• Journal of Microbiology and Biotechnology
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• 제12권4호
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• pp.544-552
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• 2002

Several microorganisms from rat and human feces and lumen fluid of cows were screened for their ability to decolorize the synthetic dyes. Consequently, a novel dye-degrading strain AB&J was isolated. Taxonomic identification including 165 rDNA sequencing and phylogenetic analysis indicated that the isolate had 99.9% homology in its 165 rDNA base sequence with Clostridium perfringens. After 27 h Incubation with the strain, brilliant blue R, bromophenol blue, crystal violet, malachite green, methyl green, and methyl orange were decolorized by about 69.3%, 97.7%, 96.3%, 97.9%, 75.1%, and 97.2%, respectively. The triphenlmethane dye, bromophenol blue, was decolorized extensively by growing Clostridium perfringens AB&J cells in liquid cultures under anaerobic condition, although their growth was strongly inhibited in the initial stage of incubation. This group of dyes is toxic, depending on the concentration used. The dye was significantly decolorized at a relatively lower concentration of below 50 $\mu g \;ml^{-1}$, however, the growth of the cells was mostly suppressed at a dye concentration of 100 $\mu g \;ml^{-1}$. The decolorization activity in cell-free extracts was much higher in cytoplasm than in periplasm and cytoplasmic membrane. Therefore, the enzyme related uptake of bromophenol blue seemed to be localized in cytoplasm. The optimal pH and temperature of bromophenol blue uptake fur decolorization activities were 7.0 and 4$0^{\circ}C$, respectively.

• Journal of Animal Science and Technology
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• 제54권6호
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• pp.435-442
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• 2012

This study was conducted to isolate and identify the lactic acid bacteria (LAB) from spent mushroom substrates (SMS) for the effective anaerobic fermentation to utilize SMS as an animal feed and to determine the optimal medium conditions for their growth. At first, a total of 23 strains were isolated from the ensiled SMS based on the LAB counts and pH tested. Then, a total of 16 strains which rapidly produce lactate and decreased the pH, were selected for a screening test. The optical density (OD), pH, and yellow clear zone were tested for the selected 16 strains. Among the strains, KU5 strain had wider yellow clear zone and lower pH and KU13 strain had higher OD at 24 hr of incubation and wider yellow clear zone compared to other strains and control strain (Lactobacillus plantarum KCCM 12116). Accordingly, KU5 and KU13 strains were finally selected. The KU5 and KU13 were identified as Lactobacillus plantarum by the 16S rRNA sequencing. The KU5 strain was named as Lactobacillus plantarum KU5, and the KU13 strain was named as Lactobacillus plantarum KU13. Lactobacillus plantarum KU5 and Lactobacillus plantarum KU13 were registered at the National Center for Biotechnology Information (NCBI). Access number of Lactobacillus plantarum KU5 was HQ542227 and that of Lactobacillus plantarum KU13 was HQ542228. The optimal medium conditions for growth of KU5 and KU13 were soybean meal 2% and formulated feed 2%, respectively.

• Journal of Microbiology and Biotechnology
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• 제24권8호
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• pp.1034-1043
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• 2014

This study determined the effect of light intensity and photoperiod on the dry cell weight and total amount of carotenoids in four isolates of purple non-sulfur bacteria obtained from shaded and exposed microhabitats of a mangrove ecosystem in Kota Kinabalu, Sabah, Malaysia. The initial isolation of the bacteria was carried out using synthetic 112 medium under anaerobic conditions (2.5 klx) at $30{\pm}2^{\circ}C$. On the basis of colony appearance, cell morphology, gram staining, motility test, and 16S rRNA gene sequencing analyses, all four bacteria were identified as Afifella marina. One of the bacterial isolates, designated as Af. marina strain ME, which was extracted from an exposed mud habitat within the mangrove ecosystem, showed the highest yield in dry cell weight ($4.32{\pm}0.03g/l$) as well as total carotenoids ($0.783{\pm}0.002mg/g$ dry cell weight). These values were significantly higher than those for dry cell weight ($3.77{\pm}0.02g/l$) and total carotenoid content ($0.706{\pm}0.008mg/g$) produced by the isolates from shaded habitats. Further analysis of the effect of 10 levels of light intensity on the growth characteristics of Af. marina strain ME showed that the optimum production of dry cell weight and total carotenoids was achieved at different light intensities and incubation periods. The bacterium produced the highest dry cell weight of 4.98 g/l at 3 klx in 72 h incubation, but the carotenoid production of 0.783 mg/g was achieved at 2.5 klx in 48 h incubation. Subsequent analysis of the effect of photoperiod on the production of dry cell weight and total carotenoids at optimum light intensities (3 and 2.5 klx, respectively) revealed that 18 and 24 h were the optimum photoperiods for the production of dry cell weight and total carotenoids, respectively. The unique growth characteristics of the Af. marina strain ME can be exploited for biotechnology applications.

