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Rodpai E.,Mahidol University | Moongkarndi P.,Mahidol University | Tungrugsasut W.,Mahidol University | Phisannoradej R.,Mahidol University | Kanarat S.,Veterinary Public Health Laboratory

The multiplex PCR (mPCR), enzyme-linked immunosorbent assay (ELISA) and immunomagnetic separation enzyme-linked immunosorbent assay (IMS-ELISA) were developed for the detection of Salmonella spp. and two specific serovars S. Typhimurium and S. Enteritidis. The detection limits of the mPCR without the pre-enriched step was at 105 cfu/ml, whereas developed immunoassays demonstrated positive results at 105, 106, and 107 cfu/ml by ELISA and at 104, 104, and 106 cfu/ml by IMS-ELISA for detecting Salmonella spp., S. Typhimurium, and S. Enteritidis, respectively. The mPCR did not produce any nonspecific amplified products when tested against other related species of bacteria. In this study, the developed IMS-ELISA gave more sensitivity but less specificity than the mPCR assay. The methods are useful for the rapid detection of salmonellae in naturally infected chicken meat in Thailand. Source

Saengthongpinit C.,Kasetsart University | Kanarat S.,Veterinary Public Health Laboratory | Sirinarumitr T.,Kasetsart University | Amavisit P.,Kasetsart University | Sakpuaram T.,Kasetsart University
Kasetsart Journal - Natural Science

The aim of this study was to use the high-resolution genotyping method of amplified fragment length polymorphism (AFLP) to investigate the genetic diversity of Campylobacter jejuni and C. coli isolates obtained from broiler farms and different stages of slaughterhouse processing. Of 490 samples, 328 isolates of Campylobacter spp. were found (328/490, 66.9%) and the isolates were identified as C. jejuni (24.1%, 118/490) and C. coli (42.8%, 210/490). The genetic fingerprint of the 314 Campylobacter isolates, as determinated by AFLP, revealed 48 AFLP strains of C. jejuni and 95 AFLP strains of C. coli. In most flocks, a broad diversity of C. jejuni and C. coli strains was found and the distribution of AFLP types changed during slaughter line processing. Some genotypes were found to be the contamination source of both species in chicken intestines and by direct contamination of chicken meat during the slaughtering and cutting processes in the slaughterhouse. AFLP fingerprinting was an effective method to discriminate between C. jejuni and C. coli strains, in which the interlinkage homology of the AFLP pattern was only 35-42%. In addition, AFLP fingerprinting could distinguish between strains that were genetically unrelated or related. Therefore, AFLP analysis was considered a suitable epidemiological tool for investigation of Campylobacter. Source

Sakai N.,Kyoto University | Mohd Yusof R.,Veterinary Public Health Laboratory | Sapar M.,Veterinary Public Health Laboratory | Yoneda M.,Kyoto University | Ali Mohd M.,University of Malaya
Science of the Total Environment

Beta-agonists and sulfonamides are widely used for treating both humans and livestock for bronchial and cardiac problems, infectious disease and even as growth promoters. There are concerns about their potential environmental impacts, such as producing drug resistance in bacteria. This study focused on their spatial distribution in surface water and the identification of pollution sources in the Langat River basin, which is one of the most urbanized watersheds in Malaysia. Fourteen beta-agonists and 12 sulfonamides were quantitatively analyzed by liquid chromatography-tandem mass spectrometry (LC-MS/MS). A geographic information system (GIS) was used to visualize catchment areas of the sampling points, and source profiling was conducted to identify the pollution sources based on a correlation between a daily pollutant load of the detected contaminant and an estimated density of human or livestock population in the catchment areas. As a result, 6 compounds (salbutamol, sulfadiazine, sulfapyridine, sulfamethazine, sulfadimethoxine and sulfamethoxazole) were widely detected in mid catchment areas towards estuary. The source profiling indicated that the pollution sources of salbutamol and sulfamethoxazole were from sewage, while sulfadiazine was from effluents of cattle, goat and sheep farms. Thus, this combination method of quantitative and spatial analysis clarified the spatial distribution of these drugs and assisted for identifying the pollution sources. © 2016 Elsevier B.V. Source

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