Mitsuaki Nishibuchi

Kyoto University

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Son Radu

Universiti Putra Malaysia

Yoshitsugu Nakaguchi

Kyoto University

R. Son

Universiti Putra Malaysia

Varaporn Vuddhakul

Prince of Songkla University

Yoke Kqueen Cheah

Universiti Putra Malaysia

Yaya Rukayadi

Universiti Putra Malaysia

Yuzuru Nakaguchi

Kindai University

Tunung Robin

Universiti Putra Malaysia

Masanori Ishibashi

Kyushu University

Yuet Ying Loo

Universiti Putra Malaysia

Cooperative Institutions

Universiti Putra Malaysia

114

Kyoto University

Prince of Songkla University

Osaka University

International Centre for Diarrhoeal Disease Research

The University of Tokyo

National Institute of Infectious Diseases

National Institute of Cholera and Enteric Diseases

Johns Hopkins University

Kyushu University

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Rapid microimmunodiffusion method with species-specific antiserum raised to purified antigen for identification of Vibrio vulnificus

Journal of Clinical Microbiology (1985)

Mitsuaki Nishibuchi Ramon J. Seidler

An antigen common to Vibrio vulnificus strains, designated VVA, was purified by ammonium sulfate precipitation, gel filtration, ion-exchange column chromatography, and preparative gel electrophoresis. The molecular weight of VVA was 64,000 when estimated by gel filtration and 40,000 when measured by denaturing polyacrylamide gel electrophoresis. Antiserum prepared against purified VVA (anti-VVA serum) did not agglutinate whole cells of V. vulnificus. Therefore, VVA was considered a possible internal antigen. By using anti-VVA serum, a microimmunodiffusion method was designed to detect the antigen VVA in bacterial cell lysates prepared from a single colony. This simple method allowed the specific identification of V. vulnificus as soon as 10 h after antigen preparation and therefore can be a useful tool in the identification of V. vulnificus from environmental or clinical specimens. VVA was not detected as a line of complete identity in some 20 other Vibrio species or in 7 other bacterial genera. VVA was present in all 63 isolates of V. vulnificus obtained from clinical and nonclinical sources.

Vibrio vulnificus

Ammonium sulfate precipitation

10.1128/jcm.21.1.102-107.1985

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Citations (10)

Clonal Diversity among Recently Emerged Strains of Vibrio parahaemolyticus O3:K6 Associated With Pandemic Spread

Journal of Clinical Microbiology (1999)

Prasanta K. Bag Suvobroto Nandi Rupak K. Bhadra Thandavarayan Ramamurthy Sujit Bhattacharya

ABSTRACT The genomes of the O3:K6 strains of Vibrio parahaemolyticus which abruptly emerged in Calcutta, India, in February 1996 and which demonstrated an unusual potential to spread and an enhanced propensity to cause infections were examined by different molecular techniques to determine clonality. No restriction fragment length polymorphism (RFLP) in the gene encoding the thermostable direct hemolysin was observed among the O3:K6 isolates of V. parahaemolyticus . Clonal diversity among the O3:K6 strains became evident by examining the RFLPs of the rrn operons and by the use of pulsed-field gel electrophoresis. Five ribotypes were distinguished among the O3:K6 strains examined, with ribotype R4 constituting the major type. Strains of O3:K6 isolated between June and August 1996 showed different pulsotypes compared to the pulsotypes of strains isolated before and after this period, indicating genetic reassortment among these strains, but those isolated between August 1996 and March 1998 showed identical or nearly similar pulsotypes. It is clear that there is a certain degree of genomic reassortment among the O3:K6 clones but that these strains are predominantly one clone.

Reassortment

Ribotyping

clone (Java method)

Molecular Epidemiology

10.1128/jcm.37.7.2354-2357.1999

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Citations (104)

Human Plasmodium knowlesi infection in Ranong province, southwestern border of Thailand

Malaria Journal (2012)

Natthawan Sermwittayawong Balbir Singh Mitsuaki Nishibuchi Nongyao Sawangjaroen Varaporn Vuddhakul

