Peripheral blood mononuclear cells (PBMCs) are the most immunologically active and readily accessible fraction of whole blood, and they initiate host immune responses following infection and vaccination. Since PBMC transcriptomes capture diverse host–pathogen interactions, they offer a promising tool to uncover the genetic drivers of viral resistance and tolerance. As a detailed case study of this principle, we examine porcine reproductive and respiratory syndrome (PRRS), which remains one of the most economically important viral diseases of swine worldwide. Following PRRS virus (PRRSV) exposure, pigs rely on two complementary defense strategies: resistance, the capacity to limit viral replication, and tolerance, the capacity to sustain performance despite infection. Since resistance and tolerance phenotypes are difficult to measure directly through experimental challenge, indirect immune-trait measurements collected after vaccination offer a practical alternative. Most transcriptomic studies of the host response to PRRSV have focused on respiratory tissues, reflecting the virus’s tropism for pulmonary macrophages. However, intramuscularly delivered modified-live PRRSV vaccine reaches the bloodstream, bypassing the lung, so PBMCs, as the frontline defense system, mount the earliest measurable innate response. This review synthesizes the current literature on PBMC transcriptome models for deciphering innate resistance and tolerance to viral disease, using PRRS as our principal worked example; presents our own approach to profiling PBMCs after PRRSV vaccination; and outlines how the field has advanced since the original candidate-gene and QTL studies of the 2010s, including the recent FDA approval of the first CD163 gene-edited PRRSV-resistant pig line, and the emergence of single-cell and multi-tissue PBMC atlases, before considering how the same PBMC-based approach could extend to other host–virus interactions.

BACKGROUND:Various studies have highlighted significant differences in developmental kinetics and sensitivity to developmental conditions between male and female bovine embryos. These differences are thought to be caused in part by the sexually dimorphic expression of genes located on the sex or autosomal chromosomes. However, little is known about the dimorphic gene expression patterns of bovine embryos at the initiation of elongation, which is one of the critical stages of development. Furthermore, to the best of our knowledge, there is little or no data available on the sexually dimorphic gene expression patterns in bovine embryos in relation to maternal environmental conditions during the initiation of elongation. Therefore, the main objective of this study was to investigate the sexually dimorphic gene expression responses of embryos to the maternal environment at the initiation of elongation in embryos developed in lactating dairy cows and nonlactating nulliparous heifers. RESULTS:Gene expression analysis showed that 159 genes including those involved in steroid biosynthesis and gastrulation were differentially expressed exclusively between male and female embryos developed in cows. Among these, 61 genes including CYP39 A1, CYP2R1 and CYP27B1 were upregulated and 98 genes including HSD17B1, HSD17B10 and aromatase (CYP19 A1) were downregulated in male embryos. Chromosomal analysis showed that 31.2% of the differentially expressed genes (DEGs) including glucose-6-phosphate dehydrogenase (G6PD) were located on the X chromosome, and 96% of those were upregulated in female embryos. Similarly, 254 genes including those involved in female sex differentiation, placenta development, transmembrane transport, and cell adhesion were differentially expressed exclusively between the male and female embryos developed in heifers. Of these, 108 genes including HSD17B11, HSD17B12, and HSD3B1 were upregulated, and 146 genes including SLC16 A9, SLC10 A1, SLC10 A3, SLC16 A5, SLC22 A23, SLC25 A43, SLC35 A2, SLC35 C1, and SLC4 were downregulated in male compared to female embryos. In addition, 17.3% of the DEGs were located on the X chromosome and 75% of the DEGs located on the X chromosome were upregulated in female embryos. On the other hand, 38 genes including SLC30 A10, SLC10 A4, ATP6 AP1, and KDM5 C showed sexually dimorphic expression patterns in day 13 bovine embryos irrespective of the maternal environment. These genes accounted for only 19% and 13% of the genes that showed sexually dimorphic expression in embryos developed in cows and heifers, respectively and the expression difference of these genes in male and female embryos was then likely influenced by the sex of the embryo. CONCLUSION:This study revealed that embryos developed in lactating cows showed sexually dimorphic expression of genes involved in various functions including steroid biosynthesis and gastrulation. In contrast, embryos developed in heifers displayed sexually dimorphic expression of genes related to placental development, female sex differentiation, and transmembrane transport. This suggests that the reproductive tract environments of cows and heifers differently affect the sex specific expression of genes in bovine embryos. A higher proportion of genes that showed sexually dimorphic expression in cow embryos were located on the X chromosome, and the majority of these genes were upregulated in female embryos. Overall, this study provides insight into genes that exhibit sexually dimorphic expression patterns in day 13 bovine embryos due to the maternal reproductive tract microenvironment or solely due to the sex of the embryo.

