Association of antioxidant defense system with semen attributes vis a vis fertility in exotic and indigenous chicken breeds.

The present study was designed to ascertain the association of antioxidant defense system with semen attributes and fertility in RIR (Rhode Island Red), PR (Punjab Red), RIR x local cross, Aseel and Kadaknath breeds. Based on sperm attributes, roosters of each breed were divided into two group i.e. G-I (exhibiting >50% sperm attributes) and G-II (exhibiting <50% sperm attributes). Motility, viability, membrane-, acrosome-, DNA-integrity and fertility differed significantly (p < 0.05) among the breeds, representing maximum in PR roosters and least in Aseel roosters. Values of sperm attributes and fertility rate were also significantly (p < 0.05) higher in G-I compared to G-II roosters in exotic as well as indigenous breeds. MDA content differed significantly (p < 0.05) in spermatozoa of five breeds. It indicated a least oxidant stress in PR and highest in Aseel. MDA concentration was significantly (P < 0.05) higher in G-II (28.36 ± 2.40-96.0 ± 6.4) than G-I (13.65 ± 1.81-52.22 ± 6.4) roosters in all breeds. Antioxidant enzyme activity varied significantly (p < 0.05) among the breeds and groups within the breeds. Significantly (p < 0.05) higher activity of four enzymes was evident in G-II as compared to G-I roosters irrespective of the breed. A moderate to strong negative correlation was perceived among LPO, SOD, GPX, catalase, GRE and sperm attributes/fertility rate. A moderate to strong positive correlation (0.21-0.92) among LPO and antioxidant enzymes revealed that with the increase in LPO, antioxidant enzymes increase too and vice versa in all breeds. Likewise, positive correlation between sperm attributes and fertility revealed that higher sperm attributes contribute to the high fertility of roosters. This is one of the first reports on complete set of antioxidant enzymes and oxidative stress in relation to sperm attributes and fertility in different five chicken breeds. A complete antioxidant enzyme system seems to modulate the oxidative stress, sperm attributes and fertility. It may be possible to use LPO as a fertility marker to select the roosters for breeding purpose in the chicken breeds.

Characterization of fertility associated sperm proteins in Aseel and Rhode Island Red chicken breeds.

This study focused on characterization of fertility associated proteins in Aseel and RIR roosters and was conducted on two generations of birds. Roosters were divided into high (>50%) and low fertility groups (<50%) based on sperm function tests and fertility rate in both the generations. Polyclonal antibodies were raised in rabbits against sperm proteins of first generation highly fertile roosters and tested for characterization of fertility associated sperm proteins in the second generation of same roosters. IgG-fraction against proteins (Anti-SP-IgG) was reacted with sperm proteins of both high and low fertile roosters of second generation on immunoblots. Sperm proteins present in highly fertile roosters were further characterized by Mass Spectrometry (MS). Use of SDS-PAGE for evaluation of sperm extracts of Aseel and RIR breeds resulted in resolution of 16 and 10 proteins on 12% acrylamide gels. Anti-SP-IgG reacted with eight and ten sperm proteins of Aseel and RIR roosters on immunoblots. The SDS-PAGE and immunoblotting analysis also indicated a variation in sperm proteins among two breeds and high/low fertile roosters. The MS analysis indicated matching of 20, 30, and 20, 25 kDa proteins (associated with high fertility rate) of Aseel and RIR roosters with immunoglobulin kappa chain variable, phospholipase A2 (PLA2), hypothetical N332-08551 partial and cystatin like partial proteins with a top score of 41, 46, 52 and 43, respectively. Considering the function and importance of matching proteins in male reproduction, these proteins may be further explored as potential markers for fertility evaluation of Aseel and RIR roosters.

Validation of immunomodulatory effects of lipopolysaccharide through expression profiling of Th1 and Th2 biased genes in Newcastle disease virus vaccinated indigenous chicken.

