• Title/Summary/Keyword: Campylobacter species

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Isolation and Identification of Pathogenic Microorganisms from Soybean Sprouts

  • Kim, Hye-Jung;Koo, Kyoung-Mo;Kim, Gi-Nahm;Lee, Dong-Sun;Paik, Hyun-Dong
    • Preventive Nutrition and Food Science
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    • v.7 no.3
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    • pp.305-309
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    • 2002
  • Raw soybean sprouts were tested for contamination with the following bacteria which have potential for pathogenesis or food spoilage : Salmonella spp., Escherichia coli O157:H7, Yersinia enterocolitica, Vibrio parahae-molyticus, Aeromonas hydrophila, Plesidomonas shigeloides, Pseudomonas aeruginosa, Staphylococcus aureus, Lis-teria monocytogenes, Bacillus cereus, Clostridium perfringens, Campylobacter jejuni, Erwinia spp., and Fusarium spp. Three of the above strains were isolated from the sprouts, and identified by morphological and biochemical methods including an API kit and ATB automated identification system. The isolate cultured in Cereus selective agar, a selective medium, was a Gram-positive, rod shaped, anaerobic spore former. The biochemical and culture tests revealed the following characteristics: catalase-positive, no growth on Simmon's citrate, NO₂ production and requirement of arginine for growth; the ATB automated identification system gave 99.8 % agreement for the identification of Bacillus cereus to the species level. The isolate cultured in Macconkey agar selective medium was Gram-negative, rod shaped and a gas former; the ATB-system gave 99.9% agreement for the identification of Aeromonas hydrophila to the species level. The isolate found in Pseudomonas isolation agar was Gram-negative, rod shaped, cytochrome oxidase-positive, a reducer of nitrates to nitrogen, and pyocyanin producer; the ATB-system gave 99.9 % agreement for the identification of Pseudomonas aeruginosa to the species level. These results indicate that the three bacteria species present in the soybean sprouts were Bacillus cereus, Aero-monas hydrophila, and Pseudomonas aeruginosa. Salmonella spp., Escherichia coli O157:H7, and Yersinia enter-ocolitica, which are associated with serious disease in humans, were not isolated from soybean sprouts examined in this study.

Detection of Campylobacter jejuni in food and poultry visors using immunomagnetic separation and microtitre hybridization

  • Simard, Ronald-E.
    • Proceedings of the Korean Society of Fisheries Technology Conference
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    • 2000.05a
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    • pp.71-73
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    • 2000
  • Campylobacter jejuni is most frequently identified cause of cause of acute diarrhoeal infections in developeed countries, exceeding rates of illness caused by both salmonella and shigilla(Skirrow, 1990 ; Lior 1994). Previous studies on campylobacter jejuni contamination of commercial broiler carcasses in u.s.(Stern, 1992). Most cases of the disease result from indirect transmission of Campylobactor from animals via milk, water and meat. In addition to Campylobactor jejuni. the closely relates species Campylobactor coli and Campylobactor lari have also been implicated as agents of gastroenteritis in humans. Campylobactor coli represented only approximately 3% of the Campylobactor isolates from patients with Campylobactor enteritis(Griffiths and Park, 1990) whereas Campylobactor coli is mainly isolated from pork(Lmmerding et al., 1988). Campylobactor jejuni has also been isolated from cases of bacteremia, appendicitis and, recently, has been associated with Guillai-Barre syndrome(Allos and Blaser, 1994; von Wulffen et al., 1994; Phillips, 1995). Studies in volunteers indicated that the infectious dose for Campylobactor jejuni is low(about 500 organisms)(Robinson, 1981). The methods traditionally used to detect Campylobactor ssp. in food require at least two days of incubation in an enrichment broth followed by plating and two days of incubation on complex culture media containing many antibiotics(Goossens and Butzler, 1992). Finnaly, several biochemical tests must be done to confirm the indentification at the species level. Therfore, sensitive and specific methods for the detection of small numbers of Campylobactor cells in food are needed. Polymerase chain reaction(PCR) assays targeting specific DNA sequences have been developed for the detection of Campylobactor(Giesendorf and Quint, 1995; Hemandex et al., 1995; Winter and Slavidk, 1995). In most cases, a short enrichment step is needed to enhance the sensitivity of the assay prior to detection by PCR as the number of bacteria in the food products is low in comparison with those found in dinical samples, and because the complex composition of food matrices can hinder the PCR and lower its sensitivity. However, these PCR systems are technically demanding to carry out and cumbersome when processing a large number of samples simutaneously. In this paper, an immunomagnetic method to concentrate Campylobactor cells present in food or clinical samples after an enrichment step is described. To detect specifically the thermophilic Campylobactor. a monoclonal antibody was adsorbed on the surface of the magnetic beads which react against a major porin of 45kDa present on the surface of the cells(Huyer et al., 1986). After this partial purification and concentration step, detection of bound cells was achieved using a simple, inexpensive microtitre plate-based hybridization system. We examined two alternative detection systems, one specific for thermophilic Campylobactor based on the detection of 23S rRNA using an immobilized DNA probe. The second system is less specific but more sensitive because of the high copy number of the rRNA present in bacterial cell($10^3-10^4$). By using specific immunomagnetic beads against thermophilic Campylobactor, it was possible to concentrate these cells from a heterogeneous media and obtain highly specific hybridization reactions with good sensitivity. There are several advantages in using microtitre plates instead of filter membranes or other matrices for hybridization techniques. Microtitre plates are much easier to handle than filter membranes during the adsorption, washing, hybridization and detection steps, and their use faciilitates the simultanuous analysis of multiple sample. Here we report on the use of a very simple detection procedure based on a monoclonal anti-RNA-DNA hybrid antibody(Fliss et al., 1999) for detection of the RNA-DNA hybrids formed in the wells.

