A search for genetic markers associated with egg production in the ostrich (Struthio camelus).

The aim of the current study was to search for genetic markers, microsatellite loci associated with laying performance in ostriches. The material consisted of two groups of ostrich hens characterized by high or low laying performance (over 75 and less than 25 eggs per season, respectively). The investigation covered 30 microsatellite loci characteristic for the ostrich (the CAU group) and led to identification of significant differences in allele and genotype frequencies between the two groups of hens considered. Out of a total of 30 microsatellite loci examined, 28 showed different alleles in relation to analyzed performance groups. In hens of high laying performance (HP group, n = 12), specific alleles occurred in 23 microsatellite loci (40 alleles of 243 identified), while in those of low egg production (LP group, n = 12), they occurred in 22 (51 alleles of 243 identified). The results indicate the usefulness of the microsatellite loci as the potential genetic markers associated with laying performance that can be applied for genetic improvement of ostrich flocks.

The use of microsatellite polymorphism in genetic mapping of the ostrich (Struthio camelus).

The aim of this study was to determine microsatellite polymorphism in ostriches and using it in creation the genetic map of the ostrich. The polymorphism analysis covered 30 microsatellite markers characteristic of ostrich, for the CAU (China Agricultural University) group. The material consisted of 150 ostriches (Struthio camelus). The 30 microsatellite loci was examined and a total of 343 alleles was identified. The number of alleles at a single locus ranged from 5 at locus CAU78 to 34 at locus CAU85. The values for the observed heterozygosity H(o) ranged from 0.467 (locus CAU78) to 0.993 (locus CAU16), whereas for the expected heterozygosity H(e)--from 0.510 (locus CAU78) to 0.953 (locus CAU85). Analyzing the individual loci, the highest PIC value, more than 0.7 was observed for: loci CAU85 (0.932), CAU64 (0.861) and CAU32, 75 (0.852), respectively. It should be noted, that the microsatellite markers used in our study were very polymorphic as evidenced by the large number of detected alleles and high rates of heterozygosity, PIC and PE as well. The analysed microsatellite markers may be used in genetic linkage mapping of ostrich, the construction of a comparative genetic map with other ratites, such as emu and rhea, and population genetics studies or phylogenetic studies of these birds.

[Expression profiling of candidate genes for abdominal fat mass in domestic chicken Gallus gallus].

The quantitative traits of mass and percentage of abdominal fat in chicken and various types of obesity in mammals are homologous and functionally similar. Therefore, the genes involved in obesity development in humans and laboratory rodents as well as those responsible for pig lard thickness could be involved in abdominal fat deposition in broilers. Expression of candidate genes FABP1, FABP2, FABP3, HMGA1, MC4R, PPARG, PPARGC1A, POMC and PTPN1 was studied in fat, liver, colon, muscle, hypophysis, and brain in chicken (broilers) using real-time PCR. Significant difference in the HMGA1 gene expression in the liver of broiler chicken with high (3.5 +/- 0.18%) and low (1.9 +/- 0.56%) abdominal fat concentration has been revealed. The expression of this gene was been shown to correlate with the amount (0.7, P < or = 0.01) and mass (0.7, P < or = 0.01) of abdominal fat. The PPARG gene expression in liver in the same chicken subsets was also significantly different. Correlation coefficients of the gene expression with the abdominal fat amount and mass were respectively 0.55 (P < or = 0.05) and 0.57 (P < or = 0.01). Based on these results, we suggest that the HMGA1 and PPARG genes are involved in abdominal fat deposition. The search for single nucleotide polymorphisms (SNPs) in the HMGA and PPARG regulatory regions could facilitate identifying genetic markers for broiler breeding according to the mass and percentage of abdominal fat.

The effect of multigenerational diet containing genetically modified triticale on immune system in mice.

