Allele‐specific PCR typing and sequencing of the mitochondrial D‐loop region in four layer breeds

This study aimed to investigate the ability of single nucleotide polymorphism (SNP) haplotypes in chicken mtDNA for presumption of the origins of chicken meat. We typed five SNPs of the D‐loop region in mtDNA by allele‐specific PCR (AS‐PCR) in 556 hens, that is 233 White Leghorn (WL), 50 Dekalb‐TX35 (D‐TX), 140 Barred Plymouth Rock (BPR) and 133 Rhode Island Red (RIR) kept in the National Institute of Livestock and Grassland Science (NILGS, Tsukuba, Japan). Five haplotypes were observed among those chickens by AS‐PCR. WL, D‐TX, BPR and RIR displayed three, two, one and four SNP haplotypes, respectively. By a combination of the haplotypes by AS‐PCR and the breeds, these chickens were classified into 10 groups. After the D‐loop was sequenced in two chickens from every group (20 individuals), 15 SNP sites (including one insertion) and eight sequence haplotypes were observed. In conclusion, haplotype variation was observed in and among the layer breeds of the NILGS. This study demonstrates that SNP haplotypes in mtDNA should be appropriate for the presumption of the origins of chicken meat.     

D‐loop sequence mitochondrial DNA variability of Sarda goat and other goat breeds and populations reared in the Mediterranean area

To provide useful knowledge on goat breed origin and history, we studied the mitochondrial DNA (mtDNA) of 69 goats from five different breeds, Camosciata delle Alpi, Maltese, Nubian, Saanen and Sarda, and one population, the Tunisian. All goats analysed displayed a moderate haplotype and nucleotide diversity. The highest was in the Sarda – the autochthonous breed reared in Sardinia. On the basis of mtDNA control region sequences, animals showed a high genetic haplotype diversity, 35 haplotypes were each represented by a single sequence and only a few haplotypes were shared among the animals. New haplotypes of goats reared in the Mediterranean area were identified and the majority of Italian goats belonged to haplogroup A. This result confirmed worldwide distribution and diversity of haplogroup A.     

Effects of dietary supplemental methionine source and betaine replacement on the growth performance and activity of mitochondrial respiratory chain enzymes in normal and heat‐stressed broiler chickens

This study aimed to evaluate the effects of dietary supplemental methionine (Met) source and betaine (Bet) replacement for Met on performance and activity of mitochondrial respiratory chain enzymes (MRCEs) in normal and heat‐stressed broiler chickens. Total of 1,200‐day‐old Ross 308 chicks were allocated to two houses, each consisted of 12 treatments, five replicates of 10 birds each with 2 × 2×3 × 2 (temperature × Met source × Met level × Bet, respectively) split‐plot factorial arrangement. Met level in the basal diets was 70% requirements (Req) that was increased to the requirement or 130% by supplemental dl ‐ or l ‐Met. Bet was or was not substituted at the rate of 30% supplemental dl ‐ or l ‐Met. Feed conversion ratio (FCR) in chicks fed 70% l ‐Met was lower than those fed 70% dl ‐Met diet during 1–10 days (p = 0.04). Broilers fed diets containing requirement or 130% Met, regardless of its source, showed higher weight gain (WG) than those received 70% Met diet during 11–42 days (p < 0.001). Feed intake (FI) of broilers fed 130% Met diet was decreased compared to other two groups during 11–42 days (p < 0.05). One hundred thirty percent Met requirement diet resulted in lower FCR comparing to other two groups during 11–42 days (p < 0.001). Heat‐stressed birds grew less than those under normal condition (p < 0.05). Broilers fed Req Met diet under normal temperature exhibited higher activities of complexes (Cox) I and III (p < 0.05). Cox I activity in heat‐stressed birds fed Bet + diet was similar to those fed Bet‐diet under normal temperature (p = 0.046). It is concluded that performance and the activities of Cox I and III were increased as the level of Met increased. Bet replacement for 30% supplemental Met resulted in similar consequences comparing to non‐Bet replacement diets on performance, but increased the activity of Cox III. l ‐Met was effective than dl ‐Met at the cellular level. High ambient temperature depressed performance and MRCE activity.     

