Performance and meat quality traits of slow-growing chickens stimulated in ovo with galactooligosaccharides and exposed to heat stress.

In vivo performance, carcass and meat quality traits of slow-growing chickens stimulated in ovo with trans galactooligosaccharides (GOS) and exposed to heat stress were evaluated. On d 12 of egg incubation, 3,000 fertilized eggs (Hubbard JA57) were divided into prebiotic group (GOS) injected with 3.5 mg GOS/egg, saline group (S) injected with physiological saline (only to assess the hatchability rate) and an uninjected control group (C). After hatching, 600 male chicks (300 from GOS and 300 from C) were housed on floor pens (6 pens/treatment, 25 birds/pen) and reared under neutral (TN) or heat stress conditions (HS, 30°C from 36 to 50 d). BW, daily feed intake (DFI), daily weight gain (DWG), feed conversion rate (FCR), and mortality were measured. At 50 d of age, 15 randomly selected birds/treatment/environmental conditions were slaughtered and the pectoral muscle (PM) was collected for analyses. Hatchability was similar among groups. BW of the newly hatched chicks was lower (P < 0.01) in GOS compared to C. Final BW, DWG, DFI, and FCR were not affected (P > 0.05) by GOS. HS reduced final BW (-12.93%, P < 0.001). During finisher phase, DFI and DWG were lower (P < 0.001) and FCR was higher (P < 0.01) in HS compared to TN. Mortality was not affected (P > 0.05) by GOS and HS. Meat from GOS chickens had a higher (P < 0.01) pH and was darker (P < 0.05) compared to C. Proximate composition, cholesterol content, fatty acid profile, and intramuscular collagen properties of PM were not affected by GOS. The HS group showed a lower (P < 0.05) content of both collagen and monounsaturated fatty acids than TN group. Significant interactions between GOS and temperature were found for FA composition. In conclusion, the differences in performance have had an impact on the responses to HS in Hubbard chickens, but not on mortality rate. GOS did not relieve the negative effect of HS on chickens' performance.

Interaction between early in ovo stimulation of the gut microbiota and chicken host - splenic changes in gene expression and methylation.

BACKGROUND: Epigenetic regulation of the gene expression results from interaction between the external environment and transcription of the genetic information encoded in DNA. Methylated CpG regions within the gene promoters lead to silencing of the gene expression in most cases. Factors contributing to epigenetic regulation include intestinal microbiota, which in chicken can be potently modified by in ovo stimulation. The main aim of this study was to determine global and specific methylation patterns of the spleen under the influence of host-microbiome interaction. RESULTS: Fertilized eggs of two genotypes: Ross 308 and Green-legged Partridgelike were in ovo stimulated on d 12 of incubation. The injected compounds were as follows: probiotic - Lactococcus lactis subsp. cremoris IBB477, prebiotic - galactooligosaccharides, and synbiotic - combination of both. Chickens were sacrificed on d 42 post-hatching. Spleen was collected, RNA and DNA were isolated and intended to gene expression, gene methylation and global methylation analysis. We have proved that negative regulation of gene expression after administration of bioactive substances in ovo might have epigenetic character. Epigenetic changes depend on the genotype and the substance administered in ovo. CONCLUSION: Epigenetic nature of microbial reprogramming in poultry and extension of issues related to host-microbiome interaction is a new direction of this research.

Genotype-dependent development of cellular and humoral immunity in the spleen and cecal tonsils of chickens stimulated in ovo with bioactive compounds.

