Estimation of breeding value, genetic parameters and maternal effects of economic traits in rural male parent line chicken using pedigree relationships in an animal model.

Breeding value (BV), genetic parameters and additive genetic, and maternal effects were evaluated on growth and production traits utilizing data from eight generations employing animal model in a rural male parent line (PD-6) chicken at ICAR-Directorate of Poultry Research, Hyderabad, India. The least squares means (LSM) for body weight (BW) and shank length (SL) up to 6 weeks of age varied significantly (p ≤ .01) among the generations and hatches. BW increased significantly (p ≤ .01) over the generations and decreased with the hatches. Sex also had a significant effect on BW and shank length except for BW at 0 day (BW0). LSM for BW (BW6) and Shank length (SL6) at 6 weeks of age were 598.84 ± 0.79 g and 74.57 ± 0.04 mm, respectively. Males recorded significantly (p ≤ .01) higher BWs and shank length. All the production traits were significantly (p ≤ .01) influenced by the generation effect. The overall LSM for age at sexual maturity (ASM), egg production at 40 weeks (EP40) and egg weight at 40 weeks (EW40) were 164.93 ± 0.23 days, 74.66 ± 0.40 eggs and 54.79 ± 0.08 g, respectively. Model 3 with additive, maternal permanent environmental and residual effects was the appropriate model for BW2, BW4, BW6, SL4 and SL6, whereas Model 4 with maternal effects was the best for BW0. The heritability estimates for BW6 and SL6 were 0.22 ± 0.02 and 0.18 ± 0.02, respectively. Model 1 with additive direct and residual effects was the best appropriate model for all the production traits. The heritability estimates of EP40 and EW40 were 0.16 ± 0.04 and 0.34 ± 0.05, respectively. BW and shank length were highly correlated with significant (p ≤ .05) positive association from different components. The correlation coefficient from direct additive component between egg production and BW40 was negative, while it was positive with less magnitude between egg production and BW20. The egg production and egg weights had a negative association at different ages. BV of SL6, the primary trait of selection, was significant (p ≤ .05) across the generations and increased linearly with an average genetic gain of 1.05 mm per generation. BV of BW6 was also significant (p ≤ .05) and increased linearly as correlated response with an average genetic response of 22.34 g per generation. BV of EP40 showed an increasing trend with a genetic gain of 0.02 eggs per generation. The EW 40 also increased linearly with an average genetic gain of 0.06 g. The average inbreeding coefficient of the population was 0.015. The study concluded that the population was in ideal status with a linearly increasing trend of average BV with negligible inbreeding over the eight generations of selection.

Variance component analysis of growth and production traits in Vanaraja male line chickens using animal model.

OBJECTIVE: A comprehensive study was conducted to study the effects of partition of variance on accuracy of genetic parameters and genetic trends of economic traits in Vanaraja male line/project directorate-1 (PD-1) chicken. METHODS: Variance component analysis utilizing restricted maximum likelihood animal model was carried out with five generations data to delineate the population status, direct additive, maternal genetic, permanent environmental effects, besides genetic trends and performance of economic traits in PD-1 chickens. Genetic trend was estimated by regression of the estimated average breeding values (BV) on generations. RESULTS: The body weight (BW) and shank length (SL) varied significantly (p≤0.01) among the generations, hatches and sexes. The least squares mean of SL at six weeks, the primary trait was 77.44±0.05 mm. All the production traits, viz., BWs, age at sexual maturity, egg production (EP) and egg weight were significantly influenced by generation. Model four with additive, maternal permanent environmental and residual effects was the best model for juvenile growth traits, except for zero-day BW. The heritability estimates for BW and SL at six weeks (SL6) were 0.20±0.03 and 0.17±0.03, respectively. The BV of SL6 in the population increased linearly from 0.03 to 3.62 mm due to selection. Genetic trend was significant (p≤0.05) for SL6, BW6, and production traits. The average genetic gain of EP40 for each generation was significant (p≤0.05) with an average increase of 0.38 eggs per generation. The average inbreeding coefficient was 0.02 in PD-1 line. CONCLUSION: The population was in ideal condition with negligible inbreeding and the selection was quite effective with significant genetic gains in each generation for primary trait of selection. The animal model minimized the over-estimation of genetic parameters and improved the accuracy of the BV, thus enabling the breeder to select the suitable breeding strategy for genetic improvement.

Molecular characterization and computational structure prediction of activin receptor type IIB in aseel and broiler chicken.

