Functional identification of an exon 1 substitution in the myostatin gene and its expression in breast and leg muscle of the Bian chicken

1. The objective of this study was to verify the functional effects of the c.234G>A substitution in the myostatin (MSTN) gene and ascertain the mechanism by which the variant affects growth traits in the Bian chicken.

2. The c.234G>A substitution was detected by PCR-RFLP analysis in the 7th-generation Bian chickens and three genotypes (AA, AG and GG) were identified. Results showed that the substitution was significantly associated with all studied growth traits, except first-d-weight, in female Bian chickens.

3. Based on these results, the substitution was used in gene-assisted selection for growth traits and thus fast-growth (AA genotype) and slow-growth (GG genotype) lines were successfully established. Significant differences in growth traits were detected between the fast-growth and slow-growth lines from 6 to 16 weeks of age. Furthermore, all slaughter traits, except leg muscle rate, were significantly different between the fast-growth and slow-growth lines.

4. Expression analysis showed that the relative expression level of MSTN in chickens with GG and AG genotypes were significantly higher than that in chickens with an AA genotype, both in breast and leg muscle. Chickens in the slow-growth line had significantly higher relative expression level of MSTN compared to chickens in the fast-growth line, both in breast and leg muscle.

5. The results suggest that the c.234G>A substitution in the myostatin (MSTN) gene negatively regulates the expression of MSTN in the Bian chicken and that it may be used in marker-assisted selection to accelerate the chicken breeding process.     

The effect of heat stress on GHR,IGF-I,ANT, UCP and COXIII mRNA expression in the liver and muscle of high and low feed efficiency female quail

1. A study was conducted to test the hypothesis that feed efficiency (FE) correlated with the expression of genes from the somatotropic axis and mitochondrial genes involved in energy production, and that the environment to which the birds are exposed influenced the expression of such genes.

2. Quails were divided into High-FE and low-FE groups and maintained in a comfortable or heat stress (HS) (38°C for 24 h) environment to evaluate changes in insulin-like growth factor I (IGF-I), growth hormone receptor (GHR), adenine nucleotide translocator (ANT), uncoupling protein (UCP) and cytochrome oxidase subunit III (COX III) messenger ribonucleic acid (mRNA) expression in liver and muscle tissues.

3. High-FE quails (0.28 g/g) presented a higher final body weight, greater weight gain and a better feed conversion ratio than low-FE birds (0.18 g/g). High-FE birds showed greater IGF-I mRNA expression in the liver and muscle and greater GHR mRNA expression in the muscle.

4. Environmental effects only affected GHR expression in the liver, with quails under comfortable conditions exhibiting greater GHR expression than quails subjected to HS.

5. There was a significant interaction between FE and environmental temperature on ANT mRNA expression in the liver. The greatest ANT mRNA expression was observed for high FE-birds that remained under comfortable conditions.

6. In the liver, UCP mRNA expression did not differ among the quails and was not affected by environment or efficiency. However, comparisons of the low- and high-FE birds revealed higher levels of UCP mRNA in the muscle of low-FE birds.

7. COX III mRNA expression in the liver was dependent on environmental temperature and FE. Higher COX III mRNA expression was observed in animals that remained under comfortable conditions, and high-FE birds exhibited higher expression levels compared to low-FE birds.

8. These results suggest a correlation between IGF-I, GHR, ANT, UCP and COX III gene expression and FE and that environmental temperature could affect the expression of some of these genes.     

Changes to daily feed intake during the laying period alters embryonic MSTN and MYOG gene expression in genetically fat and lean lines of chickens

1. Broiler breeders are unable to self-regulate feed intake to a level that permits reproductive function and restricted feeding is normal for broiler parent stock. The aim of this study was to determine whether myostatin (MSTN) and myogenin (MYOG) mRNA expression could be altered in relation to differences in body composition of the embryos and the hen’s diet.

2. A 2 × 2 factorial experiment was conducted with two dietary intake levels (100% and 75% of nutrition recommendation) and two broiler genotypes (Fat line and Lean line). Hens (384 of each line) were randomly divided at 23 weeks of age into 4 treatments, with each treatment represented by 12 replicates of 16 birds each. The experiment started when the rate of lay reached 5% and continued until 54 weeks of age.

3. There were significant effects of line and dietary intake, and interactions between them on MSTN and MYOG mRNA expression levels in embryonic tissues. The trend in MSTN mRNA expression level over the embryonic period was similar between lines. Low daily feed intake did not alter the trend in MSTN mRNA expression levels in either line, but significantly changed the peak values. MYOG mRNA expression varied by line and over time, with a delay in the Lean line. Low daily feed intake influenced the trend in MYOG mRNA expression levels in both lines and delayed its onset.     

