synovial cavity. Lubricin is a mucinous glycoprotein responsible for the boundary lubrication of articular cartilage. MSF has a significant homology to vitronectin and is composed of 12 exons. RNA was purified from human synovial fibroblasts and articular chondrocytes grown in vitro from tissue explants obtained from subjects without degenerative joint disease. RT-PCR was used with multiple complimentary primer pairs spanning the central mucin expressing exon 6 of the MSF gene and individual exons on both the N- and C-terminal sides of exon 6. Exons 2, 4 and 5 appear to be variably expressed by synovial fibroblasts and articular chondrocytes. Lubricating mucin, in the form of MSF, is expressed by both chondrocytes and synovial fibroblasts in vitro. Both lubricin and superficial zone protein (SZP), a related proteoglycan, share a similar primary structure but could differ in post-translational modifications with O-linked oligosaccharides which are predominant in lubricin and with limited amounts chondroitin and keratan sulfate found in SZP. Since most of the MSF exons are involved in the expression of lubricating mucin, a strong homology to vitronectin persists. It is therefore appropriate to consider that both SZP and lubricin occupy a new class of biomolecules termed tribonectins. Screening of a human genome bacterial artificial chromsome (BAC) library with a cDNA primer pair complimentary for exon 6 identified two clones. Both clones were complimentary for chromosome 1q25 by in situ hybridization. This same locus was previously implicated in camptodactyl-arthropathy-pericarditis syndrome (CAP) by genetic mapping. It is hypothesized that CAP, a large joint arthropathy, may be associated with ineffective boundary lubrication provided by synovial fluid.

Osmolality of liquid and defined formula diets: the effect of hydrolysis by pancreatic enzymes.

The effect of hydrolysis was determined by a pancreatic enzyme preparation (Viokase) on a series of diets available for enteral nutrition. The increase in osmolality resulting from digestion by Viokase was measured. Small amounts produced very rapid increase in osmolality in almost all diets evaluated. Neither incubation at 37 degrees C nor the addition of larger amounts of the pancreatic enzyme preparation produced further substantial increases in osmolality. The importance of proper and adequate nutritional support of acutely and chronically ill patients is discussed, and a guideline for the choice of diets, route of delivery, and concentration of diets is suggested.

Effect of respiratory acidosis on glucose homeostasis in experimental intrauterine growth retardation in rats.

Hypoglycemia and asphyxia account for a significant proportion of morbidity in the infant with intrauterine growth retardation (IUGR). The purpose of this study was to evaluate changes in glucose homeostasis in IUGR rats during acute respiratory acidosis. IUGR was produced by bilateral uterine artery ligation at 17 days of gestation in 14 pregnant rats with 23 successfully delivered pups. The normal pups (n = 31) were those whose mothers were sham operated at the same gestational period. The IUGR and normal pups were studied at 2 days of age. One group of pups was studied under room air while another was subjected to 20 min of exposure to a gas mixture of 10% O2/15% CO2, balanced with N2. Gluconeogenesis in the liver and carcass, as well as plasma glucose and catecholamines were determined before and after the exposure to the gas mixture. The results showed that the 2-day-old IUGR rats have lower body weight (P less than 0.001), liver weight (P less than 0.001), plasma glucose (P less than 0.001), and rate of gluconeogenesis (P less than 0.01) when compared with the normally grown rats. During respiratory acidosis, the normally grown rats showed an increase in plasma epinephrine (P less than 0.005) without significant change in plasma glucose and rate of gluconeogenesis. The IUGR rats on the other hand, demonstrated a decrease in rate of gluconeogenesis (P less than 0.02), an increase in plasma glucose (P less than 0.001) while the plasma epinephrine level remained unchanged. We speculate that respiratory acidosis blunted cellular metabolism in the IUGR rat resulting in decreased peripheral glucose utilization.(ABSTRACT TRUNCATED AT 250 WORDS)

Cerebral metabolic response to hypoglycemia in severe intrauterine growth-retarded rat pups.

This study was to examine the effects of hypoglycemia on the brain metabolism of severe intrauterine growth retardation (IUGR) rat pups. IUGR pups were produced by ligating one of the uterine arteries of the dams. The pups on the opposite uterine horn were used as the control. They were delivered by cesarean section on day 21 and received regular insulin 5 units/kg or equivalent volume of normal saline at 40 min of age. The plasma glucose, lactate, blood gas values and brain glucose, lactate, ATP contents were determined at 5 and 100 min of age. The IUGR pups had higher plasma and brain lactate concentrations than the control throughout the study period. They had lower plasma glucose, oxygen and pH values, brain glucose and ATP contents than control at 5 min of age. Despite insulin-induced hypoglycemia, brain ATP contents of the IUGR recovered to normal levels at 100 min of age when the oxygenation and pH improved. These data indicate that the brain energy metabolism of IUGR rat pups was suppressed by asphyxia and hypoglycemia. However, even in the continuing presence of hypoglycemia, brain energy metabolism returned to normal. The recovery is probably related to better oxygenation and utilization of alternative energy fuels, such as lactate.

Cerebral metabolic response to hypoglycemia in severe intrauterine growth-retarded rat pups.

This study was to examine the effects of hypoglycemia on the brain metabolism of severe intrauterine growth retardation (IUGR) rat pups. IUGR pups were produced by ligating one of the uterine arteries of the dams. The pups on the opposite uterine horn were used as the control. They were delivered by cesarean section on day 21 and received regular insulin 5 units/kg or equivalent volume of normal saline at 40 min of age. The plasma glucose, lactate, blood gas values and brain glucose, lactate, and ATP contents were determined at 5 and 100 min of age. The IUGR pups had higher plasma and brain lactate concentrations than the control throughout the study period. They had lower plasma glucose, oxygen and pH values, brain glucose and ATP contents than control at 5 min of age. Despite insulin-induced hypoglycemia, brain ATP contents of the IUGR recovered to normal levels at 100 min of age when the oxygenation and pH improved. These data indicate that the brain energy metabolism of IUGR rat pups was suppressed by asphyxia and hypoglycemia. However, even in the continuing presence of hypoglycemia, brain energy metabolism returned to normal. The recovery is probably related to better oxygenation and utilization of alternative energy fuels, such as lactate.