A total of 11 Vibrio cholerae isolates from 1996-1998 outbreaks in Malaysia and 4 V. alginolyticus were analyzed. Isolates were characterized by polymerase chain reaction (PCR) and Southern hybridization for the presence of the gene encoding zonula occludens toxin (zot). Screening of zot gene by PCR revealed the presence of this gene in V. cholerae and V. alginolyticus. The zot gene from one V. cholerae Ogawa isolate that was cloned in a pCR 2.1 TOPO vector was sequenced. The sequences obtained were 99% homologous to the zot gene sequence from the Gene Bank.
Bacterial resistance to various antimicrobial agents is common in area with high usage of antibiotics. In this study, the data on antimicrobial susceptibility patterns of Vibrio cholerae O1 from patients during an outbreak period was found to be high but variable rates of multidrug resistance. Thirty-two of 33 V. cholerae isolates harboured the tcp, ctx, zot and ace genes, suggesting their possible roles in the outbreak cases. We analyzed the molecular diversity of a total of 33 strains of V. cholerae O1 isolated from 33 patients between November 1997 and April 1998 using random amplified polymorphic DNA (RAPD) analysis. The 30 typable isolates could be separated into four major clusters containing 5, 17, 2 and 6 isolates, respectively. However, no particular RAPD pattern was predictive of a particular pattern of antibiotic susceptibility. The findings of this study showed that multiple clones seemed to be responsible for cases in the outbreaks in the study area.
A total of 31 strains of Vibrio cholerae O1 (10 from outbreak cases and 7 from surface water) and non-O1 (4 from clinical and 10 from surface water sources) isolated between 1993 and 1997 were examined with respect to presence of cholera enterotoxin (CT) gene by PCR-based assays, resistance to antibiotics, plasmid profiles and random amplified polymorphic DNA (RAPD) analysis. All were resistant to 9 or more of the 17 antibiotics tested. Identical antibiotic resistance patterns of the isolates may indicate that they share a common mode of developing antibiotic resistance. Furthermore, the multiple antibiotic resistance indexing showed that all strains tested originated from high risk contamination. Plasmid profile analysis by agarose gel electrophoresis showed the presence of small plasmids in 12 (7 non-O1 and 5 O1 serotypes) with sizes ranging 1.3-4.6 MDa. The CT gene was detected in all clinical isolates but was present in only 14 (6 O1 serotype and 8 non-O1 serotype) isolates from environmental waters. The genetic relatedness of the clinical and environmental Vibrio cholerae O1 and non-O1 strains was investigated by RAPD fingerprinting with four primers. The four primers generated polymorphisms in all 31 strains of Vibrio cholerae tested, producing bands ranging from < 250 to 4500 bp. The RAPD profiles revealed a wide variability and no correlation with the source of isolation. This study provides evidence that Vibrio cholerae O1 and non-O1 have significant public health implications.
The effect of heat-treatment on the internal temperature of raw cockles (Anadara granosa) and survival of their intrinsic flora of Vibrio spp. as well as of inoculated V. cholerae 0139 was examined. The cockles were purchased from markets in Malaysia and had an average weight including shells of 8.90+/-2.45 g. In one experiment heatpenetration of individual cockles was examined. Cockles weighing < 8 g (including shell) exhibited maximum internal temperatures of between 50 and 75 degrees C when heated in water at 99 degrees C for 10 s and 71-93 degrees C when heated for 30 s. Cockles weighing > 12 g exhibited maximum internal temperatures between 42 and 58 degrees C when heated in water at 99 degrees C for 10 s and 56-69 degrees C when heated for 30 s. In another experiment, heat-treatment of 10 cockles treated as a group at 99 degrees C for 10 or 30 s resulted in reduction of levels of intrinsic Vibrio spp. (enumerated directly on thiosulphate-citrate-bile salt sucrose agar; TCBS) from 5.73 to 3.15 log cfu g(-1) or below 1 log cfu g(-1), respectively. The levels of Vibrio spp. after heat-treatment decreased with an increase in numbers of cockles grouped together during treatment. In a third experiment V. cholerae 0139 was inoculated into cockles and subjected to heat-treatment at 99 degrees C for 0, 10, 15, 20, 25 or 30 s. The levels of Vibrio spp. in uninoculated, non-heat-treated cockles was 4.89 log cfu g(-1) on TCBS, and the predominant species were V. parahaemolyticus and V. alginolyticus. V. cholerae 0139 inoculated into cockles with an average weight of 13.5+/-1.90 g (including shell) decreased for samples examined immediately after heat-treatment from 6 log cfu g(-1) initially to 3.5 log cfu g(-1) after 25 s and < 1 log cfu g(-1) (TCBS) after 30 s of heat-treatment. The most probable number method by enrichment in alkaline peptone water gave in general within 1 log unit higher counts than TCBS direct enumeration. TCBS direct enumeration and MPN counts were up to 2.38 or 1.30 log units higher, respectively, for samples heat-treated for 20 s or longer and stored for 6 h at 30 degrees C before examination, than for samples heat-treated for same periods of time and examined immediately. This study shows that a mild heat-treatment of cockles for up to 25 s is inadequate to ensure a large reduction in numbers of Vibrio spp., including V. cholerae 0139.
