Silencing acetyl-CoA carboxylase A and sterol regulatory element-binding protein 1 genes through RNAi reduce serum and egg cholesterol in chicken.

Cholesterol is synthesized in chicken through de novo lipid biosynthetic pathway where two most important genes viz. SREBP1 and ACACA play immense role. To minimize cholesterol synthesis, RNAi approach was adopted and accordingly, we developed transgenic chicken possessing ACACA and SREBP1 shRNA constructs, which showed lower level of ACACA and SREBP1 in serum. The serum total cholesterol, triglycerides, HDL and LDL cholesterol was significantly lower by 23.8, 35.6, 26.6 and 20.9%, respectively in SREBP1 transgenic birds compared to the control. The egg total cholesterol and LDL cholesterol content was numerically lower in both ACACA and SREBP1 transgenic birds by 14.3 and 13.2%, and 10.4 and 13.7%, respectively compared to the control. It is concluded that the protocol was perfected to develop transgenic chicken through RNAi for knocking down the expression of ACACA and SREBP1 proteins, which minimized the cholesterol and triglycerides contents in serum and eggs.

Effect of Supplementation of Fermented Yeast Culture on Hormones and Their Receptors on Exposure to Higher Temperature and on Production Performance after Exposure in Nicobari Chickens.

Heat stress (HS) affects the production performance in chickens and causes economic loss to the producers. Most of the studies have been conducted on and for the welfare of broilers. We still lack information on the physiological parameters being affected during chronic heat stress in layers. To fill this gap, the present study evaluated the effect of heat stress (induced in the chamber) during the prelaying period (21-23 weeks) on plasma levels of the hormones leptin and ghrelin and GH and expression of the respective receptors and heat stress markers. Three groups were considered, one at room temperature (CR) and the other two groups (SH and CH) subjected to heat stress at 39°C for four hours for three weeks (21-23 weeks of age). The SH group (SH) feed was supplemented with fermented yeast culture (FYC, 700 mg/kg), whereas the CH group was devoid of it. After that, all the groups were shifted to shed under natural ambient conditions till 31 weeks of age. Studies were restricted to production performance only. Feed offered without yeast culture (CH group) had a smaller concentration of plasma hormones (P < 0.01) and increased expression fold of the hormone receptors (P < 0.01). Further, the group also presented higher liver AMP kinase enzyme, plasma MDA (malondialdehyde), and cholesterol concentrations. These changes likely explained the decrease in feed intake and the CH group's body weight and further reduced the production performance during the laying period. Supplementation with FYC to birds had an opposite effect on the above-mentioned parameters, reducing HS effects. In summary, supplementation with FYC (700 mg/kg) maintained physiological parameters as in the CR group under HS conditions and negated adverse effects on parameters both before and during laying periods.

A comparative study on the antioxidant status, meat quality, and mineral deposition in broiler chicken fed dietary nano zinc viz-a-viz inorganic zinc.

Use of nano minerals in farm animal nutrition offers considerable advantages over inorganic or organic mineral sources. But, the conventional chemical synthesis of nano minerals suffers from disadvantage of possible environmental accumulation and pollution due to the non-biodegradable materials and chemicals. This study investigated the effects of green nano-zinc (GNZ) and market nano-zinc (MNZ) with respect to the inorganic zinc (IZ) on meat quality, antioxidant status, mineral deposition, and bone development in broiler chicken. Following a 3 × 3 factorial design, nine dietary treatments were formulated by employing three levels (40, 60, and 80 ppm) and three sources (inorganic, green nano, and market nano) of zinc viz. IZ-40, GNZ-40, MNZ-40, IZ-60, GNZ-60, MNZ-60, IZ-80, GNZ-80, MNZ-80. Six replicates of broiler chicken were assigned to each treatment with eight birds in each. The birds fed 80 ppm Zinc of either GNZ or MNZ source resulted in significantly higher serum SOD, glutathione peroxidase, catalase, zinc, calcium, and phosphorus levels; increased bone dimensions, weight, total ash, phosphorus, and zinc content along with higher liver and muscle zinc concentration. The meat of chicken fed 80 ppm zinc of MNZ source followed by GNZ source has shown significantly better antioxidant (DPPH and ABTS values) status and lower lipid peroxidation (free fatty acid and TBARS values). The 80 ppm zinc of either MNZ or GNZ source resulted in significantly lower fat and cholesterol content of chicken meat compared to lower Zn levels and IZ source. This study indicated that 80 ppm dietary zinc of either MNZ or GNZ source improved the antioxidant status, and reduced the meat cholesterol, fat content, and lipid peroxidation of chicken meat along with increased bone dimensions and mineralization.

