Sublethal costs of salinity stress contribute to habitat limitation in an endangered estuarine fish
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ABSTRACT: We exposed adult delta smelt to treatments of increased salinities to quantify the transcriptomic responses involved in coping with osmotic stress 70 samples were run on 70 arrays, with 1-4 replicates for each treatment or control
Project description:We exposed adult delta smelt to varying levels of sublethal thermal stress to quantify the genes involved in their cellular stress response and identify sublethal stress thresholds Fourty-nine samples were run on fourty-nine arrays, with 3-6 replicates for each treatment or control
Project description:Ibuprofen is one of the most commonly detected pharmaceuticals in wastewater effluent; however the effects of ibuprofen on aquatic organisms are poorly understood. This study presents the transcriptome-wide response of the inland silverside, Menidia beryllina, to chronic 14 d exposures to ibuprofen. Twenty-four samples were run on twenty-four arrays, there were six replicates for each of three ibuprofen exposure concentrations and a control group.
Project description:We investigated the genomic transcriptional response of female fathead minnows (Pimephales promelas) to an acute (4 day) exposure to 0.1 or 1.0 µg/L of, 17β-trenbolone (TB), the active metabolite of an anabolic androgenic steroid used as a growth promoter in cattle and a contaminant of concern in aquatic systems. Our objectives were to investigate the gene expression profile induced by TB, define biomarkers of exposure to TB, and increase our understanding of the mechanisms of adverse effects of TB on fish reproduction. In female gonad tissue, microarray analysis using a 22K oligonucleotide microarray (EcoArray Inc., Gainesville, FL) showed 99 significantly upregulated genes and 741 significantly downregulated genes in response to 1 µg TB/L. In particular, hydroxysteroid (17β) dehydrogenase 12a (hsd17b12a), zona pellucida glycoprotein 2.2 (zp2.2), and protein inhibitor of activated STAT, 2 (pias2) were all downregulated in gonad. 4 control samples compared to 4 exposed samples, ovary.
Project description:Research on the effects of contaminants on fishes is often conducted on well-studied model test species, whose responses may be different than those of species of conservation concern. We used an oligonucleotide microarray to examine the effects of permethrin, a widely used pyrethroid pesticide, on a critically endangered fish species endemic to Northern California, the delta smelt (Hypomesus transpacificus). These results demonstrate the effects of a widely used pesticide on a sensitive fish species at concentrations below those that affect model test species. Twenty samples were run on twenty arrays, there were four replicates for each of four permethrin exposure concentrations and a control group.
Project description:Amoebic gill disease (AGD) is an ectoparasitic condition of some farm-reared marine fish and is caused by Neoparamoeba perurans. Tanks housing Atlantic salmon (Salmo salar) were inoculated with Neoparamoeba perurans and fish were sampled at 36 days postinoculation (pi.). AGD-affected gill tissue was dissected from N. perurans infected fish, and a DNA microarray was used to compare global gene expression against tissues from AGD-naive fish. To determine whether the changes in gene expression were restricted to AGD-lesions, lesion tissue from AGD-affected fish was also compared with non-lesion gill tissue dissected from the same gill arch. Samples were assessed using a DNA microarray. mRNA from lesion and non-lesion gill tissue was amplified and labeled. Six biological and two technical replicates were utilised to hybridise to 12 arrays using amplified RNA from AGD-affected lesion gill tissue with AGD-naive fish as a control. Four biological and two technical replicates were utilised to hybridise to 8 arrays using amplified RNA from AGD-affected lesion gill tissue with non-lesion tissue from the same gill arch as a control. The assignment of microarrays to treatment groups for hybridization was randomised by using a random number generator.
Project description:A 50-mer oligonucleotide microarray was designed for large-scale gene expression analysis in Z. viviparus. To measure the expression of the probes and the corresponding assembled transcripts, the pool of mRNA used for the sequencing was hybridized. 1 Sample hybridized to microarray used to evaluate the transcript assembly described in Kristiansson et al 2009.
