Identification of new genes and quantitative trait locis associated with growth curve parameters in F2 chicken population using genome-wide association study.

The markers which are correlated with the growth curve parameters help in understanding the characteristics of individual growth during the rearing of livestock. This study aimed to identify a set of biomarkers through a GWAS for growth curve parameters in crossbred chickens using the Illumnia 60K chicken SNP Beadchip. Growth data were collected from a total of 301 birds from cross of a broiler line and native chickens. Using the Gompertz-Laird model, two growth curve parameters, the instantaneous growth rate per day (L) and the coefficient of relative growth or maturing index (k), were estimated. The L and k were used to estimate five derived parameters, namely asymptotic (mature) body weight, body weight at inflection point, age at the inflection point, average growth rate and maximum growth rate. These parameters were considered as phenotypic values in the GWAS based on generalized linear models. The results of the GWAS indicated 21 significant markers, which were located near or within 46 genes. A number of these genes, such as GH, RET, GRB14, FTSJ3 and CCK, are important for growth and meat quality in chickens, and some of them are growth related in other species such as sheep and cattle (GPI, XIRP2, GALNTL6, BMS1, THSD4, TRHDE, SHISA9, ACSL6 and DYNC1LI2). The other genes are associated with developmental biological pathways. These genes are particuarly related to body weight, average daily gain and growth QTL. The results of this study can shed light on the genetic mechanism of biological functions of growth factors in broiler chickens, which is useful for developing management practices and accelerating genetic progress in breeding programs.

Supplementation effects of pomegranate by-products on oxidative status, metabolic profile, and performance in transition dairy cows.

Recent studies demonstrated a high antioxidant capacity for pomegranate components due to their rich bioactive compounds, such as conjugated fatty acids and phenolics. The objective of the present study was, therefore, to assess whether pomegranate seed or pomegranate seed pulp (peel + seed) supplementation could be effective to improve antioxidant status, and hence metabolic profile and performance in periparturient dairy cows. After a 1-wk pretreatment period, Holstein cows (primiparous n = 12, multiparous n = 18) were assigned to 3 dietary treatments from 25 d before expected calving through 25 d postcalving. The dietary treatments included (1) control (CON); (2) diet supplemented with pomegranate seeds (PS; 400 g/cow per day); and (3) diet supplemented with pomegranate seed pulp (PSP; 400 g of seeds/cow per day + 1200 g of peels/cow per day). Compared with CON, supplementation with either PS or PSP had no effects on dry matter intake, rumen fermentation, and plasma concentrations of cholesterol, total protein, globulin, and aspartate amino transferase, but enhanced plasma total antioxidant activity, and lowered triacylglycerol, free fatty acids (FFA), and ß-hydroxybutyrate at both pre- and postpartum periods. Plasma concentration of glucose, albumin, malondialdehyde (MDA) and blood superoxide dismutase (SOD) activity were not affected by dietary treatments at prepartum, whereas SOD activity increased and glucose, albumin, MDA, and FFA-to-albumin ratio decreased by feeding both by-products at postpartum period. In contrast to PS, supplementing PSP resulted in a greater decrease in plasma glucose and triacylglycerol concentration and higher increase in SOD activity. Energy- and fat-corrected milk yields were higher in cows fed PSP diet compared with those fed CON or PS diets, but content of milk fat, protein, and lactose were similar across the dietary treatments. These findings indicated that dietary pomegranate by-products supplementation, in particular PSP, could improve antioxidant status, which was associated with a decline in lipid oxidation (FFA and ß-hydroxybutyrate) and peroxidation (MDA) and an enhancement in glucose utilization as well as fat-corrected milk yield.

Effects of milk thistle meal on performance, ileal bacterial enumeration, jejunal morphology and blood lipid peroxidation in laying hens fed diets with different levels of metabolizable energy.

