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Does are usually fed ad libitum during pregnancy, a fact that can result in intrauterine overfeeding and can have detrimental effects on embryonic programming with permanent changes in the physiology and behavior of the offspring. An experiment was therefore conducted to examine the effect of does' overfeeding during two different periods of pregnancy on the behavior of kits after weaning. Fifteen does were allocated into three groups: control (M) (100% of maintenance energy requirements [MER] throughout pregnancy), O1 (129% of MER between the 7th and 19th of pregnancy), and O2 (138% of MER between the 20th and 27th day of pregnancy). At the ages of 50 and 65 days, the behavior of 24 randomly selected rabbits in home cages was recorded for three consecutive days, while an open field test that constitutes a good indicator of locomotion score and stress was conducted for another 24 rabbits. Rabbits in O1 and O2 groups had more meals in shorter duration and moved and explored their environment less than the controls ( P P P

The aim of the present study was to identify ovine genomic regions and candidate genes impacting on milk traits i.e. milk yield, fat content, protein content and lactose content in the Frizarta dairy sheep. A total number of 481 ewes genotyped with the OvineSNP50 BeadChip were used. Haplotype regression analysis along with multi-locus mixed model analysis was employed to detect genomic markers associated with the traits. A total number of 9 single nucleotide polymorphisms were found to be associated with the traits under study. Four out of the 9 significant markers resided in genomic areas previously associated with milk traits according to the SheepQTLdb ( http://www.animalgenome.org/cgi-bin/QTLdb/OA/search ). A total number of 29 positional candidate genes located within a maximum distance of 250 kb upstream and downstream of 5 significant markers were identified. Gene prioritization analysis of the positional candidate genes based on the guilt-by-association principle suggested eleven genes i.e. ITGB4, H3F3B, ACOX1, WBP2, HTR1A, GPC5, GALR2, UNC13D, PRPSAP1, SRP68 and CDK3 as the most plausible functional candidate genes for milk traits in the Frizarta dairy sheep.

In the present study, we used genomic data, generated with a medium density single nucleotide polymorphisms (SNP) array, to acquire more information on the population structure and evolutionary history of the synthetic Frizarta dairy sheep. First, two typical measures of linkage disequilibrium (LD) were estimated at various physical distances that were then used to make inferences on the effective population size at key past time points. Population structure was also assessed by both multidimensional scaling analysis and k-means clustering on the distance matrix obtained from the animals' genomic relationships. The Wright's fixation F ST index was also employed to assess herds' genetic homogeneity and to indirectly estimate past migration rates. The Wright's fixation F IS index and genomic inbreeding coefficients based on the genomic relationship matrix as well as on runs of homozygosity were also estimated. The Frizarta breed displays relatively low LD levels with r 2 and |D'| equal to 0.18 and 0.50, respectively, at an average inter-marker distance of 31 kb. Linkage disequilibrium decayed rapidly by distance and persisted over just a few thousand base pairs. Rate of LD decay (β) varied widely among the 26 autosomes with larger values estimated for shorter chromosomes (e.g. β=0.057, for OAR6) and smaller values for longer ones (e.g. β=0.022, for OAR2). The inferred effective population size at the beginning of the breed's formation was as high as 549, was then reduced to 463 in 1981 (end of the breed's formation) and further declined to 187, one generation ago. Multidimensional scaling analysis and k-means clustering suggested a genetically homogenous population, F ST estimates indicated relatively low genetic differentiation between herds, whereas a heat map of the animals' genomic kinship relationships revealed a stratified population, at a herd level. Estimates of genomic inbreeding coefficients suggested that most recent parental relatedness may have been a major determinant of the current effective population size. A denser than the 50k SNP panel may be more beneficial when performing genome wide association studies in the breed.

