The relationship between lysozyme gene polymorphism and quality changes during the storage of eggs derived from 2 commercial strains of Japanese quail.

During the storage irreversible changes occur in eggs that result in a deterioration of their quality. The most significant changes affect the albumen. One of the major proteins of albumen present in egg white is lysozyme, which protects the embryo from microorganisms. This enzyme also contributes to the qualitative characteristics of albumen. It is possible that its polymorphism also affects the quality and stability of the obtained raw material that is, table eggs. Therefore, the aim of this study was to assess the potential effect of polymorphism in the lysozyme gene and protein on the quality changes during the storage of eggs derived from 2 genetic strains of Japanese quail belonging to various utility types. Eggs from selected females of laying and meat-type breeds were stored for 14 wk. During this period the egg quality traits were evaluated 10 times. DNA was isolated from each female and all exons of the lysozyme gene had been sequenced. In total, fourteen SNPs' and one 4-bp indel mutation were identified in exons and adjacent intronic sequences, among which SNP1 (1:32140723) resulted in a substitution of lysine with glutamine (Q21K). The results showed that SNP1 (strain S22), as well as the SNP2, SNP5, SNP7, SNP8, SNP10, SNP11, SNP12 and SNP13 were significantly associated with breaking strength during egg storage in both investigated Japanese quail strains. Furthermore, a 3 haplotype blocks containing nine SNPs (2, 5, 6, 7, 8, 10, 11, 12 and 13) were identified. These blocks displayed 8 distinct haplotypes that had significant association with breaking strength at all storage time points where egg quality analyses were performed. The study also revealed significant effects of breed and storage time on the egg quality traits. These results provide new insights into the genetic basis of egg quality during storage and could be incorporated into the breeding programs involving these strains.

Effect of gentle physical contact on behavioural indicators in sheep.

The available literature provides ample evidence for the importance of gentle treatment and handling of farm animals for the establishment of relationships with humans. The present study is an attempt to verify the assumption that massage can be a tool for building sheep-human relationships and to determine the impact of this technique on the behaviour of lambs. In total, 40 3-week-old lambs (20 ewes of the synthetic prolific-meat line (BCP) and 20 Swiniarka breed ewes) were assigned to two control groups (BCP, Swiniarka) and two experimental groups (BCP, Swiniarka). The lambs from the experimental groups received modified Shantala massage. During each session, the zoophysiotherapists massaged the chest, forelimbs, abdomen, hindlimbs, back, and face always in the same way and the same order. The massage was applied three times a day (at fixed times) for 21 consecutive days. Based on the behaviour manifested by the lambs during the massage sessions, a 2-point scale was developed for assessment of the animals from the research groups. After the massage, behavioural tests with the participation of a human, a dog, and a novel object were carried out for three consecutive days. Behaviours indicating curiosity (frequency of interaction, time of direct contact, latency) and fear (frequency of climbing and attempts to escape, duration of vocalisation) were assessed. The behavioural tests revealed significant differences in the behaviour of massaged and non-massaged lambs of both breeds, which confirms that the massage has a modifying effect on the behaviour of these animals. The influence of the applied technique was reflected by an increase in curiosity and a reduced level of fear.

Effects of dietary alfalfa protein concentrate supplementation on performance, egg quality, and fatty acid composition of raw, freeze-dried, and hard-boiled eggs from Polbar laying hens.

The aim of this study was to determine the effect of addition of alfalfa protein concentrate (APC) at the dose of 15 g or 30 g per 1 kg of complete feed ration for the native Polbar breed on selected production traits and the fatty acid profile in the yolk of raw, hard-boiled or freeze-dried eggs. Laying hens were assigned randomly to 3 experimental groups, each comprising 30 birds. The control group received a standard diet without the APC addition and the experimental groups received APC, which partially replaced postextraction soybean meal. Egg laying performance, feed intake, and feed conversion ratio were controlled throughout the experiment. At 33 wk of age, 45 eggs were randomly selected for assessment of the quality of the egg content and eggshell, and 30 eggs were taken for each of the cooking and freeze-drying treatments. The fatty acid composition was determined in the yolks of hard-boiled, freeze-dried, and raw eggs. There was no effect of the APC addition on the laying performance, feed intake and feed conversion ratio, and a majority of egg quality traits. Hens fed with the APC laid eggs with a darker colored eggshell and yolk and a slightly lower breaking strength. The yolks in the eggs from hens receiving the APC addition were characterized by distinctly higher content of polyunsaturated fatty acids (PUFA). The group fed with a higher dose of APC produced eggs with a substantially lower level of saturated fatty acids (SFA). Boiling resulted in an increase in the SFA content and a decline in the level of PUFAs and carotenoids. Freeze-drying led to an increase in the total SFA content and a decrease in the level of n-3 PUFA. The APC addition to feed can replace the genetically modified soybean meal without reducing the values of production traits and egg quality and with a beneficial effect on the yolk color and fatty acid profile.