This study was conducted to evaluate the effects of different levels of milk thistle meal on performance, blood biochemical indices, ileal bacterial counts and intestinal histology in laying hens fed diets containing different levels of metabolizable energy. A total number of 200 Leghorn laying hens (Hy-Line W-36) were randomly assigned to eight experimental treatments with five cage replicates of five birds each. Dietary treatments consisted of four levels of milk thistle meal (0%, 15%, 30% and 60%) and two levels of AMEn (11.09 and 12.34 MJ/kg) fed over a period of 80 days. In vitro studies revealed that the total phenolic component of milk thistle meal was 470.64 mg gallic acid equivalent/g of the sample, and its antioxidant activity for inhibiting the 2-2-diphenyl-1-picrichydrazyl free radical and reducing ferric ions was about 21% higher than that of butylated hydroxyltoluene (p < .05). Diets containing high level of AMEn led to improved egg production (p < .05), egg weight (p < .05), egg mass (p < .01) and feed conversion ratio (p < .01). In addition, offering diets containing high energy significantly enhanced (p < .01) serum triglyceride and malondialdehyde (MDA) concentrations as well as jejunal villus height. Dietary supplementation of 3% milk thistle meal resulted in the best feed conversion ratio (p < .05), reduction of ileal Escherichia coli enumeration (p < .01) and an enhancement in the villus height-to-crypt depth ratio (p < .05). Furthermore, feeding incremental levels of this meal led to remarkable decrease in serum cholesterol, triglyceride and MDA (p < .01) concentrations while significant increase in blood high-density lipoprotein content and goblet cell numbers (p < .05). The present findings indicate that milk thistle meal with high antioxidant and antibacterial properties in laying hen diets may improve health indices and productive performance.

Effects of different levels of sanguinarine on antioxidant indices, immunological responses, ileal microbial counts and jejunal morphology of laying hens fed diets with different levels of crude protein.

This study was carried out to assess the effects of different levels of sanguinarine on antioxidant indices, immunological responses, serum biochemical parameters, ileal microbial counts and jejunal morphology of laying hens fed on diets with different levels of crude protein (CP). A total of 180 laying hens were subjected into nine dietary treatments with four cages of five birds each. Experimental treatments consisted of three levels of CP (85.0, 92.5 and 100% of Hy-Line W36 manual recommendation) and three levels of sanguinarine (0.00, 3.75 and 7.50 mg/kg) as a 3 × 3 factorial arrangement of laying hens which fed during a 70-day feeding trial. The in vitro study showed that sanguinarine exhibited sevenfold and threefold decreased antioxidant activities to inhibit 2-2-diphenyl-1-picric hydrazyl free radical as well as ferric ion reducing rather than butylated hydroxyl toluene. Although using the decremental levels of CP caused the increase in heterophil-to-lymphocyte ratio (p < 0.01), dietary administration of sanguinarine could suppress the serum cholesterol and malondialdehyde concentrations as well as heterophil-to-lymphocyte ratio (p < 0.05). Additionally, decreasing CP content resulted in the decreased percentage of albumin (p < 0.05); however, it had no negative effects on humoral immunity. Nonetheless, feeding of at least 3.75 mg/kg sanguinarine led to the remarkable increases in serum gamma globulin concentration (p < 0.01) and secondary (p < 0.05) antibody titres against sheep red blood cells. Moreover, a decline in dietary CP content led to higher villi height and crypt depth (p < 0.05; p < 0.001) and consequently decreased villi height-to-crypt depth ratio (p < 0.001) than the optimum level (100% CP). In spite of the effects of sanguinarine on the suppression of Escherichia coli and Salmonella counts (p < 0.05), it markedly enhanced villi height-to-crypt depth ratio as well as lamina propria lymphatic follicles extent, simultaneously (p < 0.001). Therefore, in spite of the detrimental effects of feeding low-CP diets on lymphocytes and serum albumin percentages, and villi height-to-crypt depth ratio, the administration of incremental levels of sanguinarine could improve cellular and humoral immunity, decrease ileal microbial counts and in turn improve the intestinal health indices in laying hens.

Genetic evaluation of calf and heifer survival in Iranian Holstein cattle using linear and threshold models.