The aim of the present study was to investigate the potential associations between polymorphisms of microsatellite markers and various traits in the Lesvos dairy sheep. In total, 350 individuals using 11 microsatellite markers were genotyped and various traits including six body anatomical measurements as well as milk yield and litter size were considered. The adaptive Least Absolute Shrinkage and Selection Operator method was employed to detect significant associations between markers’ alleles and the various phenotypic traits. In all but one microsatellite loci, rare alleles were detected. Markers OARJMP29, OARFCB128 and OARCP38 were found to be associated with milk yield. Marker OARJMP29 was also associated with body weight, girth circumference and horn length. Marker OARFCB128 was also significant for tail length, tail width and girth circumference while marker OARCP38 was significant for body length, as well. Current results shed more light in the associations between marker polymorphisms with various phenotypic traits in dairy sheep, including traits of economic importance that could potentially be implemented in a marker assisted selection program for increasing production efficiency in the Lesvos breed.

Various specific human glucocorticoid receptor (NR3C1) gene polymorphisms have been described in multiple sclerosis (MS) patients and correlated with disease progression, susceptibility and aggressiveness. Herein, we investigated the presence of gene alterations in the entire coding region of the NR3C1 in MS patients of variable clinical status (CIS, RRMS and SPMS) and the association(s) of these alterations with severity of disease (EDSS), response to glucocorticoid (GC) treatment and clinical improvement. Sixty Caucasian Greek MS patients were included. Sequencing the coding sequences and intron-exon boundaries of the NR3C1 did not reveal the presence of mutation(s) in any of the MS patients. Three previously described polymorphisms were detected: p.N363S (rs6195), p.N766N (rs6196) and c.1469-16G>T (rs6188). None of the identified alleles/genotypes were found to be associated with the severity of disease, response to glucocorticoids and disease subtypes. Known polymorphism, such as ER22/23EK that has been previously detected in MS patients, was not detected. There is a considerable ethnicity-related variation in the frequency of the NR3C1 polymorphisms. Although a genetic basis of the glucocorticoid sensitivity exists in healthy population, in the presence of chronic inflammation and abundance of cytokines--such in MS patients--other factors appear to play a more important role in GC sensitivity.

The objective of the present experiment was to investigate the implications of mild maternal undernutrition on the post-weaning behaviour of rabbits. Twenty-six dams were randomly assigned to two groups: the control group (C) that was fed ad libitum throughout pregnancy and the undernourished group (U) that was fed 75% of the recommended maintenance requirements between the 6th and 26th day of pregnancy. At the age of 50 and 65 days old, behaviour of rabbits in cages was recorded and rabbits were further subjected to an open-field test; a paradigm used as an indicator of fear and emotional distress. There were only limited differences between the examined groups in the exhibition of behaviour in the cages; values for frequencies of eating and drinking were higher in the controls compared to the prenatally undernourished rabbits (P < 0.05). Duration of drinking appeared to be higher in the group of rabbits born from undernourished does at the age of 50 days compared to the other groups (P < 0.05). Duration of resting and comfort behaviours was significantly increased at the age of 65 days in both groups (P < 0.05 and P < 0.01, respectively). In both groups, a decreased activity of rabbits during the light period was observed (as expected), whereas a reduction in frequencies of eating and drinking, and duration of locomotory and investigatory behaviours was observed at the age of 65 days (P < 0.05). Rabbits born to undernourished does displayed reduced rates of the sitting still and standing stretched behaviours compared to the controls during the implementation of the open-field test (P < 0.05). No other significant differences in the exhibition of the examined behaviours were found between the treatment groups. Although some differences were found in specific behavioural patterns, it is concluded that mild maternal undernutrition (75% of ad libitum intake) during pregnancy has generally only little effect on rabbits’ post-weaning behaviour.

One hundred and sixty-six lactating Makui breed sheeps (2–6 years old) were used to investigate the effects of two milking methods (milking with oxytocin (OT) and milking without oxytocin (HT)), ewes age and stage of lactation on milk yield and composition during 168 days of lactation and to estimate shape of lactation curve using Wood's model. Oxytocin treatment (OT) had a significant effect on the average daily milk yield (ADMY) and total milk yield (TMY) ( P P P P P P b ) parameter, ( a ) and ( c ) parameters of Wood's model were significantly ( P P P > 0.05) whereas peak time (PT), peak yield (PY) and persistency ( P ) were significant ( P P P > 0.05). In both methods, stages of lactation had a highly significant effect on milk fat, protein and SNF percentage ( P

