ABSTRACT 1. Indigenous poultry breeds, like Kadaknath and Aseel, represent invaluable genetic resources, combining unique nutritional and cultural traits. The Kadaknath breed is famed for its black, lean, iron-rich meat with high protein and antioxidant content, while Aseel is a muscular breed prized for endurance, stress tolerance and fighting abilities. Despite their economic and nutritional value, the genetic and metabolic underpinnings of their distinct traits remain underexplored. 2. This study conducted a comparative transcriptomic analysis of liver tissue from Kadaknath and Aseel chickens using RNA sequencing (RNA-seq) to uncover the molecular mechanisms driving their phenotypic diversity. The results identified 435 differentially expressed genes (DEG) linked to key processes such as lipid metabolism, oxidative phosphorylation and stress response. 3. In Kadaknath chickens, up-regulated genes, including CPT1A, PCK1, TXNRD3 and PPARGC1A, were associated with efficient triglyceride breakdown, strong antioxidant defence and optimised energy metabolism. This likely contributes to the characteristic lean, nutrient-rich meat with high protein and low fat content. 4. In contrast, Aseel birds showed elevated expression of stress response genes (MRPL18 and RPL11) and mitochondrial genes (NDUFA1, NDUFA8, NDUFB9 and PTPMT1), supporting its high energy requirements and resilience to stress, crucial for its endurance as a fighter breed. Gene co-expression network analysis highlighted critical hub genes driving these breed-specific adaptations.

Fungal and mycotoxin contamination threaten poultry health, feed safety, and global food security. Mycotoxins result in reduced poultry growth, nutrient absorption, and a compromised immune system, causing severe financial losses. The rise of antibiotic resistance due to excessive antibiotic use in poultry also necessitates alternative strategies. Total 28 strains were isolated from the gut of indigenous Aseel and broiler chickens. Strains SB1, SB2, and SD2 completely inhibited Aspergillus flavus, Aspergillus ochraceus, and Fusarium proliferatum and were assessed via spore germination and mycotoxin reduction assays. The strains significantly inhibited fungal spore germination (up to 91.7 ± 0.3%;P ≤ 0.001) and reduced aflatoxin, ochratoxin, and fumonisin production by 91.6 ± 1.2%, 93.6 ± 0.6%, and 93.0 ± 0.5%, respectively. FTIR analysis of bacterial metabolites revealed shifts in peaks at 1467-3240cm- 1, indicating the presence of various functional groups. LC-MS/MS metabolomics, supported by multivariate analysis, revealed that metabolites were enriched with antifungal and mycotoxin-reducing compounds, including phenylacetic acid. Metabolites showed zones of inhibition up to 38.0 ± 1.1mm against A. flavus, up to 36.0 ± 0.5mm against A. ochraceus, and up to 34.0 ± 0.5mm ZOI against F. proliferatum (P ≤ 0.0001). They also showed the lowest minimum inhibitory concentrations and fungicidal concentrations between 3 and 5 µLmL- 1 (P ≤ 0.001). 16S rRNA sequencing identified SB1 as Bacillus subtilis (PV569530), SB2 as Bacillus clausii (PV569531), and SD2 as Bacillus sp. (PV569532). Thus, this study highlights the microflora of indigenous Aseel chickens as a potential source of antifungal and mycotoxin-reducing compounds, offering a sustainable approach to control fungal and mycotoxin contamination in poultry feed. As this study did not involve higher animals, no clinical trial number was required.