Although differences between in vivo and in vitro derived bovine blastocysts in terms of global gene expression profiles have been reported, comparative transcriptome analyses specifically addressing the sustained impact of the oocyte source or the in vitro culture environments during maturation and post-maturation period are remain limited. Therefore, the present study aimed to investigate the specific impacts of oocyte origin and the culture environment during and after maturation on the gene expression signature at blastocyst stage. To achieve this, we utilized our recently developed technique that enables intrafollicular transfer of immature and matured slaughterhouse-derived oocytes into dominant or preovulatory follicles. The presents study identified a total of 1052 differentially expressed genes between in vitro and in vivo derived blastocysts, many of which are involved in key pathways related to protein synthesis, protein degradation and cell-cycle regulation. The majority of these genes (n = 913), particularly those associated with “ubiquitin mediated proteolysis”, “proteasome activity” as well as “cell cycle” related pathways, were differentially expressed due to the in vitro environment following oocyte maturation. Moreover, a distinct set of genes (n = 109) including DHCR7, DHCR24, HMGCR, HMGCS1 and SCD5, which are crucial for cholesterol biosynthesis and lipid metabolism, were altered in response to the in vitro environment during oocyte maturation. Notably, the origin of the immature oocyte also appeared to predetermine the later expression outline of a set of genes (n = 28), including DLD and PLAC8, which are implicated in implantation success and calf delivery. The present study provides a comprehensive overview of transcriptomic alterations and pathway disruptions resulting from the in vitro environment following oocyte maturation, offering insight into potential mechanisms underlying embryonic genome activation, DNA duplication and appropriate cell cleavage. The differential expression of genes involved in cholesterol biosynthesis and lipid metabolism due to the in vitro maturation environment may contribute to the reduced cryotolerance observed in the resulting blastocysts. Furthermore, dysregulation of specific genes as a consequence of oocyte source has implications for post-implantation developmental competence. Collectively, these findings advance our understanding of the molecular determinants affecting embryonic developmental potential. The expression signature of these pathways could therefore be used to assess the impact of various treatments and culture environments on embryonic development. In addition, the insights gained from this study could inform future strategies to improve the quality of embryos in in vitro production systems through the targeted modulation, either enhancement or inhibition, of specific genes or pathways.

Embryonic genome activation is divided into a minor and a major wave of transition to endogenous transcription. In bovines, minor genome activation begins early in the 2-cell stage and is completed by the 8-cell stage when major genome activation becomes dominant. While the activation of genes known to regulate early development have been studied extensively, genes involved in more central cellular functions have not been examined. Taking advantage of the CRISPR Cas9 system, the present study investigated the effect of knocking out the Golgi retrograde protein transporter COPA on early bovine development. After the electroporation of presumptive zygotes with Cas9 ribonucleoproteins targeting COPA exon 6, sequences of 2 (11 %) and 4-cell (16 %) embryos showed knockouts of COPA whereas 8-cell embryos and blastocysts did not, demonstrating that COPA is necessary for development to the 8-cell stage and beyond. Using a repair template containing silent mutations along the target site, COPA loss of wildtype was observed in 5 blastocysts, with successful knock-in of the template on at least one allele. This shows that an edited yet functional copy of COPA can save the developmental capacity of the embryo and demonstrates that Cas9 activity at the target region itself is not responsible for the loss of function. Together, the present study revealed that COPA is necessary for embryonic development, and that the timing of this necessity is before major genome activation onset. More generally, this study further demonstrates the utility of genome editing within reproductive biotechnology for the interrogation of gene function and early embryonic development.