Comparative study on the responses of broiler chicken to hot and humid environment supplemented with different dietary levels and sources of selenium.

Present study was carried out with the objective of investigating the role of green synthesized nano Se (GNS) in growth performance, digestibility of minerals, immunity, stress alleviation, antioxidant status, and body Se content of broiler chicken raised under hot and humid environment with respect to market nano Se (MNS) and inorganic Se. The experimental design was 3 × 3 factorial, in which three levels (0.15, 0.20, and 0.25 ppm) and three sources (inorganic, green nano, and market nano) of Se resulted in nine treatments viz. IS-0.15, GNS-0.15, MNS-0.15, IZ-0.20, GNS-0.20, MNS-0.20, IS-0.25, GNS-0.25, and MNS-0.25 (IS: inorganic Se, GNS: green nano Se, MNS: market nano Se). A total of 432 broiler chicken were divided among nine treatments with six replicates of birds per treatment (8 birds/replicate). Results of present study revealed significantly better growth performance of birds supplemented with 0.25 ppm nano Se. The supplementation of 0.25 ppm nano Se improved the immune response and lymphoid organ development of birds. Significantly higher Se and nitrogen digestibility coefficients, serum antioxidant activity and decline of Heterophil: Lymphocyte ratio and expression of HSP70 gene were observed in birds supplemented with 0.25 ppm Se and nano source of Se compared to inorganic Se. Significantly higher Se concentration in liver and breast muscle and higher serum Se concentration were observed in birds fed 0.25 ppm nano Se. The liver Se concentration was much higher than that of breast muscle. However, the nano Se synthesized by green method in this study did not differ significantly from the chemically synthesized nano Se. It was concluded that 0.25 ppm Se and nano form of Se are superior to lower levels and inorganic form of Se, respectively, in improving the immunity, growth, antioxidant status, and in stress alleviation of broiler chicken. However, GNS is equally efficient as chemically synthesized MNS.

Comparative analysis of silencing expression of myostatin (MSTN) and its two receptors (ACVR2A and ACVR2B) genes affecting growth traits in knock down chicken.

Myostatin (MSTN), a growth differentiation factor-8 regulates muscular development through its receptors, ACVR2A (Activin receptor type IIA) and ACVR2B (Activin receptor type IIB) by inhibiting cellular differentiation of developing somites during embryonic stage and diminishing myofibriller growth during post-embryonic period. The objective of this study was to compare the effect of knockdown of expression of myostatin, ACVR2A and ACVR2B genes on growth traits in chicken. The shRNAs for Myostatin, ACVR2A and ACVR2B genes were designed, synthesized and cloned in DEST vector. The recombinant molecules were transfected into the spermatozoa and transfected spermatozoa were inseminated artificially to the hens to obtain fertile eggs. The fertile eggs were collected, incubated in the incubator and hatched to chicks. Silencing of ACVR2B gene showed significantly higher body weight than other single, double and triple knock down of genes in transgenic birds. The carcass traits such as dressing%, leg muscle%, and breast muscle% were found with the highest magnitudes in birds with silencing of the ACVR2B gene as compared to the birds with that of other genes and control group. The lowest serum cholesterol and HDL content was found in ACVR2B silencing birds. The total RBC count was the highest in this group though the differential counts did not differ significantly among various silencing and control groups of birds. It is concluded that silencing of only one receptor of MSTN particularly, ACVR2B may augment the highest growth in chicken during juvenile stage. Our findings may be used as model for improving growth in other food animals and repairing muscular degenerative disorders in human and other animals.

Polymorphisms in GnRHI and GnRHII genes and their association with egg production and egg quality traits in chicken.

