Genetic effects of vascular endothelial growth factor and its receptor 2 on feather maturity in three chicken breeds

1. The goal of the current study was to evaluate the genetic effects of the vascular endothelial growth factor (VEGF) and its receptor (VEGFR-2) on feather maturity in the Qingyuan partridge chicken, Guangxi sanhuang chicken and Princess chicken.

2. Both SSCP-PCR and qPCR were employed to detect the polymorphism and gene expression of the VEGF and VEGFR-2 genes.

3. Four SNPs were identified in the VEGFR-2 gene. Exon10-A69G was associated with feather maturity (P < 0.01). Princess chickens with the genotype EF had higher feather maturity scores (P < 0.01). Higher expression levels of VEGF and VEGFR-2 were detected in the immature feather group of Qingyuan partridge chickens, especially in the skin.

4. The VEGF and VEGFR-2 genes play critical roles in feather maturity. In addition, exon10-A69G and genotype EF in the Princess chicken could potentially be utilised as genetic markers to improve efficiency in breeding.     

Bioprocess enhancement of feather degradation using alkaliphilic microbial mixture

1. The main aim of this work is to develop a robust method to generate a microbial mixture which can successfully degrade poultry feathers to overcome environmental problems.

2. Four different alkaliphilic microbes were isolated and shown to degrade poultry feathers.

3. Two of the isolates were phylogenetically identified as Lysinibacillus and the others were identified as Nocardiopsis and Micrococcus.

4. The best microbial co-culture for white and black feather degradation was optimised for pH, temperature and relative population of the isolates to achieve almost 96% of degradation compared with a maximum of 31% when applying each isolate individually.

5. The maximum activity of keratinase was estimated to be 1.5 U/ml after 3 d for white feathers and 0.6 U/ml after 4 d for black feathers in a basal medium containing feather as the main carbon source. Additionally, non-denaturing polyacrylamide gel electrophoresis showed 4 and 3 protease activity bands for white and black feather, respectively.

6. This study provides a robust method to develop potential new mixtures of microorganisms that are able to degrade both white and black feathers by applying a Central Composite Design.     

The retinoblastoma 1 gene (RB1) modulates the proliferation of chicken preadipocytes

1. The objective of this study was to reveal the role of chicken RB1 (Gallus gallus RB1, gRB1) in the proliferation of preadipocytes.

2. To measure gene expression of gRB1 in the proliferation of chicken preadipocyte, quantitative real-time PCR was used. The expression levels of gRB1 transiently increased during this process.

3. To detect the effect of gRB1 on the proliferation of chicken preadipocyte, MTT assay and cell-cycle analysis were performed. MTT assay showed that overexpression of gRB1 significantly suppressed (P < 0.05) the proliferation of chicken preadipocytes, and knockdown of gRB1 promoted the proliferation of chicken preadipocytes. Cell-cycle analysis showed that the proportion of preadipocytes in the G1 and G2 phases significantly increased (P < 0.05), and the proportion of preadipocytes in the S phase significantly decreased (P < .05) after up-regulation of the expression of gRB1. The proportion of preadipocytes in the S phase significantly increased (P < 0.05) after down-regulation of gRB1.

4. Quantitative real-time PCR was used to detect the effect of gRB1 on the expression of genes related to proliferation of chicken preadipocytes. Gene expression analysis showed that gRB1 knockdown promoted markers indicating proliferation of Ki-67 (MKi67) expression at 96 h (P < 0.05), and overexpression of gRB1 reduced MKi67 expression at 72 h (P < 0.05).

5. This study demonstrated that gRB1 inhibited preadipocyte proliferation at least in part by inhibiting the G1 to S phase transition.     

Keratin degradation by bacteria and fungi isolated from a poultry farm and plumage

1. Poultry processing generates a large quantity of feather waste. Feathers are a rich source of keratin and could be used as a feather meal in the feed industry if the keratin is degraded using suitable micro-organisms.