Plasmodium knowlesi, a simian malaria parasite, has been reported in humans in many Southeast Asian countries. In Thailand, most of the limited numbers of cases reported so far were from areas near neighbouring countries, including Myanmar. Blood samples collected from 171 Thai and 248 Myanmese patients attending a malaria clinic in Ranong province, Thailand, located near the Myanmar border were investigated for P. knowlesi using nested PCR assays. Positive samples were also investigated by PCR for Plasmodium falciparum, Plasmodium vivax, Plasmodium malariae and Plasmodium ovale, and were confirmed by sequencing the gene encoding the circumsporozoite protein (csp). Two samples, one obtained from a Thai and the other a Myanmese, were positive for P. knowlesi only. Nucleotide sequences of the csp gene derived from these two patients were identical and phylogenetically indistinguishable from other P. knowlesi sequences derived from monkeys and humans. Both patients worked in Koh Song, located in the Kawthoung district of Myanmar, which borders Thailand. This study indicates that transmission of P. knowlesi is occurring in the Ranong province of Thailand or the Kawthoung district of Myanmar. Further studies are required to assess the incidence of knowlesi malaria and whether macaques in these areas are the source of the infections.

Plasmodium knowlesi

Parasitology

Tropical Medicine

10.1186/1475-2875-11-36

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Citations (72)

Enterotoxigenicity of Vibrio parahaemolyticus with and without genes encoding thermostable direct hemolysin

Infection and Immunity (1992)

Mitsuaki Nishibuchi Alessio Fasano Robert G. Russell James B. Kaper

Vibrio parahaemolyticus produces a thermostable direct hemolysin (TDH) that has been implicated in the pathogenesis of diarrheal disease caused by this organism. However, previous studies attempting to demonstrate the contribution of the hemolysin to virulence have been inconclusive. We investigated this putative virulence factor by using an isogenic TDH-negative (TDH-) strain constructed by specifically inactivating the two copies of the tdh gene encoding TDH. The enterotoxigenicities of the parent strain (AQ3815) and the mutant strain were tested by adding sterile culture supernatants to rabbit ileal tissue mounted in Ussing chambers. The culture filtrate of the parent strain produced a significant increase in short-circuit current (Isc), compared with the change induced by the TDH- mutant. The capacity of the culture filtrate of AQ3815 to increase the Isc was reduced by neutralization with anti-TDH serum, and the return of the cloned tdh gene to the TDH- mutant restored the ability to increase the Isc. These results were corroborated by rabbit ileal loop assays in which AQ3815 caused fluid accumulation but the TDH- mutant did not. No microscopic damage was seen in mucosal tissues exposed to the culture filtrate of either strain. These results indicate that TDH has an enterotoxigenic effect on rabbit small intestine and could be responsible for the watery diarrhea seen with V. parahaemolyticus.

Hemolysin

Virulence factor

Strain (injury)

10.1128/iai.60.9.3539-3545.1992

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Citations (234)

Efficacy of household washing pre-treatments and cooking methods for reduction of Listeria monocytogenes in artificially contaminated chicken offal

Food Research (2019)

C.H. Kuan Son Radu Nor Ainy Mahyudin John Yew Huat Tang Siok Koon Yeo

Consumption of chicken offal is common and famous among Malaysians as it is often served as one of the side dishes with rice. Chicken offal can be a potential source of Listeria monocytogenes because slaughtered animals are recognized as a reservoir for foodborne pathogens. L. monocytogenes is a dangerous foodborne pathogen which can cause severe foodborne listeriosis with high fatality rate. This study aimed to determine the efficacy of different washing pre-treatment and cooking methods to reduce L. monocytogenes in artificially contaminated chicken offal. All the washing pre-treatments (dip treatment in different water sources and wash treatment with different water flow rates) showed significant reduction of the pathogen (p<0.05) when the inoculated samples were treated from 2 mins onwards. Washing the inoculated samples under the water flow rate of 2 L/min was the most effective way to reduce the number of L. monocytogenes (approximately 1.97 log reduction after washing for 10 mins). For heat treatment study, deep-frying was the most effective cooking method followed by boiling and pan-frying to reduce L. monocytogenes where all L. monocytogenes cells (7.91 log10 CFU/g) were killed within 45 s under deep-frying treatment. Overall, the study indicated that washing under running tap water (2 L/min) and deep-frying was effective in reducing and controlling the microbial populations during food preparation. The findings from this study can serve as a safe preparation step and cooking guideline. It is necessary to implement safe steps in food handling practices among food handlers to minimize the risk of foodborne infection.

