Genetic diversity of Jiangsu native chicken breeds assessed with the mitochondrial DNA D-loop region

1. The objective of this study was to determine the origin and evolution of chickens from 5 native breeds that are traditionally raised in Jiangsu Province.

2. To address this question, the complete mitochondrial DNA D-loop sequence of 149 chickens from 5 native breeds of Jiangsu Province was analysed.

3. Sequence read lengths of the native breeds were 1231 to 1232 bp, with a single-base deletion from the 859 bp site in the 1231 bp haplotype. A total of 33 variable sites that defined 19 haplotypes were identified. The average haplotype diversity and nucleotide diversity were 0.862 ± 0.017 and 0.00591 ± 0.00135.

4. Phylogenetic analysis showed that genetic structure of the mtDNA haplotypes of Jiangsu chickens are distributed across 5 clades (haplogroups): Clades A, B, C, D, and E. However, most of the individuals characterised in this study belonged to clades A and B.

5. The results of this study indicate that Jiangsu chicken populations have relatively low nucleotide and haplotype diversity and likely share 5 common maternal lineages.     

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.     

Natural resistance of Sri Lankan village chicken to Salmonella gallinarum infection

1. An experiment was conducted to compare the natural resistance of an indigenous breed of local village chickens to Salmonella gallinarum with two commercial breeds: ISA Brown and ISA White layers under experimental conditions.

2. A total of 72 chickens from each of these breeds were randomly distributed to 4 pens to provide equal numbers of two replicate pens maintained as infected and control (uninfected). All chickens in infected groups were inoculated orally with 1 × 10⁸ CFU (1 ml dose) of a field isolate of S. gallinarum, at the age of 8 and 16 weeks given over 5 consecutive days. Growth performance, clinical signs, gross pathological lesions and antibody responses were measured.

3. A significantly higher mortality was observed in the brown layers compared with the white layers, and clinical signs and mortality were absent in village chickens. However, a large number of birds with gross lesions and high antibody titres were detected in village chickens, indicating that birds had the disease subclinically. Commercial breeds had a significantly higher body weight, feed intake and feed conversion efficiency.

4. There was a significantly lower proportion of positive reactors in village chickens in the whole-blood agglutination test (35%) compared to brown (100%) and white (90%) layers even after the second inoculation. Uninfected birds were negative in all groups. The indirect enzyme-linked immunosorbent assay confirmed these observations.

5. These results suggest that the indigenous breed had superior natural resistance to S. gallinarum than the commercial breeds.     

Growth performance,carcass traits,meat chemical composition and blood serum metabolites of broiler chicken fed on diets containing flaxseed oil

1. This study evaluated the effects of diets with partial and total substitution of soya bean oil (SO) with flaxseed (linseed) oil (FO) on broiler chicken performance, carcass traits, meat chemical composition and blood serum metabolites.

2. A total of 448 one-d-old Cobb 500 broiler chicken were used. They were allotted among 4 treatments with 8 replications, using a completely randomised design, for 35 d. Four diets were compared: T1 = 100% SO (3%, 1–7 d; 4%, 8–21 d; and 5%, 22–35 d); T2 = 50% SO + 50% FO; T3 = 25% SO + 75% FO and T4 = 100% FO.

3. No significant differences were observed in body weight (BW), body weight gain (BWG), feed intake (FI), feed conversion ratio (FCR) and blood serum metabolites (total triglycerides, TRI; total cholesterol, CHO; high-density lipoprotein, HDL; low-density lipoprotein, LDL; glucose, GLU; albumin, ALB; globulin, GLO; and total proteins, TPs). Significant effects were observed for TRI, CHO, HDL, GLU, HDL, LDL, ALB and GLO with regard to the day of collection.

4. Carcass traits did not show significant differences for the treatments. No significant differences were observed for breast and drumstick chemical compositions, with the exception of drumstick fat concentration (quadratic effect).

5. In conclusion, the partial or total substitution of SO with FO did not affect growth performance, carcass traits, meat chemical composition or blood serum profile in broiler chicken. Therefore, FO can be an alternative to SO in the diet formulation for broiler chicken.     

High,low, or familiar? Nest site preferences of experienced laying hens

1. The aim of this study was to investigate which nest heights are preferred by laying hens in the absence of familiar nest locations and whether preferred nest heights are more attractive than a familiar location. In two experiments, a total of 108 hens of four different layer breeds, which were at least 50 weeks of age, were studied.

2. In the first experiment, hens were given individual free choice between nests for 1-week at four different heights (0 cm, 39 cm, 78 cm, and 117 cm above ground). Hens of the four breeds differed in their nest height preferences (P = 0.0013). However, hens of three breeds preferred ground level nests (P < 0.007) and the fourth line showed an equal preference for the ground level and level three, the latter level corresponding to the height of the nests in their home compartments.

