• Title/Summary/Keyword: Microbial detection

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Development and Application of PCR-Based Weissella Species Detection Method with recN Gene Targeted Species-Specific Primers (RecN 유전자 특이적 PCR을 이용한 Weissella 속 유산균의 검출법 개발 및 적용)

  • Lee, Myeong-Jae;Cho, Kyeung-Hee;Han, Eung-Soo;Lee, Jong-Hoon
    • Microbiology and Biotechnology Letters
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    • v.39 no.1
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    • pp.70-76
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    • 2011
  • PCR-based Weissella species-specific detection method was developed to apply for the discrimination of Korean and Chinese kimchi by detecting a Weissella species only found in Korean or Chinese kimchi. PCR primers were designed from the species-specific sequence in the recN gene of each species. The primers allowed the species-specific detection and identification of nine species in the genera Weissella, and were successfully applied to the detection of W. cibaria, W. confusa, W. koreensis, and W. soli in kimchi with 20 ng template DNA. W. cibaria, W. confusa, and W. koreensis were detected from the Korean kimchi samples tested but W. soli was not detected. However, the four species were detected from Chinese kimchi samples. PCR-based W. soli-specific detection could not be perfectly applied as the Chinese kimchi discriminating method but has significance as an approach to evaluate the potential of scientific verification method based on the difference of microbial community.

Development of SCAR Markers for the Identification of Phytophthora katsurae Causing Chestnut Ink Disease in Korea

  • Lee, Dong Hyeon;Lee, Sun Keun;Lee, Sang Yong;Lee, Jong Kyu
    • Mycobiology
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    • v.41 no.2
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    • pp.86-93
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    • 2013
  • Sequence characterized amplified region (SCAR) markers are one of the most effective and accurate tools for microbial identification. In this study, we applied SCAR markers for the rapid and accurate detection of Phytophthora katsurae, the casual agent of chestnut ink disease in Korea. In this study, we developed seven SCAR markers specific to P. katsurae using random amplified polymorphic DNA (RAPD), and assessed the potential of the SCAR markers to serve as tools for identifying P. katsurae. Seven primer pairs (SOPC 1F/SOPC 1R, SOPC 1-1F/SOPC 1-1R, SOPC 3F/SOPC 3R, SOPC 4F/SOPC 4R, SOPC 4F/SOPC 4-1R, SOPD 9F/SOPD 9R, and SOPD 10F/SOPD 10R) from a sequence derived from RAPD fragments were designed for the analysis of the SCAR markers. To evaluate the specificity and sensitivity of the SCAR markers, the genomic DNA of P. katsurae was serially diluted 10-fold to final concentrations from 1 mg/mL to 1 pg/mL. The limit of detection using the SCAR markers ranged from $100{\mu}g/mL$ to 100 ng/mL. To identify the limit for detecting P. katsurae zoospores, each suspension of zoospores was serially diluted 10-fold to final concentrations from $10{\times}10^5$ to $10{\times}10^1$ zoospores/mL, and then extracted. The limit of detection by SCAR markers was approximately $10{\times}10^1$ zoospores/mL. PCR detection with SCAR markers was specific for P. katsurae, and did not produce any P. katsurae-specific PCR amplicons from 16 other Phytophthora species used as controls. This study shows that SCAR markers are a useful tool for the rapid and effective detection of P. katsurae.

Study on Reduction of Microbial Contamination on Daruma by Combination Treatment of Strong Acidic Hypochlorous Water and Ultrasonic Waves (강산성차아염소산수와 초음파를 병용처리한 조미오징어 반가공품의 미생물 오염도 저감화에 관한 연구)