2. In this study, keratin-degrading micro-organisms were isolated from a poultry farm. The predominant organisms were identified as Bacillus subtilis, Aspergillus flavus, A. fumigatus and Trychophyton sp.

3. The isolates were inoculated into feather medium and observed for keratin degradation by measuring the protein content, free amino acids and change in pH.

4. During feather degradation by B. subtilis, the concentration of soluble protein released to the medium increased gradually and reached the maximum (433 µg/ml) during d 7 of incubation and the pH increased from the initial 6.9 to 8.4 on d 9 of incubation. Similarly, the maximum protein content of 414 µg/ml and pH of 8.5 was observed for A. fumigatus on d 21 of incubation.

5. B. subtilis and A. fumigatus showed almost the same level of keratinase activity.     

Spermatozoa DNA and plasma membrane integrity after pellet optimized processing for cryopreservation in meat type chicken breeders

1. Aim of this study was the development of an optimised cryopreservation pellet procedure for chicken semen and the assessment of DNA and membrane integrity in frozen/thawed spermatozoa in a Hubbard F15 meat type selected strain.

2. The following semen processing conditions were studied: spermatozoa working concentration (SWC), 1.5 vs 2 × 10⁹ cells/ml in pre-freezing extender; equilibration of diluted semen at 5°C, 20 vs 40 min; dimethylacetamide concentration, 6% vs 9%; dimethylacetamide equilibration time at 5°C, 1 vs 30 min; thawing at 60°C for 10 vs 50°C for 30 sec. Spermatozoa viability (EtBr exclusion procedure – stress test), mobility (Accudenz® swim-down test) and subjective motility were assessed in fresh and frozen-thawed semen.

3. The lower SWC (1.5 × 10⁹ cells/ml) and the higher dimethylacetamide concentration (9%) had positive significant effects on the recovery rate of motile (22% vs 16%) and viable spermatozoa (39 vs 34%), respectively.

4.Membrane (SYBR14-PI staining) and DNA integrity (comet assay) were assessed before and after freezing/thawing according to the optimised protocol.

5. Recovery rates of spermatozoa with undamaged plasma membrane and DNA were 41% and 76%, respectively. The distribution of spermatozoa in classes of DNA damage was also analysed and discussed.

6. It was concluded that pellet cryopreservation was a damaging process mainly for plasma membrane rather than nuclear DNA in chicken spermatozoa.     

Tall oil fatty acid inclusion in the diet improves performance and increases ileal density of lactobacilli in broiler chickens

1. Studies were conducted with tall oil fatty acids (TOFA) to determine their effect on broiler chicken performance and ileal microbiota. TOFA, a product originating from coniferous trees and recovered by fractional distillation of side-streams from pulp production, mainly comprises free long-chain fatty acids (~90%) and resin acids (~8%). Conjugated linolenic acids and pinolenic acid are characteristic fatty acid components of TOFA.

2. TOFA products at 750 mg/kg feed were tested in two 35-day broiler chicken trials, each using a wheat soya-based diet and with 12 replicate pens per treatment. In both trials, TOFA improved body weight gain at all time points (P < 0.001) and feed conversion efficiency during the first 21 days (P < 0.01). Two different dry TOFA formulations (silica carrier and palm oil coating) were tested and showed performance effects similar to liquid TOFA.

3. Ileal digesta of the broiler chickens was analysed for total eubacteria, Lactobacillus spp., Enterococcus spp., Escherichia coli and Clostridium perfringens on days 14 and 35. TOFA significantly increased total eubacteria and lactobacilli density on day 14 (P < 0.05). There was a significant positive correlation between these bacterial groups and broiler body weight on day 14 (P < 0.01).

4. A numerical reduction in C. perfringens was observed. In vitro growth inhibition studies showed that C. perfringens was strongly inhibited by 10 mg/l TOFA (P < 0.001), while common lactobacilli were resistant to >250 mg/l. The in vitro results were thus in line with in vivo observations.

5. The mechanisms behind the bacterial shifts and their role in performance improvement are unknown. Further purification of TOFA components is needed to identify the effective agents.     

