Fateful triad of reactive oxygen species, mitochondrial dysfunction and lipid accumulation is associated with expression outline of the AMP-activated protein kinase pathway in bovine blastocysts.

Low cryotolerance is considered as the major drawback of in vitro-produced bovine embryos and is frequently associated with a triad encompassing increased cytoplasmic lipid accumulation, enhanced levels of reactive oxygen species (ROS) and mitochondrial dysfunction. The aim of the present study was to explore the role of the AMP-activated protein kinase (AMPK) pathway in the process resulting such phenotypes. Comparative analysis under different environmental conditions revealed downregulation of AMP-activated protein kinase cytalytic subunit 1alpha (AMPKA1), peroxisome proliferator-activated receptor gamma coactivator 1 alpha (PGC1A) and carnitine palmitoyltransferase 1 (CPT1) genes and upregulation of acetyl-CoA carboxylase α (ACC). In contrast, the presence of fatty acids within the culture medium resulted in a distinct molecular profile in the embryo associated with enhanced levels of ROS, mitochondrial dysfunction and elevated lipid accumulation in bovine embryos. Because AMPKA1 regulates PGC1A, CPT1 and ACC, the results of the present study reveal that AMPK in active its form is the key enzyme promoting lipolysis. Because AMPK1 activity is, in turn, controlled by the AMP : ATP ratio, it is possible to speculate that excessive uptake of exogenous free fatty acids could increase cellular ATP levels as a result of the disturbed ß-oxidation of these external fatty acids and could therefore bypass that molecular feedback mechanism. Subsequently, this condition would cause enhanced generation of ROS, which negatively affect mitochondrial activity. Both enhanced generation of ROS and low mitochondrial activity are suggested to enhance the accumulation of lipids in bovine embryos.

MicroRNA-regulated molecular mechanism underlying bovine subclinical endometritis.

An impaired uterine environment triggered by the incidence of subclinical endometritis often compromises fertility in the bovine. The uterus is a dynamic organ with tight regulation of specific genes at the transcriptional and translational levels. Herein, we hypothesised that subclinical endometritis alters the expression of uterine microRNAs (miRNAs), which may result in the dysregulation of corresponding target genes and biological pathways. To test this hypothesis, we used a genome-wide RT(2) (Exiqon, Vedbaek, Denmark) miRNA PCR array consisting of 354 miRNA primers and analysed miRNA expression in uterine cytobrush samples taken from cows with and without subclinical endometritis. The results revealed aberrant expression of 23 miRNAs in cows with subclinical endometritis compared with healthy cows. Furthermore, we designed an in vitro endometrial cell culture model challenged by lipopolysaccharide (LPS) to validate the differential regulation of miRNAs in cytobrush samples. Interestingly, we observed similar expression miRNA patterns in cytobrush samples taken from cows with or without subclinical endometritis and in vitro cultured endometrial cells challenged by LPS. To trace signalling pathways and biological functions potentially controlled by the aberrantly expressed miRNAs, we filtered high-ranking target genes from miRBase and analysed them using ingenuity pathway analysis. The gene networks, canonical pathways and biological functions strikingly converged to signalling pathways that mediate inflammatory responses, cellular proliferation, cell movement, the cell cycle and apoptosis in the bovine endometrium. In addition, expression analysis of key genes from the gene networks confirmed their presence and the potential regulation of these genes by uterine miRNAs. Furthermore, luciferase assay data substantiated the primary information from bioinformatic prediction that generated potential target genes for the dysregulated miRNAs in subclinical endometritis. Together, these data suggest the potential regulatory role of uterine miRNAs in the development and progression of bovine subclinical endometritis.

Expression analysis of regulatory microRNAs in bovine cumulus oocyte complex and preimplantation embryos.