The major limitation of the widespread use of IVP derived embryos is their consistent deficiencies in vitality when compared with their ex vivo derived counterparts. Although embryo metabolism is considered a useful metric of embryo quality, research connecting mitochondrial function with the developmental capacity of embryos is still lacking. Therefore, the aim of the present study was to analyse bovine embryo respiration signatures in relation to developmental capacity. This was achieved by taking advantage of two generally accepted metrics for developmental capacity: (I) environmental conditions during development (vivo vs. vitro) and (II) developmental kinetics (day 7 vs. day 8 blastocysts). Our study showed that the developmental environment affected total embryo oxygen consumption while different morphokinetics illustrating the embryo qualities correlate with maximal mitochondrial respiration, mitochondrial spare capacity, ATP-linked respiration as well as efficiency of ATP generation. This respiration fingerprint for high embryo quality is reflected by relatively lower lipid contents and relatively higher ROS contents. In summary, the results of the present study extend the existing knowledge on the relationship between bovine embryo quality and the signature of mitochondrial respiration by considering contrasting developmental environments as well as different embryo morphokinetics.

The widespread use of cryopreserved in vitro produced (IVP) bovine embryos is limited due to their low post-warming viability compared to their ex vivo derived counterparts. Therefore, the present study aimed to analyse in detail the consequences of cryopreservation (vitrification and slow freezing) on the bioenergetic profile of the embryo and its mitochondria. To accomplish that, day 7 IVP embryos were separated in a non-cryopreserved control group (fresh, n = 120, 12 replicates) or were either slow frozen (slow frozen, n = 60, 6 replicates) or vitrified (vitrified, n = 60, 6 replicates). An in-depth analysis of the bioenergetic profiles was then performed on these 3 groups, analysing pools of 10 embryos revealing that embryo cryopreservation both via vitrification and slow freezing causes profound changes in the bioenergetic profile of bovine embryos. Noteworthy, fresh embryos demonstrate a significantly (P < 0.05) higher oxygen consumption rate (OCR) compared to vitrified and slow frozen counterparts (0.858 ± 0.039 vs. 0.635 ± 0.048 vs. 0.775 ± 0.046 pmol/min/embryo). This was found to be largely due to significantly reduced mitochondrial oxygen consumption in both vitrified and deep-frozen embryos compared to fresh counterparts (0.541 ± 0.057 vs. 0.689 ± 0.044 vs. 0.808 ± 0.025 pmol/min/embryo). Conversely, slow-frozen thawed blastocysts showed 1.8-fold (P < 0.05) higher non-mitochondrial OCR rates compared to fresh embryos. Maximum mitochondrial respiration of vitrified and slow-frozen embryos was significantly reduced by almost 1.6-fold compared to fresh embryos and the proportion of ATP-linked respiration showed significantly lower values in vitrified thawed embryos compared to fresh embryos (1.1-fold, P < 0.05). Likewise, vitrification-warming and freeze-thawing reduced reactive glycolytic capacity (1.4 fold, 1.2-fold)as well as compensatory glycolytic capacity to provide energy in response to mitochondrial deficiency (1.3-fold and 1.2-fold, P < 0.05). In conclusion, the present study has, to the best of our knowledge, identified for the first time a comprehensive overview of typical altered metabolic features of the bioenergetic profile of bovine embryos after cryopreservation, which have great potential to explain the detrimental effects of cryopreservation on embryo viability. Avoidance of these detrimental effects through technical improvements is therefore suggested to be mandatory to improve the viability of bovine embryos after cryopreservation-warming.

