Effect of supplementing vitamin E analogues on post-thaw semen parameters and fertility in chicken

1. The effect of supplementing water-soluble vitamin E analogues 6-hydroxy-2,5,7,8-tetramethylchromane-2-carboxylic acid (trolox) and butylated hydroxy toluene (BHT) was studied in two separate experiments.

2. In the first experiment, trolox was supplemented at 0.2 mM, 0.4 mM and 0.8 mM concentrations along with N-methylacetamide (MA; 12% final concentration) and semen was cryopreserved in 0.5 ml French straws. Different semen parameters and fertility were assessed from post-thaw samples.

3. Sperm motility, live sperm, and mitochondrial activity were significantly lower (P < 0.05) in cryopreserved semen. Lipid peroxidation (LPO) was significantly higher (P < 0.05) in cryopreserved semen that was reduced by trolox supplementation. The treatment containing trolox at 0.2 mM concentration produced significantly higher (P < 0.05) fertility compared to unsupplemented cryopreservation treatment.

4. In the second experiment, BHT was supplemented at 0.25 mM, 0.5 mM, and 1 mM concentrations along with MA during semen cryopreservation.

5. Sperm motility, live sperm and MTT dye reduction test were significantly lower (P < 0.05) in cryopreserved semen. These parameters declined with increasing BHT concentration. Abnormal sperm was significantly higher (P < 0.05) in the BHT supplemented treatments. The sperm chromatin dispersion (SCD) test was significantly higher (P < 0.05) in cryopreserved samples and was highest in samples supplemented with 0.5 mM and 1 mM BHT. The percentage fertility was significantly lower (P < 0.05) in cryopreserved semen and BHT supplementation did not improve fertility.

6. In conclusion, trolox supplementation at 0.2 mM concentration during semen cryopreservation improved fertility, whereas BHT supplementation resulted in a decline in post-thaw semen parameters.     

Effects of enhancing vitamin D status by 25-hydroxycholecalciferol supplementation,alone or in combination with calcium and phosphorus,on sternum mineralisation and breast meat quality in broilers

1. The aim of the present study was to examine the effects of improving vitamin D status in broiler diets by supplementary 25-hydroxycholecalciferol (25OHD₃), alone or in combination with calcium (Ca) and available phosphorus (aP), on live performance, sternum mineralisation and breast meat quality in broilers.

2. A total of 936 1-d-old Ross 308 broilers were used in the study. After gender determination at the hatchery, chicks from each sex were randomly distributed into three dietary treatments. The following dietary treatments were used in the experiment from hatch to 38 d: (1) A control diet formulated to meet all of the nutrient requirements of broiler chicks according to the management guide; (2) The control diet supplemented with 18.7–15.0 µg/kg of 25OHD₃; and (3) The control diet supplemented with 18.7–15.0 µg/kg of 25OHD₃ plus Ca + aP.

3. Improvement in vitamin D status by 25OHD₃ supplementation, alone or in combination with Ca and aP, had no effect on body weight and feed conversion ratio of broilers.

4. The serum 25OHD₃ concentration significantly increased with 25OHD₃ and 25OHD₃ plus Ca + aP supplementation (P < 0.05), whereas the ionised Ca and Mg concentrations remained unchanged.

5. Sternum absolute weight, ash content and the concentrations of Ca and P significantly increased (P < 0.01) with supplementation of 25OHD₃, alone or in combination with Ca + aP.

6. Supplemental 25OHD₃, alone or in combination with Ca + aP, slightly increased pH₂₄ (P = 0.05) and decreased (P < 0.01) squeezable water loss in breast meat, whereas it had no significant effect on lightness, yellowness and sarcoplasmic protein solubility.

7. In conclusion, the results suggested that enhancing vitamin D status by 25OHD₃ supplementation alone or in combination with Ca + aP may improve sternum structure and mineral accretion. Furthermore, supplemental 25OHD₃, even in a nutritionally complete diet, may offer an effective way to improve protein solubility in female broilers.     

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.     