Polymorphism and expression of insulin-like growth factor 1 (IGF1) gene and its association with growth traits in chicken

1. The objectives of the study were to detect polymorphism in the coding region of the IGF1 gene, explore the expression profile and estimate association with growth traits in indigenous and exotic chickens.

2. A total of 12 haplotypes were found in Cornish, control layer and Aseel breeds of chicken in which the h1 haplotype was most frequent.

3. Nucleotide substitutions among haplotypes were found at 21 positions in the IGF1 gene in which 4 substitutions resulted in non-synonymous mutations in the receptor binding domain of the IGF1 protein.

4. The haplogroup showed a significant effect on body weight at 24 and 42 d of age in the control layer line, body weight at 42 d and daily weight gain between 29 and 42 d in the control broiler line, daily weight gain between 29 and 42 d in Cornish, and body weights at 42 d as well as daily weight gain between 29 and 42 d in Aseel birds.

5. IGF1 expression varied among the breeds during embryonic and post-hatch periods. The expression among the haplogroups varied in different chicken tissues. The effect of haplogroup on myofibre number in pectoral muscle was non-significant, although there was significant variation in numbers between d 1 and d 42, and between broiler and layer lines.

6. It was concluded that the coding region of the IGF1 gene was polymorphic, expressed differentially during the pre-hatch and post-hatch periods, and haplogroups showed significant association with growth traits in chicken.     

Effect of the frizzle gene (F) on egg production and egg quality of laying hens kept in tropical villages

1. Two experiments were conducted to determine the influence of the frizzle gene (F) on the production and quality of chicken eggs kept on village farms in Ghana.

2. In the first experiment, 144 pullets, 72 each of Ff and ff pullets from F₁ birds, were compared in a randomised complete block design (RCBD), with three villages and 4 batches of hatch as blocks and the two feather genotypes (Ff and ff) as the treatments.

3. The Ff pullets were superior (P < 0.05) to their ff counterparts in terms of the number of eggs per clutch, egg mass and hen-housed and hen-d rates of lay, whereas age at first egg was lower (P < 0.05) in ff layers compared to Ff ones.

4. The eggs of the F₁ heterozygous frizzle (Ff) layers had higher values for albumen height, Haugh unit and yolk height compared with eggs from their normal feathered counterparts.

5. In the second experiment, 144 pullets, 48 each of FF, Ff and ff pullets, were compared in a RCBD, with three villages and 4 batches of hatch as blocks and the two feather genotypes (Ff and ff) as the treatments.

6. The F₂ birds with genotypes Ff and FF were better than their ff counterparts in terms of the number of eggs per clutch, egg mass, and hen-housed and hen-d rates of lay. Age at first egg was significantly lower in ff layers compared to FF and Ff ones.

7. The Haugh unit value was higher in the homozygous and heterozygous frizzles compared to the normal feathered ones.

8. The presence of the frizzle gene (F) in egg type chickens led to an improvement in egg production and egg quality traits in village chickens, and the cross-breeding scheme evaluated in this project could contribute to improved productivity.     

The occurrence of giant fibres in different muscles of two chicken genotypes

1. The occurrence of Giant Fibres (GF) in three muscles (Pectoralis major (PM), Iliotibialis lateralis and Semimembranosus) with different types of energy metabolism was studied in slow- and fast-growing chicken strains.

2. A total of 20 one-day-old Leghorn chicks (slow-growing) and 20 broiler (Ross 508) chicks (fast-growing) were reared to 100 and 45 d, respectively.

3. A small percentage of GF was seen in pre rigor muscle samples even at 3 min post mortem in both genotypes and in all muscle types studied.

4. From 3 min to 24 h post mortem GF increased both in Leghorn and broiler chickens but to a different extent according to muscle type and genotype.

5. The highest GF 24 h post mortem value was found in the PM muscles belonging to the fast-growing broiler line.

6. It was concluded that every type of muscle can develop GF, but this phenomenon is more evident in the PM especially in animals selected for increased growth rate.     