Analysis of diarrhoeal disease patterns in Malaysia from 1981-1986 suggested that infectious hepatitis ranked as the most predominant diarrhoeal disease followed by typhoid, food poisoning, dysentery and cholera. Although these five major food and water-borne diseases are still endemic in this country, diarrhoeal diseases per se no longer become an important public health problem in Malaysia. Enforcement of the cholera control program brought the incidence of the disease to a minimal. Unfortunately, this fatal form of diarrhoeal disease caused the greatest mortality compared to the others. Seasonal influence also played a part in controlling the occurrence of the disease. There was a preponderance of diarrhoeal diseases during the rainy season implicating contaminated water as a source of transmission. Although greater than half of the population has been supplied with piped water and sanitary latrines, a lot more has to be done before diarrhoeal diseases could be eliminated from this country.
The findings of a cholera epidemic in Krian district is reported. There were 77 cases and 92 carriers in the epidemic. Although the three main ethnic groups of Malays, Chinese and Indians were involved in the epidemic, the Malays constituted majority of the cases and carriers. The overall infection rate and case attack rate was higher among the younger population. The case: carrier ratio was also higher among the younger population especially among Indians. Various reasons and probable causes of the epidemic have been described briefly.
Eighty-six clinical isolates of Aeromonas hydrophila were studied for their ability to produce four exotoxins: a haemolysin active against rabbit erythrocytes, cytotoxin and enterotoxin detectable with Vero cell cultures, and the cholera toxin-like factor detected by an enzyme-linked immunosorbent assay. At least one exotoxin was produced by 80% of enteric and 96% of non-enteric isolates. The exotoxin profiles of non-enteric isolates were more restricted than those of enteric isolates, with haemolysin and cytotoxin producers preponderant. Although haemolysin and cytotoxin were produced by isolates from all sources, the enterotoxin and cholera toxin-like factor were more common amongst enteric isolates. The production of haemolysin and cytotoxin were closely related but the association between the enterotoxin and the cholera toxin-like factor was not significant.
The genome sequence analysis of a clinical Vibrio cholerae VC35 strain from an outbreak case in Malaysia indicates multiple genes involved in host adaptation and a novel Na(+)-driven multidrug efflux pump-coding gene in the genome of Vibrio cholerae with the highest similarity to VMA_001754 of Vibrio mimicus VMA223.
Eighty-four strains of Vibrio cholerae O1, O139 and non-O1/non-O139 from clinical and environmental sources were investigated for the presence of the toxin co-regulated pilus gene, tcpA, the virulence cassette genes ctxA, zot, ace and cep and also for their ability to elaborate haemolysin and protease. The ctxA and zot genes were detected using DNA-DNA hybridization while the ace, cep and tcpA genes were detected using PCR. Production of haemolysin and protease was detected using mammalian erythrocytes and an agar diffusion assay respectively. Analysis of their virulence profiles showed six different groups designated Type I to Type VI and the major distinguishing factor among these profiles was in the in vitro production of haemolysin and/or protease. Clinical O1, O139 and environmental O1 strains were similar with regard to presence of the virulence cassette genes. All environmental O1 strains with the exception of one were found to possess ctxA, zot and ace giving rise to the probability that these strains may actually be of clinical origin. One strain which had only cep but none of the toxin genes may be a true environmental isolate. The virulence cassette and colonization factor genes were absent in all non-O1/non-O139 environmental strains but production of both the haemolysin and protease was present, indicating that these may be putative virulence factors. These findings suggest that with regard to its pathogenic potential, only strains of the O1 and O139 serogroup that possess the tcpA gene which encodes the phage receptor, have the potential to acquire the CTX genetic element and become choleragenic.
This paper examines the trend and possible contributing factors for the occurrence of the food borne diseases outbreaks in Malaysia. These diseases mainly are cholera, typhoid fever, hepatitis A, dysentery and food poisoning. The outbreaks still occur sporadically in certain high risk areas throughout the country. The incidence rate of all the other three major food borne diseases steadily declined from the year 1988 to 1997 except for food poisoning and cholera. Statistic of food poisoning from the year 1996 to 1997 showed that 66.5% of the outbreak occurred in schools whereas only 0.4% originated from the contaminated food sold at various public food outlets. The school age group is always more affected than the general population. Amongst the contributing factors identified are related to unhygienic food handling practices followed by inadequate safe water supply and poor environmental sanitation. A multisectoral approach between Ministry of Health and other government agencies or private agents needs to be undertaken in the management of the food borne diseases in order to curb the incidences of food borne diseases in Malaysia.
