Eradication of avian leukosis virus subgroups J and K in broiler cross chickens by selection against infected birds using multilocus PCR.

The avian leukosis virus (ALV) is a serious threat to sustainable and economically viable commercial poultry management world-wide. Active infections can result in more than 20% flock loss, resulting in significant economic damage. ALV detection and elimination from flocks and breeding programs is complicated by high sequence variability and the presence of endogenous virus copies which show up as false positives in assays. Previously-developed approaches to virus detection are either too labor-intensive to implement on an industrial scale or suffer from high false negative or positive rates. We developed a novel multi-locus multiplex quantitative real-time PCR system to detect viruses belonging to the J and K genetic subgroups that are particularly prevalent in our region. We used this system to eradicate ALV from our broiler breeding program comprising thousands of individuals. Our approach can be generalized to other ALV subgroups and other highly genetically diverse pathogens.

Chickens productivity selection affects immune system genes.

The quantitative trait loci associated with the immune properties of chickens are of interest from the point of view of obtaining animals resistant to infectious agents using marker-assisted selection. In the process of selecting markers for genomic selection in broiler-type chickens, a non-standard genotype frequency of the RACK1 gene allele (SNP Gga_rs15788101) in the B5 line of broiler-type chicken cross Smena 8 was identified and it was suggested that this gene was involved in selection. Therefore, it was decided to investigate the available polymorphisms in the three genes responsible for the IgY titer (DMA, RACK1 and CD1B). Molecular typing of single nucleotide polymorphisms of three loci revealed an approach to fixation of the unfavorable allele of the DMA gene (SNP Gga_rs15788237), an approach to fixation of the unfavorable allele of the RACK1 gene and the prevalence of the favorable CD1B gene allele (SNP Gga_rs16057130). Analysis of the haplotypes revealed a strong linkage disequilibrium of these genes. This suggests that these genes experience selection pressure. Analysis of the protein-coding sequences of the CD1B and DMA genes of various breeds of chickens revealed a negative selection of these genes. In order to understand whether the fixation of the studied alleles is the result of artificial selection of the B5 line of the cross Smena 8, an analysis of similar loci in layer chickens Hisex White was carried out. The frequencies of the alleles at the loci of the CD1B gene (Gga_rs16057130) and the RACK1 gene (Gga_rs15788101) in the Hisex White chicken genome differ from the frequencies of the alleles obtained for chickens of the B5 line of the cross Smena 8. It can be assumed that the fixation of the allele in the DMA gene (SNP Gga_rs15723) is associated with artificial or natural selection, consistent in broilers and layers. Changes in the loci Gga_rs16057130 and Gga_rs15788101 in the B5 line of the Smena 8 chickens are most likely associated with artificial selection of broiler productivity traits, which can subsequently lead to fixation of alleles at these loci. Artificial breeding of chickens leads to degradation of the variability of genes encoding elements of the immune system, which can cause a decrease in resistance to various diseases. The study of the negative impact of selection of economic traits on immunity should provide means to mitigate negative consequences and help find ways to obtain disease-resistant animals.

[Meat quality in broilers reared in different housing systems].

Meat quality was assessed in Cobb-500 cage vs. floor-housed broilers slaughtered at 38 vs 49 days of age. Broilers (105 birds per housing system) were reared since 1 day of age in conditions of vivarium of Center for Selection and Genetics «Zagorskoye EPH¼. Fat content in breast meat was significantly higher (p<0.05) at both slaughter ages in cage-housed broilers (2.0 and 2.7% at slaughter age 38 and 49 days, respectively) compared to floor-housed (1.6 and 2.2%). Protein content in breast meat was higher in floor-caged broilers. Total collagen content in thigh meat of floor housed broilers (789.88 mg/100 g) was 1.5 fold higher compared to cage-housed (515.80 mg/100 g, p<0.05). Fatty acid profiles of meat were mostly affected by the type of meat (red vs white) and to a lesser extent by housing system and slaughter age. Water-holding capacity in red meat significantly differed between slaughter ages and between housing systems at slaughter age 38 days (р<0.05): at slaughter age 38 days water-holding capacity in red meat was 67.3 in cage-housed broilers vs. 70.1% in floor-housed; at slaughter age 49 days 74.9 vs. 76.0%, respectively. The five-point scores of sensory taste evaluation for the meat of floor-housed broilers (4.55 and 4.91 for breast meat at slaughter ages 38 and 49 days; 4.40 and 4.90 for thigh meat) were better compared to cage-housed (4.47 and 4.83 for breast meat at slaughter ages 38 and 49 days; 4.37 and 4.70 for thigh meat). The conclusion was made that meat quality estimated by a set of the relevant parameters was marginally better in floor housed broilers in compare to cage-housed.

