Application of RAPD-PCR and Phylogenetic Analysis for Accurate Characterization of Salmonella spp. Isolated from Chicken and Their Feed and Drinking Water
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Abstract
The aim of this study was the discrimination of Salmonella isolated from chicken and their feed and drinking water for the epidemiological control of salmonellosis. Totally, 289 samples, including 217 chicken cloaca swabs, 46 water, and 26 feed samples were collected from five different farms in Karbala governorate, Iraq. Conventional bacteriology tests, API 20E, Vitek 2, and serology were used for bacterial identification. Random amplified polymorphic DNA (RAPD)-polymerase chain reaction (PCR) was applied to analyze the genetic relationships among Salmonella isolates. The isolation rate of Salmonella spp. was 21.1% (61/289). While the water samples constituted the highest rate (30.4%), a rate of 21.7% was reported for the cloaca swabs, with no isolate at all from chicken feed. Vitek 2 was able to identify some isolates to the serotype level, such as S. Enteritidis, S. Paratyphi B, and S. Paratyphi C. However, the isolates were diagnosed as S. enterica by API 20E, and as S. enterica subsp. arizonae through serology. Analyzing the samples by the RAPD-PCR assay showed the presence of genetically different Salmonella spp. Dendrograms created by the GelJ software successfully delineated the genetic relationships. Therefore, RAPD-PCR can be used as a surrogate tool for the fast, reliable, and accurate detection of Salmonella in epidemiological surveys when compared with other biochemical-based identification methods.
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