Growth,carcase and meat traits and gene expression in chickens divergently selected for intramuscular fat content

1. Changes resulting from divergent selection after 5 generations for intramuscular fat (IMF) content in breast muscle in a population of purebred Beijing-You chickens were investigated. Female chickens from lines selected-up (UL) and selected-down (DL) for 5 generations and random-bred controls (CL) were studied at 120 d of age.

2. In the UL, IMF in breast muscle (4.65%) was 1.38 times that in the DL (3.36%). The IMF content in leg muscle was also higher in UL compared to DL.

3. There were significant differences in growth, carcase traits and meat quality between the two selected lines. However, ultimate pH (pHu) was lower and lightness (L*) and yellowness (b*) were higher in breast muscles of UL birds compared to DL birds. Similar trends for pHu and L* value between UL and DL were observed in leg muscle.

4. Significant mRNA expression differences of the heart fatty acid-binding protein (H-FABP) gene between UL and DL were observed in breast muscle, and a negative correlation between expression of mRNA and IMF% in breast and leg muscles pointed to H-FABP as a potential selection candidate gene.     

Neuromusgular studies of turkeys with trienbolone acetate‐induced and naturally occurring “leg weakness”

1. In vivo sciatic nerve gastrocnemius muscle preparations were made from 14‐ to 18‐week‐old normal turkeys and from those with naturally occurring or trienbolone acetate (TA)‐induced “leg weakness”.

2. Preparations from leg weakness cases displayed neither accelerated muscle fatigue nor decreased nerve conduction velocities as compared with control preparations.

3. The muscles of the TA‐treated turkeys but not of those with naturally occurring leg weakness were hyperexcitable relative to those of controls.

4. Post‐tetanic potentiation of preparations from both naturally occurring and experimentally‐induced cases of leg weakness was less than that of control preparations, both before and after partial neuromuscular blockade, significantly so at the lower pulse frequencies used.

5. It is tentatively suggested that the last finding might be of significance in explaining the clinical signs of leg weakness and might be associated with a disordered calcium metabolism.     

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.     

Effects of polymorphisms and haplotypes within the MSTN gene on duck growth trait

1. Polymorphisms of the duck MSTN gene were investigated in 413 individuals by DNA sequencing and polymerase chain reaction restriction fragment length polymorphism. Four single nucleotide polymorphisms (G129A, C324T, A981G and C1002A), with A981G and C1002A completely linked, were found in the coding region.

2. Association analysis showed that different genotypes of all the identified SNPs were significantly associated with duck growth rate from week 5, 6 and 2 for G129A, C324T and A981G (C1002A), respectively. The greatest difference in body weight was 180 g at week 9, 106 g at week 8 and 123 g at week 8, respectively, for the three SNP’s.

3. Linkage disequilibrium (LD) analysis indicated that C324T, A981G and C1002A were in strong LD. Nine main diplotypes from the reconstructed five main haplotypes were observed, and different diplotypes were significantly associated with growth rate from week 1. Birds with the h1h1 diplotype exhibited the largest body weight from week 1 onwards.

4. It was concluded that the duck MSTN gene was associated with body weight and is an important candidate gene for duck growth. traits and marker-assisted selection.     

Possible use of an indicator muscle in future breeding experiments in domestic fowl

1. Domestic fowl were chosen for different body weights in descending order of size, viz. Tetra, Rhode Island Red/Light Sussex cross, White Leghorn and Bantam. The numbers and sizes of fibres in various muscles were determined by quantitative histology, and fibre number was also estimated by DNA measurement.

2. In contrast to the situation in mammals, fibre size (diameter) was of greater importance than fibre number in determining muscle size.

3. Biochemical measurement of DNA content in a muscle was not a reliable estimate of cell number without histological verification.

4. It is suggested that the anterior latissimus dorsi muscle could be used as an indicator muscle in future selective breeding programmes for fibre number because of its fibre arrangement and accessibility.     

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.     

Changes in free amino acid concentrations in the blood,brain and muscle of heat-exposed chicks

1. An experiment was conducted to analyse the changes in free amino acid concentrations in the blood, brain and muscle of chicks in response to 15 or 30 min exposure to high ambient temperature (HT).

2. Food intake and body weight were not affected, while rectal temperature was significantly increased by short-term HT exposure.

3. Several free amino acid concentrations increased in the blood, brain and muscle even with short-term HT, whereas levels of a few amino acids declined significantly. As well as the nonessential amino acids, essential amino acids also significantly increased with exposure to HT.

