• 한국미생물·생명공학회지
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• 제24권2호
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• pp.178-184
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• 1996

Dryfilm method by using 3M Petrifilm$^{TM}$ has been examined to replace conventional agar method for isolation of microorganisms from foods. The objectives of the present study were to evaluate suitability of dryfilm method as a microbial isolation method and to determine the effect of antimicrobial agent on dryfilm for isolation of microorganisms from foods. Five different foods, milk, ground beef, fishery surimi, Takju and wheat flour were used to isolate the natural microflora in foods and the inoculated Escheri chia coli. Standard method agar (SMA, Difco) and Petrifilm$^{TM}$ aerobic count (PAC, 3M) were used to isolate total microorganisms from foods. Violet red bile agar (VRBA), brilliant green lactose bile (BGLB) broth and Petrifilm$^{TM}$ coliform count (PCC, 3M) were used to isolate coliforms from foods. E. coli broth (EC broth) and Petrifilm$^{TM}$ E. coli count (PEC, 3M) were used to isolate E. coli from foods. Acidified potato dextrose agar (APDA) and Petrifilm$^{TM}$ yeast & mold count (PYMC, 3M) were used to isolate yeasts and molds from foods. Total aerobic plate counts isolated from five different foods by SMA and PAC (3M) were riot significantly different each other at P<0.05 level and were highly correlated each other ($\geq$0.96). Mugwort extract as an antimicrobial agent did not affect microbial enumeratiion of Dryfilm. Significantly higher number of coliform colonies were formed on VRBA than PCC (3M) from ground beef, but they were not significantly different in coliform colonies from milk samples. PCC (3M) and BGLB were not significantly different for enumeration of coliforms in milk and beef samples. Significantly higher number of E. coli were isolated by EC broth than PEC from ground beef, but these were not significontly different for enumeration of E. coli from milk. Yeast and mold counts isolated from Takju and wheat flour by APDA and PYMC (3M) were not significantly different at P<0.05 level. These data indicate that dryfilm method by using 3M Petrifilm$^{TM}$ can be successively used as an alternative to conventional agar method for enumeration of microorganisms in various foods.

• Journal of Microbiology
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• 제41권3호
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• pp.262-265
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• 2003

Although enumerating bacterial cells is a fundamental step in understanding microbial ecosystems in marine environments, substantial decrease in bacterial counts with increasing sample storage time hampers the accurate estimation of bacterial biomass. We compared the variations in bacterial cell numbers caused by freezing and thawing of sample bottles or slides. Bacterial counts of seawater samples frozen only once in a sampling bottle yielded approximately 95％ of the original numbers after 90 days, whereas 80％ of the original count was obtained for samples prepared on slides. Only 67％ and 58％ of the original counts were recovered in samples repeatedly frozen and thawed in bottles or on slides, respectively. The results indicated that freezing a seawater sample in a bottle increased the consistency of the epifluorescence microscopic enumeration of bacterial cells.

• Asian-Australasian Journal of Animal Sciences
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• 제20권2호
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• pp.283-294
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• 2007

Conventional culture-based methods of enumerating rumen microorganisms (bacteria, archaea, protozoa, and fungi) are being rapidly replaced by nucleic acid-based techniques which can be used to characterise complex microbial communities without incubation. The foundation of these techniques is 16S/18S rDNA sequence analysis which has provided a phylogenetically based classification scheme for enumeration and identification of microbial community members. While these analyses are very informative for determining the composition of the microbial community and monitoring changes in population size, they can only infer function based on these observations. The next step in functional analysis of the ecosystem is to measure how specific and, or, predominant members of the ecosystem are operating and interacting with other groups. It is also apparent that techniques which optimise the analysis of complex microbial communities rather than the detection of single organisms will need to address the issues of high throughput analysis using many primers/probes in a single sample. Nearly all the molecular ecological techniques are dependant upon the efficient extraction of high quality DNA/RNA representing the diversity of ruminal microbial communities. Recent reviews and technical manuals written on the subject of molecular microbial ecology of animals provide a broad perspective of the variety of techniques available and their potential application in the field of animal science which is beyond the scope of this treatise. This paper will focus on nucleic acid based molecular methods which have recently been developed for studying major functional groups (cellulolytic bacteria, protozoa, fungi and methanogens) of microorganisms that are important in nutritional studies, as well as, novel methods for studying microbial diversity and function from a genomics perspective.