Food preparation

Foodborne pathogen

Cooking methods

Tap water

10.26656/fr.2017.4(1).268

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Citations (1)

Food Research (2017)

Rynnye Lyan V Lisha Mitsuaki Nishibuchi R. Son

Rapid genetically modified organism (GMO) screening of various food products and animal feeds using multiplex polymerase chain reaction (PCR)

10.26656/fr.2017.2

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Detection and quantification of pathogenic Vibrio parahaemolyticus in shellfish by using multiplex PCR and loop-mediated isothermal amplification assay

Food Control (2014)

Tan Turk Hsern Malcolm Yoke Kqueen Cheah Che Wan Jasimah Wan Mohamed Radzi Farihah Abu Kasim Haresh Kumar Kantilal

Multiplex

10.1016/j.foodcont.2014.08.010

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Citations (52)

Rapid detection and enumeration of pathogenic Vibrio parahaemolyticus in raw vegetables from retail outlets

International Food Research Journal (2011)

Tunung Robin Farinazleen Mohamad Ghazali Noranizan Mohd Adzahan Haresh Kumar Kantilal Yoshitsugu Nakaguchi

This study aims to determine the frequency and density of potentially pathogenic Vibrio parahaemolyticus, defned as those possessing thermostable-direct hemolysin (tdh) and/or tdh-related hemolysin (trh) genes, in raw salad vegetables at retail level in Selangor, Malaysia. A combination of Most Probable Number - Polymerase Chain Reaction (MPN-PCR) method was applied to detect the presence of tdh and/or trh gene-possessing V. parahaemolyticus and to enumerate their density in the samples. A total of 276 samples of vegetables commonly eaten raw in Malaysia (Cabbage = 30; Carrot = 31; Cucumber = 28; Four winged bean = 26; Indian pennywort = 17; Japanese parsley = 21; Lettuce = 16; Long bean = 32; Sweet potato = 29; Tomato = 38; Wild cosmos = 8) were analyzed. The samples were purchased from two supermarkets (A and B) and two wet markets (C and D). With the MPN-PCR technique, about 12.0% of the samples were positive for the presence of V. parahaemolyticus tdh-positive, with maximum densities of up to 39 MPN/g. The total frequency of V. parahaemolyticus trh-positive in the samples was 10.1%, with maximum concentration 15 MPN/g. V. parahaemolyticus tdh-positive was most prevalent in samples from Wet Market C (20.78%) and also in vegetable type Oenanthe stolonifera (Japanese parsley) with 19.0%, while V. parahaemolyticus trh- positive was predominant in samples from Wet Market D (16.7%) and was most frequent in both Oenanthe stolonifera (Japanese parsley) and Cucumis sativus (Cucumber) with 14.3% prevalence for each type. The results highlighted the fact that raw vegetables could be contaminated with virulent V. parahaemolyticus and could act as a transmission route, thus poses risk to consumers from the consumption of raw vegetables. To the author’s knowledge, this is the frst assessment of V. parahaemolyticus carrying tdh and trh genes in raw vegetables from retail outlets in Malaysia.

Most probable number

Enumeration

Cucumis

Hemolysin

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Citations (15)

2010年7~11月に世界的に有名な観光客スポット「京都」におけるデング熱やチクングニヤウイルスの媒介昆虫として働くことができるカ科種の高発生率およびその気候要因との関係

Masuda Gaku Yoshikawa Minako Jen Hideo Mizuta Shirakawa Koichi Noboru Ishikawa

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Characterisation of L. monocytogenes strains isolated from salad vegetables

International Journal of Food Safety Nutrition and Public Health (2015)

Jeyaletchumi Ponniah Tunung Robin Son Radu Yoke Kqueen Cheah Farinazleen Mohamad Ghazali

Seventy-one strains of L. monocytogenes isolated from raw salad vegetables consumed in the minimally processed state in Malaysia were characterised by serotyping, random amplified polymorphic DNA-polymerase chain reaction analysis (RAPD-PCR) and antimicrobial susceptibility testing. Based on serotyping, only one strain belonged to serotype 4 while the rest were from serotype 1. RAPD-PCR typing was carried out using two primers, i.e., OPAR-8 and GEN 1-50-09. Based on RAPD-PCR fingerprinting, the strains could be grouped into six composite profile groups and a solitary isolate. The isolates were also evaluated against 16 antibiotics. Excluding nalidixic acid and oxacillin for which L. monocytogenes has natural resistance, the highest prevalence of resistance was against cephalexin. None of the isolates were resistant against imipenem and kanamycin. The multiple antibiotic resistance index (MARI) ranged from 0.06 to 0.63. It was found that most (82%) isolates were susceptible to antibiotics commonly used in veterinary and human listeriosis treatment.

Kanamycin

10.1504/ijfsnph.2015.067562

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