Evaluation of the potential of extract of seaweed Eucheuma denticulatum as an alternative to antibiotic growth promoter in broiler chickens.

The seaweeds are in focus for their immunity and gut health-stimulating potentials in humans and farm animals, but their potential as a gut health-promoting agent and performance booster to replace antibiotic growth promoters (AGP) in broiler chicken-feed remains to be evaluated. In vivo feeding experiments were conducted on commercial broiler chickens (1-42 days post-hatch) to evaluate dried aqueous exact of red seaweed Eucheuma denticulatum (referred to as PBD 5). Each of the three test diets (basal diet with three dosing regimens of PBD5, 0.25 g kg -1 for 0-6 weeks, 0.25 g kg -1 for 0-4 weeks or 1.0 g kg -1 for 0-2 weeks), along with an AGP supplemented diet (Virginiamycin (V), 20 ppm in basal diet), and a control diet was fed to 13 pen replicates of five chicks in each. PBD5 at 1.0 g kg -1 diet for 0-2 weeks improved (P < 0.05) cumulative feed efficiency (4.65 % improvement at 28 d, and 3.74 % at 35 d) than the control and comparable to the V group and the trend in improvement persisted up to 42 d. The group fed with PBD5 @ 1.0 g kg -1 for 0-2 weeks had significantly (P < 0.05) higher serum IgG level, glutathione peroxidase levels, fat digestibility, and expression of occludin and avian beta-defensin 4 gene in the gut and a trend of increased expression of growth hormone receptor gene in the liver as compared to the control with no significant effect on body weight, phytohemagglutinin response or haemagglutination inhibition titer. At d 25 of age, fecal E. coli count was significantly (P < 0.01) lower in the seaweed extract groups and the V group as compared to the control. It can be concluded that dried aqueous extract of E. denticulatum at 1 g kg -1 diet for 0-2 weeks can be used as an alternative to antibiotic growth promoter in broiler chickens to improve feed efficiency and reduce gut pathogen load, and the improved performance was associated with increased expression of gut immunity and growth hormone receptor genes.

Carcass Characteristics, Meat Quality and Nutritional Composition of Kadaknath, a Native Chicken Breed of India.

The study was carried out to investigate the carcass and meat quality traits and nutritional profile of the meat of the Kadaknath, a unique native chicken breed in comparison with commercial broilers. The yield of the carcass, breast and giblets of the Kadaknath was lesser (p < 0.01), while that of the legs, wings, back, and neck was higher (p < 0.01) than broilers. The meat of the Kadaknath was significantly (p < 0.0001) darker (42.44, 50.92) and more yellow (6.23, 8.99) than broilers. The decline in pH of the meat was lower (p < 0.001) in the Kadaknath compared to broilers. Kadaknath meat had more protein and less fat, moisture and ash content than broilers (p < 0.01). Furthermore, it was richer (p < 0.01) in 11 amino acids, including those which are known to impart a sweet and umami taste, than the meat of broilers (3 amino acids). Both genotypes were almost similar in meeting the daily requirements of indispensable amino acids of adult human. The study concluded that the Kadaknath differed in carcass and meat quality characteristics from the broilers, and the nutritional quality of Kadaknath meat in terms of high protein and less fat and higher content of amino acids (tasty type) was better in Kadaknath meat as compared to broiler meat.

Genetic parameters of growth traits, trend of production and reproduction traits, and meat quality status of Ghagus, an indigenous chicken of India.

Indigenous chickens play a vital role in providing nutritional security and economic support for the rural and tribal people in tropical countries like India. Ghagus, an indigenous chicken breed of India, was characterized for growth, production, and reproduction (fertility and hatchability) traits over different generations and for carcass and meat quality traits. Heritability, and genetic and phenotypic correlations among growth traits were estimated using mixed model least squares and maximum likelihood methods. Estimates of heritability of growth traits such as bodyweights at different age and 8-week shank length were high (0.32 to 0.39) in initial generation and moderate to high during latest generation (0.20 to 0.42). The correlations of growth traits with 40-week egg production were positive and significant (P < 0.001). There was improvement in 8-week bodyweight (559.3 ± 4.40 g) by 177 g over the generations. The average egg production up to 40 weeks over the generations was 35.66 eggs. There was also a significant (P < 0.001) increase in egg weights over the generations. The fertility (mean 89.3%) ranged from 85.2 to 92.3%. The hatchability on fertile egg set (mean 90.8%) ranged from 86.5 to 94.1% and it was improved by 3.9% over the generations. The carcass and meat quality study revealed that this breed has the lean meat, high protein, and attractive meat color with desirable meat quality characteristics. The study concluded that the Ghagus breed with self-propagation capacity has the potential to be improved as an indigenous meat-type breed to meet the ever-increasing demand for indigenous chickens' meat in the country.

Genetic diversity at major histocompatibility complex and its effect on production and immune traits in indigenous chicken breeds of India.

