• Asian-Australasian Journal of Animal Sciences
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• v.24 no.5
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• pp.643-649
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• 2011

The objective of this study was to determine the effect of feeding direct-fed microbials (DFM) on the growth performance and prophylaxis of calf diarrhea during the pre-weaning period as an alternative to antibiotics. A multi-species DFM was formulated including three lactic acid bacteria (Lactobacillus salivarius Ls29, Pediococcus acidilactia Pa175, and L. plantarum Lp177), three Bacillus strains (B. subtilis T4, B. polymyxa T1 and SM2), one yeast, Saccharomyces boulardii, and a nonpathogenic E. coli Nissle 1917. Lactic acid bacteria and Bacillus strains were selected based on the antibacterial activity against various animal pathogens, especially pathogenic E. coli using agar diffusion methods in vitro. Test and control groups were fed milk replacer and calf starter supplemented with DFM ($10^9$ cfu each of eight species/d/head, n = 29) or with antibiotics (0.1% neomycin sulfate in milk replacer and Colistin 0.08% and Oxyneo 110/110 0.1% in calf starter, n = 15), respectively. Overall fecal score and the incidence rate of diarrhea were reduced in the DFM group compared to the antibiotics one. About 40% of calves in antibiotic group suffered from diarrhea while in DFM group only 14% showed diarrhea. There was no difference in the average daily gain and feed efficiency of two groups. The hematological levels of calves were all within the normal range with no significant difference. In conclusion, the feeding of multispecies DFM during the pre-weaning period could reduce calf diarrhea and there was no difference in the growth performance between the groups, thus showing the potential as an alternative to antibiotics.

• Microbiology and Biotechnology Letters
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• v.30 no.2
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• pp.123-128
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• 2002

A bacterium producing the extracellular xylanase was isolated from soil and has been identified as a Bacillus sp. strain. The isolate, named Bacillus sp. AMX-4, was shown to be similar to B. subtilis strain on the basis of its chemical compositions. The xylanase of culture supernatant was most active at 50℃ and pH 6.0. The additional carbon sources including monosaccharides, disaccharides, wheat bran, and rice straw increased the enzyme productivity. Especially, the maximum xylanase productivity was reached 29.2 units/ml in LB medium supplemented with 1.5％ (w/v) xylose, which was 16-folds more than that in LB medium. As the results of investigating the effects of xylose on cell growth and xylanase productivity of Bacillus sp. AMX-4, increase of xylanase production was owing to the induction of xylanase biosynthesis. It was also found that the enzyme production was in association with the growth of Bacillus sp. AMX-4.

• Journal of Microbiology and Biotechnology
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• v.8 no.4
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• pp.378-387
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• 1998

The DNA sequence immediately following the xynA gene of Bacillus stearothermophilus 236 ［about l-kb region downstream from the translational termination codon (TAA) of the xynA gene］was found to have an ability to enhance the xylanase activity of the upstream xynA gene. An 849-bp ORF was identified in the downstream region, and the ORF was confirmed to encode a novel protein of 283 amino acids designated as XaiF (xylanase-activity-increasing factor). From the nucleotide sequence of the xaiF gene, the molecular mass and pI of XaiF were deduced to be 32,006 Da and 4.46, respectively. XaiF was overproduced in the E. coli cells from the cloned xaiF gene by using the T7 expression system. The transcriptional initiation site was determined by primer extension analysis and the putative promoter and ribosome binding regions were also identified. Blast search showed that the xaiF and its protein product had no homology with any gene nor any protein reported so far. Also, in B. subtilis, the xaiF trans-activated the xylanase activity at the same rate as in E. coli. In contrast, xaiF had no activating effect on the co-expressed ${\beta}-xylosidase$ of the xylA gene derived from the same strain of B. stearothermophilus. In addition, the intracellular and extracellular fractions from the E. coli cells carrying the plasmid-borne xaiF gene did not increase the isolated xylanase activity, indicating that the protein-protein interaction between XynA and XaiF was not a causative event for the xylanase activating effect of the xaiF gene.

• Journal of Microbiology and Biotechnology
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• v.3 no.3
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• pp.146-155
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• 1993

A DNA fragment from an alkali-tolerent Bacillus sp., conferring strong promoter activity, was subcloned into the promoter probe plasmid pPL703 and the nucleotide sequence of this promoter region was determined. The sequence analysis suggested that this highly efficient promoter region containing the complex clustered promoters comprised three kinds of promoters (P1, P2 and P3), which are transcribed by $\sigma^B (formerly \sigma^{37}), \sigma^E(formerly \sigma^{29}) and \sigma^A (formerly \sigma^{43})$ RNA polymerase holoenzymes which play major rules at the onset of endospore formation, during sporulation and at the vegetative phase of growth, respectively. S1 nuclease mapping experiments showed that all three promoters had staggered transcription initiation points. The results of chloramphenicol acetyltransferase assay after the subcloning experiments also indicated that the expression of these clustered promoters was correlated with the programs of growth and endospore development. Promoter P1, P2 and P3 were preceded by 75% AT, 79% AT and 81% AT regions, respectively, and a partial deletion of AT-rich region prevented transcription from promoter P1 in vivo. Two sets of 5 -AGTGTT-3 sequences and inverted repeat sequences located around the promoter P1 were speculated as the possible cis acting sites for the catabolite repression in B. subtilis. In vivo transcripts from these sequence regions may be able to form a secondary structure, however, the possibility that a regulatory protein induced by the excess amount of glucose could be bound to such a domain for crucial action remains to be determined.