• Asian-Australasian Journal of Animal Sciences
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• 제25권1호
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• pp.114-121
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• 2012

The potential for ochratoxin A (OTA) degradation by swine intestinal microbiota was assessed in the current study. Intestinal content that was collected aseptically from swine was spiked with 100 ppb OTA and incubated for 6 and 12 h at $39^{\circ}C$. An OTA assay was conducted using the incubated samples, and it was found that 20% of the OTA toxin was detoxified, indicating the presence of microbes capable of OTA degradation. Twenty-eight bacterial species were isolated anaerobically in M 98-5 media and 45 bacterial species were isolated using nutrient broth aerobically. Screening results showed that one anaerobic bacterial isolate, named MM11, detoxified more than 75% of OTA in liquid media. Furthermore, 1.0 ppm OTA was degraded completely after 24 h incubation on a solid 'corn' substrate. The bacterium was identified by 16S rDNA sequencing as having 97% sequence similarity with Eubacterium biforme. The isolation of an OTA-degrading bacterium from the swine natural flora is of great importance for OTA biodegradation and may be a valuable potential source for OTA-degradation enzymes in industrial applications.

• 한국환경보건학회지
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• 제37권2호
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• pp.124-132
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• 2011

Biofilm and aeration tank of pilot and full RABC (rotating activated Bacillus contactor) plant were analyzed to characterize and determine bacterial community structure in food wastewater treatment system at winter. Concentration of heterotrophic bacteria and Bacillus group was $10^7$ and $10^5$ CFU/ml, respectively, at biofilm of pilot-plant while others represented $10^6$ and $10^4$ CFU/ml, respectively. Five and eight phyla were detected at biofilm of pilot- and full-plant, respectively, by 16S rDNA sequencing. Biofilm of pilot-plant was dominated by ${\beta}$-Proteobacteria (38.8%), ${\gamma}$-Proteobacteria (22.4%), and Bacteroidetes (12.2%), and the most dominant genus was Zoogloeae genus (22.4%). Candidate division TM7 (12.5%) was only detected at biofilm of full-plant and it was dominated by Bacteroidetes (33.3%), ${\gamma}$-Proteobacteria (29.2%), and ${\beta}$-Proteobacteria (20.8%). Clostridium genus specific primer set enabled to detect the sequences of Clostridium genus. These suggested that anaerobic and aerobic bacteria were coexisted even from the initial period of biofilm formation and ${\beta}$-Proteobacteria, ${\gamma}$-Proteobacteria and Bacteroidetes were major phyla in biofilm of food wastewater treatment system at winter.

• 한국유기농업학회지
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• 제20권3호
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• pp.327-343
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• 2012

In order to develop a high cellulolytic direct-fed microorganism (DFM) for ruminant productivity improvement, this study isolated cellulolytic bacteria from the rumen of Holstein dairy cows, and compared their cellulolytic abilities via DM degradability, gas production and cellulolytic enzyme activities. Twenty six bacteria were isolated from colonies grown in Dehority's artificial (DA) medium with 2% agar and cultured in DA medium containing filter paper at $39^{\circ}C$ for 24h. 16s rDNA gene sequencing of four strains from isolated bacteria showed that H8, H20 and H25 strains identified as Ruminococcus flavefaciens, and H23 strain identified as Fibrobacter succinogenes. H20 strain had higher degradability of filter paper compared with others during the incubation. H8 (R. flavefaciens), H20 (R. flavefaciens), H23 (F. succinogenes), H25 (R. flavefaciens) and RF (R. flavefaciens sijpesteijn, ATCC 19208) were cultured in DA medium with filter paper as a single carbon source for 0, 1, 2, 3, 4 and 6 days without shaking at $39^{\circ}C$, respectively. Dry matter degradability rates of H20, H23 and H25 were relatively higher than those of H8 and RF since 2 d incubation. The cumulative gas production of isolated cellulolytic bacteria increased with incubation time. At every incubation time, the gas production was highest in H20 strain. The activities of carboxymethylcellulase (CMCase) and Avicelase in the culture supernatant were significantly higher in H20 strain compared with others at every incubation time (p<0.05). Therefore, although further researches are required, the present results suggest that H20 strain could be a candidate of DFM in animal feed due to high cellulolytic ability.