Identification of quantitative trait loci affecting production and biochemical traits in a unique Japanese quail resource population.

The objective of the current study was to identify QTL associated with body weight, growth rate, egg quality traits, concentration of selected blood plasma, and yolk lipids as well as concentration of selected macro- and microelements, color, pH, basic chemical composition, and drip loss of breast muscle of Japanese quail (Coturnix japonica). Twenty-two meat-type males (line F33) were crossed with twenty-two laying-type females (line S22) to produce a generation of F1 hybrids. The F2 generation was created by mating 44 randomly chosen F1 hybrids, which were full siblings. The birds were individually weighed from the first to eighth week of age. At the age of 19 wk, 2 to 4 eggs were individually collected from each female and an analysis of the egg quality traits was performed. At slaughter, blood and breast muscles were collected from 324 individuals of the resource population. The basic chemical composition, concentration of chosen macro- and microelements, color, pH, and drip loss were determined in the muscle samples. The concentration of chosen lipids was determined in egg yolk and blood plasma. In total, 30 microsatellite markers located on chromosome 1 and 2 were genotyped. QTL mapping including additive and dominance genetic effects revealed 6 loci on chromosome 1 of the Japanese quail affecting the egg number, egg production rate, egg weight, specific gravity, egg shell weight, concentration of Na in breast muscle. In turn, there were 9 loci on chromosome 2 affecting the body weight in the first, fourth, and sixth week of age, growth rate in the second and seventh week of age, specific gravity, concentration of K and Cu in breast muscle, and the levels of triacylglycerols in blood plasma. In this study, QTL with a potential effect on the Na, K, and Cu content in breast muscles in poultry and on specific gravity in the Japanese quail were mapped for the first time.

Parental care compromises feeding in the pumpkinseed (Lepomis gibbosus).

Providing parental care is potentially costly. Costs can arise through elevated energy expenditure or from an increased risk of mortality. A cost of parental care can also occur because a parent is compromised in their ability to forage. We used pumpkinseed Lepomis gibbosus, a fish with an alternative male mating strategy, to test whether parental males differed in their feeding in comparison with females and cuckolder males. To address this question, we examined the stomach contents of female, cuckolder male, and parental male pumpkinseed during the breeding season over an entire diel cycle. We showed that parental males had a lower total weight of food in their stomachs in comparison with females, while cuckolder males did not. Parental males also had a lower weight and number of chironomids in their stomachs. The temporal pattern of feeding of parental males diverged from that of females, and they had a lower probability of pupal chironomids in their stomachs, which implies spatial segregation in foraging. Parental males had a greater probability of conspecific eggs in their stomachs than females, while the probability of egg cannibalism did not differ between cuckolder males and females. Overall, these finding meet predictions in accordance with an assumption that parental care and territoriality can compromise feeding.

Heritability of individual egg hatching success versus hen hatchability in layers.

The purpose of the study was to test the genetics of individual egg hatchability. Hatching outcome (0,1) of each of the eggs (126,430) laid during hatching seasons of 5 generations of a Rhode Island White population was analyzed with models that attributed the direct additive effect either to an egg or to a hen. A Gibbs sampling procedure, accounting for dichotomous nature of the trait, was employed for variance component estimation. The egg/embryo direct additive component was negligibly small (h(2) = 0.007) from the point of view of the accuracy of the hatchability proof. The hen direct additive component, though more than 12 times higher (h(2) = 0.087) than that of the egg, was still more than 2 times smaller than the component because of her permanent environment (p(2) = 0.221). More accurate definition of hens' environmental needs may prove to be more effective for hatching outcome improvement than increasing the accuracy of the reproduction proof, because current selection has to be performed before the individual hatchability record is known.

Calcium and salinity as selective factors in plate morph evolution of the three-spined stickleback (Gasterosteus aculeatus).

Identifying the causal factors underlying natural selection remains a key challenge in evolutionary biology. Although the genetic basis for the plate morph evolution of three-spined stickleback (Gasterosteus aculeatus) is well described, the environmental variables that form the basis for different morphs are not understood. We measured the effects of dissolved calcium and salinity on the growth of sticklebacks with different plate morphs from Scotland and Poland. There was a significant interaction of calcium with plate morph for fish from both regions, with complete morph sticklebacks growing more slowly at low calcium concentrations and low morph sticklebacks showing divergent responses to calcium concentration. A Scottish anadromous population showed evidence of local adaptation to high salinity, which was independent of plate morph. Polish and Scottish populations diverged in their response to salinity, suggesting a difference in osmotic regulation. The results implicate a role for calcium in selecting for plate morph evolution in sticklebacks, possibly as a limiting element in skeletal growth.

Towards the conservation of crucian carp Carassius carassius: understanding the extent and causes of decline within part of its native English range.