Content and distribution of different phosphorus forms in broiler chickens

To understand the background value of phosphorus in chickens, the quantitative distribution of different phosphorus forms, including total phosphorus (TP), free phosphate (FP) and phospholipid (PL), in viscera, blood and bones of broiler chickens was investigated. Results showed that phosphorus contents exhibited significant differences in different parts of chickens. TP content of breast and thigh meat was over 5.0 g/kg, in which most of the phosphorus was in the form of water‐soluble phosphates. TP content in viscera was higher than that in meat, and spleen was observed to contain the highest amount of phosphorus (10.0 g/kg). In all tested organs, FP and PL contents in liver were the highest, ranging between 1207–1989 and 81–369 mg/kg respectively. TP content in chicken bone was in the range of 52 716–136 643 mg/kg, and FP content in the bone was relatively lower than that in chicken meat.     

Gender‐specific effects of a phytogenic feed additive on performance,intestinal physiology and morphology in broiler chickens

To date, most studies published were carried out on broilers of the same sex, and possible gender‐specific effects of phytogenic substances have not been investigated so far. A 3 × 2 factorial study was performed to examine gender‐specific effects of a PFA at two dietary levels (150, 1500 ppm) on growth performance, carcass traits and gastrointestinal attributes in broiler chickens versus an untreated control group. The addition of 150 ppm of the PFA led to a downregulation of trypsinogen mRNA in pancreas compared with the control group (p < 0.05). The number of goblet cells decreased in jejunum compared with the unsupplemented group, whereby this effect was more pronounced in male birds (p < 0.05). Furthermore, higher methylamine contents compared with the control group were measured (p < 0.01). In proximal ileum, female birds, supplemented with 150 ppm PFA, had lower crypt depths than their litters in the 1500 ppm treatment (p < 0.05). In distal ileum, villus height:crypt depth ratio was higher in birds fed the PFA at 150 ppm than in the control group (p < 0.05). The 1500 ppm dosage of the PFA increased jejunal histamine concentration compared with the negative control group (p < 0.05). Jejunal histamine concentration was also affected by the interaction PFA × sex (p < 0.05). Regardless of inclusion level, total amount of biogenic amines and other microbial metabolites in digesta samples was not affected by the PFA. These results demonstrate variable, partially gender‐specific effects of the tested PFA. Although the supplementation of 150 ppm showed little effect on mRNA expression level of selected marker genes for nutrient digestion, beneficial effects on gut morphology were observed. The 10‐fold higher dosage of the PFA did not adversely affect growth performance as well as most investigated parameters compared with the control group.     

The effect of dietary turmeric on antioxidant properties of thigh meat in broiler chickens after slaughter

Two hundred and fifty‐day‐old male broiler chicks (Ross 308) were used to investigate the dietary supplementation effects of different levels of 0.0 turmeric rhizome powder (TRP) (free TRP, F.TRP), 0.25 (low TRP, L.TRP), 0.5 (medium TRP, M.TRP) and 0.75% TRP (high TRP, H.TRP) or 50 mg/kg vitamin E (VE) on antioxidant properties of thigh meat in broiler chickens after slaughter. No effect of treatment was observed for the activities of glutathione peroxidase (GPX) and superoxide dismutase (SOD) and total antioxidant status (TAS) in thigh meat (P > 0.05) but all were reduced over time (P < 0.0001). A significant treatment*time interaction was observed for thigh meat GPX activity (P = 0.02). Significant effects of treatment and time were observed for malondialdehyde (MDA) content (P < 0.05). There was no significant difference between the treatments for thigh meat MDA content at slaughter (P > 0.05). Three days after refrigerator storage, MDA content in thigh meat of M.TRP birds was lower than that of F.TRP birds (P < 0.05). Both the M.TRP and VE birds showed a lower MDA content in thigh meat as compared with F.TRP birds 7 days after storage (P < 0.05). In conclusion, dietary consumption of 5 mg/kg TRP can increases the thigh meat shelf‐life storage and quality in broiler chickens after slaughter.     

Effects of dietary supplementation with carnosine on meat quality and antioxidant capacity in broiler chickens

1. This study aimed to investigate the effects of carnosine supplementation on meat quality, antioxidant capacity and lipid peroxidation status in broiler chickens.