Calf and heifer survival are important traits in dairy cattle affecting profitability. This study was carried out to estimate genetic parameters of survival traits in female calves at different age periods, until nearly the first calving. Records of 49,583 female calves born during 1998 and 2009 were considered in five age periods as days 1-30, 31-180, 181-365, 366-760 and full period (day 1-760). Genetic components were estimated based on linear and threshold sire models and linear animal models. The models included both fixed effects (month of birth, dam's parity number, calving ease and twin/single) and random effects (herd-year, genetic effect of sire or animal and residual). Rates of death were 2.21, 3.37, 1.97, 4.14 and 12.4% for the above periods, respectively. Heritability estimates were very low ranging from 0.48 to 3.04, 0.62 to 3.51 and 0.50 to 4.24% for linear sire model, animal model and threshold sire model, respectively. Rank correlations between random effects of sires obtained with linear and threshold sire models and with linear animal and sire models were 0.82-0.95 and 0.61-0.83, respectively. The estimated genetic correlations between the five different periods were moderate and only significant for 31-180 and 181-365 (r(g) = 0.59), 31-180 and 366-760 (r(g) = 0.52), and 181-365 and 366-760 (r(g) = 0.42). The low genetic correlations in current study would suggest that survival at different periods may be affected by the same genes with different expression or by different genes. Even though the additive genetic variations of survival traits were small, it might be possible to improve these traits by traditional or genomic selection.

Expression of bovine Ecat1 gene in immature and in vitro matured oocytes as well as during early embryonic development.

Ecat1 is a maternal effect gene that is exclusively expressed in oocytes and embryonic stem cells, and has an important role in pre-implantation development. This study was designed to investigate the expression of bovine Ecat1 gene in immature and in vitro matured oocytes as well as during early embryonic development, and also Ecat1 protein localization. Samples were obtained from slaughtered animals. RNA extractions were carried out from ovary, immature and in vitro matured oocytes and also different stages of embryonic development (2-, 4-, 8- to 16-cell stages and blastocysts). RT-PCR analysis revealed the expression of Ecat1 in ovary, oocytes and embryos. Analysis in FGENESH online tool predicted three exons and one transcription start site (TSS) in Ecat1 gene, and the 3' RACE-PCR result showed that just one splice variant was amplified. By quantitative real-time PCR technique, we showed that Ecat1 transcript increased at 8- to 16-cell-stage embryos and decreased in blastocyst stage (p < 0.05). Immunofluorescence analysis showed cytoplasmic localization of Ecat1 protein in bovine oocytes. Results demonstrated bovine Ecat1 expression at protein level and also indicated that Ecat1 has a significant higher embryonic expression at 8- to 16-cell stage. This embryonic expression is probably required for further developmental stages.

Effect of LEPR, ABCG2 and SCD1 gene polymorphisms on reproductive traits in the Iranian Holstein cattle.

During the last decades, genetic selection for milk production traits has led to increased fertility and health problems in dairy cattle. The aim of this study was to investigate the impact of three polymorphisms located in the ATP-binding cassette superfamily G member 2 transporter (ABCG2), stearoyl-CoA desaturase 1 (SCD1) and leptin receptor (LEPR) genes on reproductive traits and somatic cell count (SCC). The analysis was conducted on 408 randomly selected cows. The SNPs within the genes (LEPR, ABCG2 and SCD1) were genotyped using the PCR-RFLP method. All three possible genotypes were observed for SCD1-T878C and LEPR-T945M SNPs, but not for ABCG2-Y581S SNP. LEPR-T945M and ABCG2-Y581S SNPs had no statistically significant effect on the studied reproductive traits and SCC. However, SCD1-T878C SNP were negatively and significantly related to pregnancy length, dry days and open days (p < 0.05), which lead to decreased profitability in dairy herds. The results suggest that the T878C SNP of SCD1 might be useful as a DNA marker to decrease reproductive problems and improve production traits in Iranian Holstein dairy cows.

Fine mapping quantitative trait loci under selective phenotyping strategies based on linkage and linkage disequilibrium criteria.