Background Immune traits are considered to serve as potential biomarkers for pig’s health. Medium to high heritabilities have been observed for some of the immune traits suggesting genetic variability of these phenotypes. Consideration of previously established genetic correlations between immune traits can be used to identify pleiotropic genetic markers. Therefore, genome-wide association study (GWAS) approaches are required to explore the joint genetic foundation for health biomarkers. Usually, GWAS explores phenotypes in a univariate (uv), trait-by-trait manner. Besides two uv GWAS methods, four multivariate (mv) GWAS approaches were applied on combinations out of 22 immune traits for Landrace (LR) and Large White (LW) pig lines. Results In total 433 (LR: 351, LW: 82) associations were identified with the uv approach implemented in PLINK and a Bayesian linear regression uv approach (BIMBAM) software. Single Nucleotide Polymorphisms (SNPs) that were identified with both uv approaches (n = 32) were mostly associated with immune traits such as haptoglobin, red blood cell characteristics and cytokines, and were located in protein-coding genes. Mv GWAS approaches detected 647 associations for different mv immune trait combinations which were summarized to 133 Quantitative Trait Loci (QTL). SNPs for different trait combinations (n = 66) were detected with more than one mv method. Most of these SNPs are associated with red blood cell related immune trait combinations. Functional annotation of these QTL revealed 453 immune-relevant protein-coding genes. With uv methods shared markers were not observed between the breeds, whereas mv approaches were able to detect two conjoint SNPs for LR and LW. Due to unmapped positions for these markers, their functional annotation was not clarified. Conclusions This study evaluated the joint genetic background of immune traits in LR and LW piglets through the application of various uv and mv GWAS approaches. In comparison to uv methods, mv methodologies identified more significant associations, which might reflect the pleiotropic background of the immune system more accurately. In genetic research of complex traits, the SNP effects are generally small. Furthermore, one genetic variant can affect several correlated immune traits at the same time, termed pleiotropy. As mv GWAS methods consider strong dependencies among traits, the power to detect SNPs can be boosted. Both methods revealed immune-relevant potential candidate genes. Our results indicate that one single test is not able to detect all the different types of genetic effects in the most powerful manner and therefore, the methods should be applied complementary.

Improving the immunocompetence towards pathogens represents a desirable objective of breeding strategies to increase resilience. However, the immune system is complex and the genetic foundation of the underlying components is not yet clarified. In the present study, we focused on 22 blood parameters of 1,144 Landrace (LR) and Large White (LW) piglets at the age of 6–7 weeks. The immune profiles covered immune cells, red blood cell characteristics and cytokines. Genetic parameters based on pedigree information along with possible environmental effects were estimated. Litter effects play an important role in the expression of immune parameters of their young progenies. Hence, litter impacts on the piglet's immune profile including the immune parameters of the dam itself were investigated by different models. To incorporate the complexity of the immune network, the data were further investigated with a principal component analysis. Immune traits showed low to high breed-specific heritabilities ( h 2 ). Strong positive r g were estimated among red blood cell characteristics (0.77–0.99) and among cytokines (0.48–0.99). Neutrophils and lymphocytes illustrated a high negative r g (−0.96 to −0.98). The litter impact on piglet's immunity was examined and strengthened already observed breed differences. In LR, h 2 (0.22–0.15) and litter effect ( c 2 ) (0.52–0.44) for IFN-γ decreased after statistical consideration of maternal impact. In LW, a decrease in h 2 (0.32–0.18) for IFN-γ and an increase in c 2 (0.54–0.56) were observed. Here, sufficient correlations were detected within various immune traits and functional biological networks of principal components. Most immune traits are heritable and are promising to cover global breed-specific immunocompetence in pigs. The analysis of immune traits has to be extended in order to find an optimal range and to characterize relationships between immunity and performance to gain an improved immune system without accidental losses in productivity.

