Detection of enterotoxin by targeting entFM and hblA genes in Bacillus cereus BC1 strain inoculated into ready to eat food (RTF) and drink samples using polymerase chain reaction (PCR) was conducted. The B. cereus BC1 strain was confirmed as a Bacillus diarrhoeal enterotoxin (BDE) when tested by a commercially available Enzyme-linked immunosorbent assay-BDE immunoassay (ELISA-BDE immunoassay, TECRA). In the specificity study, both enterotoxin genes were detected on chromosomal DNA of B. cereus BC1 strain by showing a specific band of 1269 bp (entFM) and 874 bp (hblA), respectively. However, none of the target genes were detected for the other 15 genomic DNA bacteria (B. cereus (ATCC 11779), B. subtilis (ATCC 6633), Campylobacter jejuni (ATCC 29428), C. coli (Jabatan Kimia Malaysia, JKM), Clostridium perfringen (ATCC 13124), Enterobacter sakazaki (ATCC 51329), Escherichia coli (ATCC 43888), E. coli (ATCC 11735), Legionella pneumophila (ATCC 33152), Listeria monocytogenes (ATCC 35967), Salmonella typhi (IMR), S. enteritidis (ATCC 13076), S. typhimurium (ATCC 14028), Shigella flexeneri (ATCC 12022) and Vibrio cholerae bengal (Institute Medical Research (IMR), Malaysia) examined. The detection limit of both genes was 0.1 ng of genomic DNA. Thus, in the presence study it is evidence that the PCR analysis targeting enterotoxin of entFM and hblA genes are suitable and useful in detecting enterotoxic B. cereus in RTFs and drinks contaminated sample.
During the study period, a total of 241 foreign workers were examined. The countries represented were Indonesia (103), Bangladesh (133), Myanmar (I), Pakistan (3) and others (1). The specimens collected were blood (238) and stool samples (173). The tests conducted on blood samples were for syphilis by RPR and TPHA, HIV, Hepatitis B, and from stool samples, enteric pathogens such as Salirzoirella spp, Shigelln spp. and Vibrio clrolerne. Table I shows the type of tests performed on the various nationalities and Table 2 the results of testing. Of the 230 blood samples tested by RPWPHA, five were positive, one from Indonesia (1.09%) and four from Bangladesh (3.79%). There was only one sample of blood out of 238 tested which was HIV positive (0.42%) and this was in an Indonesian. Twenty three workers were found to be Hepatitis B antigen positive (9.66%), 10 out of 102 (9.80%) from Indonesia and 13 out of 131 from Bangladesh (9.92%). As for the entric bacterial pathogens, only six out of 173 stool samples tested were positive, five for Saliizoilella Spp. and one for Slligdla sp. Of the five positives for Salmonella, one was from Indonesia and four from Bangladesh. The single isolate of Shigella was from Pakistan. From this pretiniinary study, it is obvious that hepatitis B is the most important problem among the workers from Indonesia and Bangladesh. The second of importance is venereal disease and enteric bacteria among Bangladesh workers. The other three national groups are too small to be analyzed. It is interesting to note that although these workers are supposed to have been screened for venereal diseases, a number of them were still found to be positive. However, we are not certain that these might not have been acquired locally. There was only one case of HIV detected but if the foreign workers continue with their pronliscuous lifestyle they are likely to pick up other sexually transmitted diseases including HIV and chlamydia1 infections. For those who were found to be stool positive for enteric pathogens, it is important to determine whether they are food-handlers as they will prove a significant risk for the spread of infections. Originally, it was intended to test blood samples for hepatitis C and E markers since the incidence in foreign countries from which the workers come are higher. However, due to the shortage of the samples, this had to be deferred. In the light that hepatitis carriage rate is the highest for the microbes tested, it is important to include these two markers in future studies.
Foodborne disease has been associated with microorganisms like bacteria, fungi, viruses and parasites. Most commonly, the outbreaks take place due to the ingestion of pathogenic bacteria like Salmonella Typhi, Escherichia coli, Staphylococcus aureus, Vibrio cholera, Campylobacter jejuni, and Listeria monocytogenes. The disease usually happens as a result of toxin secretion of the microorganisms in the intestinal tract of the infected person. Usually, the level of hygiene in the food premises reflect the quality of the food item, hence restaurant or stall with poor sanitary condition is said to be the contributor to food poisoning outbreak. In Malaysia, food poisoning cases are not rare because the hot and humid climate of this country is very suitable for the growth of the foodborne bacteria. The government is also implementing strict rules to ensure workers and owners of food premises prioritize the cleanliness of their working area. Training programme for food handlers can also help them to implement hygiene as a routine in a daily basis. A lot of studies have been done to reduce foodborne diseases. The results can give information about the types of microorganisms, and other components that affect their growth. The result is crucial to determine how the spread of foodborne bacteria can be controlled safely and the outbreak can be reduced.