Vibrio cholerae, the causative agent of cholera, is endemic in many parts of the world, especially in countries poor in resources. Molecular subtyping of V. cholerae is useful to trace the regional spread of a clone or multidrug-resistant strains during outbreaks of cholera. Current available PCR-based fingerprinting methods such as Random Amplified Polymorphic DNA (RAPD)-PCR, Enterobacterial Repetitive Intergenic Consensus Sequence (ERIC)-PCR, and Repetitive Extragenic Palindromic (REP)-PCR were used to subtype V. cholerae. However, there are problems for inter-laboratory comparison as these PCR methods have their own limitations especially when different PCR methods have been used for molecular typing. In this study, a Vibrio cholerae Repeats-PCR (VCR-PCR) approach which targets the genetic polymorphism of the integron island of Vibrios was used and compared with other PCR-based fingerprinting methods in subtyping. Forty-three V. cholerae of different serogroups from various sources were tested. The PCR-fingerprinting approaches were evaluated on typeability, reproducibility, stability and discriminatory power. Overall, Malaysian non-O1/non-O139 V. cholerae were more diverse than O1 strains. Four non-O1/non-O139 strains were closely related with O1 strains. The O139 strain in this study shared similarity with strains of both O1 and non-O1/non-O139 serogroups. ERIC-PCR was the most discriminative approach (D value = 0.996). VCR-PCR was useful in discriminating non-O1/non-O139 strains. RAPD-PCR and REP-PCR were less suitable for efficient subtyping purposes as they were not reproducible and lacked stability. The combination of the ERIC-PCR and VCR-PCR may overcome the inadequacy of any one approach and hence provide more informative data.
A novel enzyme/nanoparticle-based DNA biosensing platform with dual colorimetric/electrochemical approach has been developed for the sequence-specific detection of the bacterium Vibrio cholerae, the causative agent of acute diarrheal disease in cholera. This assay platform exploits the use of shelf-stable and ready-to-use (shelf-ready) reagents to greatly simplify the bioanalysis procedures, allowing the assay platform to be more amenable to point-of-care applications. To assure maximum diagnosis reliability, an internal control (IC) capable of providing instant validation of results was incorporated into the assay. The microbial target, single-stranded DNA amplified with asymmetric PCR, was quantitatively detected via electrochemical stripping analysis of gold nanoparticle-loaded latex microspheres as a signal-amplified hybridization tag, while the incorporated IC was analyzed using a simplified horseradish peroxidase enzyme-based colorimetric scheme by simple visual observation of enzymatic color development. The platform showed excellent diagnostic sensitivity and specificity (100%) when challenged with 145 clinical isolate-spiked fecal specimens. The limits of detection were 0.5ng/ml of genomic DNA and 10 colony-forming units (CFU)/ml of bacterial cells with dynamic ranges of 0-100ng/ml (R(2)=0.992) and log10 (1-10(4) CFU/ml) (R(2)=0.9918), respectively. An accelerated stability test revealed that the assay reagents were stable at temperatures of 4-37°C, with an estimated ambient shelf life of 200 days. The versatility of the biosensing platform makes it easily adaptable for quantitative detection of other microbial pathogens.
Introduction: Cholera epidemics can produce devastating public health outcomes. Cholera distribution is influenced by temperature, precipitation, elevation, distance to the coastline and oceanic environmental factors such as sea surface temperature, sea surface height and ocean chlorophyll concentration. The purpose of this study is to describe the spatial epidemiology of cholera in the four districts of Sabah. Methods: This is a retrospective review of 4 years (2011 to 2014) data from the districts of Kota Kinabalu, Penampang, Putatan and Papar, Sabah. All reported cases of cholera from those areas are included. Coordinates for locations of the cases are based on home addresses. SPSS v20, ArcGIS v10 and CrimeStat IV were used for data analysis and mapping. Results: Cholera showed several clustering of cases, such as in 2011 and 2014 in Kota Kinabalu. In the year 2011 and 2013, Penampang and Papar districts had the nearest neighbour index of less than 1, but p value was not significant, meaning the pattern did not appear to be significant. Nearest neighbour hierarchical clustering analysis further revealed cholera had 7 clusters, of those 6 were first order and 1 was a second order cluster. Conclusion: Cholera shows disease clustering which could mean it is due to its common point source or localised human to human transmission. Using GIS as a tool may help in surveillance and control of cholera infections.