Effect of chicken meat replacement by spent laying hen meat on physicochemical properties and sensorial characteristics of fresh sausage

1. Sausages were used to assess the effects of chicken meat replacement by spent laying hen meat on the quality parameters and sensorial properties.

2. Five formulations were developed: control (CON) containing 100% chicken meat and 4 levels of chicken meat replacement with spent laying hen meat: 25% (T25), 50% (T50), 75% (T75) and 100% (T100).

3. Appearance, texture, succulence, flavour and overall consumer acceptance were analysed. Parameters such as moisture, ash, fat and protein content, pH, cooking loss and colour parameters (lightness (L*), redness (a*), yellowness (b*)) were analysed, as well as the texture properties (hardness, elasticity, cohesiveness and chewiness).

4. Statistical analysis did not reveal significant (P > 0.05) differences in chemical composition and colour parameters among formulations. On the other hand, CON treatment showed the lowest (P < 0.05) cooking loss (20.45%) that increased as spent laying hen meat increased in the formulation (24.92% vs. 27.65% vs. 28.12% vs. 33.05%, for T25, T50, T75 and T100 batches, respectively).

5. Regarding textural parameters, T75 and T100 formulations presented higher (P < 0.05) hardness and chewiness compared to the other ones.

6. Concerning to sensorial characteristics, the T100 formulation presented the lowest average scores (P < 0.05) for all attributes studied. However, the other batches (T25, T50 and T75) did not show significant differences for appearance, texture, succulence, flavour and overall acceptability attributes compared with the CON formulation.

7. The results indicated that the substitution of up to 75% of chicken meat by spent laying hen meat did not decrease the sensory acceptance of the sausages by consumers, demonstrating that this level of substitution in sausages allows better use of spent laying hen meat.     

Prevalence and antimicrobial resistance of Arcobacter species isolated from poultry meat in Iran

1. The objective of this study was to determine the prevalence and antimicrobial resistance of Arcobacter spp. isolated from different species of retail poultry meat in Iran.

2. From August 2012 to April 2013, a total of 540 raw poultry meat samples from chicken (n = 100), turkey (n = 100), quail (n = 100), partridge (n = 80), duck (n = 50), ostrich (n = 60) and geese (n = 50) were purchased from randomly selected retail outlets in Shahrekord, Isfahan, Sari and Rasht, Iran.

3. Using culture techniques, 71 of 540 poultry meat samples (13.1%) were positive for Arcobacter spp. The highest prevalence of Arcobacter spp. was found in chicken meat (28.0%), followed by quail (12.0%), duck (11.4%), turkey (11.0%), geese (8.0%), partridge (7.5%) and ostrich (3.3%) meat. The number of A. butzleri isolated from poultry meat samples (90.1%) was significantly higher than A. cryaerophilus (7.1%) and A. skirrowii (2.8%). Significantly more poultry meat samples were found to contain Arcobacter spp. by the PCR assay than by the culture method.

4. Susceptibilities of Arcobacter isolates were determined for 14 antimicrobial drugs using the disk diffusion method. All of the 71 Arcobacter isolates tested were resistant to one or more antimicrobial agents. Resistance to cephalothin and vancomycin (95.8%) was the most common finding, followed by resistance to methicillin, azithromycin and ampicillin. All Arcobacter isolates were susceptible to gentamicin, streptomycin, tetracyclin and kanamycin.

5. The results of this study indicated the importance of poultry meat, especially chicken meat, as potential sources of Arcobacter spp. infection in people. Furthermore, the strains indicated resistance to a broad spectrum of antibiotics.     

Growth performance and meat quality of broiler chickens supplemented with Rhodopseudomonas palustris in drinking water

1. The effect of the bacterium, Rhodopseudomonas palustris, on the growth performance and meat quality of broiler chickens was investigated.

2. A total of 900-d-old Arbor Acres broilers were allocated to three experimental treatments for 6 weeks. Chicks were administered with R. palustris in drinking water as follows: (i) control group without R. palustris; (ii) treatment 1 (R1) with R. palustris of 8 × 10⁹ cells per chick per day in drinking water; (iii) treatment 2 (R2) with R. palustris of 1.6 × 10¹⁰ cells per chick per day in drinking water.