Different susceptibility to fatty liver-haemorrhagic syndrome in young and older layers and the interaction on blood LDL-C levels between oestradiols and high energy-low protein diets

1. The objective of the study was to investigate the susceptibility of young and older laying hens to fatty liver-haemorrhagic syndrome (FLHS) and to evaluate the reliability of different blood lipid fractions for predicting or diagnosing FLHS.

2. Forty young hens and 40 older hens were caged individually. Each group of hens was randomly allotted to four treatments for 21 days: either a control, an oestradiol group, a high energy-low protein diet (HELPD) group or a HELPD + oestradiol group. Blood levels of oestradiol, triglyceride (TG), cholesterol (CHOL), high density lipoprotein-cholesterol (HDL-C) and low-density lipoprotein-cholesterol (LDL-C), liver total lipids, hepatic haemorrhagic scores and productive performance were assessed.

3. In older hens, β-oestradiol increased (P < 0.05) liver total lipids, hepatic haemorrhagic scores and the incidence of FLHS but reduced (P < 0.05) productive performance; however, such changes were not observed in young hens.

4. In two groups of hens, serum TG, CHOL and HDL-C levels were increased (P < 0.001) by β-oestradiol. Hens with FLHS had higher serum TG, CHOL and HDL-C (P < 0.001) than non-FLHS birds in the older layer group of hens.

5. An interaction (β-oestradiol × HELPD) (P < 0.05) for LDL-C levels was observed in both groups of hens. In young hens, β-oestradiol induced a decrease (P = 0.004) in serum LDL-C levels but the effect was attenuated by HELPD. In older hens, HELPD caused an increase (P = 0.02) in serum LDL-C although the effect depended on the presence of β-oestradiol.

6. In conclusion, older layers were more susceptible to FLHS than young layers after oestradiol treatment. Blood TG, CHOL and HDL-C rather than LDL-C levels can be used as a prediction tool for the overall susceptibility to FLHS in older rather than young layers. There were interactions between oestradiol and HELPD on blood LDL-C levels in laying hens.     

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.     

Effects of dietary supplementation of choline and carnitine on growth performance,meat oxidative stability and carcass composition of broiler chickens fed diets with different metabolisable energy levels

1. This study was conducted to investigate the effects of two lipotropic factors (choline and carnitine) on growth performance, oxidative stability of leg and breast muscles and carcass characteristics in broiler chickens fed diets differing in metabolisable energy (ME) levels.

2. A total of 540 one-d-old Ross 308 broiler chicks were allotted to 9 experimental diets, including three ME levels (control, or 0.42 or 0.84 MJ/kg higher ME) and three types of supplemental lipotropic factors (control, 1000 mg/kg of choline or 100 mg/kg of carnitine) as a 3 × 3 factorial arrangement of treatments. Average daily feed intake (ADFI), average daily gain (ADG) and feed conversion ratio (FCR) were recorded during the starter (1–14 d of age), grower (15–28 d of age) and finisher (29–42 d of age) periods.

3. Results showed that the increase in dietary ME level had no impact on ADFI during the starter and grower periods. In the finisher period, increasing dietary ME decreased (P < 0.001) ADFI. Raising dietary ME level by 0.84 MJ/kg resulted in the greater ADG during the grower (P < 0.05) and finisher (P < 0.001) periods. Moreover, an improvement in FCR was observed with feeding the +0.84 MJ/kg diet. Dietary supplementation of lipotropic factors improved FCR values in birds fed the control and +0.84 MJ/kg diets during the grower and finisher periods (P < 0.01).

4. Dietary supplementation of both choline and carnitine increased (P < 0.05) moisture content of leg muscle, although malondialdehyde content of leg muscle was decreased (P < 0.01) in the presence of both lipotropic factors. Dietary supplementation of carnitine decreased (P < 0.01) leg fat content, and this effect was more obvious with higher ME levels, giving a significant ME × lipotrope interaction (P < 0.05). Higher dietary ME level (+0.84 MJ/kg) reduced (P < 0.05) protein content of breast muscle, but this factor was increased (P < 0.05) by dietary supplementation of choline.

5. Although dietary ME level had no marked effect on carcass yield and internal organ weight, supplemental choline increased (P < 0.01) carcass yield.

6. The results from this trial indicated that dietary supplementation with lipotropic factors can improve feed efficiency in high energy diets. In addition, oxidative stability of leg/breast muscles was improved as a result of dietary supplementation with choline or carnitine.     

