• Journal of Animal Science and Technology
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• v.62 no.2
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• pp.208-217
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• 2020

Heat stress negatively affects cattle productivity by reducing feed intake. In the present study, we assessed if the rumen microbiome composition of Hanwoo steers was altered by exposure to heat stress. Rumen samples were collected from four Hanwoo steers that were individually housed in climate-controlled chambers with 60% humidity and environmental temperatures of: 1) 15℃ (0-day group), 2) 35℃ for 3 days (3-day group), and 3) 35℃ for 6 days (6-day group). The total community DNA of samples was extracted, and 997,843 bacterial and 1,508,770 archaeal sequences were analyzed using next-generation sequencing. Assessment of the relative abundances revealed 15 major phyla of which Bacteroidetes was found to be the most dominant. After 3 days of heat stress exposure there were no significant changes in the rumen microbiome composition, except for a decrease in the Planctomycetes. However, after 6 days of heat stress exposure, we found that the relative abundance of fibrolytic Ruminococcaceae had decreased while that of lactate-producing Lactobacillaceae and amylolytic Prevotella and Ruminobacter had increased. The normal rumen microbiome of Hanwoo cattle was shown to be disrupted after 6 days of heat stress, which led to the decrease in fibrolytic bacteria that are sensitive to low pH and the increase in both lactate-producing and amylolytic bacteria. We have demonstrated that the microbiome composition of the rumen is affected by acute heat stress. Our findings may contribute to the development of different feeding strategies to restore heat stress-induced disruption of the rumen microbiome.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.52 no.4
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• pp.458-468
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• 2007

To tolerate environmentally adverse conditions such as cold, drought, and salinity, plants often synthesize and accumulate proline in cells as compatible osmolytes. ${\Delta}^1$-Pyrroline-5-carboxylate synthetase(P5CS) catalyzes the rate-limiting step of proline biosynthesis from glutamate. Two complete genes, MtP5CS1 and MtP5CS2, were isolated from the model legume Medicago truncatula by cDNA cloning and bacterial artificial chromosome library screening. Nucleotide sequence analysis showed that both genes consisted of 20 exons and 19 introns. Alignment of the predicted amino acid sequences revealed high similarities with P5CS proteins from other plant species. The two MtP5CS genes were expressed in response to high salt and low temperature treatments. Semi-quantitative reverse transcription-polymerase chain reaction showed that MtP5CS1 was expressed earlier than MtP5CS2, indicating differential regulation of the two genes. To evaluate the reverse genetic effects of nucleotide changes on MtP5CS function, a Targeting Induced Local Lesions in Genomes approach was taken. Three mutants each were isolated for MtP5CS1 and MtP5CS2, of which a P5CS2 nonsense mutant carrying a codon change from arginine to stop was expected to bring translation to premature termination. These provide a valuable genetic resource with which to determine the function of the P5CS genes in environmental stress responses of legume crops.

• Journal of Periodontal and Implant Science
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• v.38 no.sup2
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• pp.335-342
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• 2008

Purpose: Cytolethal distending toxin (CDT) considered as a key factor of localized aggressive periodontitis, endocarditis, meningitis, and osteomyelitis is composed of five open reading frames (ORFs). Among of them, the individual role of CdtA and CdtC is not clear; several reports presents that CDT is an AB2 toxin and they enters the host cell via clathrin-coated pits or through the interaction with GM3 ganglioside. So, CdtA, CdtC, or both seem to be required for the delivery of the CdtB protein into the host cell. Moreover, recombinant CDT was suggested as good vaccine material and antibody against CDT can be used for neutralization or for a detection kit. Materials and Methods: We constructed the pET28a-cdtC plasmid from Aggregatibacter actinomycetemcomitans Y4 by genomic DNA PCR and expressed in BL21 (DE3) Escherichia coli system. We obtained the antibody against the recombinant CdtC in mice system. Using the anti-CdtC antibody, we test the native CdtC detection by ELISA and Western Blotting and confirm the expression time of native CdtC protein during the growth phase of A. actinomycetemcomitans. Results: In this study we reconstructed CdtC subunit of A. actinomycetemcomitans Y4 and generated the anti CdtC antibody against recombinant CdtC subunit expressed in E. coli system. Our anti CdtC antibody can be interacting with recombinant CdtC and native CDT in ELISA and Western system. Also, CDT holotoxin existed at 24h but not at 48h meaning that CDT holotoxin was assembled at specific time during the bacterial growth. Conclusion: In conclusion, we thought that our anti CdtC antibody could be used mucosal adjuvant or detection kit development, because it could interact with native CDT holotoxin.