Project description:We report the proteomic characterization of livers from Sparus aurata exposed to cold temperatures. In this study, mimicking the winter challenge conditions, a 8 week feeding trial was carried out on gilthead sea bream juveniles reared in RAS systems at a temperature ramp made of two phases of four weeks each: a cooling phase from 18°C 8 (t0) to 11°C (t1) and a cold maintenance phase at 11°C (t2). Sparus aurata livers, after exposure to the three temperature phases (t0, t1 and t2), were collected and analyzed using a shotgun proteomics approach based on filter-aided sample preparation followed by tandem mass spectrometry, peptide identification carried out using Sequest-HT as search engine within the Proteome Discoverer informatic platform, and label-free differential analysis. Along the whole trial, sea breams underwent several changes occurring upon thermal stress in liver protein abundance. These occurred mostly during the cooling phase, when catabolic processes were mainly observed. These included protein and lipid degradation and a decrease in protein synthesis and amino acid metabolism. A decrease in protein mediators of oxidative stress protection was also seen. Liver protein profiles showed less marked changes during cold maintenance, although pathways such as the methionine cycle and sugar metabolism were significantly affected. This study provided useful hints on the dynamics and extent of the metabolic shift occurring in sea bream liver with decreasing water temperature, helping the development of feeds aimed at compensating the thermal stress encountered by fish in offshore farming conditions.
Project description:We used a reference design with a dye swap. We used 6 experimental probes grouped by treatment and sample day. "Treatment" fish consisted of rockfish exposed to forced decompression resulting in barotrauma, followed by subsequent recompression to their original depth. Control fish did not experience forced decompression. Fish were sampled at day 3, day 15 and day 31 post-decompression.
Project description:According to Mendel's laws, each parent makes an equal genetic contribution to an offspring in sexually reproducing organisms. The bipolar mitotic spindle controls the equal segregation of paternal and maternal chromosomes during the first cell division. By overexpression of a single protein, GPR-1, in the maternal strain we changed the structure of the mitotic spindle from bipolar to two monopolar spindles to segregate maternal and paternal chromosomes into different cell lineages, resulting in non-mendelian segregation for entire genomes. To follow maternal and paternal segregation of the chromosomes we used red and green histone markers respectively. By mating gpr-1-overexpressing hermaphrodites with wild-type males, mendelian F1 worms that express both markers simultaneously in all tissues and non-mendelian F1 worms that express red and green markers in different tissues will be produced representing embryos with bipolar and embryos with two monopolar spindles. Thus, we show that the rules of genetic inheritance can be changed, which may inspire the formation of a new field of synthetic zoology. Transcriptional profiling was done to investigate the differences in gene expression between mendelian and non-mendelian offspring. Approximately 60 adult worms were used per sample. Four conditions were collected: hermaphrodites of the paternal strain, hermaphrodites of the maternal strain, co-segregating (mendelian) F1 after crossing of parental strains, and (non-mendelian) F1 that segregates the paternal genotype to body wall muscle, intestine + germline and the maternal genotype to the nervous system after crossing of parental strains.
Project description:To determine the effects on the intestine gene expression of pathogen exposure to Enteromyxum leei. One fish group was exposed to E. leei-contaminated effluent (recipient group = R) . R fish (n= 66, average weight = 134 g) were placed in two replicated 200L fibre-glass tanks which were set to receive exclusively the effluent water from another tank containing 24 infected (donors = D; average weight = 127.3 g; prevalence of infection = 54%) gilthead sea bream. The D to R fish ratio was 0.8. Other 66 naïve fish were allocated in two replicated tanks (control group = CTRL) under the same conditions, but without receiving contaminated effluent. Over the course of the study, day length followed natural changes and water was heated in order to keep temperature always above 18ºC, the range was 18-23 ºC. Water was 5 µm-filtered and UV irradiated, and salinity was 37.5â°. Water flow was 10L/min and oxygen content of outlet water remained higher than 85%saturation. All fish were fed daily a commercial dry pellet diet at about 1% of body weight. Disease signs and daily mortalities were recorded throughout the experiments. The parasitic status of dead fish was checked by microscopic examination of fresh intestinal scrapings. Fish were sampled after 113 days post exposure (p.e.). Feeding was stopped one day prior to the sampling to ensure that the digestive tract was empty. Head kidney and posterior intestine were rapidly excised, frozen in liquid nitrogen, and stored at -80 °C until RNA extraction and analysis. Keywords: Treated/Untreated, confinement, cortisol, stress response, time course, microarray 28 intestine samples - Twenty eight slides were hybridised using a reference design. Three groups (control, infected and non-infected) were compared using five individual fish in each group. Each sample was hybridised twice - the second being a dye-swap of the first.