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Flavobacterium Columnare, the causative agent of columnaris disease (CD), is distributed around the world in fresh water sources, infecting freshwater finfish species. Recently, it has been identified as an emerging problem for the rainbow trout aquaculture industry in the U.S. Two live-attenuated vaccines have been commercialized, but their efficacy in rainbow trout is still not clear. The purpose of this study was to prospect genomic regions that explain large portion of the additive genetic variance of CD resistance in rainbow trout. Two important aquaculture populations were investigated: The National Center for Cool and Cold Water Aquaculture (NCCCWA) odd-year line, with resistance to bacterial cold water disease (BCWD), and the Troutlodge, Inc., May odd-year (TLUM) nucleus breeding population, which provided 54,350 and 36,265 pedigree records, in which 8,453 and 3,986 fish had CD resistance phenotype records, respectively. Fish that survived to 21 days post immersion challenge were recorded as resistant (phenotype = 2), and those that did not were rated as susceptible (phenotype = 1). Genotypes for 57k SNP (Affymetrix Axiom ®) were available for 1,185 and 1,137 fish from NCCCWA and TLUM, respectively. The SNP effects and variances were estimated using the weighted single-step genomic BLUP approach for genome-wide association (WssGBLUP), which uses pedigree, genotypes, and phenotypes from genotyped and ungenotyped animals. The weighting strategy accounted for 1 Mb moving SNP-windows. Genomic regions that explained more than 1% of the additive genetic variance were considered associated with CD resistance. A total of 13 windows located on six chromosomes were found to be associated with CD resistance in the NCCCWA population: two windows, located at 59 to 60 Mb and 61 to 62 Mb on chromosome Omy17, explained 12% and 11.33% of the genetic variance for CD resistance, respectively. In the TLUM population, a total of 16 windows located on nine chromosomes were detected. Only three similar windows (located on two chromosomes) were detected in both populations. The results suggest that CD resistance has an oligogenic architecture, and the SNP windows found to be associated with CD are not informative enough for selection decisions across populations. A moderate positive genetic correlation has been previously shown between CD and BCWD resistance in the NCCCWA population. One factor that might have contributed to detecting different QTL for CD resistance in the two populations is the five generations of selective breeding for BCWD resistance applied to the NCCCWA population in contrast to no selection pressure for disease resistance in the TLUM population. [ABSTRACT FROM AUTHOR]

The characteristic reddish-pink fillet color of rainbow trout is an important marketing trait. The gastrointestinal microbiome is vital for host health, immunity, and nutrient balance. Host genetics play a crucial role in determining the gut microbiome, and the host-microbiome interaction impacts the host's phenotypic expression. We hypothesized that fecal microbiota could be used to predict fillet color in rainbow trout. Fish were fed Astaxanthin-supplemented feed for six months, after which 16s rDNA sequencing was used to investigate the fecal microbiome composition in rainbow trout families with reddish-pink fillet coloration (red fillet group, average saturation index = 26.50 ± 2.86) compared to families with pale white fillet color (white fillet group, average saturation index = 21.21 ± 3.53). The linear discriminant analysis effect size (LEFse) tool was used to identify bacterial biomarkers associated with fillet color. The alpha diversity measure shows no difference in the red and white fillet groups. Beta diversity principal component analysis showed clustering of the samples along the white versus red fillet group. The red fillet group has enrichment (LDA score > 1.5) of taxa Leuconostoc lactis , Corynebacterium variabile , Jeotgalicoccus halotolerans , and Leucobacter chromiireducens . In contrast, the white fillet group has an enriched presence of mycoplasma , Lachnoclostridium , and Oceanobacillus indicireducens . The enriched bacterial taxa in the red fillet group have probiotic functions and can generate carotenoid pigments. Bacteria taxa enriched in the white fillet group are either commensal, parasitic, or capable of reducing indigo dye. The study identified specific bacterial biomarkers differentially abundant in fish families of divergent fillet color that could be used in genetic selection to improve feed carotenoid retention and reddish-pink fillet color. This work extends our understanding of carotenoid metabolism in rainbow trout through the interaction between gut microbiota and fillet color.

