Dynamics of norovirus genotype change and early characterization of variants in children with diarrhea in central Tunisia, 2001-2012.

Human noroviruses (HuNoVs), especially GII.4 strains, are a major cause of gastroenteritis epidemics in both children and adults. Stool samples were collected from 113 Tunisian children with acute gastroenteritis in 2001 and 2002 and were retrospectively tested for HuNoVs. Fifteen (13.2%) of the 113 samples were positive for HuNoVs, all of which were genogroup II strains, and the GII.4-2004/Hunter variant was predominant (67%). We reconstituted the temporal circulation of HuNoV strains in central Tunisia between 2003 and 2012 using HuNoV isolates reported in our previous studies. A comparative analysis showed a dynamic change in the molecular profile of the HuNoV strains over a 12-year period. We found that GII.4-2004/Hunter strains were circulating as early as June 2002 and that GIX.1[GII.P15] HuNoVs were already circulating four years before this genotype was first reported in Japan in 2006. Our data suggest that epidemic strains of HuNoV circulate for several years in the pediatric population before becoming predominant. This study suggests that children from low-income countries with poor sanitation may play a significant role in the molecular evolution of noroviruses and the global emergence of new epidemic strains.

Molecular analysis of the role of osmolyte transporters opuCA and betL in Listeria monocytogenes after cold and freezing stress.

Listeria monocytogenes is a food-borne pathogen of humans and other animals. The striking ability to survive several stresses usually used for food preservation makes L. monocytogenes one of the biggest concerns to the food industry. This ubiquity can be partly explained by the ability of the organism to grow and persist at very low temperatures, a consequence of its ability to accumulate cryoprotective compound called osmolytes. A quantitative RT-PCR assay was used to measure mRNA transcript accumulation for the stress response genes opuCA and betL (encoding carnitine and betaine transporters, respectively) and the housekeeping gene 16S rRNA. Assays were conducted on mid-exponential phase L. monocytogenes cells exposed to conditions reflecting cold and freezing stress, conditions usually used to preserve foods. We showed that expression of the two cold-adapted genes encoded the transporters of the cryoprotectants carnitine and betaine in ATCC 19115 and the food-isolated L. monocytogenes S1 is induced after cold and freezing stress exposure. Furthermore, transcriptional analysis of the genes encoding opuCA and betL revealed that each transporter is induced to different degrees upon cold shock of L. monocytogenes ATCC 19115 and S1. Our results confirm an increase in carnitine uptake at low temperatures more than in betaine after cold-shocked temperature compared to the non-stress control treatment. It was concluded the use of carnitine and betaine as cryoprotectants is essential for rapid induction of the tested stress response under conditions typically encountered during food preservation.