• Journal of Microbiology
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• v.44 no.5
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• pp.548-555
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• 2006

FaeG is the key factor in the infection process of K88ad enterotoxigenic Escherichia coli (ETEC) fimbrial adhesin. In an attempt to determine the possibility of expressing recombinant FaeG with immunogenicity for a new safe and high-production vaccine in E. coli, we constructed the recombinant strain, BL21 (DE3+K88), which harbors an expression vector with a DNA fragment of faeG, without a signal peptide. Results of 15% SDS-polyacrylamide slab gel analysis showed that FaeG can be stably over-expressed in BL21 (DE3+K88) as inclusion bodies without FaeE. Immunoglobulin G (IgG) and M (IgM) responses in pregnant pigs, with boost injections of the purified recombinant FaeG, were detected 4 weeks later in the sera and colostrum. An in vitro villius-adhesion assay verified that the elicited antibodies in the sera of vaccinated pigs were capable of preventing the adhesion of K88ad ETEC to porcine intestinal receptors. The protective effect on the mortality rates of suckling piglets born to vaccinated mothers was also observed one week after oral challenge with the virulent ETEC strain, $C_{83907}$ (K88ad, $CT^+,\;ST^+$). The results of this study proved that the adhesin of proteinaceous bacterial fimbriae or pili could be overexpressed in engineered E. coli strains, with protective immune responses to the pathogen.

• Asian-Australasian Journal of Animal Sciences
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• v.18 no.11
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• pp.1582-1588
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• 2005

This study aimed to evaluate the effects of silage additives on the fermentation qualities and residual mono- and disaccharides composition of silages. Forage Oats (Avena sativa L.) and Italian Ryegrass (Lolium multiflorum Lam.) were ensiled with glucose, sorbic acid and pre-fermented juice of epiphytic lactic acid bacteria (FJLB) treatments for 30 days. In both species grass silages, although the respective controls had higher contents of butyric acid (20.86, 33.45g $kg^{-1}$ DM) and ammonia-N/total nitrogen (100.07, 114.91 g $kg^{-1}$) as compared with other treated silages in forage oats and Italian ryegrass, the fermentation was clearly dominated by lactic acid bacteria. This was well indicated by the low pH value (4.27, 4.38), and high lactic acid/acetic acid (6.53, 5.58) and lactic acid content (61.67, 46.85 g $kg^{-1}$ DM). Glucose addition increased significantly (p<0.05) lactic acid/acetic acid, and significantly (p<0.05) decreased the values of pH and ammonia-N/total nitrogen, and the contents of butyric acid and volatile fatty acids as compared with control, however, there was a slightly but significantly (p<0.05) higher butyric acid and lower residual mono- and di-saccharides as compared with sorbic acid and FJLB additions. Sorbic acid addition showed the lowest ethanol, acetic acid and ammonia-N/total nitrogen, and highest contents of residual fructose, total mono- and di-saccharides and dry matter as well as high lactic acid/acetic acid and lactic acid content. FJLB addition had the lowest pH value and the highest lactic acid content, the most intensive lactic acid fermentation occurring in FJLB treated silages. This resulted in the faster accumulation of lactic acid and faster pH reduction. Sorbic acid and FJLB additions depressed clostridia or other undesirable bacterial fermentation, thus this decreased the water-soluble carbohydrates loss and saved the fermentable substrate for lactic acid fermentation.

• Journal of Animal Science and Technology
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• v.47 no.6
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• pp.947-954
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• 2005

This study was conducted to investigate the effects of hydrolyzed Lactobacillus supplementation with digestive enzymes treatment on the macrophage activation, the induction of nitric oxide(NO), interleukin (IL)-6 and tumor necrosis factor(TNF)-$\alpha$. The RAW 264.7 murine macrophage was exposed to porcine Lactobacillus strains which were digested with both pepsin and pancreatin. The production of NO, TNF-$\alpha$ and IL-6 in the macrophage were strain and dose-dependent, respectively. The induction of NO and cytokines were higher in both 3149 and 3146 strains compared with other Lactobacillus strains. Overall, the level of NO was observed at the lower range between 10 and 150 μg hydrolysates per ml, whereas IL-6 and TNF-$\alpha$ were observed at relatively higher concentration between 50 and 300 μg hydrolysates per ml. Lactobacillus strains which produced a high level of NO also showed a high induction of TNF-$\alpha$ and IL-6. Therefore, the present results suggest that hydrolysates of Lactobacillus strains are related to induction of several macrophage mediators, and then it could be beneficially used to modulate gastrointestinal immune function of the host. Also, the methodogly employed in this study might be useful to investigate the effects of lactic acid bacteria on gastrointestinal immunity.