MicroRNAs (miRNAs) are small endogenous molecules that are involved in a diverse of cellular process. However, little is known about their abundance in bovine oocytes and their surrounding cumulus cells during oocyte development. To elucidate this situation, we investigated the relative expression pattern of sets of miRNAs between bovine oocyte and the surrounding cumulus cells during in vitro maturation using miRNA polymerase chain reaction (PCR) array. Results revealed that a total of 47 and 51 miRNAs were highly abundant in immature and matured oocytes, respectively, compared with their surrounding cumulus cells. Furthermore, expression analysis of six miRNAs enriched in oocyte miR-205, miR-150, miR-122, miR-96, miR-146a and miR-146b-5p at different maturation times showed a dramatic decrease in abundance from 0 h to 22 h of maturation. The expression of the same miRNAs in preimplantation stage embryos was found to be highly abundant in early stages of embryo development and decreased after the 8-cell stage to the blastocyst stage following a typical maternal transcript profile. Similar results were obtained by localization of miR-205 in preimplantation stage embryos, in which signals were higher up to the 4-cell stage and reduced thereafter. miR-205 and miR-210 were localized in situ in ovarian follicles and revealed a spatio-temporal expression during follicular development. Interestingly, the presence or absence of oocytes or cumulus cells during maturation was found to affect the expression of miRNAs in each of the two cell types. Hence, our results showed the presence of distinct sets of miRNAs in oocytes or cumulus cells and the presence of their dynamic degradation during bovine oocyte maturation.

Investigation into association and expression of PLCz and COX-2 as candidate genes for boar sperm quality and fertility.

Phospholipase C zeta (PLCz) and cyclooxygenase isoenzyme type 2 (COX-2) are important in spermatogenesis, but their effect has not yet confirmed in pigs. Therefore, this study was aimed to analyse their association with sperm quality and fertility and to identify the mRNA and protein expression in boars reproductive tissues. DNA samples from 231 Pietrain (PI) and 109 Pietrain × Hampshire (PIHA) pigs with records of sperm quality [sperm concentration (SCON), motility, semen volume, plasma droplet and abnormal spermatozoa rate] and fertility (non-return rate and number of piglet born alive) traits were available. A SNP in non-coding region of PLCz g.158 A > C was associated with SCON (p < 0.05) in PIHA population while the polymorphism of COX-2 g.68 G > A in 3' UTR was not associated with any traits. For mRNA and protein expression study, a total of six boars were divided into two groups with G-I and G-II, where G-I was characterized for relatively better sperm quality. Both genes expressed higher in reproductive tissues compared with non-reproductive tissues. Phospholipase C zeta mRNA expressed higher in testis (p < 0.01), all parts of epididymis and spermatozoa from G-I, while COX-2 expressed higher in testis (p < 0.05), head and body of epididymis (p < 0.01), and spermatozoa from G-II boar. Both proteins were localized in Leydig cells and spermatozoa. These results might shed light on roles of these genes in spermatogenesis as candidate for boar sperm quality and fertility, but still the lack of association across populations should be considered.

Investigation on association and expression of ESR2 as a candidate gene for boar sperm quality and fertility.

ESR2 is involved in oestrogen-related apoptosis in cell cycle spermatogenesis but their effects have not yet confirmed in pig. Therefore, this study was aimed to investigate the association of ESR2 polymorphism with sperm quality and boar fertility traits and to analyse the ESR2 mRNA and protein expressions in boar reproductive tissues. DNA samples from 203 Pietrain (PI) and 100 Pietrain × Hampshire (PIHA) pigs with records of sperm quality [sperm concentration (SCON), motility (MOT), semen volume (VOL), plasma droplet rate (PDR) and abnormal spermatozoa rate (ASR)] and fertility [non-return rate (NRR) and number of piglet born alive (NBA)] traits were available. A SNP in coding region of ESR2 g.35547A>G in exon 5 was associated with MOT and PDR in the PI and with SCON, VOL, MOT and PDR in PIHA population. For mRNA and protein expression study, a total of six boars were divided into two groups with group I (G-I) and group II (G-II) where G-I characterized for relatively a better sperm quality according to the mean of two groups. mRNA expression was higher in brain and testis than that in all parts of epididymis. Both qRT-PCR and western blot analysis revealed that the ESR2 gene expression and protein expression were significantly higher in testis collected from G-II compared with that of G-I boars. Moreover, ESR2 protein localization in germ cell, Leydig and Sertoli cells, epithelial cells and spermatozoa was remarkable, which indicated the important role of ESR2 in spermatogenesis process. These results might shed new light on the roles of ESR2 in spermatogenesis as candidate for boar fertility, but still the lack of association across populations should be considered.

Transcriptome dynamics and molecular cross-talk between bovine oocyte and its companion cumulus cells.

