• Journal of Microbiology and Biotechnology
• /
• v.19 no.5
• /
• pp.525-529
• /
• 2009

Cytolethal distending toxins (CDTs) represent an emerging family of newly described bacterial products that are produced by a number of pathogens. The genes encoding these toxins have been identified as a cluster of three adjacent genes, cdtA, cdtB, and cdtC, plus 5 cdt genetic variants, designated as cdt-I, cdt-II, cdt-III, cdt-IV, and cdt-V, have been identified to date. In this study, a general multiplex PCR system designed to detect Escherichia coli cdts was applied to investigate the presence of cdt genes among isolates. As a result, among 366 E. coli strains, 2.7% were found to carry the cdtB gene. In addition, the use of type-specific primers revealed the presence of cdt-I, cdtIV, and cdt-V types of the cdt gene, yet no cdt-II or cdt-III strains. The presence of other virulence genes (stxl, stx2, eae, bfp, espA, espB, and espD) was also investigated using a PCR assay. Among the 10 cdtB gene-positive strains, 8 were identified as COT-producing typical enteropathogenic E. coli (EPEC) strains ($eae^+$, $bfp^+$), whereas 2 were identified as CDT-producing atypical EPEC strains ($eae^+$, $bfp^-$). When comparing the cytotoxic activity of the CDT-producing typical and atypical EPEC strains, the CDT-producing atypical EPEC strains appeared to be less toxic than the CDT-producing typical EPEC strains.

• Biomedical Science Letters
• /
• v.24 no.1
• /
• pp.1-8
• /
• 2018

${\mu}MT$ knockout mice are genetically deficient in the transmembrane domain of mu chain of the immunoglobulin M (IgM) heavy chain, resulting in the absence of mature B cells. ${\mu}MT$ knockout mice is an in vivo model system used to clarify the role of B cells in various diseases. Enteropathogenic Escherichia coli (EPEC) induces acute and chronic diarrheal disease, especially in children of developing countries. The formation of attaching and effacing (A/E) lesion is a prominent pathogenic factor in the intestinal epithelium of EPEC infection. The A/E lesion is modulated by genes located on the pathogenic island locus of enterocyte effacement (LEE) which encode a type III secretion system (T3SS) and A/E lesion-related effector proteins. Citrobacter rodentium is a murine pathogen utilized in studying the pathogenic mechanisms of EPEC in human infections. Citrobacter rodentium produce A/E lesion to attach to intestinal epithelium, thus providing a murine model pathogen to study EPEC. Several studies have investigated the pathogenesis of Citrobacter rodentium in the ${\mu}MT$ knockout mice. In this review, we introduce the ${\mu}MT$ murine model in the context of C. rodentium pathogenesis and describe in detail the role of B cells and antibodies in this disease.

• Pediatric Gastroenterology, Hepatology & Nutrition
• /
• v.20 no.2
• /
• pp.107-113
• /
• 2017

Purpose: This study clarified the bacterial pathogens currently causing acute infectious enterocolitis (AIE) in children and evaluated the clinical characteristics and ultrasonographic findings according to the different pathogens. Methods: Medical records regarding age, sex, clinical symptoms, laboratory data, identified enteropathogens, ultrasonographic findings, treatment, and outcome of 34 patients who were diagnosed with AIE via stool examination using multiplex polymerase chain reaction (PCR) or culture, were retrospectively reviewed. Results: Twenty-four patients (70.6%) were male. The mean age of the patients was $8.5{\pm}6.2$ (range, 1.1-17.1) years. Six bacterial pathogens were isolated: Salmonella species (spp.) (32.4%), Campylobacter spp. (20.6%), verotoxin-producing Escherichia coli (14.7%), Staphylococcus aureus (11.8%), Clostridium difficile (8.8%), and Shigella spp. (2.9%). Abdominal pain occurred in all patients regardless of pathogen. The patients infected with Salmonella were older than those infected with verotoxin-producing E. coli (p<0.05). C-reactive protein levels were higher in patients with Salmonella and Campylobacter infections than in those with verotoxin-producing E. coli infection (p< 0.05), the other clinical and laboratory data were indistinguishable between pathogens. Ultrasonography demonstrated diverse involvement of bowel segments according to pathogen. Wall thickening of both the ileum and the entire colon was the most common lesion site regardless of pathogen. Conclusion: Various bacterial agents cause AIE and the symptoms are diverse symptoms, however, all most children recovered spontaneously. Use of multiplex PCR on stool samples warrants improvement of its sensitivity for diagnosis of enteropathogenic bacteria. Ultrasonographic examination is useful for diagnosis of AIE; it can also detect the disease extent and severity.