1. Two candidate genes, namely, Gonadotropin releasing hormone I (GnRHI) and Gonadotropin releasing hormone II (GnRHII) play pivotal roles in ovulation and egg production in chicken. The objective of this study was to explore polymorphism in these genes and to estimate the effects of polymorphism of these two genes on egg production and egg quality traits in White Leghorn laying hens. 2. Single strand conformation polymorphism followed by sequencing was performed to detect polymorphism in these genes. 3. The coding regions of the GnRHI and GnRHII genes were found to be polymorphic. In the GnRH1 gene, 12 haplotypes were determined, of which the h1 haplotype was predominant and the h5, h9 and h11 haplotypes were the least frequent ones. In the GnRHII gene, eight haplotypes were found, of which the h1 haplotype was the most frequent and the h6 was the least frequent haplotype in the White Leghorn population. 4. The haplogroups of GnRHI had a significant effect on body weight and egg production up to 64 weeks of age, yolk content, Haugh units and egg shell parameters. The h1h2 haplogroup of the GnRHI gene showed the highest egg production, with 211.0 ± 24.3 eggs up to 64 weeks of age, while the highest yolk content and Haugh unit was found in h3h10 haplogrouped birds. The haplogroups of GnRHII had a significant effect on age at sexual maturity (ASM) where the shortest ASM was found in the h1h4 birds (147.3 ± 5.9 d) and the longest ASM was observed in the h1h3 birds (160.6 ± 23.4 d). 5. It was concluded that GnRHI and GnRHII genes are polymorphic and have a significant effect on body weight, egg production and egg quality traits in White Leghorn laying hens.

Effect of dietary manipulation and vaccination of turkey breeder hens on immunoglobulin levels of yolk, yolk sac and neonate poults.

Two hundred turkey breeder hens and 24 viable toms of 30-35 weeks age of small white variety were distributed into two treatment groups having four replicates of 25 hens and three toms in each treatment. First four replicates were offered a turkey breeder diet (Diet A) (Nutrient requirements of poultry, 1994, National Academic Press, Washington, DC) and the rest four replicates were maintained on a higher plane of nutrition (Diet B) for 8-week duration. After 6 weeks of experimental feeding, two replicates from each treatment groups were vaccinated with ND (R2 B) vaccine. Yolk sac of embryo from birds fed Diet B had a significantly higher (p < .05) IgG, IgM level and HI titre (log 2) than those fed Diet A. HI titre values of embryonic yolk sac from the vaccinated birds fed Diet B were significantly higher (p < .05) than that of the control groups. In addition, HI titre values were significantly higher (p < .05) in the day-old poults of the birds fed Diet B than that of those fed Diet A. There was significantly (p < .01) positive correlation between serum IgG and IgM of the breeder birds and day-old chicks. Similarly, there was significantly (p < .05) positive correlation between yolk IgG and IgM after 1-month experimental feeding and yolk sac IgG and IgM. Positive correlation (p < .05) also existed between yolk sac IgM and day-old chick serum IgM. Furthermore, the HI titres of breeder birds' serum at 14 days post-vaccination were positively correlated with their egg yolk after 10 and 15 days post-vaccination, yolk sac and day-old chicks. Thus, the study envisaged that a higher immunity in neonate poults from turkey breeders maintained on a higher plane of nutrition may be elicited as there was maternal transfer of antibodies from the serum of breeder birds to their offsprings through their yolk sac.

In ovo carbohydrate supplementation modulates growth and immunity-related genes in broiler chickens.