BACKGROUND: The bi-directional communication between the oocyte and its companion cumulus cells (CCs) is crucial for development and functions of both cell types. Transcripts that are exclusively expressed either in oocytes or CCs and molecular mechanisms affected due to removal of the communication axis between the two cell types is not investigated at a larger scale. The main objectives of this study were: 1. To identify transcripts exclusively expressed either in oocyte or CCs and 2. To identify those which are differentially expressed when the oocyte is cultured with or without its companion CCs and vice versa. RESULTS: We analyzed transcriptome profile of different oocyte and CC samples using Affymetrix GeneChip Bovine Genome array containing 23000 transcripts. Out of 13162 genes detected in germinal vesicle (GV) oocytes and their companion CCs, 1516 and 2727 are exclusively expressed in oocytes and CCs, respectively, while 8919 are expressed in both. Similarly, of 13602 genes detected in metaphase II (MII) oocytes and CCs, 1423 and 3100 are exclusively expressed in oocytes and CCs, respectively, while 9079 are expressed in both. A total of 265 transcripts are differentially expressed between oocytes cultured with (OO+CCs) and without (OO-CCs) CCs, of which 217 and 48 are over expressed in the former and the later groups, respectively. Similarly, 566 transcripts are differentially expressed when CCs mature with (CCs+OO) or without (CCs-OO) their enclosed oocytes. Of these, 320 and 246 are over expressed in CCs+OO and CCs-OO, respectively.While oocyte specific transcripts include those involved in transcription (IRF6, POU5F1, MYF5, MED18), translation (EIF2AK1, EIF4ENIF1) and CCs specific ones include those involved in carbohydrate metabolism (HYAL1, PFKL, PYGL, MPI), protein metabolic processes (IHH, APOA1, PLOD1), steroid biosynthetic process (APOA1, CYP11A1, HSD3B1, HSD3B7). Similarly, while transcripts over expressed in OO+CCs are involved in carbohydrate metabolism (ACO1, 2), molecular transport (GAPDH, GFPT1) and nucleic acid metabolism (CBS, NOS2), those over expressed in CCs+ OO are involved in cellular growth and proliferation (FOS, GADD45A), cell cycle (HAS2, VEGFA), cellular development (AMD1, AURKA, DPP4) and gene expression (FOSB, TGFB2). CONCLUSION: In conclusion, this study has generated large scale gene expression data from different oocyte and CCs samples that would provide insights into gene functions and interactions within and across different pathways that are involved in the maturation of bovine oocytes. Moreover, the presence or absence of oocyte and CC factors during bovine oocyte maturation can have a profound effect on transcript abundance of each cell types, thereby showing the prevailing molecular cross-talk between oocytes and their corresponding CCs.

Bovine blastocysts with developmental competence to term share similar expression of developmentally important genes although derived from different culture environments.

This study was conducted to investigate the gene expression profile of in vivo-derived bovine embryo biopsies based on pregnancy outcomes after transferring to recipients. For this, biopsies of 30-40% embryos were taken from grade I blastocysts (International Embryo Transfer Society Manual) and the remaining 60-70% of the intact embryos were transferred to recipients. Frozen biopsies were pooled into three distinct groups based on the pregnancy outcome after transferring the corresponding parts, namely those resulting in no pregnancy (NP), pregnancy loss (PL), and calf delivery (CD). Array analysis revealed a total of 41 and 43 genes to be differentially expressed between biopsies derived from blastocysts resulting in NP versus CD and PL versus CD respectively. Genes regulating placental development and embryo maternal interaction (PLAC8) were found to be upregulated in embryo biopsies that ended up with CD. Embryo biopsies that failed to induce pregnancy were enriched with mitochondrial transcripts (Fl405) and stress-related genes (HSPD1). Overall, gene expression profiles of blastocysts resulting in NP and CD shared similar expression profiles with respect to genes playing significant roles in preimplantation development of embryo. Finally, comparing the transcript signatures of in vivo- and in vitro-derived embryos with developmental competence to term revealed a similarity in the relative abundance of 18 genes. Therefore, we were able to present a genetic signature associated with term developmental competence independent of the environmental origin of the transferred blastocysts.

Mapping quantitative trait loci for innate immune response in the pig.

