a b s t r a c tThis study aimed to determine the prevalence and quantity of Salmonella spp., Salmonella Typhi and Salmonella Typhimurium in sliced fruits from hawker stalls and hypermarkets in Malaysia. Analysis was carried out using the most probable number (MPN) e multiplex polymerase chain reaction (PCR) method. The prevalence of Salmonella spp., Salmonella Typhi and Salmonella Typhimurium in 210 samples of sliced fruits examined were 23.3%, 7.6% and 3.8%, respectively with estimated quantity varying from 0 to 19 MPN/g. This study urged the authority to look into the biosafety of sliced fruits in Malaysia.
Various prevalence studies on Leptospira in animals and humans, as well as environmental samples, had been conducted worldwide, including Malaysia. However, limited studies have been documented on the presence of pathogenic, intermediate, and saprophytic Leptospira in selected animals and environments. This study was therefore conducted to detect Leptospira spp. in rats, soil, and water from urban areas of Sarawak using the polymerase chain reaction (PCR) method. A total of 107 rats, 292 soil samples, and 324 water samples were collected from April 2014 to February 2015. Pathogenic Leptospira was present in 5.6% (6/107) of rats, 11.6% (34/292) of soil samples, and 1.9% (6/324) of water samples. Intermediate Leptospira was present in 2.7% (8/292) of soil samples and 1.9% (6/324) of water samples. Saprophytic Leptospira was present in 10.3% (11/107) of rats, 1.4% (4/292) of soil samples, and 0.3% (1/324) of water samples. From this study, 76 Leptospira spp. were isolated. Based on DNA sequencing, the dominant Leptospira spp. circulating in urban areas of Sarawak are pathogenic Leptospira noguchii, intermediate Leptospira wolffii serovar Khorat, and saprophytic Leptospira meyeri, respectively. Overall, this study provided important surveillance data on the prevalence of Leptospira spp. from rats and the environment, with dominant local serovars in urban areas of Sarawak.
In the last decades, leptospirosis had gained public health concern due to morbidity and mortality rates caused by pathogenic Leptospira. The need for rapid and robust molecular typing methods to differentiate this zoonotic pathogen is of utmost importance. Various studies had been conducted to determine the genetic relatedness of Leptospira isolates using molecular typing methods. In this study, 29 pathogenic Leptospira isolates from rat, soil, and water samples in Sarawak, Malaysia, were characterized using BOX-PCR and ERIC-PCR. The effectiveness of these two methods with regard to the ease of interpretation, reproducibility, typeability, and discriminatory power was also being evaluated. Using BOX-PCR, six clusters and 3 single isolates were defined at a genetic distance percentage of 11.2%. ERIC-PCR clustered the isolates into 6 clusters and 2 single isolates at a genetic distance percentage of 6.8%. Both BOX-PCR and ERIC-PCR produced comparable results though the discriminatory index for ERIC-PCR (0.826) was higher than that for BOX-PCR (0.809). From the constructed dendrogram, it could be summarized that the isolates in this study were highly heterogeneous and genetically diverse. The findings from this study indicated that there is no genetic relatedness among the pathogenic Leptospira isolates in relation to the locality, source, and identity, with some exceptions. Out of the 29 pathogenic Leptospira isolates studied, BOX-PCR and ERIC-PCR successfully discriminated 4 isolates (2 isolates each) into the same cluster in relation to sample sources, as well as 2 isolates into the same cluster in association with the sample locality. Future studies shall incorporate the use of other molecular typing methods to make a more thorough comparison on the genetic relatedness of pathogenic Leptospira.
In Malaysia, there is an increasing number of reported leptospirosis cases which led to mortality. The infection is usually spread through the urine of infected animals and may contaminate the environmental soil and water. This study was conducted to determine the prevalence of Leptospira spp. in two national parks in Sarawak. A total of 50 soil and 60 water samples were collected from Tanjung Datu National Park (TDNP) and Bako National Park (BNP). The samples were filtered through sterile membrane filter prior to inoculation into modified semisolid Ellinghausen-McCullough-Johnson-Harris (EMJH) media added with 5-fluorouracil. The cultures were incubated at room temperature (28-30 °C) for 30 days before specific polymerase chain reaction (PCR) was conducted. PCR-positive samples were subjected to DNA sequencing. Out of the 110 environmental samples from two national parks in Sarawak, the prevalence of pathogenic, intermediate and saprophytic Leptospira was 0.9%, 5.5% and 0%, respectively. Results indicated that Leptospira borgpetersenii serovar Mini and Leptospira wolffii serovar Khorat were the pathogenic and intermediate Leptospira circulating in these study areas, respectively. Due to the ability of Leptospira to survive for months in environment, there is a risk of exposure to the public and tourists who visit these national parks. Increased awareness, continuous monitoring and effective preventive measures should be taken by local authorities to control leptospirosis outbreak.
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