EDACO,a derivative of myricetin,inhibits the differentiation of Gaoyou duck embryonic osteoclasts in vitro

1. This study determined the effects of (E)-3-(2-(4-(3-(2,4-dimethoxyphenyl)acryloyl)phenoxy)ethoxy)-5,7-dimethoxy-2-(3,4,5-trimethoxyphenyl)-4H-chromen-4-one (EDACO) on the differentiation of Gaoyou duck embryonic osteoclasts cultured in vitro.

2. Bone marrow mononuclear cells (BM-MNC) were collected from 23-d-old Gaoyou duck embryos and induced by macrophage colony-stimulating factor and receptor activator of nuclear factor κB ligand in the presence of EDACO at different concentrations (i.e. 10, 20, 40, 80 and 160 µM). Tartrate-resistant acid phosphatase (TRAP) staining and resorption ability determination were conducted.

3. Results suggested that EDACO suppressed the shaping of positive multinucleated cells and the number of TRAP-positive cells in the 20, 40, 80 and 160 μM EDACO groups was significantly decreased (P < 0.05 or P < 0.01). Besides, the absorption activity of differentiated duck embryonic osteoclasts was significantly inhibited (P < 0.05) in both 80 and 160 μM EDACO groups.

4. Overall, EDACO can inhibit the differentiation of BM-MNC into mature osteoclasts in duck embryos.1     

Influence of citrus flavonoids on laying hen performance,inflammatory immune response,egg quality and yolk oxidative stability

1. The objective of this study was to investigate the effects of dietary supplementation with natural flavonoids (naringin and hesperidin) on laying hens’ performance, cellular immunity and egg quality parameters.

2. A total of 72 individually caged laying hens were allocated into 1 of 6 treatment groups: a control (C) group that was fed with a basal diet and groups that were offered the same diet further supplemented with hesperidin at 0.75 g/kg (E1), or 1.5 g/kg (E2), or naringin at 0.75 g/kg (N1), or 1.5 g/kg (N2) or α-tocopheryl acetate at 0.2 mg/kg (VE) for 67 d.

3. Supplementation with naringin or hesperidin did not affect the performance and egg quality (P > 0.05) apart from an improvement in the yolk colour that was more orange in naringin and hesperidin groups in comparison to the controls (P-linear < 0.05). Egg yolk and plasma cholesterol levels were not affected by citrus flavonoids (P-linear > 0.05).

4. Inflammatory immune response, measured by phytohaemagglutinin skin test (PHA), was suppressed in laying hens that were fed with either naringin (P-linear < 0.05) or hesperidin (P < 0.05). Egg yolk oxidative stability was improved from the 4th d after naringin or hesperidin supplementation. This beneficial effect was comparable to that of α-tocopheryl acetate and was observed in eggs that were stored for up to 120 d.

5. In conclusion, naringin and hesperidin may favourably prolong the shelf life of eggs, appear to possess anti-inflammatory properties and could improve the yolk colour without any side effects on the performance or egg quality traits.     

Evaluation of bee venom as a novel feed additive in fast-growing broilers

1. The present study was designed to evaluate purified bee venom (BV) as an alternative to antibiotics in broiler chickens. The experimental treatment diets were formulated by adding BV into a maize-soybean meal-based diet to give 0, 10, 50, 100, and 500 μg BV per kg of diet.

2. Dietary BV quadratically improved (P < 0.05) feed conversion ratio and increased body weight gain at 1–21 d as level in diet increased. Higher BV levels lowered relative weight of spleen (linear and quadratic, P < 0.05), bursa of Fabricius (quadratic, P < 0.05), and liver (linear and quadratic, P < 0.05) at 21 d of age. Relative breast meat yields were increased quadratically at 21 d and linearly at 35 d with supplementation levels. Dietary BV increased (linear and quadratic, P < 0.05) lightness (L*) value for meat at 21 d, decreased (linear, P < 0.05) ileal villus height and narrowed (quadratic, P < 0.05) width.