Follicular fluid (FF) plays an important role during follicular development and it contains several bioactive molecules including extracellular microRNAs (ECmiRNAs) that may mediate cell-cell communication during follicular development. Yet, the distribution patterns of ECmiRNAs in FF is not well characterized. This study aims to investigate the distribution of ECmiRNAs in two major fractions, namely exosomal and non-exosomal, of bovine follicular fluid (bFF). Exosomal and non-exosomal fractions from bFF were separated using Exoquick™ exosomes precipitation kit. miRNA expression was evaluated using the human miRCURY LNA™ Universal RT miRNA PCR array system. Transmission electron microscopy and immunoblotting revealed that the isolated vesicles were exosomes. The real-time PCR-based expression analysis revealed that 516 miRNAs were detected in the exosomal fraction of bFF, while 393 miRNAs were detected in the non-exosomal fraction. Among the detected miRNAs, a total of 370 miRNAs were detected in both fractions, while 145 miRNAs and 23 miRNAs were solely detected in exosomal and non-exosomal fractions, respectively. Exploratory pathway analysis showed that the genes targeted by exosomal and non-exosomal miRNAs to be involved in MAPK, Wnt, FoxO, TGF-beta, Oxytocin, ErbB, PI3K-Akt, Neurotrophin signalling pathways which are believed to be involved in follicular development, cell proliferation, and meiotic resumption. The results of our study demonstrated that besides the exosomal fraction, non-exosomal fractions can carry a significant amount of miRNAs in bFF where the exosomal fraction carries a significantly higher number of detectable miRNAs.

At the embryonic level, CRISPR technologies have been used to edit genomes reliably and efficiently in various mammalian models, with Ribonucleoprotein (RNP) electroporation potentially representing a superior delivery method into mammalian zygotes. However, detailed insights of the interactions between varying technical settings as well as the time point of electroporation in a bovine zygote's cell cycle on developmental metrics and the frequency and type of editing events are largely unknown. The present study uncovers that increasing pulse lengths result in higher Full Edit rates, with Mosaicism in Full-Edit embryos being significantly affected by adjusting RNP-electroporation relative to zygote cell cycle. A considerable proportion of Full Edit embryos demonstrated loss-of-heterozygosity after RNP-electroporation prior to S-phase. Some of these loss-of-heterozygosity events are a consequence of chromosomal disruptions along large sections of the target chromosomes making it necessary to check for their presence prior use of this technique in animal breeding. One out of 2 of these loss-of-heterozygosity events, however, was not associated with loss of an entire chromosome or chromosomal sections. Whether analysed loss-of-heterozygosity in these cases, however, was a false negative result due to loss of PCR primer sequences after INDEL formation at the target side or indeed due to interhomolog recombination needs to be clarified in follow up studies since the latter would for sure offer attractive options for future breeding schedules.

Satellite cells take an indispensable place in skeletal muscle regeneration, maintenance, and growth. However, only limited works have investigated effects of dietary compounds on the proliferation of porcine satellite cells (PSCs) and related mechanisms. Sulforaphane (SFN) at multiple levels was applied to PSCs. The PSCs’ viability and HDAC activity were measured with a WST-1 cell proliferation kit and Color-de-Lys® HDAC colorimetric activity assay kit. Gene expression and epigenetics modification were tested with qRT-PCR, Western blot, bisulfite sequencing, and ChIP-qPCR. This study found that SFN enhanced PSC proliferation and altered mRNA expression levels of myogenic regulatory factors. In addition, SFN inhibited histone deacetylase (HDAC) activity, disturbed mRNA levels of HDAC family members, and elevated acetylated histone H3 and H4 abundance in PSCs. Furthermore, both mRNA and protein levels of the Smad family member 7 (SMAD7) in PSCs were upregulated after SFN treatment. Finally, it was found that SFN increased the acetylation level of histone H4 in the SMAD7 promoter, decreased the expression of microRNAs, including ssc-miR-15a, ssc-miR-15b, ssc-miR-92a, ssc-miR-17-5p, ssc-miR-20a-5p, and ssc-miR-106a, targeting SMAD7, but did not impact on the SMAD7 promoter’s methylation status in PSCs. In summary, SFN was found to boost PSC proliferation and epigenetically increase porcine SMAD7 expression, which indicates a potential application of SFN in modulation of skeletal muscle growth.