• Journal of Environmental Health Sciences
• /
• v.46 no.5
• /
• pp.525-531
• /
• 2020

Objectives: This study was performed to identify the epidemiological features of a food poisoning outbreak in a company cafeteria located in Chungcheongnam-do Province, Korea in June of 2020 and to suggest preventive measures for a similar incidence. Methods: A total of 84 patients with acute gastroenteritis were examined. Environmental samples were obtained from 16 food handlers, six food utensils, 135 preserved foods served over three days and nine menus, and six drinking water samples. These are analyzed to detect viruses and bacteria. Results: Ninety-four out of the 402 people who were served meals (23.4%) predominantly showed symptoms of diarrhea, and the number was over 3 times. Among the 84 patients under investigation, 17 cases (20.2%) were positive for Enteropathogenic E. coli (EPEC) and 18 cases were positive for Clostridium (C.) perfringens (21.4%). Based on the investigation, it was concluded that the main pathogens were EPEC and C. perfringens. For EPEC, it was detected in three of the food service employees and in the preserved food and curry rice. The results of pulsed field gel electrophoresis indicate that all EPEC cases are closely related except for one food service employee. Assuming that isolated EPEC originated from the preserved food, the incubation period is about 25 hours. The origin of the C. perfringens was not determined as it was not detected in the food service employees or environmental samples. Conclusions: This case suggests that food provided in group food service centers must be thoroughly managed. In addition, identifying the pathogens in preserved food is very important for tracing the causes of food poisoning, so food must be preserved in an appropriate condition. To prevent similar food poisoning cases, analyzing cases based on epidemiological investigation and sharing the results is needed.

• Pediatric Gastroenterology, Hepatology & Nutrition
• /
• v.9 no.1
• /
• pp.1-13
• /
• 2006

Purpose: Acute gastroenteritis in children is one of the frequently encountered diseases with relatively high admission rate. The aim of this study is to determine the isolation trends of common and emerging pathogens in acute gastroenteritis in children over a 12-month period in a community hospital. Methods: The study group included the children who were hospitalized to Seoul National University Boramae Hospital from April, 2003 to March, 2004 or visited outpatient clinic from April, 2003 to July, 2003 with presenting features of acute gastroenteritis. Stool specimens were obtained within 2 days after the visit and examined for the following pathogens: rotavirus, adenovirus, Salmonella, Shigella, Vibrio, pathogenic Escherichia coli (E.coli), Campylobacter and Yersinia species. Viral study was done with commercial kits for antigen detection. Identification of the bacterial pathogens was done by culture using selective media. For pathogenic E.coli, polymerase chain reaction (PCR) was done with the target genes related to the pathogenecity of enterotoxigenic E.coli (ETEC), enteropathogenic E.coli (EPEC) and enterohemorrhagic E.coli (EHEC). Results: The 130 hospitalized children and 28 outpatients were included in this study. The majority of children (>93%) were less than 6 years. Pathogens were isolated in 47% of inpatients and 43% of outpatients, respectively. Rotavirus was the most frequently identified pathogen, accounting for 42.3% of inpatients and 29.6% of outpatients. Nontyphoidal salmonella is the most commonly isolated bacterial pathogen (3.9%) in hospitalized children. Pathogenic E.coli (EPEC, ETEC) was detected in 2.1% (2/97) of inpatients and 25% (3/12) of outpatients. EHEC, adenovirus, Campylobacter, Yersinia and Shigella species were not detected in this study. Conclusion: Rotavirus is the most common enteropathogen in children with acute gastroenteritis. Nontyphoidal salmonella and pathogenic E.coli are important bacterial pathogens. Campylobacter species may not be commonly detected organism in hospitalized children with acute diarrhea.

• Journal of agricultural medicine and community health
• /
• v.30 no.2
• /
• pp.137-149
• /
• 2005