Background: The characteristic pink-reddish color in the salmonids fillet is an important, appealing quality trait for consumers and producers. The color results from diet supplementation with carotenoids, which accounts for up to 20-30% of the feed cost. Pigment retention in the muscle is a highly variable phenotype. In this study, we aimed to understand the molecular basis for the variation in fillet color when rainbow trout (Oncorhynchus mykiss) fish families were fed an Astaxanthin-supplemented diet. We used RNA-Seq to study the transcriptome profile in the pyloric caecum, liver, and muscle from fish families with pink-reddish fillet coloration (red) versus those with lighter pale coloration (white)., Results: More DEGs were identified in the muscle (5,148) and liver (3,180) than in the pyloric caecum (272). Genes involved in lipid/carotenoid metabolism and transport, ribosomal activities, mitochondrial functions, and stress homeostasis were uniquely enriched in the muscle and liver. For instance, the two beta carotene genes (BCO1 and BCO2) were significantly under-represented in the muscle of the red fillet group favoring more carotenoid retention. Enriched genes in the pyloric caecum were involved in intestinal absorption and transport of carotenoids and lipids. In addition, the analysis revealed the modulation of several genes with immune functions in the pyloric caecum, liver, and muscle., Conclusion: The results from this study deepen our understanding of carotenoid dynamics in rainbow trout and can guide us on strategies to improve Astaxanthin retention in the rainbow trout fillet., (© 2023. BioMed Central Ltd., part of Springer Nature.)

Background: In aquaculture, the proportion of edible meat (FY = fillet yield) is of major economic importance, and breeding animals of superior genetic merit for this trait can improve efficiency and profitability. Achieving genetic gains for fillet yield is possible using a pedigree-based best linear unbiased prediction (PBLUP) model with direct and indirect selection. To investigate the feasibility of using genomic selection (GS) to improve FY and body weight (BW) in rainbow trout, the prediction accuracy of GS models was compared to that of PBLUP. In addition, a genome-wide association study (GWAS) was conducted to identify quantitative trait loci (QTL) for the traits. All analyses were performed using a two-trait model with FY and BW, and variance components, heritability, and genetic correlations were estimated without genomic information. The data used included 14,165 fish in the pedigree, of which 2742 and 12,890 had FY and BW phenotypic records, respectively, and 2484 had genotypes from the 57K single nucleotide polymorphism (SNP) array., Results: The heritabilities were moderate, at 0.41 and 0.33 for FY and BW, respectively. Both traits were lowly but positively correlated (genetic correlation; r = 0.24), which suggests potential favourable correlated genetic gains. GS models increased prediction accuracy compared to PBLUP by up to 50% for FY and 44% for BW. Evaluations were found to be biased when validation was performed on future performances but not when it was performed on future genomic estimated breeding values., Conclusions: The low but positive genetic correlation between fillet yield and body weight indicates that some improvement in fillet yield may be achieved through indirect selection for body weight. Genomic information increases the prediction accuracy of breeding values and is an important tool to accelerate genetic progress for fillet yield and growth in the current rainbow trout population. No significant QTL were found for either trait, indicating that both traits are polygenic, and that marker-assisted selection will not be helpful to improve these traits in this population., (© 2023. The Author(s).)

The visual appearance of the fish fillet is a significant determinant of consumers' purchase decisions. Depending on the rainbow trout diet, a uniform bright white or reddish/pink fillet color is desirable. Factors affecting fillet color are complex, ranging from the ability of live fish to accumulate carotenoids in the muscle to preharvest environmental conditions, early postmortem muscle metabolism, and storage conditions. Identifying genetic markers of fillet color is a desirable goal but a challenging task for the aquaculture industry. This study used weighted, single-step GWAS to explore the genetic basis of fillet color variation in rainbow trout. We identified several SNP windows explaining up to 3.5%, 2.5%, and 1.6% of the additive genetic variance for fillet redness, yellowness, and whiteness, respectively. SNPs are located within genes implicated in carotenoid metabolism (β,β-carotene 15,15'-dioxygenase, retinol dehydrogenase) and myoglobin homeostasis (ATP synthase subunit β, mitochondrial ( ATP5F1B )). These genes are involved in processes that influence muscle pigmentation and postmortem flesh coloration. Other identified genes are involved in the maintenance of muscle structural integrity (kelch protein 41b ( klh41b ), collagen α-1(XXVIII) chain ( COL28A1 ), and cathepsin K ( CTSK )) and protection against lipid oxidation (peroxiredoxin, superoxide dismutase 2 ( SOD2 ), sestrin-1, Ubiquitin carboxyl-terminal hydrolase-10 ( USP10 )). A-to-G single-nucleotide polymorphism in β,β-carotene 15,15'-dioxygenase, and USP10 result in isoleucine-to-valine and proline-to-leucine non-synonymous amino acid substitutions, respectively. Our observation confirms that fillet color is a complex trait regulated by many genes involved in carotenoid metabolism, myoglobin homeostasis, protection against lipid oxidation, and maintenance of muscle structural integrity. The significant SNPs identified in this study could be prioritized via genomic selection in breeding programs to improve fillet color in rainbow trout.