• Journal of Animal Science and Technology
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• v.48 no.1
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• pp.77-90
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• 2006

The purpose of this study was to isolate cellulolytic and hemicellulolytic anaerobic bacteria inhabiting from gut of ruminants and investigate their hydrolytic enzyme activities. Extracellular CMCase activities of H-strains isolated from the rumen of a Holstein dairy cow were higher than those of D- and DC- strains from the rumen and large intestine of Korean spotted deer. Most isolated bacteria utilized more efficiently Dehority's artificial medium containing starch, glucose and cellobiose (DAS) than those in Dehority's artificial medium containing cellulose only (DAC). The results of biochemical reactions and sugar fermentation indicated that the isolated bacteria belong to one of bacterial strains of Peptostreptococcus spp., Bifidobacterium spp., Prevotela ruminicola/buccae, Clostridium beijer/butyricum and Streptococcus intermedis which are not highly cellulolytic. Activities of Avicelase, xylanase, β-D-glucosidase, α-L-arabinofuranosidase and β-xylosidase of the isolated anaerobic bacteria in DAS were higher than those in DAC. In conclusion, the results indicated the higher enzyme activities of the isolated strains cultured in DAS medium were mainly caused by their specific carbohydrate utilization for enzyme production and growth rate. The highly cellulolytic bacteria were not isolated in the present experiment. Thus further research is required to investigate characteristics of gut bacteria from Korean spotted deer.

• Microbiology and Biotechnology Letters
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• v.39 no.1
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• pp.9-19
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• 2011

The Bacillus subtilis pyrimidine biosynthetic (pyr) operon encodes all of the enzymes for the de novo biosynthesis of Uridine monophosphate (UMP) and additional cistrones encoding a uracil permease and the regulatory protein PyrR. The PyrR is a bifunctional protein with pyr mRNA-binding regulatory funtion and uracil phosphoribosyltransferase activity. To study the global regulation by the pyrR deletion, the proteome comparison between Bacillus subtilis DB104 and Bacillus subtilis DB104 ${\Delta}$pyrR in the minimal medium without pyrimidines was employed. Proteome analysis of the cytosolic proteins from both strains by 2D-gel electrophoresis showed the variations in levels of protein expression. On the silver stained 2D-gel with an isoelectric point (pI) between 4 and 10, about 1,300 spots were detected and 172 spots showed quantitative variations in which 42 high quantitatively variant proteins were identified. The results showed that production of the pyrimidine biosynthetic enzymes (PyrAA, PyrAB, PyrB, PyrC, PyrD, and PyrF) were significantly increased in B. subtilis DB104 ${\Delta}$pyrR. Besides, proteins associated carbohydrate metabolism, elongation protein synthesis, metabolism of cofactors and vitamins, motility, tRNA synthetase, catalase, ATP-binding protein, and cell division protein FtsZ were overproduced in the PyrR-deficient mutant. Based on analytic results, the PyrR might be involved a number of other metabolisms or various phenomena in the bacterial cell besides the pyrimidine biosynthesis.

• Journal of Life Science
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• v.22 no.2
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• pp.251-258
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• 2012

The purpose of this study was to investigate the probiotic properties of lactic acid bacterial strains isolated from animal feces. BCNU 9041 and BCNU 9042 isolates were assigned to Lactobacillus brevis on the basis of their physiological properties and 16S ribosomal DNA sequences analysis. They were confirmed as safe bioresources because of their non-hemolytic activities and non-production of harmful ${\beta}$-glucosidase, ${\beta}$-glucuronidase, tryptophanase, or urease. These isolates were also highly resistant to acid (at pH 2.5) and bile acids (at concentration of 0.3%, 0.6%, and 1% oxgall). In addition, they exhibited good antibacterial activity against food-borne bacteria, especially Bacillus cereus, Listeria monocytogenes, and Shigella sonnei. Furthermore, it was demonstrated that they have the highest levels of hydrophobicity and that they showed bile salt hydrolytic and cholesterol assimilation activity. These results suggest that BCNU 9041 and 9042 have good potential for application in functional foods and health-related products.