2. A total of 256 1-d-old male Arbor Acres broilers were randomly assigned to 4 treatments consisting of 8 replicates of 8 chickens each. The birds were supplied with 4 different diets: a basal diet or a basal diet supplemented with 100, 200 or 400 mg/kg carnosine, respectively. The whole experiment lasted 42 d.

3. The results showed that dietary supplementation with carnosine linearly increased the values of pH_45 min and redness and reduced drip loss of breast meat. Dietary carnosine increased the activity of antioxidant enzymes in liver, serum and breast meat and decreased the contents of lipid peroxides at 21 and 42 d of age.

4. These findings indicated that dietary supplementation with carnosine was beneficial to enhance meat quality, antioxidant capacity and decrease lipid peroxidation status of breast meat.     

In ovo carbohydrate supplementation modulates growth and immunity‐related genes in broiler chickens

A study was undertaken to investigate the role of in ovo administrated carbohydrates on the expression pattern of growth and immune‐related genes. In ovo injections (n = 400) were carried out on the 14th day of incubation into the yolk sac/amnion of the broiler chicken embryos. Expression of growth‐related genes: chicken growth hormone (cGH), insulin‐like growth factor‐I & II (IGF‐I & II) and mucin were studied in hepatic and jejunum tissues of late‐term embryo and early post‐hatch chicks. Expression of candidate immune genes: Interleukin‐2, 6, 10 and 12 (IL‐2, IL‐6, IL‐10 and IL‐12), Tumour necrosis factor‐alpha (TNF‐α) and Interferon gamma (IFN‐γ) were studied in peripheral blood monocyte cells of in ovo‐injected and control birds following antigenic stimulation with sheep RBC (SRBC) or mitogen concanavalin A (Con‐A). Glucose injection significantly increased the expression of IGF‐II gene during embryonic period and both cGH and IGF‐II in early post‐hatch period, while ribose‐injected chicks had higher expression of IGF‐II gene during embryonic stage. Enhanced mucin gene expression was also observed in fructose‐injected chicks during embryonic age. Glucose‐injected chicks had higher expression of IL‐6 or IL‐10, while those injected with fructose or ribose had higher expression of IL‐2, IL‐12 and IFN gamma. It is concluded that in ovo supplementation of carbohydrates might help in improving the growth of late‐term embryos and chicks. In ovo glucose could modulate humoral‐related immunity, while fructose or ribose might help in improving the cellular immunity in broiler chickens.     

Dietary mannanase‐hydrolyzed copra meal improves growth and increases muscle weights in growing broiler chickens

The utilization of copra meal as a feed ingredient is limited because it contains a high level of mannan. However, recent findings indicate that the effect of copra meal on growth performance in broiler chickens can be improved by the supplementation of mannanase in the diet. In the present study, we examined the effect of mannanase‐hydrolyzed copra meal (MCM) on growth performance and muscle protein metabolism in growing broiler chickens (Gallus gallus domesticus). Forty 8‐day‐old male broiler chicks were assigned to two groups (four birds in each pen, five replicates) and fed either a commercial diet (as a control diet) or a diet containing MCM at 0.2% until 22 days of age. Dietary MCM significantly increased the weights of body, breast muscle, and thighs in chickens, whereas the weights of abdominal adipose tissue and liver were not affected. Cumulative feed intake was significantly increased by MCM. Dietary MCM significantly decreased plasma 3‐methylhistidine level. The messenger RNA and protein levels of muscle protein metabolism‐related factors were not altered by MCM. These findings suggest that the growth‐promoting effect of MCM is related to the suppression of muscle proteolysis in growing broiler chickens.     

Analgesic effects of morphine and butorphanol in broiler chickens

Objective

To evaluate analgesic efficacies of morphine and butorphanol in lame broiler chickens.