The aim of the present study was to assess the presence of single nucleotide polymorphisms (SNPs) in the Growth Differentiating Factor 9 (GDF9) and Bone Morphogenetic Protein 15 (BMP15) genes, in two Greek sheep breeds i.e. Chios and Karagouniki, with high and moderate litter size (LS) i.e. number of lambs born, respectively. An examination of association(s) between the polymorphisms and LS, within breeds, was also conducted. Blood samples from 92 Chios and 96 Karagouniki ewes were collected, while repeated records on LS, were also available ( n = 239 and 259, respectively). Detection of SNPs was performed on the DNA products by PCR-RLFP analysis. Data analysis included examination of allelic and genotypic differentiation between the breeds, testing for Hardy–Weinberg Equilibrium (HWE) and association analysis between polymorphisms and LS. Genetic analysis showed that the G1 and G8 mutations of the GDF9 gene were significantly over-presented only in the highly prolific breed (Chios). The B4 mutation of the BMP15 gene was significantly over-presented only in the low prolific breed (Karagouniki). Both breeds were in Hardy–Weinberg disequilibrium (HWD), possibly due to selection pressure, population sub-structure and genetic drift. Certain polymorphisms of the G1, G4 and G8 sites of the GDF9 gene in the Chios breed were found to show heterozygote advantage with no evidence of infertility for the homozygous females. A similar trend was observed in the BMP15 gene although at no statistically significant level. In the Karagouniki breed, no significant associations between the studied polymorphisms and LS were detected.

The aim of this study was to investigate the effect of dietary probiotic inclusion level on mucin composition (monosaccharide ratio), mucosal morphometry, mucus layer staining intensity, and mucus layer thickness along the broiler intestinal tract. One-day-old male Cobb broilers were administered maize-soybean meal basal (BD) diets for 42 d and depending on the feed additive used, broilers were allocated into the following 5 experimental treatments: control C (BD, no additive), treatment P1 (10(8) colony forming units of probiotic/kg of BD), treatment P2 (10(9) cfu of probiotic/kg of BD), treatment P3 (10(10) cfu of probiotic/kg of BD), and treatment A (2.5 mg avilamycin/kg of BD). Intestinal samples from duodenum, ileum, and cecum of 14- and 42-d-old broilers were collected and analyzed. Mannose (Man) decreased linearly with increasing probiotic level in duodenum (P=0.015) and ileum (P=0.042) of 14-d-old broilers. N-Acetyl-glucosamine and galactose decreased linearly (P=0.012 and P=0.001, respectively), while fucose increased linearly (P

The effect of a dietary phytogenic feed additive (PFA) inclusion level in mucin monosaccharide composition, mucosal morphometry and mucus histochemistry along the broiler intestinal tract was studied. Cobb male broilers (n = 525) were allocated into five experimental treatments that, depending on the type of addition in the basal diet (BD), were labeled as follows: C (BD based on maize–soybean meal with no other additions), E1 (80 mg PFA/kg BD), E2 (125 mg PFA/kg BD), E3 (250 mg PFA/kg of BD) and A (2.5 mg avilamycin/kg BD). Samples from duodenum, ileum and cecum of 14- and 42-day-old broilers were collected and analyzed. In 14-day-old broilers, treatments E2 and E3 had higher (P < 0.01) duodenal mannose than treatments C, E1 and A. Ileal mannose was lower (P < 0.05) in treatment C compared with PFA treatments, and ileal galactose (Gal) was higher (P < 0.01) in treatments E2 and E3 compared with C and A. Polynomial contrast analysis with respect to PFA inclusion level showed that in 14-day-old broilers there was a linear increase (P = 0.001) in duodenal mannose and a quadratic effect (P = 0.038) in duodenal N-acetyl-galactosamine with increasing PFA level. Ileal Gal and mannose increased linearly (P = 0.002 and P = 0.012, respectively) with PFA inclusion level. There were no significant differences between treatments in mucin monosaccharide molar ratios of 42-day-old broilers. However, increasing PFA inclusion level resulted in a linear decrease of ileal fucose (P = 0.021) and cecal N-acetylgalactosamine (P = 0.036). Experimental treatments did not differ (P > 0.05) regarding duodenal villus height (Vh), crypt depth (Cd) and Vh/Cd ratio, irrespective of broiler age and the intestinal segment examined. However, increasing dietary PFA inclusion level showed a pattern of linear increase of duodenal Vh/Cd ratio in 14-day-old broilers and ileal Vh in 42-day-old broilers (P = 0.039 and P = 0.039, respectively). Alcian Blue–Periodic Acid-Schiff (pH 2.5) staining of neutral and acidic mucins showed that the staining intensity of mucus layer in villi was fragment (i.e. tip, midsection and base) dependent, whereas in crypts it was dependent both on intestinal segment (i.e. duodenum, ileum and cecum) and fragment. Finally, mucus layer thickness did not differ (P > 0.05) between treatments, yet a pattern of linear increase (P < 0.05) with PFA inclusion level was observed in the duodenum of 42-day-old broilers. In conclusion, the dietary inclusion level of PFA modulated broiler intestinal mucin composition and morphology. Further studies are required to elucidate the physiological implications of such changes in host–microflora interactions.

