Kumar Rakesh, Datta Tirtha K, Lalitha Kuttanappilly V
Microbiology, Fermentation & Biotechnology Division, Central Institute of Fisheries Technology, Cochin, India.
Animal Biotechnology Centre, National Dairy Research Institute, Karnal, India.
BMC Microbiol. 2015 Nov 3;15:254. doi: 10.1186/s12866-015-0579-1.
Seafood is not considered the natural habitat of Salmonella except the river fish, but still, the incidence of Salmonella in seafood is in a steady rise. By extending our understanding of Salmonella growth dynamics and pathogenomics in seafood, we may able to improve seafood safety and offer better strategies to protect the public health. The current study was thus aimed to assess the growth and multiplication of non-typhoidal and typhoidal Salmonella serovars on seafood and further sought to evaluate their virulence and stress genes expression while in contact with seafood at varying temperature exposure.
Salmonella enterica Weltevreden and Salmonella enterica Typhi were left to grow on fish fillets at -20, 4, room temperature (RT) and 45 °C for a period of one week. Total RNA from both Salmonella serovars were extracted and qRT-PCR based relative gene expression approach was used to detect the expression of rpoE, invA, stn and fimA genes at four different temperature conditions studied on incubation days 0, 1, 3, 5 and 7. Salmonella Weltevreden growth on seafood was increased ~4 log10 at RT and 45 °C, nevertheless, nearly 2 and >4 log 10 reduction was observed in cell count stored at 4 and -20 °C on seafood, respectively. Growth pattern of Salmonella Typhi in seafood has shown identical pattern at RT and 45 °C, however, growth was sharply reduced at 4 and -20 °C as compared to the Salmonella Weltevreden. Total RNA of Salmonella Weltevreden was in the range from 1.3 to 17.6 μg/μl and maximum concentration was obtained at 45 °C on day 3. Similarly, RNA concentration of Salmonella Typhi was ranged from 1.2 to 11.8 μg/μl and maximum concentration was obtained at 45 °C on day 3. The study highlighted that expression of invA and stn genes of Salmonella Weltevreden was >8-fold upregulated at RT, whereas, fimA gene was increasingly down regulated at room temperature. Storage of Salmonella Weltevreden at 45 °C on seafood resulted in an increased expression (>13 -fold) of stn genes on day 1 followed by down regulation on days 3, 5, and 7. Nevertheless, other genes i.e. fimA, invA and rpo remained downregulated throughout the storage period. More intense upregulation was observed for invA and stn genes of Salmonella Typhi at RT and 45 °C. Further, incubating Salmonella Weltevreden at 4 °C resulted in down regulation in the expression of rpoE, invA and stn genes. Regarding Salmonella Typhi, fimA and stn genes were upregulated on day one, in addition, an increased expression of fimA was noted on day 3. At -20 °C, there was no obvious expression of target genes of Salmonella Weltevreden and Salmonella Typhi when stored along with seafood.
Here we demonstrate that nutritional constituents and water content available in seafood has become useful growth ingredients for the proliferation of Salmonella in a temperature dependent manner. Although, it was absence of serovar specific growth pattern of non-typhoidal and typhoidal Salmonella in seafood, there was observation of diverse expression profile of stress and virulent genes in non-typhoidal and typhoidal Salmonella serovars. In presence of seafood, the induced expression of Salmonella virulent genes at ambient temperature is most likely to be impacted by increased risk of seafood borne illness associated with Salmonella.
除了河鱼外,海鲜通常不被认为是沙门氏菌的自然栖息地,但海鲜中沙门氏菌的发病率仍在稳步上升。通过深入了解沙门氏菌在海鲜中的生长动态和病原学,我们或许能够提高海鲜安全性,并提供更好的策略来保护公众健康。因此,本研究旨在评估非伤寒和伤寒沙门氏菌血清型在海鲜上的生长和繁殖情况,并进一步评估它们在不同温度暴露下与海鲜接触时的毒力和应激基因表达。
肠炎韦尔太夫登沙门氏菌和伤寒沙门氏菌在-20℃、4℃、室温(RT)和45℃的鱼片上生长一周。提取两种沙门氏菌血清型的总RNA,并采用基于qRT-PCR的相对基因表达方法,检测在培养第0、1、3、5和7天研究的四种不同温度条件下rpoE、invA、stn和fimA基因的表达。肠炎韦尔太夫登沙门氏菌在室温(RT)和45℃下在海鲜上的生长增加了约4个对数10,然而,在4℃和-20℃储存于海鲜上的细胞计数分别减少了近2个和>4个对数10。伤寒沙门氏菌在海鲜中的生长模式在室温(RT)和45℃下显示出相同的模式,然而,与肠炎韦尔太夫登沙门氏菌相比,在4℃和-20℃下生长急剧减少。肠炎韦尔太夫登沙门氏菌的总RNA范围为1.3至17.6μg/μl,在第3天45℃时获得最高浓度。同样,伤寒沙门氏菌的RNA浓度范围为1.2至11.8μg/μl,在第3天45℃时获得最高浓度。该研究强调,肠炎韦尔太夫登沙门氏菌的invA和stn基因在室温下上调>8倍,而fimA基因在室温下逐渐下调。肠炎韦尔太夫登沙门氏菌在45℃下储存于海鲜上导致第1天stn基因表达增加(>13倍),随后在第3、5和7天下调。然而,其他基因,即fimA、invA和rpo在整个储存期内仍下调。在室温(RT)和45℃下观察到伤寒沙门氏菌的invA和stn基因上调更为强烈。此外,将肠炎韦尔太夫登沙门氏菌在4℃下培养导致rpoE、invA和stn基因表达下调。对于伤寒沙门氏菌,fimA和stn基因在第1天上调,此外,在第3天观察到fimA表达增加。在-20℃下,与海鲜一起储存时,肠炎韦尔太夫登沙门氏菌和伤寒沙门氏菌的靶基因没有明显表达。
我们在此证明,海鲜中可获得的营养成分和水分含量已成为沙门氏菌以温度依赖方式增殖的有用生长成分。虽然在海鲜中未观察到非伤寒和伤寒沙门氏菌血清型的血清型特异性生长模式,但在非伤寒和伤寒沙门氏菌血清型中观察到应激和毒力基因的不同表达谱。在有海鲜存在的情况下,沙门氏菌毒力基因在环境温度下的诱导表达很可能受到与沙门氏菌相关的食源性疾病风险增加的影响。