Project description:<p>The study of antimicrobial resistance (AMR) in infectious diarrhea has generally been limited to cultivation, antimicrobial susceptibility testing and targeted PCR assays. When individual strains of significance are identified, whole genome shotgun (WGS) sequencing of important clones and clades is performed. Genes that encode resistance to antibiotics have been detected in environmental, insect, human and animal metagenomes and are known as "resistomes". While metagenomic datasets have been mined to characterize the healthy human gut resistome in the Human Microbiome Project and MetaHIT and in a Yanomani Amerindian cohort, directed metagenomic sequencing has not been used to examine the epidemiology of AMR. Especially in developing countries where sanitation is poor, diarrhea and enteric pathogens likely serve to disseminate antibiotic resistance elements of clinical significance. Unregulated use of antibiotics further exacerbates the problem by selection for acquisition of resistance. This is exemplified by recent reports of multiple antibiotic resistance in Shigella strains in India, in Escherichia coli in India and Pakistan, and in nontyphoidal Salmonella (NTS) in South-East Asia. We propose to use deep metagenomic sequencing and genome level assembly to study the epidemiology of AMR in stools of children suffering from diarrhea. Here the epidemiology component will be surveillance and analysis of the microbial composition (to the bacterial species/strain level where possible) and its constituent antimicrobial resistance genetic elements (such as plasmids, integrons, transposons and other mobile genetic elements, or MGEs) in samples from a cohort where diarrhea is prevalent and antibiotic exposure is endemic. The goal will be to assess whether consortia of specific mobile antimicrobial resistance elements associate with species/strains and whether their presence is enhanced or amplified in diarrheal microbiomes and in the presence of antibiotic exposure. This work could potentially identify clonal complexes of organisms and MGEs with enhanced resistance and the potential to transfer this resistance to other enteric pathogens.</p> <p>We have performed WGS, metagenomic assembly and gene/protein mapping to examine and characterize the types of AMR genes and transfer elements (transposons, integrons, bacteriophage, plasmids) and their distribution in bacterial species and strains assembled from DNA isolated from diarrheal and non-diarrheal stools. The samples were acquired from a cohort of pediatric patients and controls from Colombia, South America where antibiotic use is prevalent. As a control, the distribution and abundance of AMR genes can be compared to published studies where resistome gene lists from healthy cohort sequences were compiled. Our approach is more epidemiologic in nature, as we plan to identify and catalogue antimicrobial elements on MGEs capable of spread through a local population and further we will, where possible, link mobile antimicrobial resistance elements with specific strains within the population.</p>
| phs001260 | dbGaP
Project description:Polysaccharide contributes to the enrichment of antibiotic resistance genes in mariculture
Project description:Caligid copepods, also called sea lice, are common ectoparasites of wild and farmed marine fish. The salmon louse Lepeophtheirus salmonis (KrM-xyer, 1837) has emerged as a serious problem for salmon farming in the Northern hemisphere. The annual cost of sea lice to the global salmon mariculture industry has been estimated at M-^@300 million, of which the majority accounts for the cost of chemically treating the farmed salmon. The treatments available for salmonids with sea lice infestation have been limited with a large scale reliance on single products and the use of antiparasitics with similar modes of action, which when used over a long period of time can enhance the selection pressure for reduced sensitivity. Two L. salmonis laboratory strains, established from field isolates and differing in susceptibility to emamectin benzoate (EMB) were studied using a custom sea louse 15K oligonucleotide microarray and RT-qPCR. The aim of the present study was to identify differential expression of transcripts between these two strains to identify potential constitutive gene expression changes associated with reduced susceptibility to EMB. Adult male salmon lice were sampled without exposure to antiparasitic agents for the purpose of studying gene expression from unchallenged individuals. In this study changes in expression of Glutamate-gated Chloride channel (GluCl) subunits, considered the major target site for avermectin (AVM) drugs in invertebrates, was not observed, but expression changes were seen for alternative ligand-gated ion channel (LGIC) subunits that form an ion channels shown to interact with AVMs in vertebrates, but which is not traditionally considered to be a target site for AVMs in invertebrates. We hypothesise that these LGIC subunits represent additional EMB target sites in salmon lice, and that the down-regulation of these channel subunits in this EMB-resistant strain is related to the resistance phenotype.
Project description:Due to multi-generation domestication selection, farmed and wild Atlantic salmon diverge genetically, which raises concerns about potential genetic interactions among escaped farmed and wild populations and disrupts local adaptation through introgression. When farmed strains of distant geographic origin are used, it is unknown whether the genetic risks posed by escaped farmed fish will be greater than if more locally derived strains are used. Quantifying gene expression differences among divergent farmed, wild and F1 hybrids under controlled conditions is one of the ways to explore the consequences of hybridization. We compared the transcriptomes of late sac fry of a European (EO) farmed (“StofnFiskur”, Norwegian strain), a North American (NA) farmed (Saint John River, NB strain), a Newfoundland (NF) wild population with EO ancestry, and related F1 hybrids using 44K microarrays. Our findings indicate that the wild population showed greater transcriptome differences from the EO farmed strain than that of the NA farmed strain. We also found the largest differences in global gene expression between the two farmed strains. We detected fewer differentially expressed transcripts between F1 hybrids and domesticated/wild maternal strains. We also found that the differentially expressed genes between cross types over-represented GO terms associated with metabolism, development, growth, immune response, and redox homeostasis processes. These findings suggest that the interbreeding of escaped EO/NA farmed and NF wild population would alter gene transcription, and the consequences of hybridization would be greater from escaped EO farmed than NA farmed salmon, resulting in potential effects on the fitness of wild populations.