Background: One of the most important goals for the rainbow trout aquaculture industry is to improve fillet yield and fillet quality. Previously, we showed that a 50 K transcribed-SNP chip can be used to detect quantitative trait loci (QTL) associated with fillet yield and fillet firmness. In this study, data from 1568 fish genotyped for the 50 K transcribed-SNP chip and ~ 774 fish phenotyped for fillet yield and fillet firmness were used in a single-step genomic BLUP (ssGBLUP) model to compute the genomic estimated breeding values (GEBV). In addition, pedigree-based best linear unbiased prediction (PBLUP) was used to calculate traditional, family-based estimated breeding values (EBV)., Results: The genomic predictions outperformed the traditional EBV by 35% for fillet yield and 42% for fillet firmness. The predictive ability for fillet yield and fillet firmness was 0.19-0.20 with PBLUP, and 0.27 with ssGBLUP. Additionally, reducing SNP panel densities indicated that using 500-800 SNPs in genomic predictions still provides predictive abilities higher than PBLUP., Conclusion: These results suggest that genomic evaluation is a feasible strategy to identify and select fish with superior genetic merit within rainbow trout families, even with low-density SNP panels.

Background: Fish gut microbial assemblages play a crucial role in the growth rate, metabolism, and immunity of the host. We hypothesized that the gut microbiota of rainbow trout was correlated with breeding program based genetic selection for muscle yield. To test this hypothesis, fecal samples from 19 fish representing an F2 high-muscle genetic line (ARS-FY-H) and 20 fish representing an F1 low-muscle yield genetic line (ARS-FY-L) were chosen for microbiota profiling using the 16S rRNA gene. Significant differences in microbial assemblages between these two genetic lines might represent the effect of host genetic selection in structuring the gut microbiota of the host., Results: Tukey's transformed inverse Simpson indices indicated that high muscle yield genetic line (ARS-FY-H) samples have higher microbial diversity compared to those of the low muscle yield genetic line (ARS-FY-L) (LMM, χ2(1) =14.11, p < 0.05). The fecal samples showed statistically distinct structure in microbial assemblages between the genetic lines (F 1,36  = 4.7, p < 0.05, R 2  = 11.9%). Functional profiling of bacterial operational taxonomic units predicted characteristic functional capabilities of the microbial communities in the high (ARS-FY-H) and low (ARS-FY-L) muscle yield genetic line samples., Conclusion: The significant differences of the microbial assemblages between high (ARS-FY-H) and low (ARS-FY-L) muscle yield genetic lines indicate a possible effect of genetic selection on the microbial diversity of the host. The functional composition of taxa demonstrates a correlation between bacteria and improving the muscle accretion in the host, probably, by producing various metabolites and enzymes that might aid in digestion. Further research is required to elucidate the mechanisms involved in shaping the microbial community through host genetic selection.