3. Dietary BV inclusion linearly increased the concentration of secretory immunoglobulin A (sIgA) on ileal mucosa at 21 d and decreased (quadratic, P < 0.05) nitric oxide contents in serum samples at 21 d and 35 d. Total short-chain fatty acids (SCFA) in caecal digesta were reduced with increasing venom in diets at 21 d of age. None of the serum parameters except for creatinine was affected by dietary BV.

4. It was concluded that dietary BV exhibited wide range of in vivo biological properties in broiler chickens and could be incorporated into feed to promote growth and animal health.     

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.     

Egg quality,fatty-acid composition and gastrointestinal morphology of layer hens fed whole flaxseed with enzyme supplementation

1. Flaxseed is a rich source of α-linolenic acid (ALA, 18:3 n–3). Feeding flaxseed to hens can increase n–3 fatty acids (FA) in eggs. However, non-starch polysaccharides (NSP) in flaxseed decrease nutrient digestibility and can have a negative impact on egg n–3 FA incorporation. Addition of carbohydrase enzymes to flaxseed-based diets can decrease the anti-nutritive effects of NSP.

2. An experiment was conducted to investigate the effect of enzyme supplementation on FA composition and gastrointestinal morphology in hens fed flaxseed. A total of seventy-two, 51-week old brown layer hens were randomly assigned to one of the four dietary treatments (six replicates with three hens per replicate): corn-soybean based diet containing 0% flax (Control), 10% flax (Flax), Flax+0.05% enzyme (Flax+E1), or Flax+0.1% enzyme (Flax+E2) in a 120-day feeding trial.

3. Egg weight was highest in hens fed Flax+E1 (P < 0.05). Yolk weight was higher in Flax+E1 compared with the control and Flax+E2 and was not different from Flax treatment. ALA and total n–3 FA was highest in eggs from Flax+E2 hens (P < 0.05). Addition of enzyme has no effect of on docosahexaenoic acid (DHA), total long chain (>20-C FA), or n–6:n–3 FA ratio in eggs from hens fed flaxseed-based diets (P > 0.05). Over nine-fold increase in hepatic ALA was observed in the liver of hens fed flaxseed-based diets when compared with the control diet (P < 0.0001). No effect of enzyme supplementation was observed on liver ALA, DHA or long chain n–3 FA (P > 0.05). Enzyme supplementation reduced arachidonic acid, total n–6 and LC n–6 FA in liver tissue from hens fed flaxseed-based diets (P > 0.05).

4. Villi height and width was higher in the duodenum and jejunum of hens fed flax-based diets compared to the control (P < 0.05). Enzyme supplementation led to an increase in villi width in jejunum (P < 0.05) in hens fed Flax+E2 (P < 0.05). No effect of diet was observed in the crypt depth and villi height:crypt depth ratio in the jejunum (P > 0.05).

5. It was concluded that enzyme supplementation enhanced total n–3 FA deposition in eggs and liver and influence gastrointestinal morphology in layer hens fed flaxseed.     

Growth performance of crossbred naked neck and normal feathered laying hens kept in tropical villages

1. Two experiments were conducted to develop naked neck (Na/na) and normal feathered (na/na) crossbreds and compare their growth performance, linear body measurements and carcass characteristics in the first and second filial generations.

2. In the first experiment, 4 indigenous naked neck males (Na/na) were mated to 36 Lohmann commercial females (na/na) in a ratio of 1:9. The two genotypes (Na/na, na/na) were allocated randomly according to batches of hatch, sire lines and sex to three different villages.

3. In the second experiment, 10 males and 100 females of F₁ Na/na birds were selected and mated inter se in a ratio of 1:10. The three genotypes (Na/Na, Na/na and na/na) were compared in a randomised complete block design experiment, with the three villages, hatch and sex as blocks and the three genotypes as treatments. F₁ Na/na birds had significantly higher (P < 0.05) feed conversion ratio, body weight, body weight gain, linear body measurements, survivability and carcass yield than their na/na counterparts.

4. In the F₂ generation, Na/Na and Na/na birds had significantly higher (P < 0.05) feed conversion ratio, body weight, body weight gain, linear body measurements, survivability and carcass yield compared to their na/na counterparts.

5. The birds showing the naked neck phenotype appeared to show superior performance compared to normal feathered birds and could be exploited for potential utilisation in local poultry production.     