4. 3-Methylhistidine, a marker of proteolysis, significantly declined in the muscle of HT chicks, implying a reduction of protein breakdown under HT.

5. These results indicate that alteration of protein metabolism may occur in chicks even with short-term heat exposure.     

Effects of phytoestrogens on testosterone secretion by Leydig cells from Biłgoraj ganders (Anser anser)

1. The aim of this study was to examine ganders at different stages of the reproductive season and the effect of: (1) diets with high phytoestrogen content and (2) in vitro phytoestrogen treatment on testosterone secretion by isolated Leydig cells.

2. Thirty-six male Bi?goraj geese were fed control diets with low phytoestrogen content (containing grass meal) and diets with high phytoestrogens (containing alfalfa meal and soy).

3. Testes were obtained from both groups of ganders at three different times of the breeding season: peak of reproductive activity (March), second half of reproductive activity (May) and beginning of photorefractoriness (July).

4. Isolated Leydig cells were incubated with LH as well as genistein, daidzein, equol and coumestrol and the concentration of testosterone in the medium was determined by radioimmunoassay.

5. The mean weight of testes from ganders in May and July decreased relative to their weights in March, but no significant differences among experimental groups were noted.

6. No differences were observed in basal and LH-stimulated testosterone secretion by Leydig cells of ganders fed the control diets and the diets with higher phytoestrogen content. In July, LH did not stimulate testosterone secretion in either group. In vitro treatment with genistein, daidzein and equol (5 and 50?µM) inhibited basal and LH-stimulated testosterone production by Leydig cells from both groups. Coumestrol (5 and 50?µM) inhibited basal testosterone secretion only in March in the control group.

7. Dietary exposure to phytoestrogens had a slight effect on in vitro testicular secretion in ganders. In vitro treatment with phytoestrogen inhibited testosterone production by Leydig cells. Genistein showed the strongest effect and coumestrol had the weakest influence on testicular secretion.     

Molecular cloning,alternative splicing and mRNA expression analysis of MAGI1 and its correlation with laying performance in geese

1. In a previous study, a laying-related single nucleotide polymorphism (SNP) Record-106975 (A–G) was found in the intron of Membrane-associated Guanylate Kinase with Inverted Domain Structure-1 (MAGI1) gene by restriction site-associated DNA sequencing. This study was performed to further identify the function of MAGI1 on laying performance in Yangzhou geese.

2. The complete MAGI1 cDNA sequence was cloned from the ovary of the Yangzhou goose. A total of 4 alternative splice variants were identified, and the characterisation of their nucleic acid and amino acid (AA) sequences were analysed. The AA sequence of goose MAGI1 was highly conserved with other avian species.

3. MAGI1 was ubiquitously expressed in 11 tested tissues, with greater amounts of mRNA in the granulosa cell layer and ovary. GG (n = 14) and GA (n = 6) genotypes of SNP Record-106975 showed significantly lower MAGI1 mRNA levels than those of the AA genotype.

4. Over-expression and knockdown experiments revealed that MAGI1 mRNA expression was associated with caspase-3 mRNA levels in granulosa cells, and may play a role in regulating apoptosis of granulosa cells.     

An analysis of sexual size dimorphism in goose

1. Sexual size dimorphism (SSD) is a common phenomenon in animals. Rensch’s rule states that larger species generally exhibit a higher male to female body size ratio than smaller ones.

2. Domesticated animals offer excellent opportunities for testing predictions of the functional explanations of Rensch’s rule and this was tested in a meta-analysis of SSD in 38 breeds of domestic geese compared among themselves and with their wild relatives (subfamily Anserinae, 35 species).

3. Domestic geese and wild Anser species taken together supported Rench’s rule but the wild species did not.

4. The non-targeted sex selection hypothesis seems to provide the best intuitive explanation for the lack of SSD in geese.     

Effects of method of processing and chilling of Turkey carcasses on processing losses,water absorption and yields

1. The effects of wet or dry plucking, wet or dry evisceration procedures and method of chilling on processing losses and water uptake during chilling were studied using 19‐week‐old female large white turkeys.

2. Dry plucking increased weight loss due to plucking and eviscerating, and reduced chilled carcass yield.

3. Dry eviscerating increased carcass water uptake during chilling.

4. Pre‐chilling with running water increased carcass water uptake compared with chilling in ice and water alone.

5. Water uptake during chilling appeared to be increased by longer eviscerating time.     

Production traits of artificially and naturally hatched geese in intensive and free-range systems: I. Growth traits

1. This study investigated the effect of incubation type and production system on geese growth traits.

2. A total of 216 geese were either naturally (114) or artificially (102) hatched and reared in intensive or free-range production systems (4 replicates each) until 18 weeks of age.