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Application of MALDI-TOF mass spectrometry-based identification of foodborne pathogen tests to the Korea Food Standard Codex (MALDI-TOF 질량분석기를 이용한 식품중독균 확인시험 적용)

  • Ha, Miyoung;Son, Eun Jung;Choi, Eun Jeong
    • Korean Journal of Food Science and Technology
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    • v.48 no.5
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    • pp.437-444
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    • 2016
  • Rapid and reliable identification of microorganisms is important to maintain food quality and to control safety. MALDI-TOF MS-based identification methods are relatively fast and simple compared to other conventional methods including gram staining and biochemical characterization. A colony on subcultured media can be directly prepared on the analysis plate without further complex treatments. In this study, we confirmed the applicability of MALDI-TOF MS-based identification of foodborne pathogens such as Salmonella Enteritidis/Typhimurium, Staphylococcus aureus, Vibrio parahaemolyticus, Clostridium perfringens, Listeria monocytogenes, Yersinia enterocolitica, Bacillus cereus, Campylobacter jejuni, Campylobacter coli, and Cronobacter sakazakii on the Korea Food Standard Codex. MALDI-TOF MS data of the pathogenic reference strains were incorporated into a commercial MicroID (ASTA Inc.) database. Other pathogenic reference strains and seven isolates from various food samples were correctly identified to the species level by using the MicroID database. In conclusion, MALDI-TOF MS is comparable with commercial biochemical identification.

Survey on the Status of Microbial Contamination of Chicken Meats Collected from Poultry Processing Plants in Nationwide (우리나라 도계장 수거계육의 미생물학적 위생실태 조사)

  • Woo, Yong-Ku
    • Korean Journal of Microbiology
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    • v.43 no.3
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    • pp.186-192
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    • 2007
  • This study was conducted to survey the hygienic status of chicken meats on the microbial levels, which were collected from poultry processing plants located in the local provinces in nationwide including the JeJu island (n=15) in 1997. In particular, Salmonella spp., Campylobacter jejuni, and Listeria monocytogenes, which were retarded as one of the most important entero-pathogens relating to food home illness from poultry, were investigated on their isolation frequency including the other pathogens related on the food-borne illness. A total of 115 processed chickens were submitted on the present study. In general, the bacterial contamination frequency showed more or less lower $(10{\sim}100 cells)$ than those of sold on the retail and super markets and department stores because of lacking of cross-contamination incidences, depending on the total cells, Coliforms and Staphylococcal cells count. While, Salmonella species, Campylobacter jejuni, Listeria monocytogenes, and coagulase positive Staphylococcus aureus isolation frequency of chicken meats from slaughter houses were 58.3%, 37.4%, 43.5%, and 30.4%, in order. But the present microbial isolation data were a little lower levels than those of sold on the retail and super markets and famous department stores in Seoul and GyeongGi province at the same period. It seemed that the cross-contamination problems (including the human, environmental and instrumental factors) during the marketing stage (after the last processing procedure; rinsing step) had the major roles on the increasing of the microbial contamination frequency on the chicken meats after the slaughter houses.