Objectives and Methods: This study was performed to investigate the etiologic bacterial, viral and protozoal organisms for the diarrhea from hospitals in Chungnam area from January to December in 2004. Total of 787 fecal samples were collected and examined. Results and Conclusions: In test for enteropathogenic bacteria, total of 79 cases out of 787 samples from hospitals showed positive isolation. Among 79 positive samples, 27 cases were confirmed as Salmonella spp.. 20 cases as pathogenic E. coli, 18 cases as Clostridium perfringens, 6 cases as Staphylococcus aureus, 4 cases as Shigella spp. and 4 cases as Vibrio parahaemolyticus. In test for enteropathogenic virus, 190 cases out of 787 samples from hospitals showed positive reaction. Among 190 samples, 115 cases were confirmed as rotavirus, 55 cases as norovirus, 5 case as astrovirus, 4 case as rotavirus & norovirus, 3 cases as adenovirus, 2 case as rotavirus & astrovirus. In test for enteropathogenic protozoa, 6 cases out of 787 samples from hospitals showed positive result. Among 6 samples, 5 cases were confirmed as Entamoeba histolytica and 1 cases as Giardia lamblia. When we classified the positive results by the age of the patients, the highest isolation rate was noted in a group of age under 10 and over 60 for bacterial, viral and protozoal pathogens. Especially, patient below age of 5 showed high positive rate. When we classified the positive results by the time, pageathogenic bacteria were isolated throughout the year, and the highest frequency was noted in August. On the other hand, pathogenic viruses were detected more frequently during the colder season from December to April. Antimicrobial susceptibility test for the isolated bacteria resulted as follows; Salmonella strains showed high drug resistance rates against ampicillin, chloramphenicol, tetracycline, nalidixic acid, ticarcillin. Shigella strains showed high drug resistance rates against ampicillin, trimethoprim/sulfamethoxazole, chloramphenicol, tetracycline, ampicillin/sulbactam, ticarcillin. Pathogenic E. coli strains showed high drug resistance rates against ampicillin, cephalothin, gentamicin, tetracycline, nalidixic acid, ampicillin/sulbactam, ticarcillin.

• Pediatric Gastroenterology, Hepatology & Nutrition
• /
• v.27 no.2
• /
• pp.95-103
• /
• 2024

Purpose: Diarrhea is one of the leading causes of mortality in children living in developing countries. The etiology of acute diarrhea in each healthcare center varies depending on place, time, and population. This study aimed to identify pathogen patterns in human immunodeficiency virus (HIV)-infected and non-HIV children suffering from acute diarrhea, using multiplex real time reverse transcriptase polymerase chain reaction (RT-PCR), in an Indonesian tertiary hospital. Methods: This cross-sectional study was conducted at Dr. Cipto Mangunkusumo National Hospital from March 2019 to April 2020. Results: The study showed that multiplex RT-PCR results were positive in 58.9% of the specimens, with more positive results in HIV-infected children than in non-HIV-infected children (70% vs. 54.7%). Altogether 72 enteropathogens were detected from all specimens. Enteropathogens in non-HIV children with acute diarrhea consisted of bacteria (70.6%) and viruses (29.4%) with a predominance of enteroaggregative Escherichia coli (25.4%), followed by Campylobacter spp. (11.8%), enteropathogenic E. coli (9.8%), Norovirus GII (7.8%), and Clostridium difficile (7.8%). Enteropathogens in HIV-infected children consisted of viruses (57.1%), bacteria (28.6%), and parasites (14.3%) comprising Norovirus GII (24%), Cryptosporidium spp. (14.3%), Campylobacter spp. (14.3%), Norovirus GI (14.3%), and Astrovirus (14.3%). Cryptosporidium spp. was the only parasite found in this study and was found only in HIV-infected children. In non-HIV children with acute diarrhea, most pathogens were invasive bacteria, while in HIV-infected children, more viral and parasite infections occurred, primarily caused by opportunistic pathogens. Conclusion: The pattern of enteropathogens can help clinicians determine further examinations and appropriate empirical antimicrobial therapy for the patient.

• Asian-Australasian Journal of Animal Sciences
• /
• v.30 no.4
• /
• pp.576-584
• /
• 2017

Objective: To generate recombinant Bacillus subtilis (B. subtilis) engineered for expression of porcine ${\beta}-defensin-2$ (pBD-2) and cecropin P1 (CP1) fusion antimicrobial peptide and investigate their anti-bacterial activity in vitro and their growth-promoting and disease resisting activity in vivo. Methods: The pBD-2 and CP1 fused gene was synthesized using the main codons of B. subtilis and inserted into plasmid pMK4 vector to construct their expression vector. The fusion peptide-expressing B. subtilis was constructed by transformation with the vector. The expressed fusion peptide was detected with Western blot. The antimicrobial activity of the expressed fusion peptide and the recovered pBD-2 and CP1 by enterokinase digestion in vitro was analyzed by the bacterial growth-inhibitory activity assay. To analyze the engineered B. subtilis on growth promotion and disease resistance, the weaned piglets were fed with basic diet supplemented with the recombinant B. subtilis. Then the piglets were challenged by enteropathogenic Escherichia coli (E. coli). The weight gain and diarrhea incidence of piglets were measured after challenge. Results: The recombinant B. subtilis engineered for expression of pBD-2/CP1 fusion peptide was successfully constructed using the main codons of the B. subtilis. Both expressed pBD-2/CP1 fusion peptide and their individual peptides recovered from parental fusion peptide by enterokinase digestion possessed the antimicrobial activities to a variety of the bacteria, including gram-negative bacteria (E. coli, Salmonella typhimurium, and Haemophilus parasuis) and grampositive bacteria (Staphylococcus aureus). Supplementing the engineered B. subtilis to the pig feed could significantly promote the piglet growth and reduced diarrhea incidence of the piglets. Conclusion: The generated B. subtilis strain can efficiently express pBD-2/CP1 fusion antimicrobial peptide, the recovered pBD-2 and CP1 peptides possess potent antimicrobial activities to a variety of bacterial species in vitro. Supplementation of the engineered B. subtilis in pig feed obviously promote piglet growth and resistance to the colibacillosis.