3. Weights of naturally hatched goslings (NHG) were significantly higher than artificially hatched goslings (AHG) at 2 weeks (644 vs. 536 g); however, weights of AHG were significantly higher than NHG at both 6 weeks (3245 vs. 3010 g) and 18 weeks (5212 vs. 4353 g).

4. AHG had better feed conversion ratios (FCRs) than NHG (6.21 vs. 6.46 at 18 weeks). Feed consumption of naturally hatched geese was found higher in first 4 weeks when compared to artificially hatched geese and artificially hatched geese consumed more feed than naturally hatched geese after 8 weeks.

5. Production system had insignificant effects on feed consumption, FCRs, viability and mutilation rates.

6. Slipped wings were more frequent in NHG than AHG (8.32% vs. 1.68% at 6 weeks; 23.84% vs. 5.12% between 7 and 18 weeks) and in free-range production when compared to intensive production (17.88% vs. 11.08% over the course of the production period).

7. The study results indicate that both artificially and NHG can be reared in free-range production systems without any loss in performance and in deference to animal welfare.     

Effect of age and sex on the composition of muscle and skin from a commercial broiler strain

1. The effects of age and sex have been studied on the proximate composition and inorganic constituents in breast, thigh, drumstick and skin from a commercial British broiler strain (Ross 1).

2. Protein content generally increased with age in all muscles and decreased in skin. Protein content in skin of males was always higher than that in females.

3. Moisture content in all muscles decreased with age. In the skin, moisture decreased with age considerably more in females than in males.

4. Fat content increased with age in all tissues, while ash content of breast, thigh and skin decreased with age.

5. Phosphorus, chloride, magnesium and potassium all decreased with age in all the tissues. Thigh and skin calcium content were affected by age, as was the sodium content of drumstick and skin.

6. The sex effect on the inorganic constituents was variable.     

Influence of age,sex and rearing systems on Toll-like receptor 7 (TLR7) expression pattern in gut,lung and lymphoid tissues of indigenous ducks

1. The objective of the experiment was to determine the influence of age, sex and rearing system on Toll-like receptor 7 (TLR7) gene expression in gut, lung and lymphoid tissues and physiological responses to stress in male and female indigenous ducks of Tamil Nadu, India.

2. A total of 36 ducks (12 males and 24 females) were obtained from local farmers and tissue samples of gut tissues (duodenum, jejunum, ileum and caecum), lymphoid organs (spleen and bursa) and lungs were collected in RNAlater solution followed by RNA extraction.

3. After normalisation to β-actin (endogenous control) qPCR analysis identified a significant effect of age, sex and rearing system on TLR7 expression in the ducks.

4. A significant up-regulation of TLR7 expression was observed in lungs, duodenum, jejunum, ileum and caecum of sexually mature (45 wk) compared with that of immature ducks (16 wk). Among sexes, male ducks had significantly higher TLR7 expression than female ducks.

5. Age and sex interactions were significant in lungs, duodenum, jejunum and caecum. Ducks reared in an extensive housing system showed significantly higher TLR7 expression in bursa, lungs, duodenum, ileum and caecum compared to intensively reared ducks. There were no effects of age, sex and rearing systems on TLR7 expression in the spleen.

6. The heterophil-to-lymphocyte ratio and serum corticosterone were higher in ducks reared on an intensive system compared with ducks from an extensive rearing system.     

Genetic evaluation of carcass traits in Japanese quail using ultrasonic and morphological measurements

1. A study was conducted to evaluate the carcass composition of 1083 live birds using ultrasonic and morphological measurements and to estimate the genetic relationship between predicted and dissected carcass composition in Japanese quail.

2. Birds were reared for 35 d, and morphological measurements consisting of the length and width of breast muscle were recorded for all birds using a digital caliper. After slaughtering, the weight and percentage of carcass traits were measured on chilled carcasses. The dimensions of breast muscle were measured in 638 birds with an ultrasound scanner before slaughter at 35 d of age.

3. Genetic parameters from univariate and bivariate analyses were obtained by restricted maximum likelihood using ASREML software.

4. Genetic correlations between body weight at 35 d (BW35) and the percentage of carcass traits were low. Therefore, selection for BW35 may not effectively improve the yield of carcass components in Japanese quail.

5. High genetic correlations between carcass traits and ultrasonic measurements compared to morphological measurements suggest that the ultrasonic technique is a better method to improve breast weight and yield in Japanese quail.