• Journal of Microbiology
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• 제44권2호
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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.

• 한국미생물·생명공학회지
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• 제42권3호
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• pp.211-218
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• 2014

본 연구에서는 동물유래 시료들로부터 양돈 사료첨가용 유산균을 개발하기 위하여 enterotoxigenic E. coli, Salmonella Typhimurium, Clostridium perfringens 등 양돈산업에서 심각한 문제를 일으키는 인수공통 병원성 세균들과 Clostridium jejuni, Listeria monocytogenes, Staphylococcus aureus와 같은 식중독균을 강하게 저해하는 Bifidobacterium 유산균을 분리하고자 하였다. 이를 위하여 소의 반추위 내용물, 닭 창자 내용물, 돼지 분변 등의 시료들로부터 총 65주의 혐기성미생물을 분리하였다. 이 중에서 항병원세균활성이 가장 높은 4주를 선별하였는데, 이들은 16S rDNA 염기서열 분석방법에 의하여 3주의 B. boum과 1주의 B. thermophilum으로 동정되었다. 특히 B. thermophilum는 분리주들 중에서 가장 높은 돼지 장 상피세포에 대한 부착성을 보였고, 여러 인수공통병원세균들과 식중독균들에 대한 높은 항균력, 산과 담즙내성을 보여 양돈용 생균제 후보균주로서 우수한 특성을 나타내었다.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제29권1호
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• pp.48-55
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• 2003

The purpose of this study was to isolate and identify the bacteria in osteomyelitis lesion of 3 patients. Two lesions were due to the post-infection after extraction. The other was resulted from mal-fixation of both sides of mandibular angles. Pus samples were collected by needle aspiration from the lesion and examined by culture method. Bacterial culture was performed in three culture systems (anaerobic, $CO_2$, and aerobic incubator). Identification of the bacteria was performed by 16S rRNA gene cloning and nucleotide sequencing method. Our results showed that Streptococci species was predominantly isolated in both lesions of extraction socket. Only one species (Proteus vulagris) was detected in lesion of mandibular angle. This study was not sufficient to identify the causative bacteria in those osteomyelitis. However, our data may be offered the clue to solve the problem.

• Journal of Microbiology and Biotechnology
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• 제16권10호
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• pp.1518-1522
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• 2006

In the present study, it was observed how the phage-host system that is naturally reproduced in activated sludge is affected by the host inoculation. The system of Microlunatus phosphovorus and its phages was selected as the phage-host system native to an activated sludge system operated for 19 days under sequencing anaerobic-aerobic conditions with glutamate as the main carbon source. The phage-host system related to M. phosphovorus was monitored by plaque assay for the phages and by fluorescent in situ hybridization (FISH) for the bacterial host. In addition, the whole phage structure was also monitored by pulsed-field gel electrophoresis (PFGE). During the first 9 days, the phage-host system was more or less steady at approx. 9% (FISH/ DAPI) for M. phosphovorus and approx. 10,000 PFU/ml for its lytic phages. Microlunatus phosphovorus JCM9379 was inoculated into the activated sludge on day 10. Right after the inoculation, M. phosphovorus was approx. 24% (FISH/DAPI) whereas its lytic phages dropped down to approx. 500 PFU/ ml. After the host inoculation (within 9 days), however, the phage-host system eventually reverted to its original level in each population. On the other hand, the whole phage structure was not significantly changed by M. phosphovorus inoculation but stable throughout the process operation. Only the minor change that four phage groups gradually became abundant after the host inoculation was observed.