Influence of graded inclusion of white lupin (Lupinus albus) meal on performance,nutrient digestibility and ileal viscosity of laying hens

1. The aim of this study was to investigate the effect of white lupin (Lupinus albus) meal (WLM) addition on the intestinal viscosity, bird performance and nutrient utilisation of laying hens.

2. The experiment was conducted with 360 laying hens aged 21 weeks fed one of 6 treatments, including a corn-soybean meal control diet (CON) and 5 experimental diets containing 60, 120, 180, 240 and 300 g/kg WLM.

3. A linear increase in feed intake (p < 0.001) was observed with higher levels of WLM from 0 to 300 g/kg. Laying rate decreased quadratically (p < 0.05) and egg weight (at 6th, 12th and 18th weeks of the trial) decreased linearly with WLM inclusion from 0 to 300 g/kg. Birds fed 60 g/kg or more of WLM laid lighter eggs (p < 0.05) than CON hens. When 240 g/kg or more WLM was included into the diet, laying rate was affected negatively (p < 0.05).

4. As WLM increased from 0 to 300 g/kg, apparent metabolisable energy and pre-caecal digestibility of dry matter and crude protein decreased quadratically (p < 0.05). When 300 g/kg of WLM was used, there was a tendency (p < 0.1) to decrease pre-caecal starch digestibility. WLM dose exerted a quadratic effect (p < 0.05) on total sialic acid excretion. As WLM increased, the viscosity of ileal digesta linearly increased (p < 0.05).

5. In the 6th and 12th weeks of the experiment (p < 0.05), eggshell thickness decreased linearly when 240 g/kg of WLM was added. At the 6th, 12th and 18th weeks, a linear decrease in eggshell content was observed (p < 0.05) after WLM addition.

6. In conclusion, the graded inclusion of WLM into laying hens’ diets resulted in depressed performance, AME_N and eggshell quality.     

Effects of heat-treated hempseed supplementation on performance,egg quality,sensory evaluation and antioxidant activity of laying hens

1. This study was conducted to determine the effects of raw and heat-treated hempseed (HHS, Cannabis sativa L.) on performance, egg quality and antioxidant activity in laying hens.

2. A total of 108 laying hens, aged 36 weeks, were divided into three treatment groups with 12 replicates and each replicate contained three laying hens. The treatments were as follows: (1) Control (no hempseed), (2) 15% raw hempseed (RHS) in diet and (3) 15% HHS in the diet. Experiments lasted for 12 weeks.

3. Feed intake of the RHS group was lower than those of the control and HHS groups. Egg weight, egg mass, shell weight, shell surface area and shell thickness of the HHS group were significantly (P < 0.05) higher than that of the RHS group. Roche (DSM) colour fan values of the RHS group were higher than that of the HHS group (P < 0.01).

4. Palmitic, palmitoleic and oleic acids of egg yolk were significantly (P < 0.05) decreased in the RHS and HHS groups; however, linoleic, α-linolenic and docosahexaenoic acids (DHA) of egg yolk increased (P < 0.05) for both treatment diets compared to the control group.

5. Both RHS and HHS supplementation to layer diets did not influence malondialdehyde (MDA) and superoxide dismutase (SOD) activity and blood lipid profile.

6. It was concluded that HHS was superior in improving the egg quality of laying hens as compared to the RHS.     

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.     

Cryopreserving turkey semen in straws and nitrogen vapour using DMSO or DMA: effects of cryoprotectant concentration,freezing rate and thawing rate on post-thaw semen quality

1. This study was designed to identify a suitable protocol for freezing turkey semen in straws exposed to nitrogen vapour by examining the effects of dimethylacetamide (DMA) or dimethylsulfoxide (DMSO) as cryoprotectant (CPA), CPA concentration, freezing rate and thawing rate on in vitro post-thaw semen quality.

2. Pooled semen samples were diluted 1:1 (v:v) with a freezing extender composed of Tselutin diluent containing DMA or DMSO to give final concentrations of 8% or 18% DMA and 4% or 10% DMSO. The semen was packaged in 0.25 ml plastic straws and frozen at different heights above the liquid nitrogen (LN₂) surface (1, 5 and 10 cm) for 10 min. Semen samples were thawed at 4°C for 5 min or at 50°C for 10 s. After thawing, sperm motility, viability and osmotic tolerance were determined.