In fine mapping of a large-scale experimental population where collection of phenotypes are very expensive, difficult to record or time-demanding, selective phenotyping could be used to phenotype the most informative individuals. Linkage analyses based sampling criteria (LAC) and linkage disequilibrium-based sampling criteria (LDC) for selecting individuals to phenotype are compared to random phenotyping in a quantitative trait loci (QTL) verification experiment using stochastic simulation. Several strategies based on LAC and LDC for selecting the most informative 30%, 40% or 50% of individuals for phenotyping to extract maximum power and precision in a QTL fine mapping experiment were developed and assessed. Linkage analyses for the mapping was performed for individuals sampled on LAC within families and combined linkage disequilibrium and linkage analyses was performed for individuals sampled across the whole population based on LDC. The results showed that selecting individuals with similar haplotypes to the paternal haplotypes (minimum recombination criterion) using LAC compared to random phenotyping gave at least the same power to detect a QTL but decreased the accuracy of the QTL position. However, in order to estimate unbiased QTL parameters based on LAC in a large half-sib family, prior information on QTL position was required. The LDC improved the accuracy to estimate the QTL position but not significantly compared to random phenotyping with the same sample size. When applying LDC (all phenotyping levels), the estimated QTL effect were closer to the true value in comparison to LAC. The results showed that the LDC were better than the LAC to select individuals for phenotyping and contributed to detection of the QTL.

Across-family marker-assisted selection using selective genotyping strategies in dairy cattle breeding schemes.

This study investigated the potential loss expected from marker-assisted selection (MAS) when only a proportion of animals are genotyped using several selective genotyping strategies. A population resembling a commercial dairy cattle population over 25 yr was simulated, and the most informative individuals for genotyping were identified among the potential breeding candidates (young bulls and bull-dams). Two strategies were used to identify the most informative animals. The first genotyping strategy was based on selecting individuals for genotyping with predicted total genetic effect [sum of the predicted quantitative trait locus (QTL) and polygenic effects] close to the truncation point for selection. The second strategy used an index that extended the previous strategy to include the variance due to segregation of the QTL in the parents. The 2 strategies for selective genotyping were applied at the 2 different genotyping levels and compared with random selection of candidates for genotyping and complete genotyping of the potential candidates. All selective genotyping strategies at the same proportion of genotyping showed similar cumulative genetic level. The frequency of the favorable QTL allele increased faster with more animals genotyped. Extra response in total genetic effect (polygenic and QTL) was not significantly different between genotyping all candidates (100%), 20%, and 50% genotyping (except for yr 13), but all MAS strategies resulted in significantly higher response than BLUP until yr 18. With 50% (20%) genotyping of candidates for selection within a population, 95% (89%) of maximum cumulative QTL response was achieved in yr 13. All MAS schemes resulted in a 19% decrease in the rate of inbreeding compared with the BLUP scheme. Therefore, it is possible to use selective genotyping in practical dairy cattle breeding and decrease the genotyping costs with a minimal loss of response compared with complete genotyping of the potential candidates.

Combined use of phenotypic and genotypic information in sampling animals for genotyping in detection of quantitative trait loci.

Conventional selective genotyping which is using the extreme phenotypes (EP) was compared with alternative criteria to find the most informative animals for genotyping with respects to mapping quantitative trait loci (QTL). Alternative sampling strategies were based on minimizing the sampling error of the estimated QTL effect (MinERR) and maximizing likelihood ratio test (MaxLRT) using both phenotypic and genotypic information. In comparison, animals were randomly genotyped either within or across families. One hundred data sets were simulated each with 30 half-sib families and 120 daughters per family. The strategies were compared in these datasets with respect to estimated effect and position of a QTL within a previously defined genomic region at genotyping 10, 20 or 30% of the animals. Combined linkage disequilibrium linkage analysis (LDLA) was applied in a variance component approach. Power to detect QTL was significantly higher for both MinERR and MaxLRT compared with EP and random genotyping methods (either across or within family), for all the proportions of genotyped animals. Power to detect significant QTL (alpha = 0.01) with 20% genotyping for MinERR and MaxLRT was 80 and 75% of that obtained with complete genotyping compared with 70 and 38% genotyping for EP within and across families respectively. With 30% genotyping, the powers were 78, 83, 78 and 58% respectively. The estimated variance components were unbiased in EP strategies (within and across family), only when at least 30% was genotyped. To decrease the number of genotyped individuals either MinERR or MaxLRT could be considered. With 20% genotyping in MinERR, the estimated QTL variance components were not significant compared with complete genotype information but all studied strategies at 20% genotyping overestimated the QTL effect. Results showed that combining the phenotypic and genotypic information in selective genotyping (e.g. MinERR and MaxLRT) is better than using only the EPs and the combined methods can be considered as alternative approaches to decrease genotyping costs, with unbiased QTL effects, decreased sampling variance of the QTL variance component and also increased the power of QTL detection.