The safety assessment of genetically modified (GM) food and feed is performed to identify the possible effects upon animal and human health, also the long-term, multigenerational influence upon functioning of different organs and systems, such as the immune system. In this study C57BL/6J mice were fed for five consecutive generations with pellets containing 20% of conventional triticale grain (control) vs. pellets containing 20% of the transgenic triticale grain resistant to BASTA herbicide (experimental). The F5 experimental animals showed enlarged inguinal and axillary lymph nodes, but not spleens, and increased WBC counts in blood (but within the norm for Mus musculus). Immunophenotyped cell suspensions derived from spleens, inguinal and axillaris lymph nodes and PBMCs from blood showed the significant decrease in the percentage of T cells in spleen and lymph nodes and the B cells in lymph nodes and blood of the F5 experimental mice in comparison to the control F5 mice. Immunoblotting analysis of IL-2, IL-4, IL-10, IL-12, IL- 6, IFN-gamma levels in serum showed significantly increased IL-2 levels and decreased IL-6 levels in the F5-experimental mice sera. No significant changes in the levels of IgE in sera in both mice groups were observed. The obtained results indicate that multigenerational use of feeds for rodents containing the GM-triticale leads to expansion of the B cell compartment in the secondary lymphoid organs, but it is not caused by malignant processes or the allergic response.

Microsatellite markers may be ineffective in selection of laying hens for polygenic production traits.

Previous research on mapping QTL in a reference family of laying hens indicated that 5 microsatellite loci (MCW0133, MCW0170, MCW0114, MCW0139, and LEI0074) were significantly associated with genome regions affecting shell strength as well as egg and yolk weights. The aim of our investigation was to verify if those markers could be useful in selection of laying hens. The study involved 2 breeds of randomly segregating populations: Rhode Island Reds selected divergently and Green-legged Partridgenous chickens selected upwardly, over 4 generations, for the mentioned egg quality traits. The influence of marker genotype on bird performance was assessed through the prediction of breeding values using a model that distinguished the marker effect from that of the polygenic effect and by comparing breeding values between different genotypes at given marker loci. The effects of the linked QTL regions appeared too small to significantly differentiate the outcomes of classifications fitting or not fitting the marker genotype. Comparison of breeding values between microsatellite genotypes for laying and egg traits revealed that antagonistic pleiotropic effects exist between these 2 groups of traits, adding to the difficulty of accounting for marker genotypes in the selection of laying hens.

Genetic characteristics of the ostrich population using molecular methods.

A genetic analysis was performed on Polish ostriches from the 3 principal ostrich breeds: red-, blue-, and black-necks. The analysis was based on 2 molecular methods: DNA fingerprinting and microsatellites. The DNA fingerprinting patterns were obtained using the restriction enzyme HinfI and Jeffrey's 33.15 probe. The second method consisted of a PCR procedure, for which 5 VIAS-OS primers specific to the ostrich were used. The PCR products were separated on polyacrylamide gel using ALFexpress (Authomated Laser Fluorescent DNA Sequencer). The study aimed at assessing the genetic variability within and among the 3 ostrich breeds as well as evaluating the genetic distance between them, and represents the first report on the genetic characteristics of the ostrich breeds. The results obtained by both methods showed considerable compatibility, especially with regard to the relationship among the breeds analyzed. The diversity within breeds, obtained on the basis of the DNA fingerprinting analysis, proved to be low. Among the ostrich populations analyzed, the highest variability potential was observed for black-necked ostriches (the mean diversity of patterns amounted to 29.04%, whereas the mean heterozygosity was 0.30) and the lowest was observed for the red-necks. The largest genetic similarity was recorded between red- and blue-necked ostriches, but the greatest genetic distance was between the red- and black-necks. This means that the use of birds of those breeds in crosses should result in the highest heterotic effect. Both of these methods measured the genetic distance between the analyzed ostrich breeds that was expected from the geographic origin of these birds. The results obtained in the present study showed that both analytic methods used can be successfully applied when elaborating on the genetic characteristics of the ostrich.

Expression of positional candidates for shell thickness in the chicken.

Expression of 12 positional candidates for QTL affecting shell thickness at 53 wk of lay age (ST53) was investigated by real-time PCR in the distal part of chicken oviducts (uterus) with a forming eggshell. In the local chicken breed Green-legged Partridgenous, the complete cDNA CR523443 (ChEST985k21) was downregulated with ratio of means 0.49 (P < or = 0.01) in the group with low ST53 (248.6 +/- 16.62 microm) relative to the group with the highest ST53 (372.4 +/- 2.07 microm). Expression of this gene was highly correlated (0.85, P < or = 0.01) with shell thickness. No significant difference in expression between the 2 groups with thick (378.4 +/- 3.65 microm) and thin (227.8 +/- 8.99 microm) shell and no significant correlation of expression level with ST53 were detected in Rhode Island Red, which could be explained by strict selection to egg quality traits, including optimal shell thickness in this commercial layer breed. These data suggested that CR523443 was a candidate gene for QTL ST53 in the chicken.