The aim of the present study was to detect quantitative trait loci (QTL) for the serum levels of cytokines and Toll-like receptors as traits related to innate immunity in pig. For this purpose, serum concentration of interleukin 2 (IL2), interleukin 10 (IL10), interferon-gamma (IFNG), Toll-like receptor 2 (TLR2) and Toll-like receptor 9 (TLR9) were measured in blood samples obtained from F(2) piglets (n = 334) of a Duroc × Piétrain resource population (DUPI) after Mycoplasma hypopneumoniae (Mh), tetanus toxoid (TT) and Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) vaccination at 6, 9 and 15 weeks of age. Animals were genotyped at 82 genetic markers covering all autosomes. QTL analysis was performed under the line cross F(2) model using QTL Express and 33 single QTL were detected on almost all porcine autosomes. Among the single QTL, eight, twelve and thirteen QTL were identified for innate immune traits in response to Mh, TT and PRRSV vaccine, respectively. Besides single QTL, six QTL were identified by a two-QTL model, of which two for TLR9_TT were in coupling phase and one for IL10_PRRSV was in repulsion phase. All QTL were significant at 5% chromosome-wide level including one and seven at 5% genome- and 1% chromosome-wide level significance. All innate immune traits are influenced by multiple chromosomal regions implying multiple gene action. Some of the identified QTL coincided with previously reported QTL for immune response and disease resistance, and the newly identified QTL are potentially involved in the immune function. The immune traits were also influenced by environmental factors like year of birth, age, parity and litter size. The results of this work shed new light on the genetic background of innate immune response and these findings will be helpful to identify candidate genes in these QTL regions related to immune competence and disease resistance in pigs.

Suppression of the transcription factor MSX1 gene delays bovine preimplantation embryo development in vitro.

This study was conducted to investigate the effect of suppressing transcription factor gene MSX1 on the development of in vitro produced bovine oocytes and embryos, and identify its potential target genes regulated by this gene. Injection of long double-stranded RNA (LdsRNA) and small interfering RNA (siRNA) at germinal vesicle stage oocyte reduced MSX1 mRNA expression by 73 and 37% respectively at metaphase II stage compared with non-injected controls. Similarly, injection of the same anti-sense oligomers at zygote stage reduced MSX1 mRNA expression by 52 and 33% at 8-cell stage compared with non-injected controls. Protein expression was also reduced in LdsRNA- and siRNA-injected groups compared with non-injected controls at both stages. Blastocysts rates were 33, 28, 20 and 18% in non-injected control, scrambled RNA (scRNA), LdsRNA- and siRNA-injected groups respectively. Cleavage rates were also significantly reduced in Smartpool siRNA (SpsiRNA)-injected group (53.76%) compared with scRNA-injected group (57.76%) and non-injected control group (61%). Large-scale gene expression analysis showed that 135 genes were differentially regulated in SpsiRNA-injected group compared with non-injected controls, of which 54 and 81 were down- and up-regulated respectively due to suppression of MSX1. Additionally, sequence homology mapping and gene enrichment analysis with known human pathway information identified several functional modules that were affected due to suppression of MSX1. In conclusion, suppression of MSX1 affects oocyte maturation, embryo cleavage rate and the expression of several genes, suggesting its potential role in the development of bovine preimplantation embryos.

Reconstructing nonlinearities with intermodulation spectroscopy.

We describe a method of analysis which allows for reconstructing the nonlinear disturbance of a high Q harmonic oscillator. When the oscillator is driven with two or more frequencies, the nonlinearity causes intermodulation of the drives, resulting in a complicated spectral response. Analysis of this spectrum allows one to approximate the nonlinearity. The method, which is generally applicable to measurements based on resonant detection, increases the information content of the measurement without requiring a large detection bandwidth, and optimally uses the enhanced sensitivity near resonance to extract information and minimize error due to detector noise.

Expression of apoptosis regulatory genes and incidence of apoptosis in different morphological quality groups of in vitro-produced bovine pre-implantation embryos.