Comparative Study on the Epidemiology of Food-Borne Disease Outbreaks in Korea and Japan (한국과 일본의 식중독 발생 역학의 비교연구)

  • Hwang, Sun-Young;Moon, Bo-Youn;Park, Yong-Ho;Lee, Myeong-Jin;Bang, Hyeong-Ae;Rhim, Kook-Hwan;Kim, Jin-Seok;Che, Nong-Hun;Lee, Won-Chang
    • Journal of Food Hygiene and Safety
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    • v.25 no.2
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    • pp.129-132
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    • 2010
  • The epidemiology of reported food-borne disease (FBD) outbreaks from 2001 to 2008 in Korea nd Japan were compared in this study. The outbreak rate of FBD in Japan was significantly higher although the average umber of patient in each outbreak in Korea was much higher. In both countries, summer was the season when most FBD outbreaks occurred. The comparison study revealed that FBD outbreaks in spring were more frequent in Korea, and outbreaks in winter were more frequent in Japan. Almost half of FBD outbreaks were observed at restaurants in both countries while FBD outbreaks at schools and work-places in Korea were much higher than in Japan. The most frequent cause of bacterial FBDs in Korea was pathogenic Escherichia coli followed by Salmonella species. On the other hand, Campylobacter jejuni was the most frequent source of bacterial FBDs in Japan. Norovirus, which is elated to uncontrolled hand hygiene and involvement of ill food workers, was the main cause of viral FBDs in both countries. In conclusion, there are common epidemiological characteristics as well as several differences in FBD outbreaks of Korea and Japan. These are suggested to be originated from the characteristic of climate, food sources, and life styles in two countries. Establishment of stricter control and surveillance system for FBD outbreaks are required or prevention and reduction of FBD outbreaks in both countries.

Simultaneous Detection of 10 Foodborne Pathogens using Capillary Electrophoresis-Based Single Strand Conformation Polymorphism

  • Oh, Mi-Hwa;Hwang, Hee-Sung;Chung, Bo-Ram;Paik, Hyun-Dong;Han, Sang-Ha;Kang, Sun-Moon;Ham, Jun-Sang;Kim, Hyoun-Wook;Seol, Kuk-Hwan;Jang, Ae-Ra;Jung, Gyoo-Yeol
    • Food Science of Animal Resources
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    • v.32 no.2
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    • pp.241-246
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    • 2012
  • This report outlines the development of a rapid, simple, and sensitive detection system for pathogenic bacteria using a capillary electrophoresis-based, single strand conformation polymorphism (CE-SSCP) combined with PCR. We demonstrate that this method, used with primers targeting the V4 region of the16S rRNA gene, is capable of the simultaneous detection of 10 microbes that could be associated with foodborne illness, caused by animal-derived foods: Salmonella enterica, Listeria monocytogenes, Escherichia coli O157:H7, Campylobacter jejuni, Staphylococcus aureus, Bacillus cereus, Clostridium perfringens, Yersinia enterocolitica, Vibrio parahaemolyticus, and Enterobacter sakazakii. The traditional detection techniques are time-consuming and labor-intensive, due to the necessary task of separate cultivation of each target species. As such, the CE-SSCP-PCR method, that we have developed, has the potential to diagnose pathogens rapidly, unlike the traditional technique, in order to prevent foodborne illness in a much more efficient manner.

Efficacy of salivary versus subgingival bacterial sampling for the detection and quantification of periodontal pathogens