• The Korean Journal of Physiology and Pharmacology
• /
• v.22 no.3
• /
• pp.269-275
• /
• 2018

$Na^+/H^+$ exchangers (NHEs) have been shown to be involved in regulating cell volume and maintaining fluid and electrolyte homeostasis. Pooled evidences have suggested that loss of $Na^+/H^+$ exchanger isoform 8 (NHE8) impairs intestinal mucosa. Whether NHE8 participates in the pathology of infectious colitis is still unknown. Our previous study demonstrated that somatostatin (SST) could stimulate the expression of intestinal NHE8 so as to facilitate $Na^+$ absorption under normal condition. This study further explored whether NHE8 participates in the pathological processes of infectious colitis and the effects of SST on intestinal NHE8 expression in the setting of infectious colitis. Our data showed that NHE8 expression was reduced in Citrobacter rodentium (CR) infected mice. Up-regulation of NHE8 improved diarrhea symptom and mucosal damage induced by CR. In vitro, a similar observation was also seen in Enteropathogenic E. coli (EPEC) infected Caco-2 cells. Seglitide, a SST receptor (SSTR) 2 agonist, partly reversed the inhibiting action of EPEC on NHE8 expression, but SSTR5 agonist (L-817,818) had no effect on the expression of NHE8. Moreover, SST blocked the phosphorylation of p38 in EPEC-infected Caco-2 cells. Taken together, these results suggest that enhancement of intestinal NHE8 expression by SST could ameliorate the symptoms of mice with infectious colitis.

• Journal of Korean Biological Nursing Science
• /
• v.9 no.1
• /
• pp.33-48
• /
• 2007

목적 : 본 연구에서는 한국인 소아의 침 및 성인의 질부로부터 분리한 1367개의 유산 생성 균주 중 4가지의 장내 병원성 세균;Methicillin-resistant Staphylococcus aureus(MRSA), Listeria monocytogenes, Salmonella typhimurium, Escherichia coli O157에 대해 발육억제 효과를 나타내는 유산 균주를 분리하고 분리된 균주의 Probiotic으로서의 가능성을 조사하고자 하였다. 방법 : 네 가지 병원성 세균을 대상으로 agar spot test와 Catalse test를 시행하여 1367개 균주 중 최종적인 실험대상 균주를 선정하였다. 최종 선정된 유산균과 네 가지 병원성 세균을 혼합 배양하여 병원성 세균증식 억제 효과를 보았다. 또한 최종균주의 probiotic으로서의 가능성을 확인하기 위해 균을 동정하고 내산성 등 몇 가지 특성을 관찰하였다. 결과 : Agar spot test 결과 전체의 15.7%에 해당하는 215 균주가 MRSA에 대해 억제 효과를 나타내었으며 215 균주 중 9.8%에 해당하는 21 균주가 L. monocytogenes에도 동시에 억제효과를 나타냈다. 이들 균주 중 가장 탁월한 효과를 보인 Lb 1250을 실험 대상 균주로 선정하였다. Lb(Lactobacillus) 1250과 상술한 병원성 세균을 각각 혼합 배양하면 Lb 1250 존재 하에서 MRSA의 증식은 $10^4$배 억제되었으며, L. monocytogenes는 $10^3$배의 억제 효과를 보였다. 그러나 S. typhimurium, E. coli O157에 대해서는 아주 미약한 효과를 나타냈다. Lb 1250의 몇 가지 특성을 조사한 결과 Lactobacillus delbrueckii subsp. delbrueckii로 동정되었으며, 이는 $H_{2}O_2$를 잘 생산해 내는 균주이었다. 또한 이 균주는 우유에서 curd를 약하게 생성하며 이 curd의 향은 달콤하고 최종 pH가 4.6까지 감소하였으며, pH 2와 같은 강산의 조건하에서 배양한 결과 2시간 후에도 상당한 생존도를 유지하였다. 결론 : 이상의 결과에서 Lb 1250은 병원성 세균의 성장을 억제함을 관찰하였고, probiotic으로서의 이용가능성을 확인하였다.