Apoptosis occurs during early development in both in vivo- and in vitro-produced embryos, and is considered as one of the causes of embryonic loss. The objectives of this study were, therefore, investigating stage-specific expression profiles of apoptosis regulatory genes in three quality groups of in vitro-produced bovine pre-implantation embryos; and analysing the relationship between cell number and DNA fragmentation with expressions of those genes. The relative abundance of mRNA of 9 pro- (Bax, caspase-9, Bcl-xs, P53, Caspase-3 and Fas) and anti- (Bcl-w and Mcl-1) apoptotic genes was analysed. Differential cell staining and terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labelling were performed to analyse the variation in cell numbers and detect apoptotic nuclei respectively. Expression of Bax and Caspase-3 genes was significantly (p < 0.05) higher in poor quality pre-implantation embryos as compared with that of morphologically good quality embryos of the same developmental stages. Moreover, Mcl-1 expression was significantly higher in good quality immature oocytes than that in the poor quality group. Moreover, higher DNA fragmentation was evidenced in morphologically poor quality blastocysts. In conclusion, our study demonstrates that Bax, caspase-3 and Mcl-1 can be used as potential markers of embryo quality to evaluate in vitro-produced bovine embryos. Further studies are required to investigate specific molecular signatures that can be used in evaluating in vivo-derived embryos.

Invited review: Piglet survival: benefits of the immunocompetence.

Piglet mortality has a negative impact on animal welfare and public acceptance. Moreover, the number of weaned piglets per sow mainly determines the profitability of piglet production. Increased litter sizes are associated with lower birth weights and piglet survival. Decreased survival rates and performance of piglets make the control of diseases and infections within pig production even more crucial. Consequently, selection for immunocompetence becomes an important key aspect within modern breeding programmes. However, the phenotypic recording of immune traits is difficult and expensive to realize within farm routines. Even though immune traits show genetic variability, only few examples exist on their respective suitability within a breeding programme and their relationships to economically important production traits. The analysis of immune traits for an evaluation of immunocompetence to gain a generally improved immune response is promising. Generally, in-depth knowledge of the genetic background of the immune system is needed to gain helpful insights about its possible incorporation into breeding programmes. Possible physiological drawbacks for enhanced immunocompetence must be considered with regards to the allocation theory and possible trade-offs between the immune system and performance. This review aims to discuss the relationships between the immunocompetence of the pig, piglet survival as well as the potential of these traits to be included into a breeding strategy for improved robustness.

Verification of chromosomal regions affecting the innate immunity in pigs using linkage mapping.

Understanding the genetic control of innate immunity in pigswould offerthe opportunity to utilize natural variation and improve selective breeding programmes. As part of our porcine genome scan to identify quantitative trait loci (QTL) we examined immune response traits in a Duroc x Berlin miniature pig resource family (DUMI). Complement activity via classical (CH50) and alternative (AH50) pathways, antibody response to Mycoplasma hyopneumoniae (Mh), tetanus toxoid (TET) and PRRS virus (PRRSV), the complement component (C3c), and Haptoglobin serum concentration (HP) were used as phenotypes for linkage mapping. A total of 220 backcross animals were used for the QTL analysis. Blood was collected six times from each animal prior to and after vaccinations against Mycoplasma hyopneumoniae, tetanus toxoid and PRRS, respectively. Seventy-four microsatellites from 18 autosomes were used for QTL mapping. The analyses were performed treating the measurements of phenotypes at different time points as single traits. Forty-two significant and 24 highly significant QTL were detected, using the program QTL Express, for all immune traits using the single traits. Most QTL were detected on SSC3, SSC16, and SSC18. No significant F-value corresponded to data for SSC12 and SSC13.

Physiological interactions between the endocrine and immune systems shown in gene analysis in pigs.

A number of observations support the concept of important physiological interactions between the endocrine and immune systems. It could be confirmed that hormones secreted by the neuroendocrine system play an important role in communication and regulation of the cells of the immune system. Among protein hormones, this has been most clearly documented for prolactin (PRL), growth hormone (GH), and insulin-like growth factor 1 (IGF-1). A number of traits of the immune response in a Duroc x Berlin Miniature pig family (DUMI) were examined. The haemolytic complement activity in the classical complement pathway (CH50) and the alternative pathway (AH50)was examined at eight different time points in pigs that were vaccinated with different vaccines. Single nucleotide polymorphism (SNP) genotyping was employed to genotype the DUMI F2 animal for growth hormone (Gh), growth hormone releasing hormone (Ghrh), pituitary-specific transcription factor 1 (Pit1), and prolactin receptor (Prlr) loci, and also a microsatellite within insulin-like growth factor 1 (Igf1). Using a family-based association test (FBAT) program, a highly significant association of Gh, Pit1, and Prlr to AH50 (p < or = 0.01) and a significant association of Pit1 to CH50 (p < or = 0.05) were found. Using the SAS system for mixed model, a highly significant association of Gh, and Igf1 to AH50 and CH50 (p < or = 0.01) was detected, while Prlr and Ghrh had a highly significant association (p < or = 0.01) with CH50 only.