The present study was formulated to characterize and comprehend the molecular structural characteristics of ACTRIIB receptor in Aseel and control broiler (CB) populations. The full length coding sequence (1539 bp) of the receptor was amplified, cloned, sequenced and analyzed using bioinformatic tools. The physico chemical properties of protein and structural features like secondary structure, solvent accessibility and disorder regions were computed. The 3D structure was predicted by I-TASSER and evaluated by Ramachandran Plot and tools under Structural Analysis and Verification Server. The nucleotides differences between CB and Aseel were c. [156G > A; 210 T > C; 493C > T; c.520G > C; 665A > C; 686G > A; 937C > G; 1011A > C; 1130A > G; 1208 T > A; 1326 T > C; 1433 T > C]. The amino acid substitutions between CB and Aseel were p. [(Pro165Ser; Glu174Gln; Gln222Pro; Ser229Asn; His313Asp; Gln377Arg; Val403Asp; and Ile478Thr)]. While, the silent changes includes p. [(Lys53=; Glu71=; Leu337=; Asp442=)]. The molecular weight of mature protein was predicted to be 55.51 kDa and 57.80 kDa in Aseel and CB, respectively. The higher rank 3D model had a C-score of -1.60 in Aseel and - 1.41 in CB, while the estimated TM-score (0.54 ±â€¯0.14) and RMSD (5.8 ±â€¯1.2 Å) were found to be similar in Aseel and CB. Among the 512 residues, >90% were in favored region, 4.7% in allowed region and <1.5% in disallowed region in both Aseel and CB. The pattern of contact map was comparable in Aseel and CB. The Hydrogen bond plots of the Aseel and CB shared similar secondary structure pattern. The ACTRIIB protein was predicted to interact with ACVR1B, ACVR1C, INHBA, SMAD 1,2,5,7 & 9 and BMPR1A&B. Clustal and phylogenetic analysis implied that both the lines were closely related and formed a sub cluster with in avian cluster. The current research provides insights about structural and functional aspects of the receptor and also aids in understanding the evolutionary history of ACTRIIB.

Genetic polymorphism in core promoter sequence of ACTRIIB gene and association analysis with growth traits in chicken.

Molecular breeding exploiting candidate genes is burgeoning reproductive approach to improve growth traits in poultry. The activin type IIB receptor (ACTRIIB) is a negative growth regulator, modulating action of many muscle growth regulators. PCR-single-strand conformation polymorphism was employed to unravel polymorphism in promoter region of the ACTRIIB gene and delineate its association with growth traits in Aseel and control broiler (CB). Analysis of 5' promoter region (1122bp) of ACTRIIB gene identified five SNPs, that is g. [56 G > C (SNP1), 352A > C (SNP2), 580G > A (SNP3), 625C > T (SNP4) and 962C > T (SNP5)] at SMAD, paired box 7 homeodomain binding motif, GC box and bHLH-PAS type transcription factors in CB and Aseel. CB had significantly higher body weight (BW) and average daily gain (ADG) at all SNP sites, except at SNP 1. The haplotype construction resulted 8 haplotypes in CB and Aseel population. The BW and ADG differed significantly (p < .05) at all ages in CB and Aseel. The diplotypes H1H8 and H1H4 manifested higher BW and ADG, while diplotypes H3H8 and H3H7 displayed BW and ADG at each age in both lines (p < .05). Aseel exhibited higher expression of ACTRIIB gene than CB by 70.17, 4.83, 1.41, 2.38, 5.13, 1.20, 2.90, 6.53 and 11.75 times for h1h2, h1h3, h1h4, h1h6, h1h7, h1h8 h3h4, h3h7 and h3h8, respectively. The H3H8 and H3H7 diplotypes exhibited higher level of mRNA and protein than H1H8 and H1H4. The regulatory upstream region of ACTRIIB gene demonstrates high degree of genetic diversity and can be harnessed as potential marker in genetic selection programmes for increasing meat production.

In silico prediction of short hairpin RNA and in vitro silencing of activin receptor type IIB in chicken embryo fibroblasts by RNA interference.

Gene silencing by RNA interference is extensively used reverse genetic approach to analyse the implications of any gene in mammalian systems. The silencing of the Activin type IIB receptor belonging to transforming growth factor beta superfamily has demonstrated increase in muscle growth in many species. We designed five short hairpin RNA constructs targeting coding region of chicken ACTRIIB. All the shRNAs were transfected into chicken embryo fibroblast cells and evaluated their silencing efficiency by real time PCR and western blotting. Initially the computational analysis of target region and shRNA constructs was undertaken to predict sequence based features (secondary structures, GC% and H-b index) and thermodynamic features (ΔGoverall, ΔGduplex, ΔGbreak-target, ΔGintra-oligomer, ΔGinter-oligomer and ΔΔGends). We determined that all these predicted features were associated with shRNA efficacy. The invitro analysis of shRNA constructs exhibited significant (P < 0.05) reduction in the levels of ACTRIIB at mRNA and protein level. The knock down efficiency of shRNAs varied significantly (P < 0.001) from 83% (shRNA 1) to 43% (shRNA 5). All the shRNAs up regulated the myogenic pathway associated genes (MyoD and MyoG) significantly (P < 0.05). There was significant (P < 0.05) up-regulation of IFNA, IFNB and MHCII transcripts. The ACTRIIB expression was inversely associated with the expression of myogenic pathway and immune response genes. The anti ACTRIIB shRNA construct 1 and 3 exhibited maximum knock down efficiency with minimal interferon response, and can be used for generating ACTRIIB knockdown chicken with higher muscle mass.

