9High-throughput sequencing was used to reveal the highly diverse bacterial 10 populations in shrimp paste at different fermentation stages. We studied three stages of 11 fermentation and obtained 448,916 reads. Using this approach, we revealed the presence 12 of 30 phyla, 55 classes, 86 orders, 206 families and 695 genera of bacteria in the shrimp 13 paste. Shrimp paste in fermentation metaphase had a more diverse microbiota than that in 14 fermentation prophase and fermentation anaphase. Diversity appeared greatest in 15 fermentation anaphase. The four dominant phyla were Proteobacteria, Firmicutes, 16 Actinobacteria, and Bacteroidetes. The most common genera were Psychrobacter, 17 Halomonas, Bacillus, Alteribacillus, and Lactococcus. Their content varied at different 18 stages of fermentation. All the microbiome presented a variety of changes in the 19 microbial diversity of shrimp paste. 20 Importance 21 Most research on the microbial diversity of shrimp paste has focused on the shrimp 22 culture environment, or the chemical composition and sensory attributes of the paste. 23 Applied and Environmental Microbiology 2 Little research has been conducted on the microbial diversity and composition of shrimp 24 paste. The relationship between microbes and the flavor and quality of shrimp paste has 25 thus been unknown. We therefore analyzed the microbial composition and variation of 26 shrimp paste at different stages of fermentation. The dominant bacteria in fermentation 27 prophase, metaphase, and anaphase were identified. Our preliminary findings give some 28 insight into which microbes contribute to the flavor of shrimp paste and suggest how to 29 improve its flavor. In addition, our findings are relevant to optimizing the production of 30 shrimp paste and guaranteeing its quality and safety.31

Glucocorticoid (GC) signaling is an important regulator of glucose homeostasis. GC secretion also serves as an entrainment signal that coordinates circadian gene expression of peripheral circadian clocks. Pancreatic beta cells are key biological controllers of glucose homeostasis through the coordinated circadian release of insulin. However, the role of GC signaling in the regulation of beta cell circadian clocks and insulin secretion remains unknown. To address this, we first performed chromatin immunoprecipitation/sequencing (ChIP-seq) in beta cell lines (MIN6 and INS1) and identified >1,000 glucocorticoid receptor (GR)-bound regions (FDR<0.01) in response to the synthetic GC dexamethasone (DEX, 100nM). Annotation of GR-bound regions showed enrichment for pathways regulating insulin secretion and circadian rhythms. To further explore the effects of GC on circadian beta cell gene expression and function in vivo, we used C57BL/6 mice housed under either 12h/12h light-dark cycles (LD) or constant light-mediated circadian disruption (CD). Mice were subsequently administered daily intraperitoneal injections of DEX (2 mg/kg) or vehicle, designed to restore circadian secretion of GC disrupted in CD group. DEX treatment normalized circadian regulation of glucose tolerance (p<0.05 vs. vehicle) and glucose-stimulated insulin secretion (p<0.05 vs vehicle) in CD group. We next utilized RNA sequencing to assess circadian transcriptional profiles in islets isolated from LD, CD, and CD + DEX mice. Consistent with previous studies, CD resulted in abrogation of circadian islet mRNA expression, whereas DEX restored circadian rhythmicity and enhanced expression of core clock genes (e.g., Bmal1, Dbp, Nr1d1) and genes regulating beta cell identity (e.g., Ins1, Nkx6.1) and insulin secretion (e.g., Gck, Irs2). These data highlight the GC/GR axis as a direct regulator of beta cell circadian gene expression, insulin secretion and glucose tolerance. Disclosure S.K.Sen: None. L.Wang: None. M.Brown: None. J.Wu: None. Z.Wei: None. A.Matveyenko: None. Funding National Institutes of Health (DK098468)

Background: Environmental disorders of the circadian rhythms can lead to metabolism-related diseases or exacerbate pathological conditions. Non-alcoholic fatty liver disease (NAFLD) has emerged with a growing occurrence. In the present study, we attempted to indicate whether circadian clock may influence lipid deposition and the expression levels of autophagy-related genes in liver of mice. Methods: High-fat diet and abnormal light/dark cycles were employed to induce a mouse model of NAFLD with circadian rhythm sleep disorder. Herein, liver samples were obtained at ZT0, ZT4, ZT8, ZT12, ZT16, and ZT20 time-point to detect the rhythmic expressions of circadian genes, autophagy-related genes, and Rev-erbα. Results: Abnormal exposure to light aggravated lipid deposition in liver of mice and exacerbated disorders related to 24-h expression levels of clock genes, autophagy-related genes, and Rev-erbα. Besides, Rev-erbα could transcriptionally control the expression levels of autophagy-related genes. Conclusions: The long-term high-fat diet combined with abnormal light/dark cycle stimulation aggravated the development of NAFLD and disturbed the expressions levels of autophagy-related genes. An abnormal circadian expression may lead to NAFLD aggression. Besides, the abnormal expression levels of clock genes may create an association between circadian rhythm sleep disorder and autophagy.