Clostridium difficile is a spore-forming, toxin-producing, anaerobic bacterium that colonizes the human gastrointestinal tract. This pathogen causes antibiotic-associated diarrhea and colitis in animals and humans. Antibiotic-associated diseases may be treated with probiotics, and interest is increasing in such uses of probiotics. This study investigated the effect of Lactobacillus strains on the quorum-sensing signals and toxin production of C. difficile. In addition, an in vivo experiment was designed to assess whether Lactobacillus acidophilus GP1B is able to control C. difficile-associated disease. Autoinducer-2 activity was measured for C. difficile using the Vibrio harveyi coupled bioluminescent assay. Cell extract (10μg/mL) of L. acidophilus GP1B exhibited the highest inhibitory activity among 5 to 40μg/mL cell-extract concentrations. Real-time PCR data indicated decreased transcriptional levels in luxS, tcdA, tcdB, and txeR genes in the presence of 10μg/mL of cell extract of L. acidophilus GP1B. Survival rates at 5d for mice given the pathogen alone with L. acidophilus GP1B cell extract or L. acidophilus GP1B were 10, 70, and 80%, respectively. In addition, the lactic acid-produced L. acidophilus GP1B exhibits an inhibitory effect against the growth of C. difficile. Both the L. acidophilus GP1B and GP1B cell extract have significant antipathogenic effects on C. difficile.

This study was undertaken to find optimum conditions of tryptone, yeast extract, glucose, Tween 80, and incubation temperature for the growth of Lactobacillus casei YIT 9018 and to assess the effects of these factors by use of response surface methodology. A central composite design was used as an experimental design for allocation of treatment combinations. A second-order polynomial regression model, which was used at first for analysis of the experiment, had a significant lack of fit. Therefore, cubic and quartic terms were incorporated into the regression model through variable selection procedures. Effects involving incubation temperature, yeast extract, glucose, and tryptone were significant, whereas the only significant effect involving Tween 80 was the interaction effect between temperature and Tween 80. It turned out that growth of L. casei YIT 9018 was most strongly affected by the incubation temperature. Estimated optimum conditions of the factors for growth of L. casei YIT 9018 are as follows:

This study was conducted to evaluate effects of dietary protease (PR) on growth performance, nutrient digestibility, and intestinal morphology of weaned pigs. A total of 75 weaned pigs [7.06 ± 0.18 kg of average body weight (BW); 28 day old] were randomly allotted to 3 dietary treatments in a randomized complete block design (blocks = BW and sex): a diet based on corn and soybean meal to meet the requirement of crude protein (CP) as a positive control (PC; CP = 24.49%), a low protein diet as a negative control (NC; CP = 22.51%), and NC + 0.02% PR. The PR used in this study was a commercial product containing 75,000 protease units/g derived from Nocardiopsis prasina produced in Bacillus licheniformis. Pigs were fed the dietary treatments for 6 weeks and the diets containing 0.2% chromic oxide for the last week of this study. Blood, feces, ileal digesta, and ileum samples were collected from randomly selected two pigs in each pen on respective time points. Measurements were growth performance, apparent ileal digestibility (AID) and apparent total tract digestibility (ATTD) of dry matter (DM), CP, and energy, frequency of diarrhea, packed cell volume (PCV), and ileal morphology of weaned pigs. Pigs fed PC and PR had higher (p < 0.05) final BW, average daily gain (ADG), and gain to feed ratio (G:F) during overall experimental period than those fed NC. Pigs fed PC and PR had higher (p < 0.05) AID or ATTD of DM, CP, or energy than those fed NC. Moreover, pigs fed PR had higher ratio between villus height and crypt depth (p < 0.05) and number of goblet cells (p < 0.05) than those fed NC. Addition of PR decreased (p < 0.05) frequency of diarrhea for the first two weeks after weaning compared with PC and NC. In addition, pigs fed PR had lower (p < 0.05) PCV on d 14 after weaning than those fed PC and NC. In conclusion, addition of PR in nursery diets with a low protein level significantly improved growth performance, nutrient digestibility, and intestinal morphology of weaned pigs.

We investigated the probiotic properties of Lactobacillus acidophilus NS1, such as acid resistance, bile tolerance, adherence to HT-29 cells, and cholesterol assimilation activity. In an animal study, 7-wk-old male C57BL/6 mice were fed a normal diet, a high-fat diet (HFD), or an HFD with L. acidophilus NS1 (ca. 1.0×10(8) cfu/mL) for 10 wk. Total cholesterol and low-density lipoprotein (LDL) cholesterol levels were significantly lower in mice fed an HFD with L. acidophilus NS1 than in those fed an HFD only, whereas high-density lipoprotein cholesterol levels were similar between these 2 groups. To understand the mechanism of the cholesterol-lowering effect of L. acidophilus NS1 on the HFD-mediated increase in plasma cholesterol levels, we determined mRNA levels of genes involved in cholesterol homeostasis in the liver. Expression of sterol regulatory element-binding protein 2 (Srebp2) and LDL receptor (Ldlr) in the liver was dramatically reduced in mice fed a HFD compared with those fed a normal diet. When L. acidophilus NS1 was administered orally to HFD-fed mice, an HFD-induced suppression of Srebp2 and Ldlr expression in the liver was abolished. These results suggest that the oral administration of L. acidophilus NS1 to mice fed an HFD increased the expression of Srebp2 and Ldlr in the liver, which was inhibited by high fat intake, thus leading to a decrease in plasma cholesterol levels. Lactobacillus acidophilus NS1 could be a useful probiotic microorganism for cholesterol-lowering dairy products and the improvement of hyperlipidemia and hepatic lipid metabolism.

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