This research studies the morphology and morphometry traits of Aseel chickens raised in various regions of Türkiye. We used Turkish Aseel genotypes to assess the live weight and physical features of 60 female and 58 male subjects of varying ages. We measured brood length, breadth, head and beak width, neck and body length, chest width, depth, and circumference, thigh diameter, length, and depth, and leg length. Males' average live weight, chest circumference, thigh length, and comb length, which affect game efficacy, are 3.23 kg, 37.88 cm, 20.69 cm, and 40.20 mm, respectively. Although there was a statistically significant difference between the gender groups for all evaluated characteristics, there was only a statistically significant difference between the age groups for head width and chest circumference, shank depth, diameter, and length, and live weight (P<0.05). The data showed differences among different areas, particularly about the measurements of the comb, prompting researchers to propose a hypothesis suggesting a potential correlation between these changes and regional temperature disparities. We evaluated Turkish Aseels for feather, comb, eye color, markings, and comb type. Medium-weight breeds with hair, earlobes, beards, and spurs Weight and characteristics determine the Turkish Aseel breed. Comb structures were shorter, and their live weight was lower than that of other game roosters or Aseel kinds. Based on their morphology, domestic Aseel males may have a distinct genotype and subtype. Evaluating morphometric characteristics is an important aspect of the research. However, conducting additional comparative research is crucial. Genotyping studies with larger numbers are necessary for greater accuracy.

Study of Aseel chicken egg quality characteristics

The purpose of this study is to look at the seminal characteristics of short-term stored spermatozoa (at 4°C) from indigenous Aseel chickens in Bangladesh. The abdominal massage technique was utilized to collect semen from adult Aseel chickens at 3-day intervals and microscopically analyze the semen quality. The semen was then diluted with egg yolk-citrate (EYC) extender and stored at 4°C. They were then evaluated at various storage times, including Day 0 (0 hours), Day 1 (24 hours), Day 2 (48 hours), Day 3 (72 hours), Day 4 (96 hours), and Day 5 (120 hours). The findings of the study revealed that as storage time progressed, sperm motility, live spermatozoa, and the percentage of normal spermatozoa decreased. Total motility percentage significantly (P<0.05) decreased with the extended storage time in case of all individual cock. Progressive motility percentage was significantly (P<0.05) decreased from Day 0 (57.4±3.21) to Day 5 (0.11±3.21). On the contrary, non-motile sperm percentage significantly increased in Day 5 (99.3±4.85) compared to Day 1 (21.9±4.85). On the other hand, live spermatozoa were significantly (P<0.05) higher (76.68±5.06%) on Day 0 (0 h) and gradually declined day after day and finally it was 0% on day 5 (120 h). Similarly, spermatozoa abnormalities were highest at Day 5 (11.29±1.69% head, 3.33±1.42% mid-piece and 7.26±1.66% tail) and head abnormalities were significantly (P<0.05) affected by the storage time. Therefore, this study concluded that the storage duration had a substantial impact on the quality of Aseel chicken sperm, which may be used for artificial insemination (AI). The outcomes of this study may be valuable for the conservation and enhancement of the indigenous Aseel chicken breed in Bangladesh through the use of AI. J. of Sci. and Tech. Res. 6(1): 65-72, 2024

Microanatomical insight into Reticuloepithelial cells in the Thymus of TANUVAS Aseel Chicken

Immunomodulatory activity of Withania somnifera against Newcastle disease in Aseel chicken

Effects of Dietary Energy Levels on Performance and Gene Expression in Indigenous Aseel Chickens

Principal component analysis of morpho-biometric traits in Indigenous Aseel chicken

This study aimed to assess the genetic diversity, maternal origin, and population structure of Turkish Aseel chickens. The research was conducted on Aseel roosters and chicks older than one year. DNA was extracted from 96 Turkish Aseel feather samples collected from various regions of Türkiye for the study. Sample DNAs were amplified using specific primers for the D-loop region of the chicken mitochondrial DNA (mtDNA). The sequences were produced using the amplified DNA samples. The mtDNA D-loop regions were compared with the sequences of the same genetic area from other chicken breeds. Turkish Aseel chickens provided 34 haplotypes and 18 genuine polymorphisms across 41 distinct regions that were particular to the breed. According to the findings of the phylogenetic tree, Turkish Aseel chickens tend to establish clusters of haplogroups among themselves. Although most Turkish Aseel samples clustered individually into three clades, at least one sample has been discovered with five distinct clades. According to the phylogenetic tree and the Median Joining Network, it can be concluded that while many of the haplogroups of Turkish Aseel cluster among themselves, they have diverse maternal origins and lack breed-specific maternal lineages. This research is the first extensive study to examine genetic variability in Turkish Aseel chickens. Considering the lack of similar data on other Aseel varieties raised in different counties, the current work is a pioneering study with data on the genetic characterization of the Aseel breed.