Pulmonary hypertension and right ventricular failure in broiler chickens reared at high altitude is affected by dietary source of n‐6 and n‐3 fatty acids

The present study evaluated the development of pulmonary hypertension and right ventricular failure in broiler chickens reared at high altitude (2100 m) as affected by dietary intake of n‐3 and n‐6 fatty acid sources. Flax oil and soy oil were used as sources of n‐3 and n‐6 fatty acids, respectively, either with or without α‐tocopheryl acetate. A total of 192 day‐old broiler chicks (Ross 308) were used in a completely randomized design using isoenergetic and isonitrogenous experimental diets. Results showed that dietary flax oil significantly (p < 0.05) improved feed conversion ratio during 21–42 days of age. However, body weight gain did not significantly differ among the experimental groups in entire trial. Birds received flax oil had significantly higher serum concentration of nitric oxide (NO) but they had lower serum concentration of malondialdehyde when compared with their counterparts fed with soy oil. Liver and abdominal fat weights were significantly (p < 0.05) reduced by substitution of soy oil for flax oil. The right‐to‐total ventricle weight ratio (RV/TV) and mortality from pulmonary arterial hypertension (PAH) were significantly (p < 0.05) decreased in birds that received flax oil. In conclusion, n‐3 fatty acids could significantly reduce RV:TV and PAH mortality in birds by increasing circulatory level of NO and suppressing hepatic lipogenesis.     

Mitochondrial D‐loop analysis reveals low diversity in Mangalica pigs and their relationship to historical specimens

The genetic relationship between 195 Mangalica and 79 non‐Mangalica pigs was studied using mitochondrial D‐loop SNP genotyping. Altogether, 35 polymorphic sites and 27 haplotypes were identified. Of the haplotypes, eight and 16 are Mangalica and non‐Mangalica specific, respectively, while three contain both Mangalica and non‐Mangalica individuals. Genetic distance values and phylogenetic analysis indicate that Mangalica individuals are very closely related, and five haplotypes represent approximately 92% of the Mangalica pigs involved in the study, thus determining the major maternal lineages. In contrast to previous microsatellite studies, individuals of Mangalica could not be distinguished as three separate breeds using mtDNA genotyping. Comparing modern and archaeological mtDNA sequences revealed that present day Mangalica is related to pigs that lived in the Carpathian basin where postulated ancestors of Mangalica also lived. This is the first DNA‐based genetic evidence to support the described breeding history of Mangalica.     

Effects of 25‐hydroxycholecalciferol with two D3 vitamin levels on production and immunity parameters in broiler chickens

This study was performed in Ross 308 chickens aged 1–21 days and aimed to evaluate whether the addition of 25‐hydroxycholecalciferol (25(OH)D₃) to broiler chicken diets affects their growth performance and immunity. A completely random 2 × 2 factorial arrangement was used with two levels of vitamin D₃ and the absence or presence of 25(OH)D₃, corresponding to four treatments based on sorghum + soya bean diets: (i) 200 IU of vitamin D₃/kg of feed (Diet 1) (NRC, 1994 ), (ii) Diet 1 + 69 μg of 25(OH)D₃/kg of feed (Diet 2), (iii) 5,000 IU of vitamin D₃/kg of feed (Diet 3) and (iv) Diet 3 + 69 μg of 25(OH)D₃/kg of feed (Diet 4). Each treatment was conducted with six replicates of 10 chickens each. Water and feed was supplied ad libitum. The results showed significantly increased growth and tibia ash (p < .05) in the birds fed 5,000, IU of vitamin D₃/kg + 25(OH)D₃. Additionally, the cellular immune response increased significantly (p < .05) in both treatments with added 25(OH)D_3. Based on the results obtained under the current test conditions, the addition of 25(OH)D₃ at a rate of 69 μg/kg to diets containing vitamin D₃ improved the cellular immune response and mineral deposition in the bones of broilers aged 1–21 days. Because these parameters are very important in modern poultry farming, these results indicate that supplementation with 25(OH)D₃ should improve broiler production.     

Hypophagic and dipsogenic effect of the 5‐HT1A receptor agonist 8‐OH‐DPAT in broiler chickens

The effects of the 5‐HT_1A receptor agonist 8‐OH‐DPAT on food and water intake in male broiler chickens were investigated. The injection of 25 or 50 μg/kg of 8‐OH‐DPAT 15 min before refeeding in fasted animals produced a decrease in food intake. No effect was observed in drinking. The injection of 25 or 50 μg/kg of the 8‐OH‐DPAT 60 min after the start of refeeding did not produce any significant modification in food intake. No effect on drinking was recorded. The agonist 8‐OH‐DPAT injected 15 min before water presentation in water‐deprived chickens, produced an increased drinking 60 min after the presentation of water. No effect on food intake was observed. The results show that the effect on food intake of the agonist 8‐OH‐DPAT in fasted–refed broiler chickens was similar to those observed in mammals and layer‐strain chickens. However, the agonist did not alter significantly the food intake when the broilers were fed 60 min before the injection. These results are contrary to the observed effects in mammals and in layer‐strain chickens. Probably, the selection for rapid growth rate in broilers causes modifications in the feeding control pattern. The comparison between broilers and layers strain may be a useful tool to elucidate the complex mechanisms involved in food and water intake regulation in chickens.     