• Restorative Dentistry and Endodontics
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• v.28 no.4
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• pp.303-313
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• 2003

It has been documented that periodontopathic bacteria are also implicated in endodontic infections. 168 rDNA gene-directed PCR was to examine the prevalence of periodontopathic bacteria including Actinobacillus actinomycetemcomitans (Aa), Prevotella intermedia (Pi), Prevotella nigrescens (Pn), Porphyromonas gingivalis (Pg), Porphyromonas endodontalis (Pe), and Treponema denticola (Td) in the root canals of 36 endodontically infected teeth having apical lesions with or without clinical symptoms like pain, swelling, and fistula. 1. In 36 infected root canals, most frequently detected bacterial species was Pg (61.1%), followed by Td (52.8%) and Pe (38.9%). 2. Of 36 infected root canals, Aa was detected in 6 canals (16.7%) of the teeth, all of which showed clinical symptoms. 3. Of 36 infected root canals, Pi and Pn were found in 4 03.9%) and 5 (33.3%), respectively. Notably, prevalence of Pn in the symptomatic teeth was 50.0%. 4. One of black-pigmented anaerobic bacteria (BPB) including Pi, Pn, Pe, and Pg was detected in all of the teeth that showed pain or especially swelling but not fistula. It was, however, found that prevalence of BPB in the asymptomatic teeth or the teeth with fistula was only 40%. 5. Pe and Pg were detected in the teeth regardless of the presence or absence of symptoms. 6. Td was detected in the teeth regardless of the presence or absence of symptoms. High prevalence of BPB in the symptomatic teeth but low in the asymptomatic teeth suggests that BPB may play an important role in the pathogenesis of periapical lesions.

• Mycobiology
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• v.41 no.4
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• pp.234-242
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• 2013

A total of 62 bacterial isolates were obtained from Gomsohang mud flat, Mohang mud flat, and Jeju Island, Republic of Korea. Among them, the isolate CNU114001 showed significant antagonistic activity against pathogenic fungi by dual culture method. The isolate CNU114001 was identified as Bacillus amyloliquefaciens by morphological observation and molecular data analysis, including 16SrDNA and gyraseA (gyrA) gene sequences. Antifungal substances of the isolate were extracted and purified by silica gel column chromatography, thin layer chromatography, and high performance liquid chromatography. The heat and UV ray stable compound was identified as iturin, a lipopeptide (LP). The isolate CNU114001 showed broad spectrum activity against 12 phytopathogenic fungi by dual culture method. The semi purified compound significantly inhibits the mycelial growth of pathogenic fungi (Alternaria panax, Botrytis cinera, Colletotrichum orbiculare, Penicillium digitatum, Pyricularia grisea and Sclerotinia sclerotiorum) at 200 ppm concentration. Spore germ tube elongation of Botrytis cinerea was inhibited by culture filtrate of the isolate. Crude antifungal substance showed antagonistic activity against cucumber scleotiorum rot in laboratory, and showed antagonistic activity against tomato gray mold, cucumber, and pumpkin powdery mildew in greenhouse condition.

• Korean Journal of Medicinal Crop Science
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• v.18 no.6
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• pp.429-438
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• 2010

The use of cellulolytic lactic acid bacteria in new method to prepare high nutrition complementary foods was investigated. For the screening of cellulolytic lactic acid bacteria, more than 1,150 bacterial colony were isolated from diluted infant feces samples. A typical strain which appeared the most excellent cellulolytic activities was identified novel acidophilic Enterococcus sp. TO-94 through the results of morphological, biochemical and chemotaxonomic characteristics and 16S rDNA sequencing. The optimal lactic acid fermentation conditions of Omija(Schizandra chinensis Baillon) by Enterococcus sp. TO-94 were as follows: pH and temperature were 3.0 and $37^{\circ}C$, respectively, and fermentation time was 20hrs. The fructose and glucose were major free sugar and the contents were 5.83 and 4.30 mg/g after fermentation, respectively. The contents of lactic acid and acetic acid were 9.84 mg/g and 2.08 mg/g after fermentation, respectively. The vitamin $B_1$, $B_2$, niacin, folic acid and C were major vitamin in the fermented broth, the contents were 1.5~3 times higher than those of initial fermentation time. Also, the contents of polyphenol and anthocyanine were 3.8 and 1.2 times higher than those of initial fermentation time.