Background: Genetic improvement of fillet quality attributes is a priority of the aquaculture industry. Muscle composition impacts quality attributes such as flavor, appearance, texture, and juiciness. Fat and moisture make up about ~ 80% of the tissue weight. The genetic architecture underlying the fat and moisture content of the muscle is still to be fully explored in fish. A 50 K gene transcribed SNP chip was used for genotyping 789 fish with available phenotypic data for fat and moisture content. Genotyped fish were obtained from two consecutive generations produced in the National Center for Cool and Cold Water Aquaculture (NCCCWA) growth-selective breeding program. Estimates of SNP effects from weighted single-step GBLUP (WssGBLUP) were used to perform genome-wide association (GWA) analysis to identify quantitative trait loci (QTL) associated with the studied traits., Results: Using genomic sliding windows of 50 adjacent SNPs, 137 and 178 SNPs were identified as associated with fat and moisture content, respectively. Chromosomes 19 and 29 harbored the highest number of SNPs explaining at least 2% of the genetic variation in fat and moisture content. A total of 61 common SNPs on chromosomes 19 and 29 affected the aforementioned traits; this association suggests common mechanisms underlying intramuscular fat and moisture content. Additionally, based on single-marker GWA analyses, 8 and 24 SNPs were identified in association with fat and moisture content, respectively., Conclusion: SNP-harboring genes were primarily involved in lipid metabolism, cytoskeleton remodeling, and protein turnover. This work provides putative SNP markers that could be prioritized and used for genomic selection in breeding programs.

Background: Growth is a major economic production trait in aquaculture. Improvements in growth performance will reduce time and cost for fish to reach market size. However, genes underlying growth have not been fully explored in rainbow trout., Results: A previously developed 50 K gene-transcribed SNP chip, containing ~ 21 K SNPs showing allelic imbalances potentially associated with important aquaculture production traits including body weight, muscle yield, was used for genotyping a total of 789 fish with available phenotypic data for bodyweight gain. Genotyped fish were obtained from two consecutive generations produced in the NCCCWA growth-selection breeding program. Weighted single-step GBLUP (WssGBLUP) was used to perform a genome-wide association (GWA) analysis to identify quantitative trait loci (QTL) associated with bodyweight gain. Using genomic sliding windows of 50 adjacent SNPs, 247 SNPs associated with bodyweight gain were identified. SNP-harboring genes were involved in cell growth, cell proliferation, cell cycle, lipid metabolism, proteolytic activities, chromatin modification, and developmental processes. Chromosome 14 harbored the highest number of SNPs (n = 50). An SNP window explaining the highest additive genetic variance for bodyweight gain (~ 6.4%) included a nonsynonymous SNP in a gene encoding inositol polyphosphate 5-phosphatase OCRL-1. Additionally, based on a single-marker GWA analysis, 33 SNPs were identified in association with bodyweight gain. The highest SNP explaining variation in bodyweight gain was identified in a gene coding for thrombospondin-1 (THBS1) (R 2  = 0.09)., Conclusion: The majority of SNP-harboring genes, including OCRL-1 and THBS1, were involved in developmental processes. Our results suggest that development-related genes are important determinants for growth and could be prioritized and used for genomic selection in breeding programs.

Filet quality traits determine consumer satisfaction and affect profitability of the aquaculture industry. Soft flesh is a criterion for fish filet downgrades, resulting in loss of value. Filet firmness is influenced by many factors, including rate of protein turnover. A 50K transcribed gene SNP chip was used to genotype 789 rainbow trout, from two consecutive generations, produced in the USDA/NCCCWA selective breeding program. Weighted single-step GBLUP (WssGBLUP) was used to perform genome-wide association (GWA) analyses to identify quantitative trait loci affecting filet firmness and protein content. Applying genomic sliding windows of 50 adjacent SNPs, 212 and 225 SNPs were associated with genetic variation in filet shear force and protein content, respectively. Four common SNPs in the ryanodine receptor 3 gene (RYR3) affected the aforementioned filet traits; this association suggests common mechanisms underlying filet shear force and protein content. Genes harboring SNPs were mostly involved in calcium homeostasis, proteolytic activities, transcriptional regulation, chromatin remodeling, and apoptotic processes. RYR3 harbored the highest number of SNPs ( n = 32) affecting genetic variation in shear force (2.29%) and protein content (4.97%). Additionally, based on single-marker analysis, a SNP in RYR3 ranked at the top of all SNPs associated with variation in shear force. Our data suggest a role for RYR3 in muscle firmness that may be considered for genomic- and marker-assisted selection in breeding programs of rainbow trout.