A study was undertaken to investigate the role of in ovo administrated carbohydrates on the expression pattern of growth and immune-related genes. In ovo injections (n = 400) were carried out on the 14th day of incubation into the yolk sac/amnion of the broiler chicken embryos. Expression of growth-related genes: chicken growth hormone (cGH), insulin-like growth factor-I & II (IGF-I & II) and mucin were studied in hepatic and jejunum tissues of late-term embryo and early post-hatch chicks. Expression of candidate immune genes: Interleukin-2, 6, 10 and 12 (IL-2, IL-6, IL-10 and IL-12), Tumour necrosis factor-alpha (TNF-α) and Interferon gamma (IFN-γ) were studied in peripheral blood monocyte cells of in ovo-injected and control birds following antigenic stimulation with sheep RBC (SRBC) or mitogen concanavalin A (Con-A). Glucose injection significantly increased the expression of IGF-II gene during embryonic period and both cGH and IGF-II in early post-hatch period, while ribose-injected chicks had higher expression of IGF-II gene during embryonic stage. Enhanced mucin gene expression was also observed in fructose-injected chicks during embryonic age. Glucose-injected chicks had higher expression of IL-6 or IL-10, while those injected with fructose or ribose had higher expression of IL-2, IL-12 and IFN gamma. It is concluded that in ovo supplementation of carbohydrates might help in improving the growth of late-term embryos and chicks. In ovo glucose could modulate humoral-related immunity, while fructose or ribose might help in improving the cellular immunity in broiler chickens.

Microarray analysis of gene expression in parthenotes and in vitro-derived goat embryos.

The present work was carried out to investigate the global gene expression profile to search differentially expressed candidate transcripts between parthenogenetic and in vitro-fertilized (IVF) caprine morula. For this study, total RNA was isolated from diploid parthenogenetic and IVF embryos, and complementary DNA was synthesized. Microarray and relative real-time polymerase chain reaction analysis were performed to check global gene expression profile and validation, respectively. According to the microarray analysis, the total number of upregulated (UR) and downregulated (DR) genes was 613 and 220, respectively in diploid parthenogenetic morula as compared with IVF morula. The number of genes showing about two-, two- to five-, five- to 10-, 10- to 20-, and above 20-fold UR and DR genes was 147, 229, 122, 59, and 56 and 94, 73, 18, 13, and 22, respectively. Five UR genes validated (PTEN, PHF3, CTNNB1, SELK, and NPDC1) and all of them were significantly higher in parthenotes, which was in accordance with microarray results, whereas the expression of DR (AURKC and KLF15) genes were downregulated in parthenotes as observed in microarray results but the difference was not significant (P < 0.05). In conclusion, our findings demonstrate differential expression of a large number of genes in parthenotes compared with IVF embryos, which may be the reason for aberrant parthenogenetic embryo development in caprine species.

Effect of actin polymerization inhibitor during oocyte maturation on parthenogenetic embryo development and ploidy in Capra hircus.

This study was designed to observe the effect of cytochalasin B (CCB) concentrations on ploidy and early development of parthenogenetic embryos in a caprine species. Caprine oocytes were matured in the presence of different concentrations of CCB (5, 10, 15, and 20 µg/ml) and activated by 7% ethanol followed by incubation with 2 mM DMAP. For embryos fertilized in vitro, oocytes were matured in maturation medium without CCB. The cleavage rate and further embryo development were significantly higher (P < 0.05) when oocytes were treated in this way. The percentage of embryos showed higher diploid values in 15 µg/ml CCB (83.66 ± 1.13), followed by 20 (72.22 ± 1.22), 10 (68.57 ± 1.17), and 5 µg/ml (62.00 ± 2.48). These results indicate that CCB with a concentration of 15 µg/ml in maturation medium can be used for the production of diploid parthenogenetic embryos in the caprine species.

Effect of in ovo threonine supplementation on early growth, immunological responses and digestive enzyme activities in broiler chickens.

1. The effects of injecting threonine in ovo on early growth, some immunological responses and the activity of digestive enzymes of broiler chicks were investigated. Fertile eggs were distributed into 6 groups, each of 60. These were: untreated control, sham control, 10, 20, 30 or 40 mg threonine. Threonine was dissolved in 0.5 ml sterile saline and inoculated into the yolk sac of the 14-d-old embryo through the narrow end of the egg. 2. The ratio of chick to egg weight was 1.6% higher in the group given 30 mg threonine and at 28 d of age chicks receiving threonine were 29 to 79 g heavier than untreated controls. 3. Food conversion ratio until 7 d after hatching was improved in those chicks receiving 10, 20 or 40 mg threonine but there was no significant effect on the activities of amylase, pepsin or trypsin. 4. The humoral response to sheep red blood cells was significantly greater in those groups receiving 10, 20 or 30 mg threonine supplementation than in untreated controls. 5. The response to phytohaemagglutinin-P, a measure of the cell-mediated immune response, was not affected, however. 6. It is concluded that injections of 20 to 30 mg threonine into yolk sac can improve post-hatching growth and humoral responses of broiler chicks.