Inflammation is regulated by epigenetic modifications, including DNA methylation and histone acetylation. Sulforaphane (SFN), a histone deacetylase (HDAC) inhibitor, is also a potent immunomodulatory agent, but its anti-inflammatory functions through epigenetic modifications remain unclear. Therefore, this study aimed to investigate the epigenetic effects of SFN in maintaining the immunomodulatory homeostasis of innate immunity during acute inflammation. For this purpose, SFN-induced epigenetic changes and expression levels of immune-related genes in response to lipopolysaccharide (LPS) stimulation of monocyte-derived dendritic cells (moDCs) were analyzed. These results demonstrated that SFN inhibited HDAC activity and caused histone H3 and H4 acetylation. SFN treatment also induced DNA demethylation in the promoter region of the MHC-SLA1 gene, resulting in the upregulation of Toll-like receptor 4 (TLR4), MHC-SLA1, and inflammatory cytokines' expression at 6 h of LPS stimulation. Moreover, the protein levels of cytokines in the cell culture supernatants were significantly inhibited by SFN pre-treatment followed by LPS stimulation in a time-dependent manner, suggesting that inhibition of HDAC activity and DNA methylation by SFN may restrict the excessive inflammatory cytokine availability in the extracellular environment. We postulate that SFN may exert a protective and anti-inflammatory function by epigenetically influencing signaling pathways in experimental conditions employing porcine moDCs.

Dynamic changes in microRNAs in oocyte and cumulus cells before and after maturation may explain the spatiotemporal post-transcriptional gene regulation within bovine follicular cells during the oocyte maturation process. miR-20a has been previously shown to regulate proliferation and differentiation as well as progesterone levels in cultured bovine granulosa cells. In the present study, we aimed to demonstrate the function of miR-20a during the bovine oocyte maturation process. Maturation of cumulus-oocyte complexes (COCs) was performed at 39°C in an humidified atmosphere with 5% CO2 in air. The expression of miR-20a was investigated in the cumulus cells and oocytes at 22 h post culture. The functional role of miR-20a was examined by modulating the expression of miR-20a in COCs during in vitro maturation (IVM). We found that the miR-20a expression was increased in cumulus cells but decreased in oocytes after IVM. Overexpression of miR-20a increased the oocyte maturation rate. Even though not statistically significant, miR-20a overexpression during IVM increased progesterone levels in the spent medium. This was further supported by the expression of STAR and CYP11A1 genes in cumulus cells. The phenotypes observed due to overexpression of miR-20a were validated by BMP15 supplementation during IVM and subsequent transfection of BMP15-treated COCs using miR-20a mimic or BMPR2 siRNA. We found that miR-20a mimic or BMPR2 siRNA transfection rescued BMP15-reduced oocyte maturation and progesterone levels. We concluded that miR-20a regulates oocyte maturation by increasing cumulus cell progesterone synthesis by simultaneous suppression of BMPR2 expression.

Transcription factors (TFs) are known to be involved in regulating the expression of several classes of genes during folliculogenesis. However, the regulatory role of TFs during oxidative stress (OS) is not fully understood. The current study was aimed to investigate the regulation of the TFs in bovine granulosa cells (bGCs) during exposure to OS induced by H2O2 in vitro. For this, bGCs derived from ovarian follicles were cultured in vitro till their confluency and then treated with H2O2 for 40 min. Twenty-four hours later, cells were subjected to various phenotypic and gene expression analyses for genes related to TFs, endoplasmic reticulum stress, apoptosis, cell proliferation, and differentiation markers. The bGCs exhibited higher reactive oxygen species accumulation, DNA fragmentation, and endoplasmic reticulum stress accompanied by reduction of mitochondrial activity after exposure to OS. In addition, higher lipid accumulation and lower cell proliferation were noticed in H2O2-challenged cells. The mRNA level of TFs including NRF2, E2F1, KLF6, KLF9, FOS, SREBF1, SREBF2, and NOTCH1 was increased in H2O2-treated cells compared with non-treated controls. However, the expression level of KLF4 and its downstream gene, CCNB1, were downregulated in the H2O2-challenged group. Moreover, targeted inhibition of NRF2 using small interference RNA resulted in reduced expression of KLF9, FOS, SREBF2, and NOTCH1 genes, while the expression of KLF4 was upregulated. Taken together, bovine granulosa cells exposed to OS exhibited differential expression of various transcription factors, which are mediated by the NRF2 signaling pathway.