• 미생물학회지
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• 제45권2호
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• pp.208-214
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• 2009

장류에서 식중독 미생물인 Bacillus cereus의 계수 방법상 문제점을 해결하기 위하여, B. cereus 선택 배지인 mannitol-egg york-polymyxin B (MYP)에서 자란 약 1,500개의 집락들을 형태적 차이에 따라 12종으로 분류하였다. 생화학 특성 및 16S rRNA 유전자의 염기서열 분석 결과 집락 주위로 환을 형성하는 직경 11~16 mm 크기의 집락들은 B. cereus 와 B. cereus subsp. cytotoxis로 동정되었고, 환을 형성하지 않아 B. cereus 계수에 포함시키지 않았던 직경 5 mm 크기의 분홍색 집락은 B. cereus로, 6.5 mm 크기의 이중환을 형성하여 B. cereus로 계수하기가 모호하였던 붉은색 집락은 B. cereus로 동정되었다. 나머지 세 종들은 Enterococcus sp., Brevibacillus sp., B. subtilis로 밝혀졌다. B. cereus로 동정된 9종 모두에서 적어도 한 종류 이상의 B. cereus 독소 유전자들을 함유하고 있었지만 다른 형태들에서는 발견되지 않았다.

• 한국식품위생안전성학회지
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• 제23권3호
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• pp.191-197
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• 2008

희아리 고춧가루에 특징적인 표지자를 찾기 위하여 GC/MASS분석, 고춧가루 표면 분석, 미생물 계수법을 수행하였다. 희아리 고춧가루는 정상 고춧가루에서는 발견되지 않은 분자량 204의 1,2,3,5,6,7,8,$8\alpha$-octahydro-1,$8\alpha$-dimethyl-7-(1-methylethenyl)-naphthalene과, 분자량 204의 2-isopropenyl-$4\alpha$,8-dimethyl-1,2,3,4,$4\alpha$,5,6,$8\alpha$-octahydronaphthalene을 함유하고 있었다. 그러나 희아리 시료간의 함량 차이가 10배 이상으로 컸고, 낮은 함량을 지닌 희아리로 불량고춧가루를 제조 시 검출 기준 이하로 떨어지는 문제점이 있었다. 희아리의 전자 현미경 사진은 정상고추의 매끈한 표면과는 다르게 가루 표면이 거칠고 수많은 미생물이 자라고 있음을 보였다. 한편 희아리는 10%(w/w) 희아리가 혼합된 불량고춧가루 경우에도 정상고춧가루와 확실히 구분되는 세균 및 효모/곰팡이 균수의 증가를 나타냈다. 결과적으로 이 3가지 기술에 의한 정성 정량적인 결과를 상호 보완적으로 분석하는 경우 불량고춧가루 판정은 가능하다고 생각된다.

• Journal of Microbiology and Biotechnology
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• 제29권8호
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• pp.1240-1247
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• 2019

This study was conducted to examine the inactivation effect of the combined treatment of high hydrostatic pressure (HHP; 400 MPa for 1, 3, and 5 min) and cationic surfactant washing (0.05% benzethonium chloride, BEC) against Listeria monocytogenes inoculated on fresh-cut broccoli (FCB). Washing with BEC at concentrations exceeding 0.05% resulted in 2.3 log-reduction of L. monocytogenes counts on FCB, whereas HHP treatment had approximately 5.5-5.6 log-reductions regardless of the treatment time. Scanning electron microscopy corroborated microbial enumeration, revealing that the combined treatment was more effective in removing L. monocytogenes from FCB than individual treatment with HHP or BEC. Color and total glucosinolate content were maintained after the combined treatment, although the hardness of the FCB slightly decreased. The results clearly suggest that the combined treatment of HHP and BEC washing has potential value as a new sanitization method to improve the microbial safety of FCB.

• Environmental Engineering Research
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• 제21권2호
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• pp.211-218
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• 2016

Cheonggye-cheon is a partly restored urban stream located in central Seoul. We monitored fecal contamination using three different fecal indicators, total coliforms (TC), fecal coliforms (FC) and E. coli, to assess differences in each indicator on days of varying weather conditions. Presumptive TC, FC and E. coli colonies were identified by their 16S rRNA sequences. The results showed that enumeration of E. coli provided a better reflection of fecal contamination of the stream than TC and FC. The main sources of contamination were the inflow of fecal-polluted groundwater from the vicinity of a subway line and two inflowing streams. The fecal contamination was worsened on days with heavy rain because untreated sewage from a collecting facility flowed into the stream. Moreover, growth potential of fecal indicator (E. coli) in situ induced by algal exudates was measured. Our results suggest that an appropriate standard based on E. coli rather than TC and FC should be established for improving water quality management strategies of Cheonggye-cheon in the future.