[Antibiotic-free poultry production based on innovative nutritional programs with the involvement of probiotics].

EU banned antibiotic growth promoters (AGP) for farm animals and poultry since 2006 in relation to the problem of drug resistance. This requires alternative products for equally efficient prevention and treatment of certain alimentary poultry diseases. One of the most actual trends is the development of innovative nutritional strategies for poultry providing an effective symbiosis between the host and its intestinal microbiota. The study presented was aimed at the comparative evaluation of productivity, nutritive value of meat, and composition of intestinal microbial populations in broiler chicks fed different diets (corn - soybean meal, wheat - sunflower cake, barley - sunflower cake) supplemented with AGP or a probiotic (cellulolytic and lactic microorganisms). In three trials straight-run Cobb 500 broilers reared from 1 to 36 days of age were fed these diets supplemented with bacitracin from 1 to 29 days of age (control) or probiotic preparation from 1 to 36 days of age (70 birds per dietary treatment in each trial). There were no differences in live bodyweight and carcass yield between the treatments in all three trials. In the two trials with sunflower cake, protein content in breast meat was significantly higher by in birds fed probiotic in compare to birds fed AGP (by 10.0 and 6.8%, p<0.05), while fat content in thigh meat was lower by 12.0% (p<0.05) and 14.1% (p<0.01), respectively. Content of amino acids in meat did not differ. Vitamin content in the poultry meat of the experimental groups was significantly higher compared to control (p<0.001). The resulting concentrations of bacitracin in meat in control treatments (no more than 0.02 U/g) did not exceed local legislative limitations. The substitution of the probiotic for AGP beneficially affected the composition of bacterial populations in the duodenum and cecae determined using T-RFLP analysis. It was concluded that the supplementation of diets with probiotic allows to produce antibiotic-free broiler meat without detrimental effects on the productive performance.

Metagenomic bacterial community profiles of chicken embryo gastrointestinal tract by using T-RFLP analysis.

Thirty microbial phylotypes of microorganisms were found in the gastrointestinal tract of chicken belonging to the Hajseks White breed, and 38 phylotypes were found in the gastrointestinal tract of chicken belonging to the Hajseks Brown breed. The microbiome of the gastrointestinal tract of the chicken embryos of the Hajseks White breed was dominated by the typical representatives of avian intestinal microflora--bacteria of the family Enterobacteriaceae (47.3%), orders Actinomycetales (13.6%) and Bifidobacteriales (20.6%), and the family Lachnospiraceae (1.1%). The microbiome of the gastrointestinal tract of the chicken embryos of the Hajseks Brown breed was dominated by the pathogenic bacteria of the order Rickettsiales (94.8%). The metagenome of gastrointestinal tract of both breeds also contained a small number of genes of unidentified bacteria.

Broiler Cecal Microbiocenoses Depending on Mixed Fodder.

Molecular genetic techniques (NGS sequencing and quantitative PCR) were used to determine the composition of the cecal bacterial community of broiler chickens fed with different mixed fodder. The Cecal microbiome exhibited taxonomic diversity, with both typical inhabitants of avian intestine belonging to the families Clostridiaceae, Eubacteriaceae, and Lactobacillaceae and to the phylum Bacteroidetes, and new un- identified taxa, as well as bacteria of the families Lachnospiraceae and Ruminococcaceae, which were previ- ously considered restricted to the rumen microflora. Contrary to traditional concepts, enterococci and bi- fidobacteria were among the minor components of the community, lactate-fermenting species were absent, and typical avian pathogens of the genus Staphylococcus were detected but seldom. Members of the family Suterellaceae and the genus Gallibacterium, which are responsible for avian respiratory infections, were also detected. Significant fluctuations of abundance and composition of microbial groups within the cecal com- munity and of the parameters of broiler productivity were found to occur depending on the feed allowance. Cellulose content in the feed had the most pronounced effect on the composition aid structure of bacterial communities. Decreased cellulose content resulted in a decrease of bacterial abundance by-aii order of mag- nitude and in increased ratios of members of the phylum Bacteroidetes and the family Clostridiaceae, which possess the enzymes degrading starch polysadcharides. Abundance of the normal inhabitants of avian intes- tine belonging to the genus Ldctobacillus and the order Bacillales decreased, while the share of Escherichia and members of the family Sutterellaceae increased, including some species capable of causing dysbiotic changes in avian intestine. No significant change in abundance of cellulolytics of the families Ruminococca- ceae, Lachnospiraceae, and Eubacteriaceae was observed.

[Efficacy of various methods of a reporter gene transfer to chicken embryonic cells].

The methods of transfection ofa plasmid with a reporter gene involving DNA injection into chicken embryonic cells were studied. The parameters of the efficient transfection of chicken blastodermal cells with a foreign gene have been determined (20-24 and up to 40% in culture and embryos, respectively). A high efficiency of transfection of primordial germ cells isolated from the gonads has been obtained after DNA injection into the dorsal aorta of 2.5-day-old chicken embryos.