Effect of condensed tannins in rations of lactating dairy cows on production variables and nitrogen use efficiency.

The objective of this study was to evaluate the effect of supplemented condensed tannins (CT) from the bark of the Black Wattle tree (Acacia mearnsii) on production variables and N use efficiency in high yielding dairy cows. A feeding trial with 96 lactating German Holstein cows was conducted for a total of 169 days, divided into four periods. The animals were allotted to two groups (control (CON) and experimental (EXP) group) according to milk yield in previous lactation, days in milk (98), number of lactations and BW. The trial started and finished with a period (period 1 and 4) where both groups received the same ration (total-mixed ration based on grass and maize silage, ensiled sugar beet pulp, lucerne hay, mineral premix and concentrate, calculated for 37 kg energy-corrected milk). In between, the ration of EXP cows was supplemented with 1% (CT1, period 2) and 3% of dry matter (DM) (CT3, period 3) of a commercial A. mearnsii extract (containing 0.203 g CT/g DM) which was mixed into the concentrate. In period 3, samples of urine and faeces were collected from 10 cows of each group and analyzed to estimate N excretion. Except for a tendency for a reduced milk urea concentration with CT1, there was no difference between groups in period 2 (CON v. CT1; P>0.05). The CT3 significantly reduced (P<0.05) milk protein yield, the apparent N efficiency (kg milk N/k feed N) and milk urea concentration; but total milk yield and energy-corrected milk yield were not affected by treatment. Furthermore, as estimated from 10 cows per group and using urinary K as a marker to estimate the daily amount of urine voided, CT3 caused a minor shift of N compounds from urine to faeces, as urea-N in urine was reduced, whereas the N concentration in faeces increased. As an improvement in productivity was not achieved and N use efficiency was decreased by adding the CT product it can be concluded that under current circumstances the use in high yielding dairy cows is not advantageous.

Genetic aspects concerning drip loss and water-holding capacity of porcine meat.

The amount and distribution of water inside the meat has a considerable influence on its properties. High losses of fluid in the form of drip may affect financial output, nutritional value, consumer appeal and/or technological properties of porcine meat. Therefore, a deeper insight into the traits water-holding capacity (WHC) and drip is favourable on behalf of producers, industry and consumers. Similar to most meat quality traits, WHC and drip loss (DRIP) are moderately heritable. The genetic correlation between these two traits is high. Correlation to other meat quality traits, such as pH value, cooking loss, reflectance, etc. is existent as predictable. Two major genes are known, RYR1 on chromosome 6 and RN on chromosome 15, to influence meat quality in general and WHC in particular. Furthermore, a number of candidate genes exist, e.g. phosphoglycerate mutase 2. Within the variety of quantitative trait loci (QTL) experiments, a number of QTL have been identified. QTL for DRIP and/or WHC have been found on chromosome 1, 2, 4, 5, 6, 11, 13, 14, 15, 18; for cooking loss on 7, 14 and18, and for pH value on nearly all chromosomes. Recently, a QTL study for meat quality and body composition traits in a Duroc-Pietrain (DUPI) resource population has been conducted at the University of Bonn, Germany. Four QTL for DRIP were identified on chromosomes 2, 3, 5 and 18. The QTL regions are in agreement with previously published QTL for this and other related traits. Further research and finemapping has begun and candidate genes located within the QTL regions are currently under investigation. Combination and comparison of results should lead to deeper insights in the genetic background of meat quality and DRIP.

Candidate gene markers for sperm quality and fertility of boar.

Candidate genes gonadotropin releasing hormone receptor (GNRHR), prolactin (PRL), prolactin receptor (PRLR), follicle-stimulating hormone beta (FSHB), luteinizing hormone beta (LHB), follistatin (FST), inhibin alpha (INHA), inhibin beta A (INHBA) and inhibin beta B (INHBB) were investigated for their association with sperm quality traits of sperm concentration (SCON), motility (MOT), semen volume per ejaculate (VOL), plasma droplets rate (PDR), abnormal sperm rate (ASR) and fertility traits of non return rate (NRR) and number of piglets born alive (NBA). The experimental material included 356 boars of Pietrain (PI) and Pietrain x Hampshire (PI x HA). Analysis of variance revealed significant association of GNRHR with MOT (P = 0.0161), PDR (P = 0.0048) and ASR (P = 0.0201), INHBA was found to have significant effects on PDR (P = 0.0318) and ASR (P = 0.0067), INHBB was significant (P = 0.0360) for SCON trait. FSHB, FST, INHA, PRL, PRLR and LHB had no significant effects on any trait in this experiment.