3. The results showed that, compared with that of control, both groups of R. palustris treatment increased daily weight gain and improved feed conversion ratio of broiler chickens significantly during the whole growing period of 6 weeks.

4. Both total and glutamic acid contents of chicken breast fillet in R. palustris treatment R2 were higher, while the fat content was lower, than those of the control group. Furthermore, R. palustris treatments also improved sensory attributes of chicken breast fillet.

5. As a probiotic providing rich nutrients and biological active substances, R. palustris administration in drinking water displayed a growth promoting effect and improved meat quality of broiler chickens.     

Feather pecking behaviour in White Leghorns,a genetic study

1. Genetic variables of feather pecking (FP) behaviour in a 1993 commercial pure line of White Leghorns were estimated at the age of 6, 38 and 69 weeks.

2. Heritability estimates of performing FP were 0.05 ± 0.06, 0.14* ± 0.07 and 0.38** ± 0.12 for 6, 38 and 69 weeks respectively for sum of pecks (PECKS) and 0.13* ± 0.07, 0.13* ± 0.07 and 0.35** ±0.12 for sum of bouts (BOUTS).

3. Heritability estimates of receiving FP were not significantly different from 0 except at 6 weeks (0.15* ±0.07 and the average of the 3 age classes (AVG) (0.22** ± 0.09) for PECKS and at 6 weeks (0.15* ±0.07) using BOUTS.

4. Genetic correlations of performing FP among age classes were in general high and significant. This was not the case with receiving FP.

5. Plumage cover at 51 weeks had a negative genetic correlation with performing FP at 69 weeks and AVG, but not with receiving FP. No phenotypic correlations were significant between plumage and FP.

6. Body weight at 51 weeks had a negative genetic correlation with performing FP at AVG.

7. Heritability estimates for performing and receiving FP at 6 weeks correspond to those in the literature. No estimates have previously been reported on feather pecking at 38 weeks or 69 weeks.

8. Selection of birds with no or a very low tendency to perform feather pecking should, on the basis of our results, be feasible.     

Effects of different dietary sources and levels of selenium supplements on growth performance,antioxidant status and immune parameters in Ross 308 broiler chickens

1. Although different impacts of various sources of selenium (Se) on chicken performance have been largely studied, there is a lack of comparative experiments studying the effects of these sources on the immune system and antioxidant indices of broiler tissues. The aim of this study was to examine the effects of various sources and levels of dietary Se supplements on performance, antioxidant status and immune parameters in Ross 308 broiler chickens.

2. A total of 1200 1-d-old male broilers (Ross × Ross 308) were divided into 8 treatments with 6 replicate pens and 25 birds per pen. This experiment was conducted as a completely randomised design with a 4 × 2 factorial arrangement. Main factors included Se sources as sodium selenite (SS), Se-enriched yeast (SY), DL-selenomethionine (SM) and nano-selenium (NS) and levels at 0.1 or 0.4 mg/kg Se.

3. Dietary supplementation of organic Se sources significantly improved average daily gain (ADG), gain: feed ratio and European production efficiency factor (P < 0.05) compared to birds fed on diets supplemented with inorganic source. In addition, ADG was increased in response to increased level of supplemental Se. Based on contrast comparison, there were significant differences in these parameters between organic versus inorganic sources of Se. However, there was no difference between contrast comparisons of NS versus SM and SY.

4. Total anti-sheep red blood cell (SRBC) and Immunoglobulin G (IgG) titres and hypersensitivity were enhanced by increasing supplemental concentration of Se and using organic sources of Se rather than SS (P < 0.05).

5. Oxidation resistance assessment of tissues demonstrated that supplementation of organic sources of Se and increase in supplemental concentration of Se ameliorated glutathione peroxidase activity, total antioxidant capacity and malondialdehyde formation (P < 0.05). Mostly, there were significant differences between organic versus inorganic sources of Se in oxidation resistance.