The genetic diversity at major histocompatibility complex (MHC) in indigenous chicken breeds of India (Ghagus and Nicobari) in comparison with the White Leghorn (WLH) breed was investigated by genotyping the MHC-linked LEI0258 marker. Altogether 38 alleles and 96 genotypes were observed among three breeds. The observed and effective alleles were highest in Ghagus (23, 8.3) followed by Nicobari (14, 3.2) and WLH (10 and 2.2) breeds. The size of alleles ranged from 193 to 489 bp in Ghagus, 193 to 552 bp in Nicobari and 241 to 565 bp in the WLH breed. The number of private alleles was also highest in Ghagus (18) followed by Nicobari (8) and WLH (5) breeds. The most frequent allele was 261 bp in WLH (66 %), 343 bp in Nicobari (50.4 %) and 309 bp in the Ghagus (28.15 %) breed. Observed and expected heterozygosities were highest in Ghagus (0.83, 0.88) followed by Nicobari (0.58, 0.68) and WLH (0.53, 0.54). The genetic distance (Nei) between Ghagus and Nicobari breeds (2.24) was higher as compared to that of Ghagus and WLH (1.23) and that between Nicobari and WLH breeds (0.89). Association analysis revealed significant influence of MHC alleles on body weight, egg production in Ghagus and WLH breeds and antibody titres to Newcastle disease vaccine in the Nicobari breed.

Polymorphisms in pattern recognition receptor genes of indigenous and White Leghorn breeds of chicken.

Functional polymorphisms in pattern recognition receptors (PRRs) modulate innate immunity and play a crucial role in resistance or susceptibility to diseases. The present study was carried out to explore polymorphic patterns in the coding sequences of PRR genes TLR3, TLR1LA (TLRs), MDA5, LGP2 (RLRs) and NOD1 (NLR) in chicken breeds of India, namely Ghagus (GH), Nicobari (NB) and the exotic White Leghorn (WLH) breed. Out of 209 SNPs observed in five genes among three breeds, 117 were synonymous (Syn) and 92 were non-synonymous (NS) SNPs. In TLR genes the highest polymorphism was observed in NB (16, 28) compared to GH (14, 16) and WLH (13, 19) breeds. In the MDA5 gene the highest polymorphism was observed in GH (12) compared to NB (eight) and WLH (four) breeds. However, an almost similar level of polymorphism was observed in the LGP2 gene among the three breeds. In the NOD1 gene, the highest polymorphism was observed in NB (27), followed by WLH (11) and GH (10) breeds. The overall highest number of SNPs was observed in NB (90), followed by GH (62) and the WLH (57) breed. With regard to variation in polymorphism among different classes of PRRs, the study revealed the highest polymorphism in TLRs compared to NOD1 and the RLR class of PRRs. Further, the domain locations of various Syn and NS SNPs in each PRR among the three breeds were identified. In silico analysis of NS SNPs revealed that most of them had a neutral effect on protein function. However, two each in TLR1LA and LGP2 and one in the MDA5 gene were predicted to be deleterious to protein function. The present study unravelled extensive polymorphism in the coding sequences of the TLR and NLR class of PRR genes, and the polymorphism was higher in indigenous chicken breeds.

Toll-like receptors gene expression in the gastrointestinal tract of Salmonella serovar Pullorum-infected broiler chicken.

Salmonella enterica serovar Pullorum causes substantial mortality in chicks as well as results in persistent infection and vertical transmission in layer birds. An effective innate immune response in the early stages of infection could reduce bacterial colonization and mortality in chicks and persistency of infection in later stages. Toll-like receptors (TLRs), important components of innate immune response, plays a pivotal role in early recognition of pathogen as well as in the initiation of robust and specific adaptive immune response. In the present study, we quantified the expression levels of chicken TLRs (1LA, 1LB, 2A, 2B, 3, 4, 5, 7, 15, and 21) mRNA by quantitative real-time PCR in the gastrointestinal (GI) tissues (duodenum, jejunum, ileum, and cecum) of 3-day-old broiler chicks after 24 h of oral infection with S. enterica serovar Pullorum. We found significant upregulation of TLRs (TLR2, TLR4, TLR21) mRNA expressions in GI tract tissues after S. Pullorum infection. The exceptions were for TLR3 and TLR15 with decrease in the expression levels in the jejunum after infection. TLR4 gene expression was significantly (P < 0.05) upregulated in the duodenum and ileum of infected chicks. Gene expression for some of the TLRs (TLR1LA, ILB, 2B, and TLR5) remained unchanged after infection with S. Pullorum in all the GI tissues studied. Most substantial change in gene expression was found for TLR21, being significantly (P < 0.05) upregulated in all the tissues investigated. The differential expression levels of TLRs shed light on tailored innate immune response induced by S. Pullorum during the early stages of infection in chicks.

Expression analysis of turkey (Meleagris gallopavo) toll-like receptors and molecular characterization of avian specific TLR15.