Can multi-subpopulation reference sets improve the genomic predictive ability for pigs?

In most countries and for most livestock species, genomic evaluations are obtained from within-breed analyses. To achieve reliable breeding values, however, a sufficient reference sample size is essential. To increase this size, the use of multibreed reference populations for small populations is considered a suitable option in other species. Over decades, the separate breeding work of different pig breeding organizations in Germany has led to stratified subpopulations in the breed German Large White. Due to this fact and the limited number of Large White animals available in each organization, there was a pressing need for ascertaining if multi-subpopulation genomic prediction is superior compared with within-subpopulation prediction in pigs. Direct genomic breeding values were estimated with genomic BLUP for the trait "number of piglets born alive" using genotype data (Illumina Porcine 60K SNP BeadChip) from 2,053 German Large White animals from five different commercial pig breeding companies. To assess the prediction accuracy of within- and multi-subpopulation reference sets, a random 5-fold cross-validation with 20 replications was performed. The five subpopulations considered were only slightly differentiated from each other. However, the prediction accuracy of the multi-subpopulations approach was not better than that of the within-subpopulation evaluation, for which the predictive ability was already high. Reference sets composed of closely related multi-subpopulation sets performed better than sets of distantly related subpopulations but not better than the within-subpopulation approach. Despite the low differentiation of the five subpopulations, the genetic connectedness between these different subpopulations seems to be too small to improve the prediction accuracy by applying multi-subpopulation reference sets. Consequently, resources should be used for enlarging the reference population within subpopulation, for example, by adding genotyped females.

Evaluation of carcass composition of intact boars using linear measurements from performance testing, dissection, dual energy X-ray absorptiometry (DXA) and magnetic resonance imaging (MRI).

The objective of this study was to investigate non-invasive imaging methods to update the used regression equation for stationary tested boars. A total of 94 boars were examined. 20 boars were dissected to provide the reference LMP. Performance data (PD) from right carcasses were available from all groups. The left carcasses were studied by MRI & DXA. Based on the reference LMP and the MRI & DXA data, regression equations for LMP were developed. The estimates for LMP based on MRI & DXA data were used to calculate new regression equations for entire male carcass halves based on linear PD. Further 33 PD sets served as independent sample, which was included in a Monte Carlo simulation for imputing the missing reference LMPs (n=74) and discussing the accuracy of the results. The LMP regression equation based on the combined MRI & DXA data is as accurate as the former regression equation, but needs only three instead of seven variables.

Comparison of different methods to assess the composition of pig bellies in progeny testing.

The objective of the study was to validate methods that assess the belly composition of stationary tested progenies of Piétrain boars. In German performance test stations, there are currently three methods of determining belly compositon in use: 1) a regression equation that contains different carcass characteristics, such as fat thickness and muscle area; 2) planimetric analysis of video or digital images acquired at the cut between the 13th and 14th ribs; and 3) estimation of the belly composition using ultrasound data from a three-dimensional ultrasound image produced an online carcass-grading system. Validation of these techniques was performed on 400 carcasses of stationary-tested Piétrain and Piétrain-sired crossbred pigs, which were slaughtered at a mean carcass weight of 85 and 97 kg. Magnetic resonance imaging (MRI) served as a reference to determine the lean content of the bellies. The correlation to MRI lean content ranged from 0.71 to 0.81, and corresponding correlation values were 0.62 to 0.64 for the digital imaging technique, and 0.53 to 0.59 for the AutoFOM online carcass-grading system. An increase in precision was achieved when information from digital imaging and linear carcass measures were included in the regression equation. Accuracy of the AutoFOM system does not seem to be sufficient to assess the belly composition for the special breeds in performance testing. However, extracting and combining 127 AutoFOM-base recordings into modified equations using partial least squares techniques yielded an improvement in the prediction accuracy for all tested breed and/or weight groups.