Prebiotics, probiotics, and synbiotics, delivered in ovo influence the colonization and development of the peripheral immune system in poultry. This study aimed to investigate the influence of the host genotype (broiler chickens [Ross 308] and old native Polish breed Green-legged Partridgelike [GP] chickens) on the number of B and T cells in the spleen and cecal tonsils (CT). The solution of a bioactive compound was injected in ovo on day 12 of egg incubation: prebiotics (galactooligosaccharides [GOS]), probiotics (Lactococcus lactis subsp. cremoris IBB477), and synbiotics (GOS + L. lactis). The samples were collected on day 7, day 21, and day 42 after hatching (n = 8). The number of Bu-1+ (B) cells, CD4+ cells, and CD8+ cells in the spleen and CT was estimated using immunohistochemistry. The number of germinal centers (GC) was determined in the spleen. In broilers, probiotics increased (P < 0.05) the number of CD4+ cells in the CT on day 7. On day 21, prebiotics raised (P < 0.01) the number of cells involved in cellular immunity in the CT (CD4+ and CD8+ cells) and spleen (CD8+ cells). On day 42, it was synbiotics that stimulated the colonization of both the CT and spleen by B cells, but colonization of the spleen only by CD4+ and CD8+ cells. In GP chickens, synbiotics enforced the cellular immunity (CD4+ or CD8+ cells) in the spleen at all time points. Synbiotics also stimulated the GC appearance on day 21 and day 42. In GP chickens, the influence of bioactive compounds on colonization of the CT was very limited. In broilers, we determined pronounced and age-dependent effects of prebiotics and synbiotics on the number of B and T cells in both the CT and spleen. In GP chickens, the most potent compound was synbiotics, which stimulated cellular immunity in the spleen but not in the CT. However, given the long-term effects on adaptive immune cells, synbiotics were the most potent compounds in both chicken genotypes.

Impact of galactooligosaccharides delivered in ovo on mitigating negative effects of heat stress on performance and welfare of broilers.

Galactooligosaccharides (GOS) delivered in ovo improve intestinal health of broiler chickens. This study aimed to demonstrate the impact of in ovo stimulation with GOS prebiotic on day 12 of egg incubation on performance and welfare traits in broiler chickens. The incubating eggs were divided into 3 groups, based on the substance injected in ovo: 3.5 mg of GOS dissolved in 0.2 mL physiological saline (GOS), 0.2 mL physiological saline (S), or uninjected controls (C). Constant heat stress (HS) was induced on days 32 to 42 post-hatch by increasing environmental temperature to 30°C. Thermoneutral (TN) animals were kept at 25°C. The performance (body weight [BW], daily feed intake [DFI], daily weight gain [DWG], and feed conversion rate [FCR]) were measured and mortality was scored for starter (days 0 to 13), grower (days 14 to 27), and finisher (days 28 to 42) feeding phases. Rectal temperature was scored on days 32 to 42. Food-pad dermatitis (FPD) was scored post-mortem (day 42). GOS increased (P < 0.01) BW on day 42 (2.892 kg in GOS vs. 2.758 kg in C). Heat stress significantly reduced (P < 0.01) final BW (2.516 kg in TN vs. 3.110 kg in HS). During finisher phase, DFI was significantly higher in GOS vs. C (173.2 g vs. 165.7 g; P < 0.05). FCR calculated for the entire rearing period (days 0 to 42) ranged from 1.701 in C to 1.653 in GOS (P < 0.05). GOS improved FCR in HS animals during finisher phase (P < 0.05). Rectal temperature of GOS chickens under HS reached 42.5°C 1 day earlier than C and S (P < 0.05), which suggests that those birds recovered earlier from the high environmental temperature. Heat stress increased (P < 0.05) mortality about 5 times compared to TN during finisher phase (from 1.59% in TN to 7.69% in HS). GOS decreased FPD in TN conditions by 20% (no lesions in 81% in GOS vs. 60% in C). GOS delivered in ovo mitigated negative effects of HS on performance and welfare in broiler chickens.

Effect of galactooligosaccharides delivered in ovo on meat quality traits of broiler chickens exposed to heat stress.

A study was carried out to evaluate meat quality traits in fast-growing chickens stimulated in ovo with trans-galactoolighosaccarides (GOS) and exposed to heat stress. On day 12 of egg incubation, 3,000 fertilized eggs (Ross 308) were divided into prebiotic group (GOS) injected with 3.5 mg GOS/egg, saline group (S) injected with physiological saline, and control group (C) uninjected. After hatching, 900 male chicks (300 chicks/treatment) were reared in floor pens in either thermoneutral (TN; 6 pens/group, 25 birds/pen) or heat stress conditions (HS, 30°C from 32 to 42 D; 6 pens/group, 25 birds/pen). At 42 D of age, 15 randomly chosen birds/treatment/temperature were slaughtered and the pectoral muscle (PM) was removed for analyses. Data were analyzed by GLM in a 3 × 2 factorial design. In ovo treatment had no effect on PM weight, pH, water-holding capacity, and shear force. GOS and S birds had lighter (L*, P < 0.01) PM than C group, whereas the latter showed a higher (P < 0.05) yellowness index (b*) compared to S group. Proximate composition, cholesterol, and intramuscular collagen properties were not affected by treatment. As for fatty acid composition, only total polyunsaturated fatty acids (PUFA) content and n-6 PUFA were slightly lower in GOS group compared to S. Heat stress had a detrimental effect on PM weight (P < 0.01) and increased meat pH (P < 0.01). PM from HS chickens was darker with a higher b* index (P < 0.05) and had a higher (P < 0.01) lipid content and a lower (P < 0.05) total collagen amount. Total saturated fatty acids (SFA), monounsaturated fatty acids (MUFA), and PUFA were similar among groups. Significant interactions between factors were found for fatty acid composition: GOS decreased (P < 0.01) SFA and increased (P < 0.05) MUFA contents in HS birds. In conclusion, in ovo injection of GOS could mitigate the detrimental effect of heat stress on some meat quality traits.