4. In the second experiment, hens from the four breeds were given a choice between ground level nests and nests at a familiar location, i.e. at the same location as in their home compartment. Hens of all strains preferred the familiar nest location (P = 0.002) and preferences did not differ between strains (P = 0.77).

5. Laying hens seem to prefer nests at ground level in the absence of a familiar nest. However, if possible, experienced 50 week old hens continue to use a familiar nest location instead of a ground nest location. The results are discussed with respect to a potential primary preference that may be modifiable by experience and with respect to possible relevance in commercial housing.     

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.     

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.     

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.     

Embryonic development in the ostrich (Struthio camelus) during the first 7 days of artificial incubation

1. Early development of ostrich embryos was investigated in relation to time of egg collection and genotype.

2. A total of 321 ostrich eggs were collected during the 2008 and 2009 breeding seasons and the development of the embryo for up to the first 168 h of incubation was described and analysed. A sample of the incubated eggs was weighed and opened daily to investigate developmental changes.

3. In fresh eggs, the blastoderm contained a round, translucent dark area pellucida (AP) in the centre, with a surrounding thin white ring, likely to be the beginning of the area opaca (AO). Fresh eggs were considered infertile if the blastoderm was absent and instead numerous white droplets were present surrounded by vacuoles.

4. The average blastoderm area of a fresh fertile egg was 15.8 mm², increasing to 143.3 mm² after 2 d of incubation. By 72 h of incubation the area vasculosa (AV) was discernible in the posterior half of the blastoderm.

5. At 48 h of incubation the blastoderm area in eggs from the South African Black genotype (SAB) × Zimbabwean Blue genotype (ZB) crosses (104.5 ± 18.6 mm²) was lower than the pure SAB (141.0 ± 10.5 mm²), ZB (161.7 ± 13.5 mm²) and ZB × SAB crosses (166.1 ± 14.2 mm²).

6. Embryo length was 5.01 mm after 72 h of incubation and 14.5 mm after 168 h of incubation. At 168 h of incubation AV lengths for both ZB × SAB (53.2 mm) and SAB × ZB crosses (54.1 mm) were longer than in embryos from the pure breeds.

7. Results from this study can be put to practical use when determining whether eggs are infertile or fertile and also in investigating the age of early embryonic mortalities.     

Identification of chromosome instability in Japanese quail (Coturnix japonica)

1. A study of the incidence of chromosome instability in the Japanese quail as assessed by sister chromatid exchange (SCE) and fragile site identification in chromosomes was conducted in two parent breeds and their F1 and F2 generations.

2. The mean incidence of SCEs was 6.02 ± 0.45 and the frequency of fragile sites was 1.17 ± 0.79.

3. There were moderately negative correlations of 0.51–0.64 between chromosome instability and fertility in the F1 and 0.10–0.23 in the F2. The hatch of fertilised eggs was negatively correlated with the number of SCE in male (0.31) and female (0.33) F1 and was lower in P (0.18 and 0.19, respectively), whereas the correlations were similar for the number of fragile sites in both generations (0.51–0.62).     

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.     

Depletion of florfenicol and florfenicol amine residues in chicken eggs

1. The aim of this study was to develop a suitable method for the analysis of florfenicol (FF) and its metabolite florfenicol amine (FFA) in chicken eggs and to determine FF and FFA residue depletion in eggs of laying hens.

2. The analytes were extracted from yolk, albumen and whole egg by phosphate buffer and ethyl acetate. Following purification, samples were analysed by high-performance liquid chromatography.

3. Fifty laying hens were divided into 5 groups, and each hen received doses of 20 mg/kg FF: Group 1 (received a single oral dose by gavage); Group 2 (a single intramuscular dose); Group 3 (a single subcutaneous dose); Group 4 (multiple oral doses for 3 d) and Group 5 (multiple oral doses for 5 d).

4. Limits of detection and of quantitation values were 1.94 and 6.45 g/10⁹ g (ppb) for FF, respectively, and 0.48 and 1.58 ppb for FFA, respectively. Relative standard deviation values of intra-day and inter-day variation below 11% also confirmed the usefulness of the method for analysing FF and FFA in eggs.

5. From the first day of both oral and parenteral administration, FF and FFA were detected at 0.1% and 0.08% of dosage, respectively, and 57% of the drugs were eliminated from the egg yolk. Elimination time of FF was 8 d in Groups 1, 2 and 3; 9 d in Group 4 and 10 d in Group 5.     