  • Lee, Yoonsub;Hong, Yoojin;Kim, Bome;Lee, Dajung;Kim, Sungtae;Rhyu, In-chul
    • Journal of Periodontal and Implant Science
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    • v.50 no.6
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    • pp.358-367
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    • 2020
  • Purpose: The aim of this study was to investigate the efficacy and validity of subgingival bacterial sampling using a retraction cord, and to evaluate how well this sampling method reflected changes in periodontal conditions after periodontal therapy. Methods: Based on clinical examinations, 87 subjects were divided into a healthy group (n=40) and a periodontitis group (n=47). Clinical measurements were obtained from all subjects including periodontal probing depth (PD), bleeding on probing (BOP), clinical attachment loss (CAL), and the plaque index. Saliva and gingival crevicular fluid (GCF) as a subgingival bacterial sample were sampled before and 3 months after periodontal therapy. The salivary and subgingival bacterial samples were analyzed by reverse-transcription polymerase chain reaction to quantify the following 11 periodontal pathogens: Aggregatibacter actinomycetemcomitans (Aa), Porphyromonas gingivalis (Pg), Tannerella forsythus (Tf), Treponema denticola (Td), Prevotella intermedia (Pi), Fusobacterium nucleatum (Fn), Pavimonas micra (Pm), Campylobacter rectus (Cr), Prevotella nigrescens (Pn), Eikenella corrodens (Ec), and Eubacterium nodatum (En). Results: Non-surgical periodontal therapy resulted in significant decreases in PD (P<0.01), CAL (P<0.01), and BOP (P<0.05) after 3 months. Four species (Pg, Tf, Pi, and Pm) were significantly more abundant in both types of samples in the periodontitis group than in the healthy group. After periodontal therapy, Cr was the only bacterium that showed a statistically significant decrease in saliva, whereas statistically significant decreases in Cr, Pg, and Pn were found in GCF. Conclusions: Salivary and subgingival bacterial sampling with a gingival retraction cord were found to be equivalent in terms of their accuracy for differentiating periodontitis, but GCF reflected changes in bacterial abundance after periodontal therapy more sensitively than saliva.

Immediate effect of Nd:YAG laser monotherapy on subgingival periodontal pathogens: a pilot clinical study

  • McCawley, Thomas K.;McCawley, Mark N.;Rams, Thomas E.
    • Journal of Periodontal and Implant Science
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    • v.52 no.1
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    • pp.77-87
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    • 2022
  • Purpose: This pilot study assessed the immediate in vivo effect of high peak pulse power neodymium-doped yttrium aluminum garnet (Nd:YAG) laser monotherapy on selected red/orange complex periodontal pathogens in deep human periodontal pockets. Methods: Twelve adults with severe periodontitis were treated with the Laser-Assisted New Attachment Procedure (LANAP®) surgical protocol, wherein a free-running, digitally pulsed, Nd:YAG dental laser was used as the initial therapeutic step before mechanical root debridement. Using a flexible optical fiber in a handpiece, Nd:YAG laser energy, at a density of 196 J/cm2 and a high peak pulse power of 1,333 W/pulse, was directed parallel to untreated tooth root surfaces in sequential coronal-apical passes to clinical periodontal probing depths, for a total applied energy dose of approximately 8-12 joules per millimeter of periodontal probing depth at each periodontal site. Subgingival biofilm specimens were collected from each patient before and immediately after Nd:YAG laser monotherapy from periodontal pockets exhibiting ≥6 mm probing depths and bleeding on probing. Selected red/orange complex periodontal pathogens (Porphyromonas gingivalis, Tannerella forsythia, Prevotella intermedia/nigrescens, Fusobacterium nucleatum, Parvimonas micra, and Campylobacter species) were quantified in the subgingival samples using established anaerobic culture techniques. Results: All immediate post-treatment subgingival biofilm specimens continued to yield microbial growth after Nd:YAG laser monotherapy. The mean levels of total cultivable red/orange complex periodontal pathogens per patient significantly decreased from 12.0% pretreatment to 4.9% (a 59.2% decrease) immediately after Nd:YAG laser monotherapy, with 3 (25%) patients rendered culture-negative for all evaluated red/orange complex periodontal pathogens. Conclusions: High peak pulse power Nd:YAG laser monotherapy, used as the initial step in the LANAP® surgical protocol on mature subgingival biofilms, immediately induced significant reductions of nearly 60% in the mean total cultivable red/orange complex periodontal pathogen proportions per patient prior to mechanical root instrumentation and the rest of the LANAP® surgical protocol.

A Study on Dose-Response Models for Foodborne Disease Pathogens (주요 식중독 원인 미생물들에 대한 용량-반응 모델 연구)