[Amino acid and lipid metabolism in embryos of flesh fowl with different yolk mass].

It has been proposed that variations in relative yolk mass in a population of flesh fowl be used as a model of development of nidicolous and nidifugous birds. During development of the eggs with a high proportion of yolk, an excess of lipids is cleaved at a higher rate and oxidized until day 17 of incubation, while in the embryos developing from the eggs with a low relative yolk mass, amino acids are intensely cleaved during the period preceding the hatching. Significant differences in the body content of cystine were found in 17-day embryos and upon hatching, thus suggesting a delayed activity of the genes encoding keratins in the group corresponding to the seminidicolous type according to the egg content of lipids. These biochemical differences question the widespread concept on the occurrence of dichotomy by the end of embryogenesis and beginning of neonatal growth of nidifugous and nidicolous birds.

[Altricial and precocial developmental features in modern meat-type chicks].

Variability of egg weight, egg yolk content, neonatal growth rate and relationships of these parameters were studied in meat-type chicks. As it had been established the level of variability in neonatal growth traits was greater than variability of the egg morphology parameters. Egg weight had stronger influence on the chicks' neonatal growth rate than egg yolk content did. Low egg size was associated with limited neonatal growth rate variability, declined chick weight at hatching and increased relative growth rate throughout four days post hatch. Comparison of egg morphological parameters in two species having the same female definitive body weight--meat-type domestic fowl (precocial type) and brown pelican (altricial type) has shown, that, in contrary to predicted on the basis of avian developmental typology, egg weight to female body ratio was greater in brown pelican, egg yolk content was equal in both species.

[Eggs fertilizability in domestic hens Gallus gallus domesticus as a function of the yolk surface area]. / Zavisimost' oplodotvoriaemosti iaitz domashnikh kur Gallus gallus domesticus ot ploshchadi poverkhnosti zheltka.

The increase of egg mass and reliable decrease of egg fertilizability were observed when the mass surface area of yolk (egg) increased. The results obtained suggest that, in addition to the number of viable spermatozoa penetrating across the perivitelline membrane within 15-20 min after ovulation, the probability of fertilization depends on the area of egg surface, which approximately corresponds to the area of perivitelline membrane. Apparently, the ratio of receptors' numbers and spermatozoa, which contact with them on the germ disc surface, to their number on the rest part of perivitelline membrane decreases with the increase of yolk size. The decreased egg fertilizability concomitant with the increased area of perivitelline membrane suggests that the egg size is one of the factors of fertility of the female gametes as concerns both variability of the egg composition and age.

Exclusion of broiler breeder females from male feeders with a male only grill. 1. Behavioral and technical aspects of design and development.

A series of experiments was conducted in order to develop a feeder grill that would allow broiler breeder males, but not females, to eat. Females are typically excluded from male feeders by increasing the height of the feeder. However, male feeder heights that exclude most females increase the time required for males to consume a given amount of feed 19 to 60%. Typical male feeder height from floor to feeder pan lip in commercial practice is 55 cm, which is about the distance from floor to the head of the male in an erect posture. Females measure about 40 cm in an erect posture. Because the necks of males (8.0 cm) are longer than those of females (6.3 cm), the lip of male feeder pans was extended horizontally 12.7 cm with a wire mesh on the assumption that females could not stretch their necks enough to reach the feed. However, the females learned quickly to perch on the extended lip and eat. Total exclusion of females from the male feeder, regardless of feeder height, was achieved by placing a horizontal upper mesh 5.6 to 10.2 cm above the extended lip, and connecting the upper mesh to the lower extended lip of the feeder with vertical bars spaced 5.1 cm apart. The horizontal upper mesh prevented perching by females and the 5.1-cm spacing of the vertical bars allowed males but not females to reach the feed by inserting their heads and necks up to their shoulders.(ABSTRACT TRUNCATED AT 250 WORDS)

[3-Hydroxybutyrate dehydrogenase in the liver mitochondria of rats and chickens]. / 3-Oksibutiratdegidrogenaza v mitokhondriiakh pecheni krys i kur.

The activity of mitochondrial 3-hydroxybutyrate dehydrogenase (EC 1.1.1.30) in rat and chicken liver was found to be comparable with the activity of electron transport chain of rat liver mitochondria. This activity is absent in chicken liver mitochondria, which are devoid of the 3-hydroxybutyrate oxidase activity. Both types of mitochondria have nearly identical respiration parameters but respond differently to Mg2+. It was assumed that chicken liver mitochondria are characterized by a low rate of fatty acids oxidation due to the absence of 3-hydroxybutyrate dehydrogenase in these organelles.