Project description:Maintaining marine salinities in inland aquaculture systems can be cost-prohibitive. As such, there has been an emphasis on the identification of marine species that can be reared at low salinities. The Florida pompano (Trachinotus carolinus) is a popular sportfish that can survive in salinities as low as 5 ppt and has been identified as a potential target species for commercial production. Vibrio spp., bacterial pathogens ubiquitous in aquaculture systems, are capable of quickly decimating aquaculture populations. However, there is limited data available on the effects of salinity influences on Vibrio susceptibility in juvenile Florida pompano. This study investigated the influence of lowered salinity on the immune response of juvenile Florida pompano exposed to Vibrio across a 14-day period through assessment of transcriptomic changes, fatty acid biosynthesis, and identification of internal and external signs of vibriosis. During the experimental period, there were no differences in osmoregulatory or immune related gene expression, no obvious signs of vibriosis, and no changes in fatty acid biosynthesis in fish exposed to 8, 12, 20, or 28 ppt seawater. These results suggest juvenile Florida pompano are resilient to salinity changes and exhibit baseline molecular machinery capable of responding to pathogenic challenges across a range of salinities, making them strong candidates for rearing in inland aquaculture.
Project description:To identify cAMP-dependent lncRNAs, we treated DIV16 primary hippocampal neurons with 50uM Forskolin or equal volume DMSO for 30 minutes prior to RNA isolation using Trizol and Total RNA sequencing. We submitted samples to Scripps Florida Genomics Core for library preparation and sequencing.
Project description:To identify cAMP-dependent and mGluR1/5-dependent RNAs, we treated DIV16 primary hippocampal neurons with 50uM Forskolin, 30uM DHPG, or equal volume DMSO for 30 minutes prior to RNA isolation using Trizol and Total/small RNA sequencing. We submitted samples to Scripps Florida Genomics Core for library preparation and sequencing.
Project description:Carbapenem-resistant Acinetobacter baumannii (CRAB) is a critical nosocomial pathogen with limited treatment options. Although antibiotic resistance in CRAB is well-characterized, its interactions with host immunity and the contribution of outer membrane vesicles (OMVs) to pathogenesis remain poorly understood. We examined a clinical CRAB isolate and compared it with the reference strain A19606. Antimicrobial susceptibility testing revealed complete resistance of CRAB to commonly used antibiotics in clinical practice, while A19606 remained susceptible to most agents. In murine intranasal infection models and bone marrow-derived macrophages, CRAB induced significantly stronger activation of inflammatory signaling pathways and elevated levels of pro-inflammatory cytokines relative to A19606. Transcriptomic analysis of infected lung tissue identified differentially expressed genes, enriched for inflammatory response pathways. proteomics showed upregulated proteins in CRAB related to secretion systems. OMVs characterization revealed that CRAB-derived OMVs highly enriched in proteins associated with periplasmic and outer membrane spaces, and more potent in triggering macrophage inflammatory signaling. CRAB displays expansive antibiotic resistance and enhanced pro-inflammatory potential mediated in part by unique OMVs properties. Targeting OMVs formation or host immune modulation may represent effective strategies for combating CRAB infections.
Project description:Background: The Scylla paramamosain is a very important aquaculture crustacean species in the southeast coastal areas of China including Shantou. For the past few years, mud crab cultured in Niutianyang of Shantou suffered from serious diseases, especially the bacterial diseases (such as Vibrio parahaemolyticus). In eukaryotes, small RNAs can regulate gene expression in post-transcription to act on host-pathogen interaction system. Aims: V.parahaemolyticus isolated from Shantou Niutianyang crab culture area was injected to S.paramamosains to carry out an essential analysis on global miRNA expression in diverse tissues between two groups by the Illumina Solex deep sequencing technology. Methodology:To examine the relationship between mud crab miRNA expression and the bacterial pathogen, we collected mixed two pools of equal amounts of RNA from 7 different mud crab tissues (mesenteron, heart, liver, gill, brain, muscle and blood) and sequencing by Illumine/Solexa deep sequencing technology under normal conditions and during infection with V.parahaemolyticus. The high throughput sequencing resulted in 19,144,358 and 18,559,070 raw reads corresponding to 17,496,577 and 16,888,096 high-quality mappable reads for the normal and infected mixed pools, respectively. Stem-loop RT-qPCRs were used to confirm the microRNAs expression in different tissues of two pools. The results show that miRNAs might play a key role in regulating gene expression during mud crab S.paramamosain infection with V.parahaemolyticus. Conclusions: We identified a large number of miRNAs during the mud crab Scylla paramamosain infection with V.parahaemolyticus, some of which are differentially expressed between the treatments and the controls. The study provides an opportunity for further understanding of small RNA function in the regulation of molecular response and gives us clues for further studies of the mechanisms of V.parahaemolyticus infection in mud crab.
Project description:Crab is one of the major source for V. parahaemolyticus outbreak among aquatic products in Northeast Asian due to improperly cooking and wound infection by mishandling. However, there is no report on whole genome sequence of V. parahaemolyticus isolated from contaminated crab, thus no information is available for major virulence factors about V. parahaemolyticus obtained from crab. Therefore, the analysis of transcriptome of isolated V. parahaemolyticus from crab products are necessary to investigate potential risk of foodborne illness by contaminated products.
Project description:We report the application of DNA sequencing technology for high-throughput sequencing of mix bis-PCR products totally 38 based on bisulfate treated DNA from human, chimpanzee, gibbon, macaque and crab eating macaque profrontal cortex tissues. Mix bisulfate PCR products from 1 tissues, 23 individula humans, 2 individual chimpanzees, 1 individual gibbons, 7 individual rhesus macaques and 5 crab eating macaques were sequenced by using MiSeq