  • Park, Myoung Su;Cho, June Ill;Lee, Soon Ho;Bahk, Gyung Jin
    • Journal of Food Hygiene and Safety
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    • v.29 no.4
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    • pp.299-304
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    • 2014
  • The dose-response models are important for the quantitative microbiological risk assessment (QMRA) because they would enable prediction of infection risk to humans from foodborne pathogens. In this study, we performed a comprehensive literature review and meta-analysis to better quantify this association. The meta-analysis applied a final selection of 193 published papers for total 43 species foodborne disease pathogens (bacteria 26, virus 9, and parasite 8 species) which were identified and classified based on the dose-response models related to QMRA studies from PubMed, ScienceDirect database and internet websites during 1980-2012. The main search keywords used the combination "food", "foodborne disease pathogen", "dose-response model", and "quantitative microbiological risk assessment". The appropriate dose-response models for Campylobacter jejuni, pathogenic E. coli O157:H7 (EHEC / EPEC / ETEC), Listeria monocytogenes, Salmonella spp., Shigella spp., Staphylococcus aureus, Vibrio parahaemolyticus, Vibrio cholera, Rota virus, and Cryptosporidium pavum were beta-poisson (${\alpha}=0.15$, ${\beta}=7.59$, fi = 0.72), beta-poisson (${\alpha}=0.49$, ${\beta}=1.81{\times}10^5$, fi = 0.67) / beta-poisson (${\alpha}=0.22$, ${\beta}=8.70{\times}10^3$, fi = 0.40) / beta-poisson (${\alpha}=0.18$, ${\beta}=8.60{\times}10^7$, fi = 0.60), exponential (r=$1.18{\times}10^{-10}$, fi = 0.14), beta-poisson (${\alpha}=0.11$, ${\beta}=6,097$, fi = 0.09), beta-poisson (${\alpha}=0.21$, ${\beta}=1,120$, fi = 0.15), exponential ($r=7.64{\times}10^{-8}$, fi = 1.00), betapoisson (${\alpha}=0.17$, ${\beta}=1.18{\times}10^5$, fi = 1.00), beta-poisson (${\alpha}=0.25$, ${\beta}=16.2$, fi = 0.57), exponential ($r=1.73{\times}10{-2}$, fi = 1.00), and exponential ($r=1.73{\times}10^{-2}$, fi = 0.17), respectively. Therefore, these results provide the preliminary data necessary for the development of foodborne pathogens QMRA.

Etiology and Clinical Manifestation of Acute Gastroenteritis in Children (소아 급성 위장관염에서의 원인과 임상양상)

  • Im, Ik-Jae;Lee, Mee Jeong;Chung, Eun Hee;Yu, Jeesuk;Chang, Young Pyo;Park, Woo Sung;Park, Kwisung;Song, Nak Soo;Baek, Kyung Ah;Cha, Yune Tae
    • Pediatric Infection and Vaccine
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    • v.13 no.2
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    • pp.147-155
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    • 2006
  • Purpose : The purpose of this study is to evaluate epidemiological data of pathogens obtained from stool exams and compare them with the clinical course in pediatric patients with symptoms of acute gastroenteritis. Methods : Subjects were selected from patients presenting with symptoms of acute gastroenteritis who visited the outpatient clinic or who were admitted to the Dankook University Hospital from December of 2004 to December of 2005. Stool exams for 17 pathogens was performed. RT-PCR was used to detect norovirus and enzyme-linked immunoabsorbant assay (ELISA) was used to detect rotavirus, adenovirus and astrovirus in the subjects stool samples. Ten different species of bacteria(Salmonella spp., Shigella spp., Clostridium perfrigens, Campylobacter spp., Escherichia coli, Vibrio spp., Staphylococcus aureus, Bacillus cereus, Yersinia spp., and L. monocytogenes) were each selectively cultivated and enzyme immunoassays(EIA) was used to test for antigens for C. parvum, E. histolytica and G. lamblia. Retrospective chart review was performed for comparisons of clinical manifestations. Results : A total of 215 subjects was selected and of these 89 cases(41.4%) showed positive results for at least one pathogen. Male to female ratio was 1.3:1. Age distribution showed 4 cases less than one month(4.5%), 4 cases from 1~2 months(4.5%), 24 cases from 3~12 months(26.7%), 47 cases form 13~48 months(52.8%), 10 cases greater than 48 months (21.2%). Viruses showed the greatest proportion of cases with 68 subjects(77.5%), of these rotavirus being the most commonly reported in 50 cases. Bacteria was identified in 26 cases (29.2%), of these nontyphoidal salmonella was noted in 10 cases. Protozoa followed with 21 cases(23.6%), of these C. parvum was noted in 11 cases and G. lamblia was noted in 10 cases. Mixed infections with more than two pathogens were seen in 22 cases(24.7%), of these viral infection with accompanying parasitic infection was seen in 12(54.5%) cases. Conclusion : In this study we examined various pathogens known to cause acute gastroenteritis in children. Further studies for various pathogens can provide useful information for management of the acute gastroenteritis.

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