Toll-like receptors (TLRs) constitute a multi-gene family, which plays a pivotal role in sensing invading pathogens by virtue of conserved microbial patterns. TLR repertoire of chicken and zebra finch has been well studied. However TLR family of other avian species is yet to be characterized. In the present study, we identified TLR repertoire of turkey, characterized avian specific receptor TLR15 in turkey and profiled the TLRs expressions in a range of tissues of turkey poults. All ten TLR genes orthologous to chicken TLR repertoire were found in turkey. Turkey TLR genes showed 81-93 % similarity at amino acid level to their chicken counter parts. Phylogenetic analysis confirmed the orthologous relationship of turkey TLRs with chicken and zebra finch TLRs. Open reading frame of turkey TLR15 was 2,607 bp long encoding 868 amino acids similar to that of broiler chicken and showed 92.4, 91.1 and 69.5 % identity at amino acid levels with chicken, Japanese quail and zebra finch TLR15 sequences respectively. Overall TLR expression was highest for TLR4 and lowest for TLR21. TLR1A, 2A, 2B and 21 were significantly higher in liver than other tissues investigated (P < 0.01). TLR3 expression was significantly higher in bone marrow (BM) and spleen in comparison to other tissues studied (P < 0.01). Furthermore, no significant differences in the expression levels of TLR1B, 4, 5, 7 and 15 genes were detected among the tissues studied. Our findings contribute to the characterization of innate immune system of birds and show the innate preparedness of young turkey poults to a range of pathogens.

Differential gene expression of antimicrobial peptides ß defensins in the gastrointestinal tract of Salmonella serovar Pullorum infected broiler chickens.

Salmonella enterica serovar Pullorum causes substantial mortality in chicks as well as results in persistent infection and vertical transmission in layer birds. An effective innate immune response in the early stages of infection could reduce bacterial colonization and mortality in chicks and persistency of infection in later stages. ß Defensins (AvBDs) are now considered as one of the key components of innate immunity in avian species. In the present study, we quantified the mRNA expression levels of AvBDs (1-14) by real-time PCR in the gastrointestinal (GI) tissues (duodenum, jejunum, ileum and caecum) of 3-day-old broiler chicks after 24 h of oral infection with Salmonella Pullorum. Quantitative real-time PCR analysis revealed significant (P < 0.05) upregulation of AvBD3, 4, 5, 6 and 12 and a significant (P < 0.05) down regulation in the expressions of AvBD10, 11, 13 and 14 in one or few GI tissues, while no significant changes were observed for AvBD1, 2, 7, 8 and 9 gene expressions in any of the GI tissues investigated upon infection with S. Pullorum. Most substantial change in gene expression was found for AvBD5, being significantly (P < 0.01) upregulated in most of the GI tissues investigated. The differential expression levels of ß defensins shed light on tailored innate immune response induced by S. Pullorum during the early stages of infection in chicks.

Toll-like receptor mRNA expression, iNOS gene polymorphism and serum nitric oxide levels in indigenous chickens.

Toll-like receptor (TLR) family is one of the important members of innate immune system that recognizes conserved microbial patterns and induces innate immune response. They also act as a link to adaptive immune response. Nitric oxide (NO) is a multi-functional mediator with diverse physiological and immunological roles. In the present study TLR mRNA expression in heterophils, serum nitric oxide level and iNOS (inducible Nitric Oxide Synthase) gene polymorphism were investigated in cockerels of two Indian native chicken breeds, Aseel and Kadaknath. TLR (4 and 5) mRNA expression as quantified by real time RT-PCR revealed Kadaknath males expressed significantly (P < 0.01) higher TLR4 mRNA than Aseel males. iNOS gene polymorphism analyzed by PCR-RFLP method revealed difference in allele frequency. Kadaknath males had higher allele B frequency (0.81) than Aseel males (0.56). However, there were no genotype and breed effect on serum nitric oxide level. Based on the present study we conclude that Kadaknath has comparatively higher innate immunity levels than Aseel, however further investigations are needed.

Differential expression of Toll-like receptor mRNA in White Leghorn and indigenous chicken of India.

In the present experiment, the expression profile of Toll-like receptor mRNA in indigenous and pure line chickens was studied. The expression of TLR3, TLR4, TLR5 and TLR7 were quantified in heterophils of Aseel, Kadaknath, Naked neck, Dwarf and White Leghorn lines by Quantitative Real-time PCR. White Leghorns expressed significantly (P < 0.01) higher levels of TLR3 mRNA compared to other lines. TLR4 and TLR5 mRNA were significantly highly expressed in Kadaknath line. Among the TLRs investigated TLR5 was more expressed in all lines studied. TLR7 was highly expressed in indigenous chicken Aseel and Kadaknath than other lines. Dwarf chicken expressed significantly (P < 0.01) lower levels of all TLRs investigated. On the basis of the present study we conclude that the differential expression of TLR mRNA in the heterophils of indigenous and other chicken breeds might contribute to their variable disease resistance/susceptibility.