• 한국식품과학회지
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• 제38권2호
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• pp.232-236
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• 2006

건강한 성인의 분변에서 분리한 내산성 및 내담즙성이 우수한 B. longum BO-11을 김치 제조용 스타터로 이용하여 그 생존 능력과 김치 발효에 미치는 영향에 대해 조사하였다. 발효과정 중 bifidobacteria의 계수는 BS 배지를 사용하였으며 20일의 저장 기간 동안 균수가 약간 증가하였다. 일반 미생물은 bifidobacteria 비첨가군과 비슷하였으며 bifidobacteria 첨가군의 경우 산의 생산이 더 많았다. 김치 맛은 일반 김치보다 bifidobacteria 첨가 김치가 더 우수하였다. 이러한 결과는 bifidobacteria를 이용한 기능성 김치의 상업화 가능성을 보여준다.

• 한국식품위생안전성학회지
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• 제27권2호
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• pp.169-175
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• 2012

본 연구는 농산물로부터 S. aureus분리에 적합한 배지를 선발하고자 난황첨가 mannitol salt agar (MSA), 난황과 tellurite과 첨가된 Barid-Parker agar (BPA), 토끼혈장이 첨가된 Barid-Parker agar (BPA+RPF), 3M petrifilm Staph Express count plates (Petriflm) 및 CHROMagar Staphylococcus aureus (CSA)의 민감도, 특이도, 손상된 세포에 대한 회복능, 농산물로부터 회수율에 대하여 비교 실험을 하였다. 민감도와 특이도를 조사하기 위하여 S. aureus 21주, Stapylococcus spp. 20주, 그 외 식중독균 33주를 각 선택배지에 접종하였으며 손상된 세포에 대한 회복능은 조사를 위하여 열($60^{\circ}C$, 90초), 산(1% lactic acid, 10분), 저온($-20^{\circ}C$, 1시간) 처리하였다. 또한 양상추, 토마토, 고추에 S. aureus를 4.0 log CFU/g농도로 접종하고 각 선택배지에 도말하였다. 민감도는 BPA+RPF(100%) = CSA(100%)=petrifilm(100%) > MSA(90.5%) > BPA(90.5%), 특이도는 BPA+RPF(100%) = CSA(100%) > MSA(84.6%) > BPA(75.0%) > petrifilm(67.3%) 순으로 나타났다. 한편, 손상된 세포의 회복능과 농산물에서 회수율은 5종의 배지에서 차이가 없었다. 따라서 농산물 중 S. aureus를 분리할 시 BPA+RPF와 CSA를 사용하는 것이 가장 효과적일 것으로 판단된다.

• 한국토양비료학회지
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• 제21권2호
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• pp.135-140
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• 1988

제주도전토양(濟州道田土壤)의 미생물(徵生物)의 종류(種類)와 분포(分布)가 토양(土壤)의 종류(種類) 및 작물재배방법(作物栽培方法)에 따라서 어떻게 상이(相異)한가를 알고저 실내시험(室內試驗)한 결과(結果)를 요약(要約)하면 다음과 같다. 1. 세균수(細菌數)는 비화산회토양(非火山灰土壤)의 윤작재배지토양(輪作栽培地土壤)에서 많고 방사상균(放射狀菌)과 사상균(絲狀菌)은 화산회토양(火山灰土壤)의 단재배지토양(單栽培地土壤)에서 많은 결과(結果)를 보였다. 따라서 세균(細菌)에 대한 사상균(絲狀菌)의 비율(比率)은 비화산회토양(非火山灰土壤)의 윤작재배지토양(輪作栽培地土壤)에서 현저(顯著)히 적었다. 2. 세균(細菌)의 분리동정결과(分離同定結果) 세균(細菌)의 종류(種類)는 대부분(大部分)이 단간(短稈)의 Gram 음성균(陰性菌)이 주종(主種)을 이루었으며 전체(全體) 세균(細菌) 중(中) Rhizobium spp와 Flavobacterium spp. 가 차지하는 비율(比率)이 가장 높았다. 3. 사상균(絲狀菌)의 종류(種類)는 주로 Aspergillus spp. Fusarium spp. 및 Penicillium spp가 가장 많았으며 특(特)히 비화산회토양(非火山灰土壤)의 윤작재배지(輪作栽培地)에서 현저(顯著)하였다. 4. 제주도전토양(濟州道田土壤)의 미생물수(徵生物數)와 종류(種類)는 육지(陸地)의 경제작물재배토양(經濟作物栽培土壤)의 미생물수(徵生物數)에 비하여 세균(細菌)은 10~100배정도(倍程度) 그리고 사상균수(絲狀菌數)는 2배정도(倍程度) 적었다. 그리고 세균(細菌)과 사상균(絲狀菌)의 종류(種類)가 현저(顯著)히 단순(單純)하였다.