Antioxidant supplementation of low‐protein diets reduced susceptibility to pulmonary hypertension in broiler chickens raised at high altitude

A reduced‐protein diet (designated as RPD) was prepared and its effects on growth performance and the development of pulmonary hypertension syndrome (PHS) were evaluated in broiler chickens compared to a normal‐protein diet (designated as NPD) or to the RPD supplemented with CoQ₁₀ alone (30 mg/kg) or in combination with vitamin E (30 mg/kg CoQ₁₀ + 100 mg/kg vitamin E). The RPD had 30 g/kg less crude protein compared to the NPD. A total of 208 1‐day‐old male broilers (Ross 308 strain) were used in a 42‐day trial. Serum concentrations of uric acid (UA) and nitric oxide (NO) significantly (p < 0.05) declined when chickens fed on the RPD. However, supplementing RPD with the antioxidants significantly (p < 0.05) increased the serum NO concentration. Although serum malondialdehyde (MDA) concentration was significantly (p < 0.05) higher in the RPD than the NDP, supplementing RPD with CoQ₁₀ and CoQ₁₀ + VE decreased serum MDA concentration to similar levels found in the NPD. Significant overexpression in GPX1 gene observed in the heart and lungs of broilers fed on the RPD, which was effectively restored by supplementation of CoQ₁₀. The right to total ventricular weight ratio (RV:TV) was significantly (p < 0.05) increased in birds fed the RPD, which concurred with an increase in mortality from pulmonary hypertension syndrome (PHS). However, a significant decline in mortality from PHS was observed when birds on RPD received CoQ₁₀ or CoQ₁₀ + VE. In conclusion, antioxidant supplementation effectively improves pulmonary hypertensive response in broiler chicken fed of reduced‐protein diets.     

Effects of thermo‐resistant non‐starch polysaccharide degrading multi‐enzyme on growth performance,meat quality,relative weights of body organs and blood profile in broiler chickens

This research was conducted to study the performance and carcass parameters of broiler chickens fed diets supplemented with heat‐treated non‐starch polysaccharide degrading enzyme. A total of 432 one‐day old Ross 308 broiler chickens were allocated to five treatments: (i) CON (basal diet), (ii) E1: CON + 0.05% multi‐enzyme, (iii) E2: CON + 0.1% multi‐enzyme, (iv) E3: CON + 0.05% thermo‐resistant multi‐enzyme and (v) E4: CON + 0.1% thermo‐resistant multi‐enzyme, each treatment consisted of six replications and 12 chickens in each replication. The chickens were housed in three floor battery cages during 28‐day experimental period. On days 1–7, gain in body weight (BWG) improved by feeding the diets supplemented with thermo‐resistant multi‐enzyme. On days 7–21 and 1–28, chickens fed the diets containing thermo‐resistant multi‐enzyme showed improved (p < 0.05) BWG and feed conversion ratio (FCR) compared to CON group. Supplementing the diets with multi‐enzyme or thermo‐resistant multi‐enzyme affected the percentage of drip loss on d 1 (p < 0.05). Drip loss percentage on days 3 and 5 and also meat colour were not affected significantly. Supplementing the diets with multi‐enzyme or thermo‐resistant multi‐enzyme did not affect the relative weights of organs but compared to CON group, relative weight of breast muscle increased and abdominal fat decreased (p < 0.05). Among measured blood constituents, chickens fed supplemented diets with thermo‐resistant multi‐enzyme showed higher (p < 0.05) IgG. Counts of red and white blood cells and lymphocyte percentage were not affected. In conclusion, the results demonstrated that supplementing pelleted diets with thermo‐resistant multi‐enzyme improved performance of broiler chickens.