Gene expression profiling of hereditary exencephaly in chickens.

In this preliminary study, differentially expressed genes were investigated in cranial tissues from chickens with hereditary exencephaly using cDNA microarrays containing 1,152 genes and expressed sequence tags (ESTs). Genes showing twofold or greater differences at P < 0.05 between affected and normal cranial cells were considered to be candidates for hereditary exencephaly in chicken. Eighteen ESTs (11 known genes/homologues) were upregulated and 108 ESTs (51 known genes/homologues) were downregulated. The EST AL584231 (ROS006C9), orthologous to human MTHFD1, a known candidate gene for human neural tube defects (NTDs), was expressed at the same level both in normal and affected chicken cranial tissues. ESTs AL584253 (ROS006F7, thioredoxin reductase 1) and AL585511 (ROS024H9, thioredoxin), both involved in NTD pathogenic pathways in mice, were downregulated and had mean ratios of 0.41 and 0.04 for expression in affected vs. normal cells respectively. Expression differences of these two ESTs were confirmed by quantitative real-time polymerase chain reaction. These data indicate that ESTs AL584253 and AL585511 are candidates for hereditary exencephaly in chickens.

Cytogenetic study of some tissues and age-related changes in cell proportions in a goat-sheep chimera.

Different percentages of cells with a female sheep or male goat karyotype were found in kidney (12.0% vs. 88.0%) and lung (42.6% vs. 57.4%) cell cultures from a 10-year-old chimera. Skin biopsies from patches with goat hair or sheep wool showed different, age-related goat-to-sheep fibroblast ratios. Karyotypic analysis of lymphocytes obtained from peripheral blood of the chimera at 6 and 10 yr of age showed no chimerism. Two weeks after birth, however, lymphocytes with both sheep (54,XX) and goat (60,XY) karyotypes were apparent in the blood of this chimera. Twenty percent of the blood cells examined at 2 wk had a caprine karyotype; this proportion declined with time, until it was totally eliminated at age 6.

A cytogenetic study of cows from a highly industrial or an agricultural region.

An examination was made of the frequency of chromosomal aberrations and sister-chromatid exchange in the blood lymphocytes of cows from an industrial region (aluminium smelting plant, lignite quarries, briquette factory, electric plants, mechanical plants. The control group consisted of cows from an agricultural region. In cows from the industrial region a significantly higher level of chromosomal aberrations was observed than in cows from the agricultural region (5.82 +/- 0.46 vs. 2.11 +/- 0.31 per 100 cells). In older, 7-12-year-old cows from the industrial region a slightly higher level of chromosomal aberrations was observed, and a significantly higher level of gaps than in younger cows. The frequency of SCE in the blood lymphocytes of cows from the industrial region was also higher and reached 7.47 +/- 2.33 per cell, while in cows from the agricultural region it was only 6.38 +/- 1.88, but these differences were not significant statistically. However, significant differences were observed in the number of SCE between the age groups within the industrial region P < or = 0.001) in favour of the younger animals. The results obtained indicate that the industrial pollution emitted into the environment may have a genotoxic character. Thus cytogenetic examination of cattle may be a useful test for monitoring industrial pollution.

Cytogenetic analysis of experimental interspecies goat-sheep chimera.

Chromosomal analysis was carried out on blood lymphocytes, skin fibroblasts, and germinal cells of an interspecies goat-sheep chimera. This chimera was produced by aggregation of blastomeres of goat and sheep embryos. A cell chimerism 54,XX/60,XY was found in blood lymphocytes and skin fibroblasts. At birth the percentage of lymphocytes with karyotype 54,XX (sheep) amounted to 80% and with karyotype 60,XY (goat) to 20%. With age the percentage of lymphocytes with chromosome complement 54,XX increased, so that at 18 months it was 94% sheep and 6% goat. At the same age, in skin fibroblasts the percentage of cells with goat karyotype reached 25%. Analysis of germinal cells showed in spermatogonia the presence of only karyotype 60,XY and in primary spermatocytes of 29 autosomal bivalents and the sex bivalent XY.