The present study aimed to analyze the characteristics of Aseel chicken eggs, specifically focusing on size, quality, and their impact on hatchability percentage. The study was conducted in Sindh, Pakistan, with a total of 114 eggs selected from three different talukas in district Tando Allahyar. Parameters such as egg weight, specific gravity, egg shell, and egg thickness were recorded and analyzed. The data was further examined using rank-Spearman analysis to determine the correlation between these parameters. The results indicated significant differences (p < 0.05) among the selected talukas, with Tando Allahyar and Jhando Mari talukas showing higher values for egg weight, eggshell thickness, and specific gravity. Additionally, a significant (p < 0.05) and moderate to positive correlation was observed among all parameters. These findings suggest that the variations in results across different areas may be attributed to management practices and genetic differences in Aseel chicken populations within the talukas

Background: The present study was undertaken to assess the probiotic supplement through feed to enhance performance of TANUVAS Aseel chicken. Methods: Three hundred one month old TANUVAS Aseel chicks were supplied to the farmers of three groups (T1, T2 and T3), (Irrespective of number of birds seven replication in each treatment) under backyard system of rearing, all the chicks were fed concentrate feed along with foraging. T1 was control, T2 was Probeads EC supplement (5 beads/day/bird/oral) and T3 was probiotic supplement (1 g per kg of body weight). Result: The average body weight, daily weight gain, feed conversion efficiently, livability, cost benefit analysis were significantly higher in T2 (Probeads EC supplement) (P less than 0.05) compared to T3 and control (T1) groups.

Aseel birds are known for their hardiness, thermotolerance, pugnacity, and robustness. However, they are highly susceptible to Avian Influenza (AI) and Newcastle Disease (ND) viruses which cause huge mortalities. Present study was designed to identify polymorphisms in exon-14 of Mx1, and 5’UTR & intron-7 of ROBO2 gene associated with antibody response to AIV (H9N2) and ND virus respectively in adult Aseel birds. After screening of 500 birds for HI-based antibody response against both viruses, two separate set of 40 birds, one for each virus, were selected on the basis of maximum divergence in their antibody response. The sequence data of exon-14 of Mx1 gene revealed 3 SNP, and that of ROBO2 gene showed 2 SNPs in intron-7 and 1 in 5’UTR. The genotypic frequencies of identified variants were tested for goodness-of-fit and only Mx_810 showed significant difference. The results of association analysis revealed a non-synonymous SNP (G>A), Mx1_567, significantly associated with pre- (P<0.01) and post-vaccination (P<0.01) antibody response against AIV. The LSD results showed that homozygous mutant (AA), and heterozygous (AG) genotypes had significantly greater values of pre- and post-vaccination antibody response compared with wild-type genotype. However, no SNP in the ROBO2 gene was found significantly associated with antibody response.

The present study was conducted to evaluate the dietary effects of two seaweeds (Sargassum wightii and Gracilaria corticata) on different physiological parameters of TANUVAS Aseel chicken. A total of 30 TANUVAS Aseel chicks, day old were allocated into three groups (Each with 10 chicks) viz., one control group (C1) and two treatment groups (T1 and T2). The control group (C1) were fed with commercial grower feed alone and the treatment group (T1) had control diet with 5% Sargassum wightii and the treatment group (T2) had control diet mixed with 5% Gracilaria corticata. Parameters were recorded for a continuous period of 16 weeks in 4 weeks interval. The growth performance was found to be non-significant (P ≥ 0.05) during 0 day of feed trials and highly significant (P ≤ 0.01) during 4th, 8th, 12th and 16th week of feed trails. Haematological parameters during 12th and 16th week of feed trials showed non-significant (P ≥ 0.05) difference exist in lymphocyte, monocyte and eosinophil count and a highly significant (P ≤ 0.01) difference exist in platelets count between the control (C1) and treatment groups (T1 and T2). Serum biochemical parameters during 12th and 16th week of feed trials showed non-significant (P ≥ 0.05) difference in Uric acid, creatinine, ALT, calcium and cholesterol level and a highly significant (P ≤ 0.01) difference in Total protein, albumin, AST, glucose, triglycerides, phosphorus, magnesium, potassium and chloride level between the control (C1) and treatment groups (T1 and T2). However non-significant (P ≥ 0.05) difference exists in globulin and sodium content only during 16th week of feed trials. It could be noted that 5% inclusion of red seaweed, Gracilaria corticata in commercial grower feed as a feed supplement to TANUVAS Aseel chicks had a better body weight gain than 5% inclusion level of brown seaweed, Sargassum wightii in commercial grower feed. Based on the haematological and serum biochemical analysis, the supplementation of 5% inclusion level of seaweeds in this trial did not pose any threat to the physiological well-being of TANUVAS Aseel chicken.