An experiment was conducted with the objective to study the interaction between breed and diet on conjugated linoleic acid (CLA) and fatty acids profile (FA) content of milk fat of four pure of dairy sheep breeds (Awassi, Lacaune, Friesland and Chios). All sheep used in this study were kept in one flock under the same management. During the winter months (from October to March) all sheep were kept indoors and fed with alfalfa hay and concentrates without any grazing and from April onwards were grazing native pastures without any supplementary feed. A total of 160 individual milk samples were taken from the four breeds at two sampling times: in January (n = 80) and in May (n = 80), when the ewes were in average, on 62 and 185 days in milk. These milk samples were analyzed for FA profile. The results of this experiment showed that the sheep breed had no effect on milk FA profile or on atherogenicity index (AI). The only significant effect was on Δ−9 desaturase activity index expressed as C18:1/C18:0. On the contrary, the diet affected significantly the FA profile of all ewes milk fat, with pasture to cause lower proportions of saturated and higher proportions of unsaturated FA, compared with those of sheep kept and fed indoors. The interaction between breed and diet was significant for the FA C10:0, C14:0, C15:0, C16:0, C17:1, C18:1, C18:2n6c, C18:2n6t, C18:3n6 and CLA and for FA groups SCFA, MCFA, LCFA and MUFA. The S/U FA ratio and the Δ−9 desaturase indexes (C14:1/C14:0 and C18:1/C18:0) were also affected by the breed × diet interaction. In conclusion, pasture feeding when compared with supplementary diet, induce large variation in the milk FA profile. In addition pasture increases FA in milk fat which has potential interest for human health. Breed has small effect on milk FA profile, but it can be ignored as it is shown from the interaction breed × diet.

Heritability coefficients as well as animals’ breeding values (BVs) were estimated on an average information algorithm for raw milk yield (MY), log transformed (LMY), square root transformed (SMY), adjusted for mean (CM) and adjusted for standard deviation (CSD) heterogeneity of the herd–year–lactation (HYL) classes, MY. The data included 4100 lactation records from 2551 ewes of the Lesbos dairy sheep kept in 44 herds on the homonymous island. Employment of data transformation (log or square root) and adjustment for mean heterogeneity did not result in homogenous variances across the HYL classes. Only adjustment of records by regressing the standard deviation (S.D.) of the HYLi class on population mean S.D. resulted in homogenous variances. Heritability estimates (h2) for MY, LMY and SMY were found to be of same magnitude (h2=0.19) while h2 for CM was higher (h2=0.20). Adjustment of MY for heterogeneous S.D. decreased the heritability estimate (h2=0.16), as a result of proportionally higher reduction of the additive genetic than of the residual variance. Heritability coefficient has also been estimated for two production (low and high) and two S.D. (low and high) levels of the HYL classes. Heritability estimates between low and high production levels were found of comparable magnitude (0.18 vs. 0.22, respectively) while they were clearly discriminated between low and high S.D. level (0.09 vs. 0.29, respectively). Product–moment (Pearson) and rank (Spearman) correlations and proportion of animals selected before and after adjustment were also estimated between estimated BVs on MY, CM and CSD records. Correlation estimates were generally high (r=0.88–0.98) when all sires and all ewes of the population were considered. Heterogeneity of variance had, however, a remarkable impact on ranking and thus of animals selected in 5% top sires and dams. Correction for heterogeneity of S.D. caused a great proportion, i.e. 25% and 32% of top sires and dams, respectively, to be excluded from selection when compared to the homogenous variance scenario. However, selection on a wider basis, e.g. sires with positive BVs and 20% of best dams, resulted in lower proportions of animals excluded. This was also the case for the effect of mean heterogeneity on the proportions of top sires and ewes selected. Investigation of the sources of heterogeneity of the HYL S.D.s showed that factors such as herd, lactation number, number of observations and production level are associated with heterogeneity of variance. Production year as well as genetic level did not significantly affect heterogeneity of variance (P>0.05). These above factors explained about 53% of the variation of S.D.s of the HYL classes.