• Journal of Animal Science and Technology
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• v.54 no.2
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• pp.133-139
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• 2012

A bacterium, identified as $Bacillus$ $amyloliquefacies$, CNL-90 using 16S rDNA analysis, was isolated from malt grain. The optimal activities of its ${\alpha}$-amylase and protease were observed at pH 6 and $60^{\circ}C$, and at pH 6 and $50^{\circ}C$, respectively although their activities remained stable at pH 7 and $40^{\circ}C$for ${\alpha}$-amylase and at pH 7 and $50^{\circ}C$ for protease. After solid-state fermentation of $B.$ $amyloliquefacies$, CNL-90 on wheat bran for 72hr or 144hr, the ${\alpha}$-amylase and protease activities were 170,000 and 290,000 units/kg, and 290,000 and 310,000 units/kg, respectively. The viable bacterial cell counts were $1.5{\times}10^9$ CFU/g and $2.2{\times}10^9$ CFU/g at 72hr and 144hr of the solid-state fermentation, respectively. A feeding trial with a total of 127 piglets was also conducted. The animals were divided into two groups: an experimental group fed with the fermented product (63 piglets) and a control group (64 piglets). The growth rate of the experimental group was 6.66% higher than that of the control group (P<0.05). The results of this study indicate that the ${\alpha}$-amylase and protease from $B.$ $amyloliquefacies$, CNL-90 can be used for industrial applications due to their activity in production of carbohydrate hydrolysates.

• Asian-Australasian Journal of Animal Sciences
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• v.16 no.11
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• pp.1666-1672
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• 2003

The objective of this study was to evaluate the potential of an alumninosilicate clay, marketed under the trade name Biotite V, to improve growing-finishing pig performance and to determine its effects on nutrient digestibility and excretion. Sixty crossbred pigs (22.3${\pm}2.7kg$, Camborough 15 Line female${\times}$Canabred sire) were assigned on the basis of sex, weight and litter to one of four dietary treatments in a $2{\times}4$ (two sexes and four treatments) factorial design experiment. The experimental diets were based on barley and soybean meal and contained 0, 0.25, 0.5 or 0.75% biotite during the growing period (22.3-60.5 kg) and 0, 0.5, 1.0 or 1.5% biotite during the finishing period (60.5-110.3 kg). Each pig was allowed access to its own individual feeder for 30 min twice daily (07:00 and 15:00 h). Individual pig body weight, feed consumption and feed conversion were recorded weekly. The pigs were slaughtered at a commercial abattoir when they reached an average weight of 110.3 kg. Carcass weight was recorded and dressing percentage calculated. Carcass fat and lean measurements were obtained with a Destron PG 100 probe between the 3rd and 4th last ribs, 70 mm of the midline. Total tract digestibility coefficients for dry matter, energy, nitrogen and phosphorus were determined using three males and three females per treatment starting at an average weight of $52.2{\pm}3.8kg$. These pigs were housed under identical conditions as those used in the growing stage and were fed the same diets modified only by the addition of 0.5% chromic oxide as a digestibility marker. Over the entire experimental period (22.3-110.3 kg), daily gain was unaffected (p>0.05) by the inclusion of biotite in the diet. There was a cubic response for feed intake (p=0.06) and a quadratic response (p=0.07) for feed conversion due to biotite. Feeding biotite produced no significant (p>0.05) linear or quadratic effects on any of the carcass traits measured. Dry matter digestibility decreased linearly (p=0.02) with increasing levels of biotite in the diet. However, digestibility coefficients for energy, nitrogen and phosphorus were unaffected (p<0.05) by biotite inclusion. Lactobacilli and enterobacteria numbers were unaffected by inclusion of biotite while Salmonella was not detected in any of the fecal samples. The overall results of this experiment indicate that biotite inclusion did not reduce fecal excretion of nitrogen or phosphorus and failed to improve nutrient digestibility. Neither growth rate nor carcass quality was improved while a modest improvement in feed conversion was observed at lower levels of inclusion. Based on the results of this experiment, it would be difficult to justify the routine inclusion of biotite in diets fed to grower-finisher pigs. Whether or not a greater response would have been obtained with pigs of a lower health status is unknown.