A GeXP multiplex, RT-PCR assay was developed and optimized that simultaneously measures expression of a suite of immune-relevant genes in rainbow trout ( Oncorhynchus mykiss ), concentrating on tumor necrosis factor and interleukin-1 ligand/receptor systems and acute phase response genes. The dataset includes expression values for drpt , il11a , il1b1 , il1b2 , il1b3 , il1r-like-1 (e3-5), il1r-like-1 (e9-11), il1r1-like-a , il1r1-like-b , il1r2 , saa , tnfa1 , tnfa2 , tnfa3 , tnfrsf1a , tnfrsf1a-like-a , tnfrsf1a-like-b , tnfrsf5 , and tnfrsf9 . Gene expression was measured at four time-points post-challenge in both a resistant line (ARS-Fp-R) and a susceptible line (ARS-Fp-S) of rainbow trout. In addition, fish body weight, spleen index and the Flavobacterium psychrophilum load are reported. These data are an extension of information presented and discussed in " Proinflammatory cytokine and cytokine receptor gene expression kinetics following challenge with Flavobacterium psychrophilum in resistant and susceptible lines of rainbow trout ( Oncorhynchus mykiss )" (Kutyrev et al., 2016) [1].

Flavobacterium psychrophilum (Fp) is the causative agent of bacterial cold water disease (BCWD) which causes appreciable economic losses in rainbow trout aquaculture. We previously reported development of a genetic line, designated ARS-Fp-R that exhibits higher survival relative to a susceptible line, designated ARS-Fp-S, following either laboratory or natural on-farm challenge. The objectives of this study were to determine the temporal kinetics of gene expression between experimentally-challenged ARS-Fp-R and ARS-Fp-S fish and the correlation between gene expression and pathogen load. We developed a GeXP multiplex RT-PCR assay to simultaneously examine expression of immune-relevant genes, concentrating on tumor necrosis factor and interleukin-1 ligand/receptor systems and acute phase response genes. Spleen tissue was sampled at 6 h, 24 h, 48 h and 144 h post-challenge and pathogen load quantified by qPCR. Transcript abundance of cytokine genes tnfa1, tnfa2, tnfa3, il1b1, il1b2, il11a; acute phase response genes saa and drtp1; and putative cytokine receptors il1r1-like-b, il1r2, tnfrsf1a, tnfrsf9, tnfrsf1a-like-b increased following challenge while the transcript abundance of il1r-like-1 and tnfrsf1a-like-a decreased compared to PBS-injected line-matched control fish. Principal component analysis identified transcript levels of genes il1r-like-1 and tnfrsf1a-like-a as exhibiting differential expression between genetic lines. In summary, Fp i.p. injection challenge elicited a proinflammatory cytokine gene expression response in the spleen, with ARS-Fp-R line fish exhibiting modestly higher basal expression levels of several putative cytokine receptors. This study furthers the understanding of the immune response following Fp challenge and differences in gene expression associated with selective breeding for disease resistance., (Published by Elsevier Ltd.)

Sexual maturation occurs at the expense of stored energy and nutrients, including lipids; however, little is known regarding sex effects on nutrient regulatory mechanisms in rainbow trout prior to maturity. Thirty-two, 14-month-old, male and female rainbow trout were sampled for growth, carcass yield, fillet composition, and gene expression of liver, white muscle, and visceral adipose tissue. Growth parameters, including gonadosomatic index, were not affected by sex. Females had higher percent separable muscle yield, but there were no sex effects on fillet proximate composition. Fillet shear force indicated females produce firmer fillets than males. Male livers had greater expression of three cofactors within the mTOR signaling pathway that act to inhibit TORC1 assembly; mo25, rictor, and pras40. Male liver also exhibited increased expression of β-oxidation genes cpt1b and ehhadh. These findings are indicative of increased mitochondrial β-oxidation in male liver. Females exhibited increased expression of the mTOR cofactor raptor in white muscle and had higher expression levels of several genes within the fatty acid synthesis pathway, including gpat, srebp1, scd1, and cd36. Female muscle also had increased expression of β-oxidation genes cpt1d and cpt2. Increased expression of both fatty acid synthesis and β-oxidation genes suggests female muscle may have greater fatty acid turnover. Differences between sexes were primarily associated with variation of gene expression within the mTOR signaling pathway. Overall, data suggest there is differential regulation of gene expression in male and female rainbow trout tissues prior to the onset of sexual maturity that may lead to nutrient repartitioning during maturation.