Epididymal sperm shows higher cryoresistance than ejaculated sperm. Although the sperm proteome seems to affect cell cryoresistance, studies aiming at identifying proteins involved in sperm freezing-tolerance are scarce. The aims of this study were to investigate differences of sperm freezability and proteome between epididymal and ejaculated sperm in three mountain ungulates: Iberian ibex, Mouflon and Chamois. Sperm samples were cryopreserved in straws by slow freezing. Tandem mass tag-labeled peptides from sperm samples were analyzed by high performance liquid chromatography coupled to a mass spectrometer in three technical replicates. The statistical analysis was done using the moderated t-test of the R package limma. Differences of freezability between both types of sperm were associated with differences of the proteome. Overall, epididymal sperm showed higher freezability than ejaculated sperm. Between 1490 and 1883 proteins were quantified in each species and type of sperm sample. Cross species comparisons revealed a total of 76 proteins that were more abundant in epididymal than in ejaculated sperm in the three species of study whereas 3 proteins were more abundant in ejaculated than epididymal sperm in the three species of study (adjusted P < 0.05; |log2| fold-change > 0.5). Many of the proteins that were associated with higher cryoresistance are involved in stress response and redox homeostasis. In conclusion, marked changes of sperm proteome were detected between epididymal and ejaculated sperm. This work contributes to update the sperm proteome of small ruminants and to identify candidate markers of sperm freezability.

The present study was conducted to investigate the potential adverse effect of Pb on pregnant Sprague-Dawley rats and their fetuses after maternal exposure, on gestational days (GD) 7-16. The possible protective role of taurine (TA), administered throughout the gestation period (GD 1-20) against Pb toxicity, was also evaluated. Pregnant rats were divided into four groups: Group 1 (control) was given distilled water; Group 2 was exposed to Pb (250 ppm) in drinking water (GD 7-16), whereas Group 3 received TA (50 mg/kg/day) by oral gavage (GD 1-20); Group 4 was exposed to Pb (GD 7-16), whereas pretreated with TA from GD 1 till the end of the gestation period. After termination on GD 20, maternal and embryo-fetal outcomes were evaluated. Blood samples were collected for hematological and biochemical parameters assessment. The results showed that, Pb induced a significant reduction in the maternal body weight, weight gain, uterine and placental weight, in addition to a high incidence of abortion and fetal resorption. Meanwhile, fetuses demonstrated decreased body weight and length, with a high rate of mortality as well as external and skeletal abnormalities. Additionally, Pb induced severe hematological and biochemical alterations in both dams and fetuses. The toxicity of Pb was further emphasized by placental histopathological examination and hepatic DNA fragmentation. Pretreatment with TA greatly attenuated the impact of Pb on both maternal and fetal parameters. Moreover, TA alleviated the incidence of placental damage and hepatic DNA fragmentation. The results highlight the potential prophylaxis role of TA against maternal and developmental Pb toxicity.