• Journal of Microbiology
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• v.44 no.2
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• pp.155-161
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• 2006

Comparative analysis of microbial communities in a sequencing batch reactor which performed enhanced biological phosphorus removal (EBPR) was carried out using a cultivation-based technique and 16S rRNA gene clone libraries. A standard PCR protocol and a modified PCR protocol with low PCR cycle was applied to the two clone libraries of the 16S rRNA gene sequences obtained from EBPR sludge, respectively, and the resulting 424 clones were analyzed using restriction fragment length polymorphisms (RFLPs) on 16S rRNA gene inserts. Comparison of two clone libraries showed that the modified PCR protocol decreased the incidence of distinct fragment patterns from about 63 % (137 of 217) in the standard PCR method to about 34 % (70 of 207) under the modified protocol, suggesting that just a low level of PCR cycling (5 cycles after 15 cycles) can significantly reduce the formation of chimeric DNA in the final PCR products. Phylogenetic analysis of 81 groups with distinct RFLP patterns that were obtained using the modified PCR method revealed that the clones were affiliated with at least 11 phyla or classes of the domain Bacteria. However, the analyses of 327 colonies, which were grouped into just 41 distinct types by RFLP analysis, showed that they could be classified into five major bacterial lineages: ${\alpha},\;{\beta},\;{\gamma}-$ Proteobacteria, Actinobacteria, and the phylum Bacteroidetes, which indicated that the microbial community yielded from the cultivation-based method was still much simpler than that yielded from the PCR-based molecular method. In this study, the discrepancy observed between the communities obtained from PCR-based and cultivation-based methods seems to result from low culturabilities of bacteria or PCR bias even though modified culture and PCR methods were used. Therefore, continuous development of PCR protocol and cultivation techniques is needed to reduce this discrepancy.

• Microbiology and Biotechnology Letters
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• v.20 no.3
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• pp.289-294
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• 1992

Twenty-four bacterial strains among alkalophillic bacteria isolated from soil samples were examined for the presence of type II restriction endonuclease in aerobic culture. One strain was found to contain specific enzyme to cleave lambda DNA. The characteristics of this microorganism is the ability to grow well in alkalophilic and high temperature condition, that is at pH 10.3 and $50^{\circ}C$. This strain was tentatively identified to Bacillus alkaloPhilus subsp. halodurans when morphological, physiological and biochemical characteristics were examined. The enzyme was purified from crude extract by streptomycin sulfate, ammonium sulfate precipitation, which was followed by DEAE-cellulose and phosphocellulose ion exchange column chromatography, and the subunit molecular weight was about, 32,000 daltons by polyacrylamide gel electrophoresis containing 0.1% SDS.

• Microbiology and Biotechnology Letters
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• v.42 no.4
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• pp.339-346
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• 2014

A bacterial strain was isolated from homemade doenjang (Korean fermented soybean paste) as a producer of the extracellular ${\beta}$-galactosidase, capable of hydrolyzing lactose to liberate galactose and glucose residues. The isolate YB-1414 has been identified as Bacillus licheniformis on the basis of its 16S rDNA sequence, morphology and biochemical properties. The production of ${\beta}$-galactosidase by B. licheniformis YB-1414 reached maximum levels of 6.2 U/ml in culture medium containing wheat bran (1%) and yeast extract (2.5%) as carbon and nitrogen sources, respectively. Particularly, the insoluble fraction was more effective for ${\beta}$-galactosidase production than the soluble extract of wheat bran. The enzyme exhibited maximum activity for hydrolysis of para-nitrophenyl-${\beta}$-D-galactopyranoside (pNP-${\beta}Gal$) under reaction conditions of pH 6.0 and $55-60^{\circ}C$. Its hydrolyzing activity for pNP-${\beta}Gal$ was drastically decreased by the addition of low concentrations of galactose, but only slightly decreased by glucose, with 85% of maximal activity in the presence of 400 mM glucose.

• The Journal of Natural Sciences
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• v.14 no.2
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• pp.83-91
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• 2004

Recently phage K11 lysozyme was cloned and characterized in our lab. The K11 lysozyme was identified to have dual functions. It not only cuts a peptidoglycan bond in bacterial cell wall but also acts as an inhibitor of K11 RNA polymerase. It has been known that the T7 lysozyme binds specifically to T7 RNA polymerase and inhibits transcription. The dual activities of K11 lysozyme are atreeable to the case of T7 phage lysozyme and RNA polymerare. In order to identify the binding magnitude of K11 lysozyme with K11 RNA polymerase, yeast two-hybrid system was used. K11 phage lysozyme gene was introduced into pLexA plasmid and used as a prey. Also, K11 phage RNA polymerase gene was introduced into pJG4-5 and used as a bait. The binding between K11 lysozyme and K11 RNA polymerase was demonstrated by expression of reporter genes such as lacZ and leu2.