BACKGROUND AND AIM: Newcastle disease (ND) is considered one of the most important poultry diseases with chicken morbidity and mortality rates up to 100%. Current vaccination programs allow the use of live attenuated vaccines in the field to protect against the disease, which alone is inefficient and requires repeat booster doses. Toll-like receptor agonists (e.g., lipopolysaccharide [LPS]) as adjuvants are the ones, most extensively studied and have shown to be very promising in delivering a robust balanced immune response. In the present study, we have evaluated the potential of LPS to elicit a strong immune response with respect to the elicitation of both Th1 (cell-mediated) and Th2 (humoral) immune arms. MATERIALS AND METHODS: A total of 72 apparently healthy 1-day-old indigenous unvaccinated chicks were randomly divided into six experimental Groups A to F (n=12). At 8-week of age chicks in Group A, C, and E were vaccinated with live attenuated La Sota strain ND vaccine along with LPS, bovine serum albumin, and normal saline solution, respectively, and those in Group B, D, and E were kept separately without vaccination. Sampling was done on days 0, 1, 3, 7, 14, 21, 35, and 60 after vaccination. After vaccination and respective adjuvant application, Th1 and Th2 cytokine expression were measured in mRNA of both blood and tissue samples. RESULTS: The results were validated by, hemagglutination inhibition and enzyme-linked immunosorbent assay tests, to check for the humoral as well as cell-mediated immune response in blood serum levels. The results showed an increase in mRNA expression of the Th1 biased cytokines in Group A (LPS+NDV) as compared to the control groups. Similar mRNA expression pattern was seen in blood as well as tissue samples. Validation of results also indicates an increase in Cell-mediated Immunity as well as a humoral immune response in Group A (LPS+NDV). CONCLUSION: The results of the study provided enough evidence to consider LPS as a potential vaccine adjuvants candidate against ND in chicken.

Biocomputational analysis of evolutionary relationship between toll-like receptor and nucleotide-binding oligomerization domain-like receptors genes.

AIM: The active domains (TIR and NACHT) of the pattern recognition receptors (PRRs: Toll-like receptors [TLRs] and nucleotide-binding oligomerization domain [NOD]-like receptors [NLR], respectively) are the major hotspots of evolution as natural selection has crafted their final structure by substitution of residues over time. This paper addresses the evolutionary perspectives of the TLR and NLR genes with respect to the active domains in terms of their chronological fruition, functional diversification, and species-specific stipulation. MATERIALS AND METHODS: A total of 48 full-length cds (and corresponding peptide) of the domains were selected as representatives of each type of PRRs, belonging to divergent animal species, for the biocomputational analyses. The secondary and tertiary structure of the taurine TIR and NACHT domains was predicted to compare the relatedness among the domains under study. RESULTS: Multiple sequence alignment and phylogenetic tree results indicated that these host-specific PRRs formed entirely different clusters, with active domains of NLRs (NACHT) evolved earlier as compared to the active domains of TLRs (TIR). Each type of TLR or NLR shows comparatively less variation among the animal species due to the specificity of action against the type of microbes. CONCLUSION: It can be concluded from the study that there has been no positive selection acting on the domains associated with disease resistance which is a fitness trait indicating the extent of purifying pressure on the domains. Gene duplication could be a possible reason of genesis of similar kinds of TLRs (virus or bacteria specific).

Study of vascular supply of lunate and consideration applied to Kienböck disease.

The purpose of this study was to assess vascularity of the lunate by number of foramina and radiography of vessels of the wrist. The genesis of lunatomalacia requires some vascular risk and mechanical predisposition. The findings were correlated with the cause of Kienböck's disease. The vascular foramina were more than two in 91.33% of the lunate. The lunate had consistent dorsal and palmar branches from radial artery. The additional branches from anterior interosseous artery in 72.22% and a branch of palmar inter carpal arch in 69.44% cases contributed in arterial anastomosis on palmar aspect of lunate. The dorsal blood supply was found by anterior interosseous artery in 85.71% of specimens and dorsal branch from dorsal intercarpal arch in 50% of specimens. The blood supply of lunate comes along with various ligaments which may be disrupted due to trauma or strain leading to avascular necrosis. The present observations are suggestive of rich blood supply of lunate in comparison of other investigations. Therefore Kienböck's disease is less common in northern India.