Background : The vasoprotective effects of estrogens are known to be mediated by their respective estrogen receptors (ER) alpha (ERα) and beta (ERβ), which are present on the vascular wall. The amino-terminal part of the ERα appears to be important; genetic alterations in this region have been associated with arterial hypertension. This region has not been studied in atherosclerotic disease. In the present study, we examined the association between coronary artery disease (CAD) and alterations of the NH 2 -terminal part of ERα coding region. Methods : Genomic DNA was isolated from 50 healthy men and 40 men with CAD confirmed by coronary angiography. The coding sequences of exons 1 and 2 were amplified by polymerase chain reaction (PCR) and analyzed by either denaturing gradient gel electrophoresis (DGGE) or single stranded conformational polymorphism (SSCP), or both, sequencing and restriction fragment length polymorphism (RFLP), as appropriate. In the same subjects, biochemical and vascular parameters were also determined by using the appropriate methodology. Results : In exon 1, the codon 10 polymorphism was detected in both patients and healthy men either in heterozygous or homozygous form. The codon 87 polymorphism was detected mainly in homozygous form and only five individuals were heterozygotes. No mutations were found in exon 2. Statistical analysis of the allele distribution for either codon 10 or 87 between patients and healthy men showed no significant difference. In patients, the biochemical parameters were not statistically significantly different between ERα codon 10 genotypes or alleles. However, there was a clear effect of the TCT/TCT genotype and TCT allele on the vascular parameters whereas the right internal carotid artery (RICA) intima-media thickness was significantly associated with TCT/TCT genotype and TCT allele. Conclusions : We conclude that ERα genotypes play no role in the incidence of CAD disease, however, ERα codon 10 may be a genetic factor controlling some vessels' angiographic complications.

The aim of this study was to test the usefulness of artificial neural networks (ANNs) for predicting lactation as well as test-day milk yield(s) in Chios dairy sheep on the basis of a few (2–4) available test-day records at the beginning of a lactation period. The ANN employed was a neural network-like system with some advantages over other ANNs. No selection of learning coefficients, of the number of hidden layers or of the number of neurons in the layers was required. The effect on the network's predictive ability of the number of records used in the training phase, the number of input variables (i.e. test-day records) and data preprocessing was investigated. Input variables were the county, herd, lactation, lambing month, litter size, milk yield recorder, test day and days in milk (after lambing) when the first milk sample was obtained. Various criteria of goodness of prediction of lactation as well as of test-day yields were used, including Pearson and rank correlations between observed and predicted yields; the average difference between observed and predicted yields; the difference between their standard deviations; the standard deviation of differences between observed and predicted yields, and the ratio between it and the observed mean value. The average difference between observed and predicted yields was generally statistically non-significant (P

Lactation records of the Skopelos dairy goat population were used to assess the importance of various environmental effects and to estimate genetic parameters for 90-day milk yield (9OMY) and total milk yield (TMY) on untransformed and on log scale. Average 9OMY, TMY and days of milking were 164.7, 239.2 kg and 187 days, respectively. Herd, production year, lambing month, birth type, lactation number, the herd by year, year by month, year by lactation number interactions and days of milking were found statistically significant for TMY. Heritability estimates were 0.15 and 0.14 for 9OMY and TMY, respectively. Higher heritabilities of 0.24 and of 0.28 were estimated on the log scale for 90MY and TMY, respectively. Genetic and phenotypic correlations between 9OMY and total milk yield were found high, of 0.95 and 0.80, respectively.