6. Overall, dietary supplementation of 0.4 mg/kg Se from an organic source resulted in better production performance and immune system response. Moreover, minimum formation of malondialdehyde in broiler tissue was observed in birds fed on diets supplemented with SM at 0.4 mg/kg.

7. It can be concluded that SM is more effective than other sources of Se in reducing lipid oxidation.     

Genetic diversity of mtDNA D-loop sequences in four native Chinese chicken breeds

1. To explore the genetic diversity of Chinese indigenous chicken breeds, a 585 bp fragment of the mitochondrial DNA (mtDNA) region was sequenced in 102 birds from the Xichuan black-bone chicken, Yunyang black-bone chicken and Lushi chicken. In addition, 30 mtDNA D-loop sequences of Silkie fowls were downloaded from NCBI. The mtDNA D-loop sequence polymorphism and maternal origin of 4 chicken breeds were analysed in this study.

2. The results showed that a total of 33 mutation sites and 28 haplotypes were detected in the 4 chicken breeds. The haplotype diversity and nucleotide diversity of these 4 native breeds were 0.916 ± 0.014 and 0.012 ± 0.002, respectively. Three clusters were formed in 4 Chinese native chickens and 12 reference breeds. Both the Xichuan black-bone chicken and Yunyang black-bone chicken were grouped into one cluster. Four haplogroups (A, B, C and E) emerged in the median-joining network in these breeds.

3. It was concluded that these 4 Chinese chicken breeds had high genetic diversity. The phylogenetic tree and median network profiles showed that Chinese native chickens and its neighbouring countries had at least two maternal origins, one from Yunnan, China and another from Southeast Asia or its surrounding area.     

Effect of gender on breast and thigh turkey meat quality

1. The influence of gender on chemical composition, physicochemical parameters, fatty acid profile, amino acid and mineral composition of turkey breast and thigh meat was studied in order to assess nutrient requirements.

2. Chemical composition showed that only intramuscular fat in breast meat was significantly affected by gender (p < 0.05). The results showed a higher percentage of intramuscular fat in male samples, almost double the amount found in females (0.73% vs. 0.38%).

3.For meat colour parameters, only a* showed different results between sexes, with male samples (breast: p < 0.01; thigh: p < 0.001) having the highest values.

4. Fatty acid profiles showed that medium chain unsaturated fatty acids were the most abundant. The significant differences (p < 0.05) found in both breast and thigh muscle could be linked to a difference in metabolism between males and females.

5.There were higher levels of C16:1n-7 in females (breast: p < 0.001; thigh: p < 0.01) compared with male muscle sample (5.05 vs. 2.67 g/100 g in breast and 4.95 vs. 3.27 g/100 g in thigh). Nutritional indices (n-6/n-3 and thrombogenic index) were more favourable in female samples demonstrating that female turkeys had better fatty acid profile than the others.

6. Turkey meat is an important source of dietary amino acids, and female samples had the highest contents both of essential and non-essential amino acids. Furthermore, gender had a numeric effect (p > 0.05) on amino acid composition.

7. Mineral composition showed that Na, Zn and Fe were the minerals most affected by turkey gender.     

Effect of Melissa officinalis supplementation on growth performance and meat quality characteristics in organically produced broilers

1. A trial was conducted to study the effect of Melissa officinalis supplementation on organic broiler performance and meat chemical, microbiological, sensory and nutritional quality.

2. Male and female day-old Ross 308 chicks were fed on a standard commercial diet containing 0, 2.5, 5 or 10 g/kg feed ground M. officinalis for 84 d before slaughter.

3. Weight gain and feed conversion ratio were significantly improved in the broilers receiving either 5 or 10 mg M. officinalis/kg feed.

4. Inclusion of M. officinalis did not affect muscle chemical and fatty acid composition.

5. On the basis of microbiological and sensory experimental data and subsequent extension of meat shelf life, M. officinalis did not reduce the microbial populations of the meat, but was effective in limiting lipid oxidation.