3. Cryosurvival of turkey sperm was affected by DMSO concentration. Freezing rate affected the motility of sperm cryopreserved using both CPAs, while thawing rates showed an effect on the motility of sperm cryopreserved using DMA and on the viability of sperm cryopreserved using DMSO. Significant interactions between freezing rate × thawing rate on sperm viability in the DMA protocol were found.

4. The most effective freezing protocol was the use of 18% DMA or 10% DMSO with freezing 10 cm above the LN₂ surface and a thawing temperature of 50°C. An efficient protocol for turkey semen would improve prospects for sperm cryobanks and the commercial use of frozen turkey semen.     

Hydrogenated oil decreases tissue concentrations of n‐6 polyunsaturated fatty acids and may contribute to dyschondroplasia in broilers

1. In a factorial design of dietary treatments, male Ross broilers were given diets containing soyabean oil, hydrogenated soya‐bean oil (as a source of trans‐fatty acids) or feed fat with either 0 or 300 μg of added D‐biotin/kg.

2. Growth to 28 d was not influenced by the dietary treatments.

3. Length of tibiotarsal bones was reduced (P<0.05) and severity of leg bone cartilage lesions, characteristic of dyschondroplasia, was highest (P<0.05) in broilers fed on diets containing hydrogenated soyabean oil.

4. Feeding hydrogenated soyabean oil lowered (P< 0.05) the concentrations of C₂₀:_4n6 and the ratios of C_20:4n6/C_18:2n6 in liver and growth plate cartilage.

5. Growth plate cartilage from birds affected with dyschondroplasia contained lower proportions of prostaglandin precursor fatty acids compared with normal growth plate.

6. It is speculated that an inhibition of prostaglandin biosynthesis brought about by the presence of trans‐fatty acids might contribute to the occurrence of lesions similar to dyschondroplasia.     

The effect of tonicity of storage media for fowl semen on the occurrence of neck‐bending spermatozoa,fertility and hatchability 1

1. Semen of White Leghorn and Rhode Island Red males was stored for 24 h in diluents hypertonic (460 mOsm/kg H₂O) and isotonic (340 mOsm/kg H₂O) to cock seminal plasma.

2. Compared with the fertility results with semen that had been stored in the hypertonic diluent or was fresh, the fertility of the White Leghorns was not affected after storage in the isotonic diluent; a decrease (P < 0.lb05) was observed, however, using Rhode Island Red semen and isotonic diluent.

3. Fresh RIR semen contained 2.lb31% “neck‐bent spermatozoa” (NBS) which was increased to 4.lb23% and 5.lb76% after dilution in hypertonic and isotonic diluents respectively and stored for 24 h. It is doubtful whether this increase (P < 0.lb05) is the sole reason for the lowered fertility obtained with this breed after storage in the isotonic diluent.     

In vitro initiation of the acrosome reaction in the emu (Dromaius novaehollandiae)

1. An assessment of the efficiency of the acrosome reaction (AR) provides an important predictor of the fertilizing potential of semen and for diagnosis of the causes of infertility. A standardized protocol was therefore developed for initiation of the acrosome reaction in emu spermatozoa in vitro, and the role of CaCl₂ or perivitelline membrane (PVM) proteins in determining the outcome of the reaction was investigated.

2. The acrosome reaction (assessed by FITC-PNA) was successfully induced in live spermatozoa by incubation for 2 min in NaCl-TES medium supplemented with 5 mM CaCl₂. The maximum response was 32% live acrosome-reacted spermatozoa (LAR) achieved after 10 min incubation.

3. Compared to the outcome with 5 mM CaCl₂ or PVM protein alone, the response was significantly better with a combination of PVM protein and CaCl₂.

4. A significant variation in the percentage of LAR spermatozoa among individual males was observed. No treatment affected the percentage of dead acrosome-reacted spermatozoa.

5. The results emphasize the important role played by both PVM proteins and Ca²⁺ in the in vitro initiation of the acrosome reaction.