Aseel chicken is very famous in Sindh for their fighting behavior, delicious meat quality with adaptability of tropical and subtropical climatic conditions and as attractive household bird.Present study was conducted on Aseel chicken raised in three different taluka Jhando Mari, Chambar and Tando Allahyar city of district Tando Allahyar, Sindh, Pakistan with 60 male and 40 female Aseel birds.The results of the study based on phenotypic diversity showed that maximum feather color and feather pattern was observed on neck and breast (34%) grey black and Dark brown (49%), plain (34%) and (55%) penciled type.The result for wing bow and wing bar feather color and pattern percentage was observed maximum orange/pale brown (34%) and (27%), plain (34%) and (45%) irregular pattern was recorded.The results for wing bay saddle feather color percentage of Aseel chicken was maximum found greenish black (26%) and (36%) of pale organe/pale brown (62%) and (45%) plain type pattern.The result for tail feather color percentage of Aseel chicken revealed that maximum color of feather was observed greenish black (42%) and (5%) of plain type pattern was recorded.

In this study, comparative analysis of skeletal muscle transcriptome was carried out for four biological replicates of Aseel, a fighter type breed and Punjab Brown, a meat type breed of India. The profusely expressed genes in both breeds were related to muscle contraction and motor activity. Differential expression analysis identified 961 up-regulated and 979 down-regulated genes in Aseel at a threshold of log2 fold change ≥ ±2.0 (p adj<0.05). Significantly enriched KEGG pathways in Aseel included metabolic pathways and oxidative phosphorylation, with higher expression of genes associated with fatty acid beta-oxidation, formation of ATP by chemiosmotic coupling, response to oxidative stress, and muscle contraction. The highly connected hub genes identified through gene network analysis in the Aseel gamecocks were HNF4A, APOA2, APOB, APOC3, AMBP, and ACOT13, which are primarily associated with energy generating metabolic pathways. The up-regulated genes in Punjab Brown chicken were found to be related to muscle growth and differentiation. There was enrichment of pathways such as focal adhesion, insulin signaling pathway and ECM receptor interaction in these birds. The results presented in this study help to improve our understanding of the molecular mechanisms associated with fighting ability and muscle growth in Aseel and Punjab Brown chicken, respectively.