Effects of feeding raw or reconstituted high tannin red sorghum on nutrient utilisation and certain welfare parameters of broiler chickens.

1. A feeding trial was conducted on 360 1-d-old chicks from 0 to 6 weeks of age to assess the effect of processed high tannin red sorghum in the diet of broiler chickens on nutrient utilisation and certain welfare parameters. 2. Each of 9 dietary treatments was allotted to 4 groups (replicates) of chicks in a completely randomised design. The treatments were a maize-soy based standard broiler diet (control, RS(0)) and eight test diets formulated by incorporating either raw red sorghum (RS(25), RS(50), RS(75), RS(100)) or reconstituted red sorghum (RS(25)(R), RS(50)(R), RS(75)(R), RS(100)(R)) replacing 25, 50, 75 or 100% of maize from the control diet. 3. The tannin content was reduced from 23 to 16 g/kg in reconstituted red sorghum. 4. Utilisations of nitrogen, calcium and phosphorus retention were similar in all the dietary groups. 5. The cell-mediated immune response measured as footpad index (FPI) value of birds given raw red sorghum was significantly higher than in control and reconstituted red sorghum groups. Similarly, comparatively better humoral response measured as HA titre value was observed in raw red sorghum based groups than in reconstituted ones. 6. Plasma albumin, globulin, protein, glucose, calcium, phosphorus, SGOT, SGPT and uric acid levels did not differ significantly due to dietary treatments. 7. Mild histopathological changes were observed in liver and kidney tissues of birds given raw red sorghum. 8. From the present study, it is concluded that (1) the reconstitution of high tannin red sorghum resulted in about 30% reduction in its tannin concentration; (2) the feeding of reconstituted sorghum based diets to broiler chickens did not exert any appreciable influence on nutrient utilisation, blood biochemicals and enzymes and gross pathological changes; and (3) the birds fed on raw red sorghum exhibited higher immuno-responsiveness in comparison to their reconstituted counterparts.

Effect of microbial phytase on production performance of White Leghorn layers fed on a diet low in non-phytate phosphorus.

1. An experiment with 150 White Leghorn layers was conducted to examine the effect of microbial phytase supplementation of low non-phytate phosphorus (NPP) diets on egg production, eggshell quality, bone mineralisation and retention of nutrients at 32-48 weeks of age. 2. Four isonitrogenous and isocaloric diets were formulated to contain 1.2, 1.8, 2.4 and 3.0 g NPP/kg diet with the two lowest NPP (1.2 and 1.8) supplemented with microbial phytase (Biofeed Phytase, India) at 500 FTU per kg diet. Each diet was offered ad libitum to 5 replicates of 5 layers throughout the experiment. 3. Body weight gain was reduced significantly in the layers fed on the 1.2 g/kg NPP diet as compared to those given diets containing 1.8-3.0 g/kg. Addition of phytase to the 1.2 g/kg diet significantly enhanced the body weight and was comparable with those given diets containing 1.8-3.0 g/kg NPP. 4. No additional advantage resulted from enhancing the NPP levels beyond 1.8 g/kg or adding phytase to a diet containing 1.8 g/kg NPP. 5. Hen d egg production, food intake, food efficiency, shell weight, shell thickness, shell strength and tibia strength followed the same trends as above. However, adding phytase to the 1.8 g/kg NPP diet significantly enhanced tibia ash. Egg weight, specific gravity and Haugh units were influenced by neither NPP concentration nor phytase supplementation. 6. Adding phytase to the 1.2 g/kg NPP diet significantly enhanced nitrogen and phosphorus retention. 7. It was concluded that addition of 500 FTU of microbial phytase/kg diet can allow the reduction of NPP content to 1.2g/kg in the layer diet, eliminate inorganic phosphorus supplementation and results in significant reduction of nitrogen and phosphorus excretion without affecting the production performance of layers.