Transcriptome modulation by in ovo delivered Lactobacillus synbiotics in a range of chicken tissues.

Synbiotics are the bioactive compounds that synergistically combine effects of prebiotics and probiotics. In poultry, synbiotics can be used to reprogram animal's intestinal microbiota upon perinatal in ovo injection on day 12 of eggs incubation. Optimally composed synbiotic delivered in ovo efficiently stimulates the host's intestinal microflora, which in turn exerts beneficial effects on the host and improves its physiological functions. The aim of the study was to estimate long-term changes in the chicken transcriptome after a single in ovo administration of two different synbiotics. On day 12 of eggs incubation, 5850 eggs of broiler chicken were distributed to experimental groups and injected with synbiotic 1 (S1)- Lactobacillus salivarius with galactooligosaccharides (GOS) or synbiotic 2 (S2)- Lactobacillus plantarum with raffinose family oligosaccharides (RFO). On day 21 post-hatching cockerels were sacrificed and immunological (cecal tonsils and spleen), intestinal (jejunum) and metabolic (liver) tissues were collected (n = 5). Isolated RNA served as a template for the whole-transcriptome analysis using GeneChip Chicken Gene 1.1. ST Array Strip (Affymetrix). Data analysis was performed using Affymetrix Expression Console and Transcriptome Analysis Console software, Venn diagrams, DAVID and CateGOrizer. The highest number of Differentially Expressed Genes (DEG) was detected in cecal tonsils (160 DEG) after S1 in ovo injection, and in liver (159 DEG) after S2 injection. The influence of S1 on transcriptome modulation was demonstrated by a strong activation of the genes taking part in the pathways related to metabolism and immune response in cecal tonsils. S2 injection led to modulation of the gene expression associated with metabolic and developmental signaling pathways in the liver. Obtained results let us conclude that synbiotics delivered in ovo have significant impact on chicken transcriptome and their effect depends on the composition of the bioactive compound.

Avian model to mitigate gut-derived immune response and oxidative stress during heat.

The tissue injury at the early stages of the heat stress response triggers release of inflammatory and oxidative agents from intestinal content into the milieu of the body. Intestinal homeostasis (i.e., eubiosis) improves the barrier function and mitigates the gut-derived influx of endotoxins. In this study we have analyzed the mitigating role of embryonic stimulation of the gut homeostasis in chickens on immune and oxidative responses to heat. The animal trial was conducted on broiler chickens. The treatment included a single in ovo injection of the galactooligosaccharides (GOS) prebiotic into the air cell of the egg on day 12 of incubation. Control eggs were in ovo injected with the same volume of sterile physiological saline. After hatching, birds were raised in group pens (6 pens/group, 25 birds/pen). Short-term, mild heat stress was induced on day 32 post-hatching by increase in the ambient temperature above the thermal comfort (30 °C for 8.5 h). The spleen was harvested from randomly selected individuals. The relative gene expression study was conducted with RT-qPCR. The two gene panels were analyzed: (1) immune response genes (IL-1ß, IL-2, IL-4, IL-6, IL-12p40 and IL-17) and (2) stress response genes (HSP25, HSP70, HSP90, BAG3, CAT and SOD). Data were evaluated by the analysis of variance in a 2 × 2 factorial design that included in ovo treatment and ambient temperature as factors. We have found that the immune-related and stress-related gene expression signatures were triggered in animals subjected to heat but with unbalanced intestinal flora (i.e., dysbiotic, without in ovo stimulation with GOS). These animals had increased expression of the genes involved in the immune responses (IL-4 and IL17) and stress responses (HSP25, HSP70, HSP90, CAT and SOD) to short-term heat stress that indicated presence of inflammatory and oxidative mediators (P < 0.05). The individuals that were in ovo stimulated with GOS did not mount the anti-inflammatory or antioxidative responses. Heat shock proteins (HSP25 and HSP70) were increased in both groups challenged with heat, which indicated their role in adaptation to heat.