Immunization is the process of making individuals immune. Childhood immunization is a common process for various aliments, but adult immunization in the Indian scenario is obscure. Officially, India has been declared polio-free, which is an achievement despite cultural, political, economic, geographic, and so many other factors. The changing demographics of adult, geriatric population and growing cost of health-care maintenance are a concern in developing countries like India. Thus, promoting healthy lifestyle needs prevention, early detection, and management of various diseases and disorders. Certainly, prevention in adults is yet to be tapped completely, so that goal of 100% prevention can be achieved. Various fraternities of medical association have come up with guidelines for adult immunization schedules in India. The present paper reviews infectious diseases such as anthrax, chikungunya, cholera, dengue, influenza, and malaria in this section of the review. We humbly request all health-care professionals and educators to educate the mass for adult immunization. So that, cost involved for treatment and workforce for the management of diseases can be better utilized in some other needed areas.
Introduction: Cholera is an infection caused by the bacterium Vibriocholerae that causes severe watery diarrhoea, which can lead to dehydration. Beet-roothas been shown to induce favourable outcomes and holds promise as an economic, practical natural dietary intervention in cholera. Methods: Systematic search of peer-reviewed literature on beet-root ORS and carrot ORS use for acute diarrhoea in below 6-year-old. We identified 3 studies for abstraction. Results: A study had compared a commercial carrot/rice-based ORS A (Na 52 mmol/L) and two glucose-based ORS B (Na 55 mmol/L) and C (Na 90 mmol/L). Fluid intake, faecal and urine output and absorption of fluid was measured in 161 infants and children (3-48 months old) during the first 48 hours after admission. The number of stools (p < 0.01) and the mean faecal output (p < 0.05) per kg body weight were significantly lower in group A. Children in group A also had significantly (p < 0.01) greater fluid absorption (mean 464 ml/kg) than in groups C (312 ml/kg) and B (140 ml/kg). Conclusions: While there are some promising results, this analysis indicates that the need for further investi- gation into approaches to increasing beet-root ORS and carrot ORS use.
Past major flooding events for the state of Johore, Malaysia were recorded in 1926, 1967, 1968 and 1971. However, major meteorological phenomena that hit Johore on the 19th December 2006 (first wave) and the 12th January 2007 (second wave) were claimed to be the worst flood disaster in Johore in a 100 years. All eight districts were affected displacing 157,018 and 155,368 population during the first and the second wave event respectively. The Johore Health Department deployed substantial number of medical and health personnel to deal with the Johore flood crisis. Flood-related data were collected on daily basis between 19th December 2006 and 19th February 2007 using spreadsheet format from Flood Operational Rooms located at respective District Health Offices. Among flood victims 34,530 were found to have non-communicable diseases and 19,670 with communicable diseases. No major food- and water-borne disease outbreaks, such as cholera and typhoid, were reported in Johore. High success of public health measures was depending on the workforce of medical and health personnel on the ground. On the other hand, voluntary services offered by non-governmental organisations (NGOs), private sector and other volunteers should be well coordinated without compromising regulatory and ethical requirements. Crisis guidelines and plan of actions shall be updated so that they would be more relevant to the crises encountered on the ground.
Food borne diseases like cholera, typhoid fever, hepatitis A, dysentery and food poisoning occur as the results of ingestion of foodstuffs contaminated with microorganisms or chemical. The true incidence of food borne disease in Malaysia is unknown, however the incidence is low ranging from 1.56 to 0.14 cases per 100,000 population and the food poisoning cases is on the rise as the evident by the incident rate of 62.47 cases per 100,000 population in 2008 and 36.17 in 2009. The rapid population growth and demographic shift toward ageing population, changing eating habit such as consumption of raw or lightly cooked food, long storage of such food, lack of education on basic rules of hygienic food preparation and food trading without appropriate microbiological safety procedure become contributing factors for food borne diseases. Food borne disease in Malaysia is in the rise and the direct and indirect cost management of FBD will become one of the most common issues to face by the government. The world is spending millions and millions in cost of treatment due to food borne diseases. The information on this paper was collected via findings of previous journals, data and statistics from the MOH of Malaysia and WHO websites. As a result, authors found that the prevention and management of the food borne disease outbreak needs to be addressed seriously.
A total of 32 clinical strains of Vibrio cholerae, including members of the 01 and 0139 serogroup
were collected from Klang, Selangor; Penang Island; Samarahan, Sarawak and Miri, Sarawak in Malaysia. In general, all the isolates except the 0139 serotype expressed low resistance to all the antibiotics tested with their Multiple Antibiotic Resistance (MAR) indices ranged from 0.10 to 0.48. The presence of ctx gene that encoded the cholera toxin was confirmed in all these clinical isolates by polymerase chain reaction. The results from the RAPD-PCR were analyzed using the RAPDistance software (Version 1.04). From the dendrogram generated, two main groups were observed which were subdivided into two clusters each. The Selangor’s isolates and the 0139 Penang’s isolates formed one group whereas the Samarahan, Sarawak isolates and the Miri, Sarawak isolates made up the other group, thus delineating their different sources of origin based on their geographical location.