  • Chung, Won-Hee;Ko, Jun-Soo;Shin, Il-Shik
    • Journal of Food Hygiene and Safety
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    • v.30 no.2
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    • pp.166-172
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    • 2015
  • This study was performed to develop treatment method for reducing microbial contamination on Daruma (a semi-processed product of seasoned and dried squid) by combination of strong acidic hypochlorous water (SAHW) and ultrasonic waves (UW). The available chlorine concentration, oxidation reduction potential (ORP) and pH of SAHW were $69.67{\pm}0.58ppm$, $1071.33{\pm}4.16mV$ and 2.79, respectively. The 1.49 log CFU/g of viable cell count and 1.32 log CFU/g of Staphylococcus aureus was reduced, and Escherichia coli was reduced below detection limit when the Daruma was treated with 20 times (w/v) of sodium hypochlorite solution (SHS) for 120 min. The 3.62 log CFU/g of viable cell count and 3.22 log CFU/g of Staphylococcus aureus was reduced, and Escherichia coli was reduced below detection limit when the Daruma was treated with 20 times (w/v) of SAHW for 120 min. The antibacterial effects of SAHW were stronger than those of SHS at same available chroline concentration. SAHW treatment after washing strongly alkalic electrolyzed water (SAEW) showed better bactericidal effects than SAHW treatment only. The 4.0 log CFU/g of viable cell count was reduced, S. aureus was reduced below regulation limit (Log 2.0 CFU/g), and E. coli was reduced below detection limit when the Daruma was treated with 20 times (w/v) of SAHW for 90 min after washing with 20 times (w/v) of SAEW for 60 min. The viable cell number was reduced below detection limit and S. aureus was reduced below regulation limit when the Daruma was treated with 20 times (w/v) of SAHW for 60 min in ultrasonic washer. E. coli was reduced below detection limit when the Daruma was treated with 20 times (w/v) of SAHW for 10 min in ultrasonic washer. These results suggest that combination of SAHW and UW may be a good technique to reduce the microbial contamination in daruma.

Intestinal Parasitic Infections and Environmental Water Contamination in a Rural Village of Northern Lao PDR

  • Ribas, Alexis;Jollivet, Chloe;Morand, Serge;Thongmalayvong, Boupha;Somphavong, Silaphet;Siew, Chern-Chiang;Ting, Pei-Jun;Suputtamongkol, Saipin;Saensombath, Viengsaene;Sanguankiat, Surapol;Tan, Boon-Huan;Paboriboune, Phimpha;Akkhavong, Kongsap;Chaisiri, Kittipong
    • Parasites, Hosts and Diseases
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    • v.55 no.5
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    • pp.523-532
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    • 2017
  • A field survey studying intestinal parasites in humans and microbial pathogen contamination at environment was performed in a Laotian rural village to identify potential risks for disease outbreaks. A parasitological investigation was conducted in Ban Lak Sip village, Luang Prabang, Lao PDR involving fecal samples from 305 inhabitants as well as water samples taken from 3 sites of the local stream. Water analysis indicated the presence of several enteric pathogens, i.e., Aeromonas spp., Vibrio spp., E. coli H7, E. coli O157: H7, verocytotoxin-producing E. coli (VTEC), Shigella spp., and enteric adenovirus. The level of microbial pathogens contamination was associated with human activity, with greater levels of contamination found at the downstream site compared to the site at the village and upstream, respectively. Regarding intestinal parasites, the prevalence of helminth and protozoan infections were 68.9% and 27.2%, respectively. Eight helminth taxa were identified in fecal samples, i.e., 2 tapeworm species (Taenia sp. and Hymenolepis diminuta), 1 trematode (Opisthorchis sp.), and 5 nematodes (Ascaris lumbricoides, Trichuris trichiura, Strongyloides stercoralis, trichostrongylids, and hookworms). Six species of intestinal protists were identified, i.e., Blastocystis hominis, Cyclospora spp., Endolimax nana, Entamoeba histolytica/E. dispar, Entamoeba coli, and Giardia lamblia. Questionnaires and interviews were also conducted to determine risk factors of infection. These analyses together with a prevailing infection level suggested that most of villagers were exposed to parasites in a similar degree due to limited socio-economic differences and sharing of similar practices. Limited access to effective public health facilities is also a significant contributing factor.

Comparison of detecting methods and the relationship between tissue and blood for enrofloxacin and metabolic ciprofloxacin residues in broiler chickens (육계에서 조직과 혈액의 enrofloxacin 및 대사성 ciprofloxacin 잔류 상관관계 조사 및 검사방법간의 비교)