Prebiotics and synbiotics - in ovo delivery for improved lifespan condition in chicken.

Commercially produced chickens have become key food-producing animals in the global food system. The scale of production in industrial settings has changed management systems to a point now very far from traditional methods. During the perinatal period, newly hatched chicks undergo processing, vaccination and transportation, which introduces a gap in access to feed and water. This gap, referred to as the hatching window, dampens the potential for microflora inoculation and as such, prevents proper microbiome, gastrointestinal system and innate immunity development. As a consequence, the industrial production of chickens with a poor microbial profile leads to enteric microbial infestation and infectious disease outbreaks, which became even more prevalent after the withdrawal of antibiotic growth promoters on many world markets (e.g., the EU).This review presents the rationale, methodology and life-long effects of in ovo stimulation of chicken microflora. In ovo stimulation provides efficient embryonic microbiome colonization with commensal microflora during the perinatal period. A carefully selected bioactive formulation (prebiotics, probiotics alone or combined into synbiotics) is delivered into the air cell of the egg on day 12 of egg incubation. The prebiotic penetrates the outer and inner egg membranes and stimulates development on the innate microflora in the embryonic guts. Probiotics are available after the mechanical breakage of the shell membranes by the chick's beak at the beginning of hatching (day 19). The intestinal microflora after in ovo stimulation is potent enough for competitive exclusion and programs the lifespan condition. We present the effects of different combinations of prebiotic and probiotic delivered in ovo on day 12 of egg incubation on microflora, growth traits, feed efficiency, intestinal morphology, meat microstructure and quality, immune system development, physiological characteristics and the transcriptome of the broiler chickens.We discuss the differences between in ovo stimulation (day 12 of egg incubation) and in ovo feeding (days 17-18 of egg incubation) and speculate about possible future developments in this field. In summary, decades of research on in ovo stimulation and the lifelong effects support this method as efficient programming of lifespan conditions in commercially raised chickens.

Influence of different prebiotics and mode of their administration on broiler chicken performance.

In the post-antibiotics era, prebiotics are proposed as alternatives to antibiotic growth promoters in poultry production. The goal of this study was to compare in ovo method of prebiotic delivery with in-water supplementation and with both methods combined (in ovo+in-water) in broiler chickens. Two trials were conducted. Trial 1 was carried out to optimize the doses of two prebiotics, DN (DiNovo®, extract of beta-glucans) and BI (Bi2tos, trans-galactooligosaccharides), for in ovo delivery. The estimated parameters were hatchability and bacteriological status of the newly hatched chicks. Prebiotics were dissolved in 0.2 ml of physiological saline, at the doses: 0.18, 0.88, 3.5 and 7.0 mg/embryo; control group (C) was injected in ovo with 0.2 ml of physiological saline. Trial 2 was conducted to evaluate effects of different prebiotics (DN, BI and raffinose family oligosaccharides (RFO)) delivered in ovo, in-water and in a combined way (in ovo+in-water) on broiler chickens performance. The results of the Trial 1 indicated that the optimal dose of DN and BI prebiotics delivered in ovo, that did not reduce chicks' hatchability, was 0.88 mg/embryo (DN) and 3.5 mg/embryo (BI). Both prebiotics numerically increased number of lactobacilli and bifidobacteria in chicken feces (P>0.05). In Trial 2, all prebiotics (DN, BI and RFO) significantly increased BW gain compared with the C group (P<0.05), especially during the first 21 days of life. However, feed intake and feed conversion ratio were increased upon prebiotics delivery irrespective of method used. Injection of prebiotics in ovo combined with in-water supplementation did not express synergistic effects on broilers performance compared with in ovo injection only. Taken together, those results confirm that single in ovo prebiotics injection into the chicken embryo can successfully replace prolonged in-water supplementation post hatching.