Project description:Gills of teleost fish represent a vital multifunctional organ; however, they are subjected to environmental stressors, causing gill damage. Gill damage is associated with significant losses in the Atlantic salmon aquaculture industry. Gill disorders due to environmental stressors are exacerbated by global environmental changes, especially with open-net pen aquaculture (as farmed fish lack the ability to escape those events). The local and systemic response to gill damage, concurrent with several environmental insults, are not well investigated. We performed field sampling to collect gill and liver tissue after several environmental insults. Using a 44K salmonid microarray platform, we aimed to compare the transcriptomes of pristine and moderately damaged gill tissue. The gill damage-associated biomarker genes and associated qPCR assays arising from this study will be valuable in future research aimed at developing therapeutic diets to improve farmed salmon gill health.
Project description:Understanding the molecular mechanisms of feed efficiency is an important step toward sustainability of salmonids aquaculture. In this study, the liver and white muscle proteomes of efficient (EFF) and inefficient (INEFF) Chinook salmon (Oncorhynchus tshawytscha) farmed in sea water were investigated by liquid chromatography-tandem mass spectrometry (LC-MS/MS) approach. In total, 2,746 liver and 702 white muscle quantified proteins were compared between 21 EFF and 22 INEFF fish. Protein synthesis was enriched in both liver and white muscle of the EFF group while conversely, pathways related to protein degradation (amino acid catabolism and proteolysis, respectively) were the most affected processes in the liver and white muscle of INEFF fish. The SOM in the INEFF group was significantly higher than EFF fish showing INEFF fish probably was the dominant group. The INEFF group (dominant) suffered stress and shifted to consume energy through protein catabolism. As the first study, the results provide a preliminary picture of the fundamental molecular landscape of feed efficiency in Chinook salmon farmed in sea water
Project description:Caligid copepods, also called sea lice, are common ectoparasites of wild and farmed marine fish. The salmon louse Lepeophtheirus salmonis (KrM-xyer, 1837) has emerged as a serious problem for salmon farming in the Northern hemisphere. The annual cost of sea lice to the global salmon mariculture industry has been estimated at M-^@300 million, of which the majority accounts for the cost of chemically treating the farmed salmon. The treatments available for salmonids with sea lice infestation have been limited with a large scale reliance on single products and the use of antiparasitics with similar modes of action, which when used over a long period of time can enhance the selection pressure for reduced sensitivity. The aim of the present study was to identify transcripts whose expression correlated to emamectin benzoate (EMB) susceptibility, or those genes regulated in response to EMB exposure. Two L. salmonis laboratory strains, established from field isolates and differing in susceptibility to EMB were studied using a custom sea louse 15K oligonucleotide microarray and RT-qPCR. Adult male sea lice were sampled from both strains after 1 and 3 hours of aqueous exposure to 0.2 M-5g mL-1 emamectin benzoate, 0.01% PEG300 or sea water. Bioinformatic analysis identified that in the absence of drug treatment, a large number of genes were significantly down regulated in the louse strain hyposensitive to EMB. EMB exposure had marked effects on gene expression in the EMB susceptible strain, but caused little changes in EMB hyposensitive lice. We therefore suggest that transcriptional responses induced by EMB exposure may not be responsible for reduced susceptibility to this antiparasitic compound, but may involve genes that are constitutively expressed in EMB tolerant salmon louse strains.
Project description:Hypoxia is an important environmental stressor in aquatic ecosystems, with increasingly impacts on global biodiversity. Yellow catfish is an economically important farmed fish in China, which has increased dramatically. We investigated the response of hybrid yellow catfish to hypoxia under experimental conditions and focused on the analysis of the differential expression patterns of specific genes associated with hypoxia response by RNA-seq and qPCR analysis. A total of 1556 genes were captured significantly differentially expressed, and were categorized into immune response and energy metabolism. Functional enrichment analysis revealed the NLR signaling pathway play pivotal roles in hypoxia tolerance and resistance. Our study provides important insights into the physiological acclimation, immune response and defense activity of hybrid yellow catfish under hypoxia challenge.
Project description:Norway is the largest producer and exporter of farmed Atlantic salmon (Salmo salar) worldwide. Skin disorders correlated with bacterial infections represent an important challenge for fish farmers due to the economic losses caused. Little is known about this topic, thus studying the skin-mucus of Salmo salar and its bacterial community depict a step forward in understanding fish welfare in aquaculture. In this study, we used label free quantitative mass spectrometry to investigate the skin-mucus proteins associated with both Atlantic salmon and bacteria. In addition, the microbial temporal proteome dynamics during 9 days of mucus incubation with sterilized seawater was investigated, in order to evaluate their capacity to utilize mucus components for growth in this environment.