  • Sung, Myung-Suk;Kim, Mi-Suk;Seo, Hee-Jin;Bae, Dong-Rok;Hwang, Ji-Young;Kim, Soon-Tae;Cho, Jong-Suk;Park, Hong-Je;Hong, Sung-Hee;Kim, Gyung-Dong;Jang, Seong-Jun;Yun, Mun-Jo
    • Korean Journal of Veterinary Service
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    • v.36 no.4
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    • pp.311-320
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    • 2013
  • The purpose of this study was to evaluate detecting methods and the relationship between tissues and blood for enrofloxacin and metabolic ciprofloxacin residues in broiler chickens. Two groups of broiler chickens were administrated via the drinking water with $50{\mu}g/mL$ and $100{\mu}g/mL$ of enrofloxacin for 5 days, respectively. The concentration of enrofloxacin and metabolic ciprofloxacin in tissues (muscle and kidney) and blood were measured during administration period (for 5 days) and withdrawal period (for 12 days) by high performance liquid chromatography (HPLC) method. Also, all samples were conducted for screening of residues by microbial method using E. coli for quinolone detection and immuno-chromatography method using Smart kit. The relationship between tissues (muscle and kidney) and blood for enrofloxacin and metabolic ciprofloxacin residues in broiler chickens was followed : The levels of enrofloxacin and metabolic ciprofloxacin residues in muscle and kidney were higher 2.9~3.2 folds, 3.6~3.8 folds more than the residues levels in blood, respectively. These results support we can predict the residues in muscle and kidney from the residues in blood. In comparison of detecting methods for antibiotic residues, microbial method using E. coli for quinolone detection and immuno-chromatography method using Smart kit could detect positive reaction at similar or lower concentration than violative concentration of enrofloxacin and metabolic ciprofloxacin in chicken tissues. These results support what two screening methods are useful for screening of quinolone detection in chickens.

Evaluation of Selective Media for Isolation of Staphylococcus aureus from Agricultural Products (농산물 중 Staphylococcus aureus의 분리를 위한 선택배지 평가)

  • Kim, Se-Ri;Lee, Seo-Hyun;Seo, Min-Kyoung;Kim, Won-Il;Park, Kyeong-Hun;Yun, Hye-Jeong;Yoon, Yo-Han;Yoo, Soon-Young;Ryu, Kyoung-Yul;Yun, Jong-Chul;Kim, Byung-Seok
    • Journal of Food Hygiene and Safety
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    • v.27 no.2
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    • pp.169-175
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    • 2012
  • Five kinds of selective media, such as mannitol salt agar (MSA), Baird-Parker agar (BPA), Baird-Parker supplemented with rabbit plasma fibrinogen (BPA+RPF), CHROMagar Staphylococcus aureus (CSA), and Petrifilm Staph Express count system (Petrifilm), were compared to recommend the optimum selective media for isolation of Staphylococcus aureus from agricultural products. Seventy four target and non target bacteria were inoculated on five selective media to analyze sensitivity and specificity. In the recovery test of injured S. aureus cells, S. aureus was exposed to acid (1% lactic acid for 10 min), heat ($60^{\circ}C$ for 90s), and cold ($-20^{\circ}C$ for 1h) conditions. And artificially contaminated agricultural products (iceberg lettuce, green pepper, and cherry tomato) was enumerated on five selective media. The sensitivity of BPA+RPF, CSA, Petrifilm, MSA, and BPA were 100%, 100%, 100%, 90.5%, 90.5%, respectively. In addition, the specificity of BPA+RPF, CSA, MSA, BPA and Petrifilm were 100%, 100%, 84.6%, 75.0%, 67.3%, respectively. However, no difference among five selective media was observed in recovery on injured S. aureus cell and enumeration from agricultural products. This results suggest that BPA+RPF and CSA are the optimum media for detection of S. aureus from agricultural products.

Virulence Profile and Antimicrobial Resistance of Escherichia coli from Flies Captured from Agricultural Environment (농업환경에 서식하는 파리에서 분리된 E. coli의 병원성 유전자 및 항생제 내성 조사)

  • Yun, Bohyun;Jang, Youn Jung;Kim, Yeon Rok;Kim, Hwang-Yong;Kim, Won-Il;Han, Sanghyun;Kim, Se-Ri;Ryu, Jae-Gee;Kim, Hyun Ju
    • Journal of Food Hygiene and Safety
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    • v.32 no.2
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    • pp.147-153
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    • 2017
  • The purpose of this study was to isolate Escherichia coli from flies and to assess pathogenic genes and antibiotic resistance of the isolates. A total of 188 flies were captured in agricultural environment including fruits farms (n = 19), fermented soybean farms (n = 9), municipal waste (n = 46), livestock farms (n = 66), slaughterhouses (n = 38), and manure ground (n = 10). E. coli isolates of captured flies were tested for pathogenic gene and antibiotic resistance using PCR methods and VITEK2 systems. As a result, E. coli from 63% (119/188) of the captured flies has been detected, and the detection rate of E. coli was the highest (89%, 31/34) in flies captured at particular slaughterhouse. Of the 34 isolates, 94% (32/34) were pathogenic gene (ST gene) positive. Twenty-six percent (31/119) of the E. coli isolates were observed being resistant to one or more antibiotics. Markedly, one of E. coli isolates from Livestock farms was resistant to 7 antibiotics including ampicillin, ampicillin/sulbactam, cefazolin, cefotaxime, gentamicin, levofloxacin, and trimethoprim/sulfamethoxazole. In addition, it was ESBL positive. The results of the present study may suggest a risk of transmission of pathogenic and antimicrobial resistant bacteria from flies to livestock environment Therefore, it may need to prevent introducing flies into the agricultural production environment for safe food production.