The increasing incidence of extended-spectrum β-lactamase (ESBL)-producing Escherichia (E.) coli in backyard chicken farming in Pakistan is of serious concern. This study aimed to assess the prevalence, antimicrobial resistance patterns and risk factors associated with ESBL avian pathogenic E. coli (APEC) isolated from backyard chickens in the Jhang district, Punjab, Pakistan. In total, 320 cloacal swabs were collected from four breeds of backyard chicken (Aseel, Golden, Misri and Necked Neck). ESBL E. coli were phenotypically identified using double disc synergy test (DDST) and corresponding genes were confirmed by multiplex polymerase chain reaction (mPCR). Out of the 320 samples, 164 (51.3%) were confirmed as E. coli, while 74 (45.1%) were characterized as ESBL E. coli. The frequency of isolation of ESBL E. coli was highest in Aseel chickens (35.1%). Of the 164 confirmed E. coli, 95.1%, 78.6%, 76.8%, 71.3%, 70.1%, 68.9%, 60.4% and 57.3% were resistant against tylosin, doxycycline, cefotaxime, enrofloxacin, colistin, trimethoprim/sulfamethoxazole, chloramphenicol and gentamicin, respectively. The ESBL gene types detected and their corresponding proportions were blaCTX-M (54.1 %, 40/74), blaTEM, (12.2%, 9/74) and co-existence (blaCTX-M and blaTEM) were shown in 33.8% (25/74). The blaCTX-M gene sequence showed homology to blaCTX-M-15 from clinical isolates. The mean multiple antibiotic resistance index (MARI) was found to be higher among ESBL E. coli (0.25) when compared to non-ESBL E. coli (0.17). Both free-range husbandry management system (p = 0.02, OR: 30.00, 95% CI = 1.47-611.79) and high antimicrobial usage in the last 6 months (p = 0.01, OR: 25.17, 95% CI = 1.81-348.71) were found significantly associated with isolation of ESBL-producing E. coli in the tested samples using binary logistic regression analysis. This study confirmed the potential of backyard chickens as a reservoir for ESBL E. coli in the Jhang district, Punjab, Pakistan.

A study was conducted to assess the effect of feeding different crude protein (CP) levels with isocaloric metabolizable energy (ME) diets on growth performance, carcass traits, and myostatin (MSTN) gene expression of Aseel chicken during 0 to 16 weeks of age. A total of two hundred and ten day-old Aseel chickens were randomly allotted to seven dietary treatment groups. Each group had thirty chicks distributed into three replicates of ten chicks in each. Experimental diets were formulated to have varying levels of CP, viz. 18.5, 19.0, 19.5, 20.0, 20.5, 21.0, and 21.5%, with isocaloric energy of 2800 kcal ME/kg diets of mash feed fed to birds in a completely randomized design. Different CP levels had a significant effect (P < 0.05) on the body weight gain (BWG) of Aseel chicken. At the end of 16 weeks of age, the group fed 21% CP gained 223.53 g more than the lowest CP (18.5%)-fed group. The different CP levels did not significantly (P > 0.05) influenced the feed intake of all treatment groups, but numerically highest feed intake was observed in the lowest CP (18.5%)-fed group. However, significant differences in feed efficiency (FE) appeared from the 13th week only with the 21.0% CP-fed group showing the best FE until the 16th week (3.86 to 4.06). The maximum dressing % (70.61) was observed by the 21% CP-fed group. The CP 21% diet down-regulated the MSTN gene expression in breast muscle tissue to 0.07 folds when compared to the diet of CP 20%. The best economical coordinates for maximum performance for Aseel chicken appeared to be CP of 21% and ME of 2800 kcal/kg to achieve the best FE of 3.86 at the earliest age of 13 weeks. In conclusion, 21% CP in an isocaloric diet of 2800 kcal ME/kg, in Aseel chickens, would be optimum to improve the growth performance at maximum in terms of BWG and FE up to 16 weeks of age.

Qualitative and quantitative morphological characterization of Aseel chicken

Aseel, a popular breed of native chicken, characterized by its pugnacity, fighting strength and royal gait is being used to create crosses for domestic chicken production. However, information on its growth models is scanty. An experiment was conducted to evaluate different non-linear models and to find out best fitting model in Aseel, being maintained at Central Avian Research Institute, Izatnagar, Bareilly. Data on body weights from 12-weeks of age to 20-weeks of age at biweekly intervals were recorded on a random bred single-hatched flock. Owing to the non-linear characteristic of growth, three non-linear models namely, Gompertz, Bertalanffy and Logistic models were evaluated. Goodness of fit for all the models were checked using coefficient of determination (R2), adjusted coefficient of determination (Adj-R2), mean square error (MSE), mean absolute error (MAE) and Akaike information criterion (AIC). The Bertalanffy model most accurately characterized the growth trend in males, females and pooled sex data. The study revealed that this model may be used to ascertain the average body weights in Aseel chicken under random mating. The investigation has generated baseline data on growth modelling of random bred groups and may be used in similar investigations on other native chicken breeds.