• Asian-Australasian Journal of Animal Sciences
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• v.11 no.5
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• pp.510-521
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• 1998

The effects of two types of protein, soybean meal (SBM) and fish meal (FM); and two types of energy supplements, corn flour (CF) and paper pulp (PP), on intake of guinea grass (Panicum maximum), fibre digestion and microbial activities in four Merino rams with an average weight of $54.4{\pm}4.5kg$ were studied. Each animal was fitted with a ruminal cannula and a duodenal cannula at the proximal position. The animals were fed twice daily with chopped guinea grass (5 cm) ad libitum and one of the four dietary supplements: 170 g FM+268 g PP; 170 g FM+268 g CF; 200 g SBM+200 g PP or 200 g SBM+200 g CF. All the supplements were mixed with 100 g molasses. In sacco and in vivo digestibilities, digesta flow rates, fermentation and microbial population were studied in a $4{\times}4$ Latin square design with a $2{\times}2$ factorial arrangement of dietary treatments. The effects of energy or protein sources were not significant on grass intake of sheep. The potential degradabilities of NDF and ADF were not significantly affected by any of the supplements. However, the energy and protein sources had significant efects on disappearance rate of NDF and ADF. The disappearance rate of both NDF and ADF were significantly (p < 0.05) higher in animals fed PP when compared to animals fed CF. Animals fed FM also showed significantly (p < 0.03) higher disappearance rate of ADF than those fed SBM. Animals fed PP showed better digestion in the rumen and total tract. Total flow of NDF and ADF through the duodenum was not significantly affected by the various supplements. The mean rumen pH values (5.8-6.1) were not significantly different among the four different diets. The concentration of rumen ammonia was significantly (p < 0.0001) higher in animals fed SBM (235-266.4 mg N/L) supplement than in animals fed FM (174.9-179.7 mg N/L), while total VFA concentration was not significantly affected by both energy and protein supplements. Mean values of total VFA ranged from 72.5-82.3 mM. Molar proportions of acetate, propionate and butyrate were typical of a roughage type fermentation. Molar proportion of acetate was significantly (p < 0.0001) higher in sheep fed PP when compared to sheep fed CF. Animals fed FM had higher total viable bacterial counts, while animals fed CF showed higher protozoal numbers. Proportions of cellulolytic bacteria were only slightly higher in animals fed SBM or PP.

• Asian-Australasian Journal of Animal Sciences
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• v.20 no.11
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• pp.1699-1704
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• 2007

This experiment was carried out to evaluate the effects of adding encapsulated-glucose, glucose, sorbic acid or prefermented juice of epiphytic lactic acid bacteria (FJLB) on the fermentation quality and residual mono- and disaccharide composition of Italian ryegrass (Lolium multiflorum Lam) silages. The additive treatments were as follows: (1) control (no addition), (2) encapsulated-glucose addition at 0.5% for glucose, (3) glucose addition at 1%, (4) sorbic acid addition at 0.1%, (5) FJLB addition at a theoretical application rate of $2.67{\times}10^5$ CFU (colony forming unit) $g^{-1}$, on a fresh weight basis of Italian ryegrass. Although control and encapsulated-glucose treatments had higher contents of butyric acid (33.45, 21.50 g $kg^{-1}$ DM) and ammonia-N/Total nitrogen (114.91, 87.01 g $kg^{-1}$) as compared with the other treated silages, the fermentation in all silages was clearly dominated by lactic acid. This was well indicated by the low pH (4.38-3.59), and high lactic acid/acetic acid (4.39-22.97) and lactic acid content (46.85-121.76 g $kg^{-1}$ DM). Encapsulated-0.5% glucose and glucose addition increased lactic acid/acetic acid, and significantly (p<0.05) decreased ammonia-N/total nitrogen, and the contents of butyric acid and total volatile fatty acids (VFAs) as compared with the control. However, there were higher butyric acid and lower residual mono-and di-saccharides on the two treatments as compared with sorbic acid and FJLB addition, and their utilization efficiency of water soluble carbohydrates (WSC) was lower than that of both sorbic acid and FJLB additions. Sorbic acid addition showed the lowest content of ethanol and ammonia-N/total nitrogen, and the highest content of residual fructose and total mono-and disaccharides as well as the higher lactic acid/acetic acid value. Sorbic acid addition decreased the loss of mono-and disaccharides, and inhibited the activity of clostridial and other undesirable bacteria, and greatly increased the utilization efficiency of fermentable substrates by epiphytic LAB. FJLB addition had the lowest pH value and the highest lactic acid content among all additive treatments, with the most intensive lactic acid fermentation occurring in FJLB treated silage. This resulted in the faster accumulation of lactic acid and faster pH reduction. Sorbic acid and FJLB addition depressed clostridia or other undesirable bacterial fermentation which decreased the WSC loss and saved the fermentable substrate for lactic acid fermentation.