Identification of candidate genes and mutations in QTL regions for immune responses in chicken.

There are two categories of immune responses - innate and adaptive immunity - both having polygenic backgrounds and a significant environmental component. In our study, adaptive immunity was represented by the specific antibody response toward keyhole limpet hemocyanin (KLH); innate immunity was represented by natural antibodies toward lipopolysaccharide (LPS) and lipoteichoic acid (LTA). Defining genetic bases of immune responses leads from defining quantitative trait loci (QTL) toward a single mutation responsible for variation in the phenotypic trait. The goal of the reported study was to define candidate genes and mutations for the immune traits of interest in chicken by performing an association study of SNPs located in candidate genes defined in QTL regions. Candidate genes and SNPs in QTL regions were selected in silico. SNP association was based on a custom SNP panel, GoldenGate genotyping assay (Illumina) and two statistical models: random mixed model and CAR score. The most significant SNP for immune response toward KLH was located in the JMJD6 gene located on GGA18. Four SNPs in candidate genes FOXJ1 (GGA18), EPHB1 (GGA9), PTGER4 (GGAZ) and PRKCB (GGA14) showed association with natural antibodies for LPS. A single SNP in ITGB4 (GGA18) was associated with natural antibodies for LTA. All associated SNPs mentioned above showed additive effects.

The reversal of cognitive, but not negative or positive symptoms of schizophrenia, by the mGlu2/3 receptor agonist, LY379268, is 5-HT1A dependent.

mGlu(2/3) receptor agonists were shown to possess an antipsychotic-like potential in animal studies. Recent clinical investigations revealed that their antipsychotic potential might also manifest in humans. LY379268, the group II mGlu receptor orthosteric agonist, was previously shown to exhibit antipsychotic-like action in animal models of schizophrenia. However, the mechanism of its action is not fully recognized. Here, we decided to investigate the involvement of 5-HT1A receptors in the LY379268-induced antipsychotic effects. We used models of positive, negative and cognitive symptoms of schizophrenia, such as MK-801- and amphetamine-induced hyperactivity tests, DOI-induced head twitches, social interaction and novel object recognition. LY379268 was active in a wide range of doses (0.5-5 mg/kg), depending on the paradigm. The effects of the drug were not antagonized by 5-HT(1A) antagonist, WAY100635 (0.1 mg/kg) in the models of positive and negative symptoms. Conversely, in the novel object recognition test, which exerts cognitive disturbances, the action of LY379268 was antagonized by WAY100635. Concomitantly, the action of a sub-effective dose of the drug was enhanced by the administration of a sub-effective dose of 5-HT(1A) agonist, (R)-(+)-8-Hydroxy-DPAT. Altogether, we propose that the antipsychotic-like action of group II mGlu receptors' agonist is 5-HT(1A) independent in context of positive and negative symptoms, while the action toward cognitive disturbances seems to be 5-HT(1A) dependent.

Insights into the genetic history of Green-legged Partridgelike fowl: mtDNA and genome-wide SNP analysis.

The Green-legged Partridgelike (GP) fowl, an old native Polish breed, is characterised by reseda green-coloured shanks rather than yellow, white, slate or black commonly observed across most domestic breeds of chicken. Here, we investigate the origin, genetic relationships and structure of the GP fowl using mtDNA D-loop sequencing and genome-wide SNP analysis. Genome-wide association analysis between breeds enables us to verify the genetic control of the reseda green shank phenotype, a defining trait for the breed. Two mtDNA D-loop haplogroups and three autosomal genetic backgrounds are revealed. Significant associations of SNPs on chromosomes GGA24 and GGAZ indicate that the reseda green leg phenotype is associated with recessive alleles linked to the W and Id loci. Our results provide new insights into the genetic history of European chicken, indicating an admixed origin of East European traditional breeds of chicken on the continent, as supported by the presence of the reseda green phenotype and the knowledge that the GP fowl as a breed was developed before the advent of commercial stocks.

Influence of in ovo prebiotic and synbiotic administration on meat quality of broiler chickens.