Background Morphological evaluation of embryos has been used to screen embryos for transfer. However, the repeatability and accuracy of this method remains low. Thus, evaluation of an embryo’s gene expression signature with respect to its developmental capacity could provide new opportunities for embryo selection. Since the gene expression outline of an embryo is considered as an aggregate of its intrinsic characteristics and culture conditions, we have compared transcriptome profiles of in vivo and in vitro derived blastocysts in relation to pregnancy outcome to unravel the discrete effects of developmental competence and environmental conditions on bovine embryo gene expression outlines. To understand whether the gene expression patterns could be associated with blastocyst developmental competency, the global transcriptome profile of in vivo (CVO) and in vitro (CVT) derived competent blastocysts that resulted in pregnancy was investigated relative to that of in vivo (NVO) and in vitro (NVT) derived blastocysts which did not establish initial pregnancy, respectively while to unravel the effects of culture condition on the transcriptome profile of embryos, the transcriptional activity of the CVO group was compared to the CVT group and the NVO group was compared to the NVT ones. Results A total of 700 differentially expressed genes (DEGs) were identified between CVO and NVO blastocysts. These gene transcripts represent constitutive regions, indel variants, 3′-UTR sequence variants and novel transcript regions. The majority (82%) of these DEGs, including gene clusters like ATP synthases, eukaryotic translation initiation factors , ribosomal proteins, mitochondrial ribosomal proteins, NADH dehydrogenase and cytochrome c oxidase subunits were enriched in the CVO group. These DEGs were involved in pathways associated with glycolysis/glycogenesis, citrate acid cycle, pyruvate metabolism and oxidative phosphorylation. Similarly, a total of 218 genes were differentially expressed between CVT and NVT groups. Of these, 89%, including TPT1 , PDIA6 , HSP90AA1 and CALM, were downregulated in the CVT group and those DEGs were overrepresented in pathways related to protein processing, endoplasmic reticulum, spliceasome, ubiquitone mediated proteolysis and steroid biosynthesis. On the other hand, although both the CVT and CVO blastocyst groups resulted in pregnancy, a total of 937 genes were differential expressed between the two groups. Compared to CVO embryos, the CVT ones exhibited downregulation of gene clusters including ribosomal proteins, mitochondrial ribosomal protein, eukaryotic translation initiation factors, ATP synthases, NADH dehydrogenase and cytochrome c oxidases. Nonetheless, downregulation of these genes could be associated with pre and postnatal abnormalities observed after transfer of in vitro embryos. Conclusion The present study provides a detailed inventory of differentially expressed gene signatures and pathways specifically reflective of the developmental environment and future developmental capacities of bovine embryos suggesting that transcriptome activity observed in blastocysts could be indicative of further pregnancy success but also adaptation to culture environment.

Pig production depends on a health and performance balance. An approach to improve intestinal health is the oregano essential oil (OEO) supplementation within a conventional diet. Intestinal integrity regulating effects, for example gene expression, of some feed ingredients are important key factors for that balance. We hypothesized that OEO affects the expression of genes associated with pigs' intestinal integrity. In four trials, a total of 86 pigs have been used. From weaning, the 'treated' group (n = 42) was additionally fed an oregano flavour additive [1500 mg/kg (7.5% pure OEO)] within the basal diet. The 'control' group (n = 44) was kept under identical environmental conditions, except the OEO. At age of 6 months, pigs were slaughtered with an average weight of 111.1 ± 10.9 kg. In addition to automatically generated 'Fat-o-Meter' (AutoFOM) data, carcass quality factors have been measured manually. Valuable cuts of meat, such as ham and belly, were significantly reduced in the OEO group. Effects of OEO on pigs' haematologic parameters were very limited. For transcriptome analysis, the most interesting microarray expression results have been listed in a table (topTable). Selected genes were technically validated by qPCR. As a result, few significant differences in animal development and meat quality have been found between the OEO treated and the control group. Depending on OEO supplementation, we found 93 differently regulated genes in the jejunal tissue (70 up, 23 down) and 60 in the ileal tissue (48 up, 12 down). Just three genes (GRIN3B [glutamate ionotropic receptor NMDA type subunit 3B], TJP1/ZO-1 [tight junction protein ZO-1] and one uncharacterized gene) were affected by OEO both in jejunum and ileum. qPCR validation revealed AKT serine/threonine kinase 3 (AKT3), Interferon (IFN) -ε, -ω, tight junction protein (TJP1)/ZO-1 (ZO-1) to be upregulated in the jejunum and C-C motif chemokine ligand 21 (CCL21) was upregulated in the ileum of pigs that were supplemented with OEO. OEO supplementation had limited effects on pigs' performance traits. However, we were able to demonstrate that OEO alters the expression of genes associated with adaptive immune response in pigs' small intestine. These findings help to explain OEOs' beneficial impact on pigs' intestinal integrity.