The extent and causes of crucian carp Carassius carassius decline were assessed during an initial study of c. 25 ponds in north Norfolk, eastern England, U.K., which was then replicated (a validation study) on another c. 25 ponds in an adjacent area. Of these ponds, c. 40 are known to have contained C. carassius during the 1970s-1980s. In the initial and validation studies, C. carassius were found in only 11 of these ponds, yielding declines of 76% (five of 21 ponds) and 68% (six of 19 ponds), respectively (72% decline overall). Non-native cyprinids, including goldfish Carassius auratus and common carp Cyprinus carpio and their hybrids with C. carassius, were observed in 20% of the ponds. Causes of C. carassius local extinction from 21 ponds were confidently determined as desiccation due to drought, terrestrialization and habitat deterioration, hybridization and competition with non-native cyprinids, agricultural land reclamation and predation (after the introduction of pike Esox lucius). This study led to C. carassius being designated as a Biodiversity Action Plan (BAP) species in the county of Norfolk, the first formal conservation designation for the species in the U.K. The C. carassius BAP plan aims to halt the decline of this much overlooked species through reintroductions and selective stocking of suitable ponds within the native range of the species.

A quantitative trait locus for a primary antibody response to keyhole limpet hemocyanin on chicken chromosome 14--confirmation and candidate gene approach.

A QTL involved in the primary antibody response toward keyhole limpet hemocyanin (KLH) was detected on chicken chromosome 14 in the experimental population, which was created by crossing commercial White Leghorn and a Polish native chicken breed (green-legged partridgelike). The current QTL location is a validation of previous experiments pointing to the same genomic location for the QTL linked to a primary antibody response to KLH. An experimental population was typed with microsatellite markers distributed over the chicken chromosome 14. Titers of antibodies binding KLH were measured for all individuals by ELISA. Statistical models applied in the Grid QTL Web-based software were used to analyze the data: a half-sib model, a line-cross model, and combined analysis in a linkage disequilibrium and linkage analysis model. Candidate genes that have been proposed were genotyped with SNP located in genes exons. Statistical analyses of single SNP associations were performed pointing out 2 SNP of an axis inhibitor protein (AXIN1) gene as significantly associated with the trait of an interest.

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.

The delay of anoikis due to the inhibition of protein tyrosine dephosphorylation enables the maintenance of normal rat colonocyte primary culture.

Apoptosis induced by detachment of cells from the extracellular matrix (anoikis) appears to be one of the main obstacles in attempts to establish long-term primary culture of normal colonocytes. In the present study, the dynamics of molecular events related to apoptosis of isolated normal rat colonocytes was investigated. The whole colonic crypts were isolated using collagenase/dispase digestion technique. DNA fragmentation typical for the apoptosis and the apoptotic morphology of cells were observed already at the end of their isolation. Considerable increase in caspase-3 activity was noted during the first two hours of cell cultivation. Delaying of apoptosis by treatment of cells with sodium orthovanadate, the specific protein tyrosine phosphatase inhibitor, was found to be possible. It may facilitate long-term culture of intestinal epithelial cells.

Normal colonocytes in primary culture--an experimental model for molecular pharmacology and biology of large intestine.

A simple method of whole intestinal crypts isolation from rat's colonic tissue has been developed. Culture of viable epithelial cells (colonocytes) was obtained from intact crypts using method providing colonocytes for apoptosis. Satisfactory results have been obtained if the crypts were isolated using collagenase I. Under conditions applied, spontaneous release of the colonocytes took place. Liberated cells underwent almost immediate adhesion to microcarrier surface. The primary culture of normal colonocytes indicating metabolic activity for long time (> 10 days) has been obtained.

Cholesterol content and fatty acid composition of ostrich meat as influenced by subspecies.

Two muscles, the m. gastrocnemius and m. iliofibularis, were excised from the left legs of six Red Neck (Struthio camelus massaicus) and six Blue Neck (Struthio camelus australis) ostriches and analysed for lipid and cholesterol content and fatty acid composition. Total lipid (1.43 g 100g ) and cholesterol content (65.63 mg 100g ) did not differ (ps> 0.05) in any muscle between subspecies. Although the percentage of some individual fatty acids differed (p < 0.05), the total percentage of saturated fatty acids and total monounsaturated fatty acids was similar (p > 0.05) between subspecies in both muscles. Although the percentage total polyunsaturated fatty acids was higher (p < 0.05) in Blue Necks (23.78%) than in Red Necks (23.65%) in the m. gastrocnemius, but not (p > 0.05) in the m, iliofibularis, the difference of 0.13% is probably of no practical significance.

Phagocytosis by somatic cells from dry cow secretion.

Independently of medium in which the process occurred, serum or PBS, phagocytosis and killing of Staphylococcus aureus by somatic cells from dry cow secretion were significantly higher at the early dry period than at the steady state period. Total bacterial survival was highly correlated with phagocytosis and with intracellular survival. Correlations between phagocytosis and intracellular survival were much lower. Percentage of S. aureus phagocytosed after incubation in bovine blood serum showed highly significant variation among samples of cells isolated from secretion of different cows at the early dry period and significant variation among samples of cells isolated from different cows at the steady state period.