Heterometal-Coordinated Monomeric Concanavalin A at pH 7.5 from Canavalia ensiformis

  • Chung, Nam-Jin;Park, Yeo Reum;Lee, Dong-Heon;Oh, Sun-Young;Park, Jung Hee;Lee, Seung Jae
    • Journal of Microbiology and Biotechnology
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    • v.27 no.12
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    • pp.2241-2244
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    • 2017
  • The structure of concanavalin A (ConA) has been studied intensively owing to its specific interactions with carbohydrates and its heterometal ($Ca^{2+}$ and $Mn^{2+}$) coordination. Most structures from X-ray crystallography have shown ConA as a dimer or tetramer, because the complex formation requires specific crystallization conditions. Here, we reported the monomeric structure of ConA with a resolution of $1.6{\AA}$, which revealed that metal coordination could trigger sugar-binding ability. The calcium coordination residue, Asn14, changed the orientation of carbohydrate-binding residues and biophysical details, including structural information, providing valuable clues for the development and application of detection kits using ConA.

Molecular Detection of Human Enteric Viruses in Urban Rivers in Korea

  • Lee, Cheong-Hoon;Kim, Sang-Jong
    • Journal of Microbiology and Biotechnology
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    • v.18 no.6
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    • pp.1156-1163
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    • 2008
  • We performed RT-nested PCR to study the distribution of human enteric viruses in urban rivers in Korea. During 2002-2003, water samples were collected from four rivers in Gyeonggi Province, South Korea. Among 58 samples, 45 (77.6%), 32 (55.2%), 12 (20.7%), 2 (3.4%), 4 (6.9%), and 4 (6.9%) showed positive results with adenoviruses (AdVs), enteroviruses (EVs), reoviruses (ReVs), hepatitis A viruses (HAVs), rotaviruses (RoVs), and sapoviruses (SVs), respectively. According to the binary logistic regression model, the occurrence of each enteric virus, except ReVs and HAVs, was not statistically correlated with the water temperature and levels of fecal coliforms (P<0.05). AdVs were most often detected; only 4 samples (6.9%) were negative for AdVs while positive for other enteric viruses in the studied sites. Our results indicated that monitoring human enteric viruses is necessary to improve microbial quality, and that AdVs detection by PCR can be a useful index for the presence of other enteric viruses in aquatic environments.

Detection of Equipment Faults at Sequencing Batch Reactor Using Dynamic Time Warping (동적시간와핑을 이용한 연속회분식 반응기의 장비고장 감지)

  • Kim, Yejin
    • Journal of Environmental Science International
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    • v.25 no.4
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    • pp.525-534
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    • 2016
  • The biological wastewater treatment plant, which uses microbial community to remove organic matter and nutrients in wastewater, is known as its nonlinear behavior and uncertainty to operate. Therefore, operation of the biological wastewater treatment process much depends on observation and knowledge of operators. The manual inspection of human operators is essential to manage the process properly, however, it is impossible to detect a fault promptly so that the process can be exposed to improper condition not securing safe effluent quality. Among various process faults, equipment malfunction is critical to maintain normal operational state. To detect equipment faults automatically, the dynamic time warping was tested using on-line oxidation-reduction potential (ORP) and dissolved oxygen (DO) profiles in a sequencing batch reactor (SBR), which is a type of wastewater treatment process. After one cycle profiles of ORP and DO were measured and stored, they were warped to the template profiles which were prepared already and the distance result, accumulated distance (D) values were calculated. If the D values were increased significantly, some kinds of faults could be detected and an alarm could be sent to the operator. By this way, it seems to be possible to make an early detecting of process faults.