The effect of, and associations between, loin muscle morphology and pork quality indicator traits were assessed at three cooked temperatures in loin chops from 38 purebred Berkshire and 52 purebred Landrace swine. Three loin chops from each pig were randomly assigned to cooked temperature treatments of 62, 71, or 79 degrees C and loin tenderness was assessed as Warner-Bratzler shear force (WBSF). Cooked temperature (P<0.001), breed (P<0.001) and breed x cooked temperature (P<0.001) effects influenced loin chop WBSF, whereby WBSF increased as cooked temperature increased. Chops from Landrace pigs had greater WBSF at each cooked temperature compared with chops from Berkshire pigs. Chops from Landrace pigs became less tender with increasing cooked temperature, whereas chops from Berkshire pigs became less tender only when cooked to 79 degrees C. In loins from Landrace pigs, Minolta a* at 62 degrees C (R(2)=0.07), and average muscle fiber diameter at 71 degrees C and 79 degrees C (R(2)=0.07 and 0.24, respectively), contributed to WBSF variation. In contrast, for loins from Berkshire pigs, loin ultimate pH and intramuscular fat percentage accounted for 27% and 30% of the variation in WBSF at 62 degrees C and 71 degrees C, respectively, and loin ultimate pH accounted for 7% of variation in WBSF at 79 degrees C. Results suggest that loins from Berkshire pigs have properties that resist toughening at greater cooked temperatures and that associations between quality measures and loin tenderness differ between Landrace and Berkshire pigs., (2009 Elsevier Ltd. All rights reserved.)

The purpose of the present study was to compare Rendement Napole (RN) classification from glycolytic potential (GP) and DNA techniques, and to study the effect of the RN gene on performance, carcass, muscle quality, and sensory traits. Progeny (N = 118) from the mating of 15 purebred Hampshire sires to York x Landrace females were classified for RN gene status using the GP of the loin and polymerase chain reaction-restriction fragment length polymorphism sequence methodology. Females mated in the study (N = 32) were considered normal (rn+/rn+) based on a loin GP measurement taken on samples collected by live press biopsy. Progeny were randomly selected for harvest within a litter for each sire. Observed mean, standard deviation, and range of progeny loin GP values were 132.2, 30.7, and 70.0 to 193.0 micromol/g, respectively. The GP data were not normally distributed. Peak numbers of observations occurred between 120 and 129 micromol/g and 160 to 169 micromol/g. Pigs with a loin GP of > 150 micromol/g were classified RN-/rn+ based on the observed valley between the peak values, resulting in 37 pigs classified as RN-/rn+ and 81 pigs classified as rn+/rn+. Using DNA procedures, 81 RN-/rn+ and 37 rn+/rn+ pigs were observed. All classification errors occurred when GP values were < or = 150 micromol/g, with 30 of 44 and 14 of 44 classification errors occurring when loin GP values were between 121 and 150 micromol/g and 70 and 120 micromol/g, respectively. Gene effects, based on DNA results, were evaluated using mixed-model procedures with fixed effects of DNA genotype and gender, and random sire and litter effects. No RN genotype differences for growth rate,10th-rib backfat, or loin muscle area were observed. Loins from the RN-/rn+ pigs had significantly (P < 0.05) lower ultimate pH (0.16 units), greater GP (50.3 micromol/g), greater drip loss (0.93%), paler objective color (L*, 1.66 units), paler visual color (0.31 units), and lower firmness (0.61 units) scores. Additionally, loins from RN-/rn+ pigs had significantly (P < 0.05) lower marbling scores (0.68 units) and intramuscular fat content (0.25%) and greater cooking loss (2.51%). Cooked moisture, juiciness score, and mechanical and sensory tenderness measures did not differ between genotypes. The GP-based classification did not correctly classify RN genotype in the present study, emphasizing the importance of the direct DNA analysis for estimation of gene frequency and effects. The DNA-based genotype results clearly indicate the RN- allele has negative effects on muscle quality measures.