Single-tube real-time multiple allele-specific PCR for genotyping chicken Mx gene G2032A SNP

1.?A non-synonymous, single nucleotide polymorphism (G to A) at position 2032 (Ser 631 Asn) of the chicken Mx gene has been demonstrated to be related to resistance to antiviral activity. This study developed a new real-time PCR-based allelic discrimination assay for the rapid genotyping of the chicken Mx gene G2032A SNP. The distribution of the Mx gene G2032A SNP genotypes and the allele frequencies of A and G alleles among different chicken breed populations were screened with the use of this method.

2.?We combined previously described allele-specific PCR and SYBR Green I-based real-time PCR melting curve analysis with a novel primer design strategy. A pair of outer nested primers was designed to amplify a fragment containing the SNP site, and two 3′-specific, allele-specific primers were combined with the outer primers to amplify SNP-specific fragments. Genotypes were identified based on the characteristic melting temperature of the SNP-specific fragments.

3.?Genotyping assignments were successfully performed on samples from 8 chicken breeds, which were analysed by agarose gel electrophoresis of the PCR products and compared with results obtained from the direct sequencing of the outer primer amplicon. Five native breeds from Southern China carried a relatively higher frequency of the resistant A allele than the three commercial chicken lines.

4.?This single-tube real-time multiplex allele-specific PCR assay is rapid, reliable, sensitive and easy to perform. It is appropriate for high-throughput sample analysis in large population-based Mx SNP genotyping studies.     

Improved early performance of turkey poults given an Aloe ferox leaf extract with bentonite

1. AT-402 (20 g/kg), a proprietary blend of Aloe ferox leaf extract and calcium bentonite, had no adverse effect on poult body weights at 3 weeks of age.

2. Lower body weights and lower feed consumption were observed in 4-week-old poults given 10 and 20 g/kg of AT-402 compared to poults given 0 or 5 g/kg.

3. Floor-reared poults, given either 5 or 10 g/kg AT-402 to 4 weeks of age, were significantly heavier than controls (0 g/kg AT-402), and feed conversion ratios for AT-402 given poults were improved.

4. Xylose uptake in 5 g/kg AT-402-fed poults was significantly greater than in poults given AT-402 at 0 and 10 g/kg, reflecting increased body weights of 5 g/kg AT-402-fed poults.

5. Delayed access by poults to the AT-402 until 10 d of age also improved 4-week body weights, suggesting that the AT-402 might improve performance as soon as it is given.

6. AT-402 at 5 g/kg was most efficacious as demonstrated by improved body weights and feed conversions.     

Polymorphisms of the prolactin gene and their association with egg production traits in two Chinese domestic ducks

1. Prolactin (PRL) as a polypeptide hormone which plays a crucial role in egg production traits.

2. Polymorphisms of the PRL gene were analysed with DNA sequencing and polymerase chain reaction-single-strand conformation polymorphism methods in two Chinese domestic laying duck breeds (Jinding, n = 400, Youxian, n = 400, respectively).

3. The results showed that one polymorphism was detected (A-412G) in intron 1 of the PRL gene, with three genotypes: AA, AG and GG. Association analysis showed that the ducks with the GG genotype had significantly greater egg production and egg weight than those with AG and AA genotype (p < 0.05). Hence, the 412A > G polymorphism of the PRL gene in intron 1 is a potentially valuable genetic marker for laying duck breeding programmes.     

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.     

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.     

Research note: methodology for high-quality RNA extraction from poultry whole blood for further gene expression analysis

1. There are no published methods for RNA isolation from avian whole blood where nucleated red blood cells prevent the use of established mammalian protocols. The aim of this study was therefore to develop a protocol for total RNA extraction using avian whole blood by defining the effect of anticoagulants and sample purification protocols on RNA yield and quality.

2. Blood collections from the cutaneous ulnar or medial metatarsal veins of birds yielded adequate blood volume (2–3 ml) draws. The experiment was a 2 × 2 × 3 factorial arrangement of treatments, with two levels of DNase (0 and TURBO DNA-free Kit), two levels of Cleanup (0 and RNeasy MinElute Cleanup Kit), and three anticoagulants (no anticoagulant, EDTA, or sodium citrate).

3. RNA was isolated successfully by adding TRIzol LS to 0.25 ml of chicken whole blood at 3:1 ratio. From 0.125 ml of avian whole blood, 2–3 µg of RNA with RNA integrity number values of 7.75 was successfully isolated with the TRIzol LS extraction and an RNeasy MinElute Cleanup Kit.

4. This reliable protocol can be used to extract high yield and quality of total RNA from a small amount of whole blood.     

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.