• The KIPS Transactions:PartA
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• v.16A no.3
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• pp.143-152
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• 2009

One of the main issues in comparative genomics is to study chromosomal gene order in one or more related species. For this purpose, the whole genome alignment is usually applied to find the horizontal gene transfer, gene duplication, and gene loss between two related genomes. Also it is well known that the novel visualization tool with whole genome alignment is greatly useful for us to understand genome organization and evolution process. There are a lot of algorithms and visualization tools already proposed to find the "gene clusters" on genome alignments. But due to the huge size of whole genome, the previous visualization tools are not convenient to discover the relationship between two genomes. In this paper, we propose AlignScope, a novel visualization system for whole genome alignment, especially useful to find gene clusters between two aligned genomes. This AlignScope not only provides the simplified structure of genome alignment at any simplified level, but also helps us to find gene clusters. In experiment, we show the performance of AlignScope with several microbial genomes such as B. subtilis, B.halodurans, E. coli K12, and M. tuberculosis H37Rv, which have more than 5000 alignment pairs (matched DNA subsequence).

• Food Science and Preservation
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• v.5 no.3
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• pp.299-304
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• 1998

Screening for antibacterial activities with microorganisms related to the food putrefaction by ethanol extract from Chrysanthemum petals widely used for the traditional wine production, and antibacterial activities of each concentration and minimum inhibitory concentration(MIC) from the ethanol extract were researched. Antibacterial activity of ehanol extract for B. subtilis was higher in C. boreale than in C. Morifolium, but that of E. coli was higher in C. molifolium than in C. boreale. C. boreale was higher than C. morifolium in the antibacterial activity of ehanol extract and MIC of ehanol extract from C. boreale was 60-70${\mu}\ell$/ml. Ethanol extract from C. boreale was higher Gram(-) than Gram(+) in the antibacterial activities, but Gram(-), Gram(+) were greatly inhibited on growth in 100${\mu}\ell$ concentration.

• KSBB Journal
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• v.14 no.6
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• pp.682-687
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• 1999

Virginiae butanolide C(VB-C) is one of the butyrolactone autoregulators, which triggers the production of virginiamycin in Streptomyces virginiae. In order to investigate the function of VB-C as inducer in other strains, Streptomyces erythraeus was used as a test strain(parent). VB-C binding receptor gene was introduced into S. erythraeus(transformant) and the production of VBs and specific VB-C binding protein were analysed in parent and transformant. When 300ng/ml of the synthetic VB-C was added at 0, 20, 44 h cultivation of the parent and at 44 h cultivation of the transformant, the initial production times a antibiotics were shortened by more than 8 and 6 h, respectively. The transformant showed strong antibiotic activity against B. subtilis. These results suggest that the VB-C might have an ability to induce the production of secondary metabolites in S. erythraeus.

• Asian-Australasian Journal of Animal Sciences
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• v.21 no.9
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• pp.1312-1317
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• 2008

The objective of the present trial was to determine the effect of a thermostable probiotic containing Bacillus licheniformis and B. subtilis on health and production parameters of fattening rabbits from weaning until slaughter. In a rabbitry with average post-weaning mortality of 5-9%, 1,680 rabbits were supplied with: a) a basic feed, or b) the same basic feed supplemented with probiotic from the 4th day postweaning (41st day of age) up to 88th day of age. Clinical signs, microbiological status and growth performance were recorded for two distinct fattening periods, growing and finishing. A significant decrease in mortality of probiotic-treated rabbits when compared to the controls was observed during the growing and entire fattening periods. Within these periods, E. coli and C. perfringens - but not P. multocida - were isolated at a lower frequency from probiotic-treated rabbits (p<0.05). Compared to the control animals, probiotic-treated rabbits were 54 g and 123 g heavier at the end of the growing and finishing phases, respectively, and had significantly higher average daily gain and better feed conversion ratio (p<0.05).

• Korean Journal of Microbiology
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• v.38 no.4
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• pp.318-321
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• 2002

To induce recombinant protein under phosphate restricted conditions such as soil, we have constructed the expression vector (pEAAP) with phoA gene promoter of Enterobacter aerogenes. To construct the pEAAP, deletion of the T7 promoter and lac operator from pET-22b(+) by BglII-XhoI digestion and addition of the phoA gene promoter (containing the pho box) were performed. To test pEAAP as an expression vector controled by phosphate limitation, pEAPHY1 was constructed with the phytate gene (Bsa-phy1) of Bacillus subtillis var. amyloliquefaciens (KCTC 8913P). Under the phosphate-limitation condition, CK-PHY1 ( Escherichia coli JM109 was transformed with pEAPHY1) expressed the 41 kD Bsa-Phy1 . Also CK-PHY1 formed the clear zone in solid medium containing phytate as a sole phosphate source.

• Journal of the Korean Society of Food Science and Nutrition
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• v.32 no.7
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• pp.1059-1065
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• 2003

Antimicrobial activity of Mokdan bark (Paeonia suffruticosa $A_{NDR}$) was investigated. Methanol extract of dried Mokdan was fractionated to hexane, chloroform, ethylacetate, butanol and aqueous fraction. Ethylacetate fraction among these fractions showed the highest inhibitory effect on the microorganisms such as L. monocytogenes, and E. coli at 500 $\mu\textrm{g}$/disc. Ethylacetate fraction was further fractionated into 3 fractions by silica gel column and thin layer chromatography (TLC). The results showed that ethylacetate fractions No. 1 and 2 had the highest antimicrobial activity. They were mixed again, reseparated, and 3 fractions were obtained. Among them, No. 1 had the highest inhibitory effect on the microorganisms, No. 1 fraction was identified as isobutyl isopentanoate by HPLC, and GC-MS.