In vivo cell-mediated immune, hemagglutination inhibition response, hematological and biochemical values in native vs. exotic chicken breeds.

Birds (364) of both sexes, 11-wk-old, belonging to 2 native (Brown Nicobari and Ghagus) breeds and 1 exotic breed (Dahlem Red) were evaluated for cell-mediated immune response (CMI) by phytohemagglutinin-P (PHA-P), hemagglutination inhibition (HI) assay against Newcastle disease virus (NDV) antigen (LaSota stock virus), flow cytometric analysis of CD8+ cytotoxic T lymphocytes (CTLs), and hematology and biochemical assays. The cutaneous basophil hypersensitivity response PHA-P% increase in wattle thickness (mm) was highest in Ghagus (431.14 ± 22.56) which differed significantly with that of Brown Nicobari (269.1 ± 22.66) and Dahlem Red (218.42 ± 22.30). Sex-wise observation showed that females are having significantly higher response than males. Hemagglutination inhibition test was performed to determine the serum antibodies against Newcastle disease (ND) virus. Brown Nicobari showed highest HI antibody titer than Ghagus and Dahlem Red to similar vaccination program after booster NDV dose. Flow cytometry assay revealed significantly higher CTLs proliferation in Brown Nicobari than Ghagus and Dahlem Red. Moreover, CTLs were found to be higher in control group than the treatment group. Other hematological parameters (103/µL) significant difference was found in white blood cell count between Dahlem Red (38.41 ± 1.03) with that of Brown Nicobari (35.28 ± 1.04) and Ghagus (34.57 ± 1.04) in treatment groups. Same trend was observed in the Lymphocyte treatment group. However, in Granulocyte treatment group, Brown Nicobari (11.04 ± 0.35) was found to be significantly different from Dahlem Red (8.68 ± 0.34) and Ghagus (9.27 ± 0.35). Correlations between body weight at 11 wk of age and CMI, HI, cytotoxic T cell were -0.093, 0.047, and -0.036, respectively. Egg weight was found to be positively correlated with that of chick weight. Serum biochemical values showed that Dahlem Red was having significantly higher creatinine levels compared to Ghagus. Triglycerides level was also significantly higher in Ghagus compared to Dahlem Red. No significant breed effect was observed for alkaline phosphate, aspartate transaminase, and alanine transaminase. Cholesterol and total serum protein levels were significantly higher in Dahlem Red compared to Brown Nicobari.

Expression Profiling of Activin type IIB Receptor During Ontogeny in Broiler and Indigenous Chicken.

Augmenting the meat production is among the primary breeding objective of genetic selection programs in poultry production. However, the knowledge about the expression of genes regulating muscle growth at the molecular level is inadequate. Activin type IIB receptor (ACTRIIB) has been reported to play vital role in the negative regulation of muscle growth by binding to multiple members of transforming growth factor-ß superfamily. The present investigation was carried out to comprehend the trend of ACTRIIB messenger RNA in pectoralis major muscle during embryonic (E5-20) and post embryonic age (days 1, 14, 28, and 42) in both Control Broiler (CB) and Aseel by using Real-time PCR. The expression profile of ACTRIIB gene displayed a similar trend in CB and Aseel, however Aseel showed significantly (P < 0.001) higher transcription throughout the period. The fold change in expression of ACTRIIB in Aseel relative to CB varied from 3.94 to 14.72 folds and 3.28 to 7.14 folds during embryonic and post embryonic age, respectively. ACTRIIB exhibited its peak on E7, E11, and E16 during embryonic age, which coincides with the formation of primary and secondary muscle fibers in both lines. While at the time of post-embryonic age, ACTRIIB showed highest transcription on day 1 and lowest transcription on day 28 in both CB and Aseel. Within each line, the expression of ACTRIIB differed significantly (P < 0.001) between days in the course of embryonic and post-embryonic period. ACTRIIB gene expression had significant (P < 0.05) effect on all carcass traits except neck weight. Our results suggest that Aseel expressed higher levels of ACTRIIB transcript than CB. The study inferred that expression pattern of ACTRIIB was analogous in both CB and Aseel, which might imply that molecular mechanisms underlying muscle development and regulation are comparable in nature.