A trial was conducted to evaluate the effect of in ovo injection of prebiotic and synbiotics on growth performance, meat quality traits (cholesterol content, intramuscular collagen properties, fiber measurements), and the presence of histopathological changes in the pectoral muscle (PS) of broiler chickens. On d 12 of incubation, 480 eggs were randomly divided into 5 experimental groups treated with different bioactives, in ovo injected: C, control with physiological saline; T1 with 1.9 mg of raffinose family oligosaccharides; T2 and T3 with 1.9 mg of raffinose family oligosaccharides enriched with different probiotic bacteria, specifically 1,000 cfu of Lactococcus lactis ssp. lactis SL1 and Lactococcus lactis ssp. cremoris IBB SC1, respectively; T4 with commercially available synbiotic Duolac, containing 500 cfu of both Lactobacillus acidophilus and Streptococcus faecium with the addition of lactose (0.001 mg/embryo). Among the hatched chickens, 60 males were randomly chosen (12 birds for each group) and were grown to 42 d in collective cages (n = 3 birds in each 4 cages: replications for experimental groups). Broilers were fed ad libitum commercial diets according to age. In ovo prebiotic and synbiotic administration had a low effect on investigated traits, but depend on the kind of bioactives administered. Commercial synbiotic treatment (T4) reduced carcass yield percentage, and the feed conversion ratio was higher in T3 and T4 groups compared with other groups. The abdominal fat, the ultimate pH, and cholesterol of the PS were not affected by treatment. Broiler chickens of the treated groups with both slightly greater PS and fiber diameter had a significantly lower amount of collagen. The greater thickness of muscle fibers (not significant) and the lower fiber density (statistically significant), observed in treated birds in comparison with those of the C group, are not associated with histopathological changes in the PS of broilers. The incidence of histopathological changes in broiler chickens from examined groups was low, which did not affect the deterioration of meat quality obtained from these birds.

Quantitative trait loci associated with the humoral innate immune response in chickens were confirmed in a cross between Green-Legged Partridgelike and White Leghorn.

Natural antibodies (NA) create a crucial barrier at the initial steps of the innate humoral immune response. The main role of NA in the defense system is to bind the pathogens at early stages of infection. Different pathogens are recognized by the presence of highly conserved antigen determinant [e.g., lipopolysaccharide (LPS) in gram-negative bacteria or lipoteichoic acid (LTA) in gram-positive bacteria]. In chickens, a different genetic background of NA binds LPS and LTA antigens, encoded by different QTL. The main objective of this work was to confirm known QTL associated with LPS and LTA NA. For this purpose a chicken reference population was created by crossing 2 breeds: a commercial layer, White Leghorn, and a Polish indigenous chicken, Green-Legged Partridgelike. The chromosomal regions analyzed harbored to GGA3, GGA5, GGA6, GGA8, GGA9, GGA10, GGA14, GGA15, GGA18, and GGAZ. The data collected consisted of the NA titers binding LPS and LTA (determined by ELISA at 12 wk of age) as well as the genotypes (30 short tandem repeat markers; average of 3 markers/chromosome, collected for generations F(0), F(1), and F(2)). The analyses were performed with 3 statistical models (paternal and maternal half-sib, line cross, and linkage analysis and linkage disequilibrium) implemented in GridQTL software (http://www.gridqtl.org.uk/). The QTL study of humoral innate immune response traits resulted in the confirmation of 3 QTL associated with NA titers binding LPS (located on GGA9, GGA18, and GGAZ) and 2 QTL associated with NA titers binding LTA (located on GGA5 and GGA14). A set of candidate genes within the regions of the validated QTL has been proposed.

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.

Validation of the QTL on SSC4 for meat and carcass quality traits in a commercial crossbred pig population.

Porcine chromosome 4 harbours many quantitative trait loci (QTL) affecting meat quality, fatness and carcass composition traits, detected in resource pig populations previously. However, prior to selection in commercial breeds, QTL identified in an intercross between divergent breeds require confirmation, so that they can be segregated. Consequently, the objective of this study was to validate several QTL on porcine chromosome 4 responsible for meat and carcass quality traits. The experimental population consisted of 14 crossbred paternal half-sib families. The region of investigation was the q arm of SSC4 flanked by the markers S0073 and S0813. Regression analysis resulted in the validation of three QTL within the interval: Minolta a* loin, back fat thickness and the weight of trimmed ham. The results were additionally confirmed by factor analysis. Candidate genes were proposed for meat colour, which was the most evident QTL validated in this study.