ABSTRACT
A radiation-resistant bacterial isolate from gamma-radiation-processed (5 kGy) semidried prawns was identified as a new strain of Macrococcus caseolyticus and was designated as M. caseolyticus (A) on the basis of morphological and biochemical characterization and 16S rRNA sequencing. DNA-DNA hybridization studies with M. caseolyticus DSM 20597(T) further confirmed the isolate as M. caseolyticus. Major fatty acids present in M. caseolyticus (A) were C14:0, C16:1ω11c, and C18:1ω9c, whereas C15:0anteiso, C16:0iso, and C18:0iso were absent. The closest match for the isolate, as per fatty acid methyl ester analysis, was M. caseolyticus DSM 20597(T). However, the similarity index was significantly low (0.112), which indicates that the isolate could be a new strain of M. caseolyticus. The decimal reduction dose (D10) for M. caseolyticus (A), M. caseolyticus JCSC5402, and Staphylococcus aureus MTCC96 was 1.18, 0.607, and 0.19 kGy, respectively. This is the first report on radiation resistance of M. caseolyticus. Macrococcus caseolyticus (A) is more resistant to gamma and UV radiation stress than are M. caseolyticus JCSC5402 and S. aureus MTCC96; however, it is sensitive to heat as well as desiccation stress.
Subject(s)
Crustacea/microbiology , Shellfish/microbiology , Staphylococcaceae/isolation & purification , Staphylococcaceae/radiation effects , Animals , Crustacea/radiation effects , Fatty Acids/analysis , Fatty Acids/metabolism , Food Irradiation , Gamma Rays , Phylogeny , Shellfish/radiation effects , Staphylococcaceae/genetics , Staphylococcaceae/metabolismABSTRACT
A rapid nested PCR assay for detection of Salmonella from food was developed. The sensitivity of the assay developed was comparable to the traditional culture based methods with an advantage in reduction of assay time. The assay procedure with artificially contaminated samples was able to detect as low as 4CFU Salmonella/25g of food samples (sprout, carrot, cucumber and poultry meat). With two synthetic primers of 26 mer TS11 and 25 mer TS4, a 1.2kb fragment was amplified which served as a template for amplification of final 375bp product using TS11 and TS5 primers. No non-specific amplification from the native microbial flora of food samples was observed. The reaction generates a single band specific to Salmonella which allows the analyst to interpret data at ease and without any confusion. Enriched broth serves as template for the reaction which removes labour intensive DNA isolation procedures. In case of artificially contaminated samples, 6h enriched lactose broth can serve as template. However, for market samples where the organisms are under environmental stress, it is desirable to use template from Rappaport Vasiliadis medium. The assay also employes internal amplification control, which is amplified into a 300bp fragment and thus serves as positive control for the reaction and any possibility of false negative due to inhibitory action of food components on PCR reaction can be ruled out.
Subject(s)
Food Microbiology , Polymerase Chain Reaction/methods , Salmonella Food Poisoning/prevention & control , Salmonella/isolation & purification , Consumer Product Safety , DNA, Bacterial/isolation & purification , Food Contamination/analysis , Salmonella/genetics , Sensitivity and SpecificityABSTRACT
Many virulence phenotypes of Salmonella enterica are encoded by genes located on pathogenicity islands. Based on genome analysis, it is predicted that Salmonella pathogenicity island (SPI)-8 is restricted to Salmonella serovars Typhi and Paratyphi A, and SPI-10 to Salmonella serovars Typhi, Paratyphi, Enteritidis, Dublin and Gallinarum. This study was conducted to investigate the distribution of SPI-8 and SPI-10 among Salmonella isolates from sprouts, fish, water and blood. A total of 110 Salmonella isolates and 6 Salmonella serovars from the Microbial Type Culture Collection, Chandigarh, India, were screened. All isolates belonging to Salmonella serovars Washington, Enteritidis and Paratyphi A had both SPI-8 and SPI-10. All Salmonella serovar Typhi isolates from water and blood had both SPI-8 and SPI-10, whereas isolates from fish contained only SPI-8. SPI-8 and SPI-10 were also detected in only 3 out of 42 isolates belonging to Salmonella serovar Typhimurium. Both SPI-8 and SPI-10 were absent in Salmonella serovars Worthington, Dublin, Paratyphi B and Paratyphi C. These results contradict the predictions from Salmonella genome sequences available in GenBank and indicate that SPI-8 and SPI-10 are widely distributed among Salmonella serovars and that virulence factors other than those on SPI-8 and SPI-10 may be responsible for host specificity. This is the first report on the distribution of SPIs in Salmonella isolates from India.