Current perspectives on ruminant sperm freezability: Harnessing molecular changes related to semen quality through omics technologies

The recent advances in sperm cryopreservation transcend cryobanking and other assisted reproductive technologies. Since its discovery, cryopreservation has contributed positive impacts on animal breeding as well as in genetic exchange, improvement, and conservation efforts. However, cryoinjury and variabilities in cryopreservation outcomes remain as key challenges to sperm cryobiology. The present work explored the molecular bases for such freezability differences and freezing-thawing injuries in the ruminant sperm. Relevant biomarkers identified in the seminal plasma and the spermatozoa were highlighted, including lipids, proteins, metabolites, transcripts, and genes. Specific molecular mechanisms concerning sperm structures and functions were also examined relative to their association to cryotolerance, and spermiogram or seminogram modifications following cryopreservation procedures. Current conflicts and gaps in the knowledge base on ruminant spermatozoa were also emphasized. Further investigation of these areas using the available breakthrough molecular tools such as omics technologies is therefore proposed to improve, optimize, or even predict the overall quality of frozen-thawed ruminant semen towards reproductive efficiency

Mammalian sperm capacitation: In vivo and in vitro juxtaposition

The development of assisted reproductive technologies (ART) in mammalian species such as in vitro embryo production (IVEP) has the potential to provide great benefits for significant population increase, improve genetic performance and advancement, and reduce transmission of venereal diseases. Correspondingly, in vitro capacitation of sperm is also paramount, related to the ability of sperm to fertilize oocytes, and was created to imitate in vivo conditions in the female reproductive tract. Amid in vitro capacitation developments, studies on how far in vitro capacitation has progressed in mimicking in vivo scenes have not been thoroughly reviewed as a comparative form. Therefore, the present study outlined the series of alterations in mammalian sperm capacitation during their journey in the female reproductive tract by exploring and juxtaposing processes under in vivo and in vitro conditions. Several essential aspects that become gaps between in vivo and in vitro were also identified and elaborated comprehensively in this systematic literature review. We noted that although in vitro capacitation procedures in certain mammalian species have made promising progress and improvements, it is still poorly successful in other species like horses. Our findings further postulated that the occurrence of cryocapacitation, the high ratio of capacitated sperm/oocyte required for successful fertilization, and the incidence of polyspermy cause capacitation under in vitro settings is less efficient and not yet fully comparable to in vivo. This work is therefore proposed several aspects that need to be bettered from in vitro milieu to make it analogous to in vivo environments in modulating sperm capacitation

Effect of leukemia inhibitory factor polymorphism on litter size traits in Thai commercial pig breeds

Leukemia inhibitory factor (LIF) is a crucial candidate gene that impacts on implantation process. In this study, the effects of the porcine LIF polymorphism on litter size traits were elucidated in Thai commercial pig populations. Genotyping of three single nucleotide polymorphisms (SNPs) of the porcine LIF gene was detected in coding and 3ˊ-untranslated regions. The porcine LIF c.*24C>T was segregating in Large White, Landrace, and Large White × Landrace (LW × LR) crossbred sows. No polymorphisms at two non-synonymous SNPs loci (LIF c.28C>A and LIF c.161A>G) were found in this study. The porcine LIF c.*24C>T was significantly associated with the total number born (TNB), the number born alive (NBA), and the number of piglets weaned alive (NWA) traits in Large White and Landrace sows. Moreover, the porcine LIF c.*24C>T was associated with the NBA and NWA traits in LW × LR crossbred sows. The favorable LIF c.*24C allele was positively correlated with the litter size traits. These findings indicated that the polymorphism of the porcine LIF gene was associated with litter size traits and confirms the significance of porcine LIF as a candidate gene for litter size traits in pig breeding. Thus, the porcine LIF gene could be used for improving prolific traits in these Thai commercial pig populations

Efficiency of Fresh and Fermented Banana Stem in Low Protein Diet on Nutrient Digestibility, Productive Performance and Intestinal Morphology of Crossbred Pig ((Thai native x Meishan) x Duroc)

Banana stem is a common feed component for raising pigs in mountainous Southeast Asia. However, its nutritive value and digestibility are low. This study was carried out to investigate the effects of unfermented and fermented banana stems on crossbred pigs concerning nutrient digestibility, productive performance, and intestinal morphology. Initially, an in vitro ileal digestibility test was performed for the following feedstuffs: fresh banana stem (BS), fermented banana stem (FBS), concentrate (C), fresh banana stem + concentrate (BSC), and fermented banana stem + concentrate (FBSC). For the 120-day experiment, 16 crossbred pigs were divided into two groups and fed with BSC and FBSC. They were placed in individual cages and subsequently moved to metabolic cages for seven days to determine apparent total tract digestibility (ATTD). Finally, all pigs were slaughtered and their small intestines were analyzed for intestinal morphology. The results show that pure fresh and fermented banana stems had low digestibility. However, their digestibility increased by 50% when mixed with concentrate. Crossbred pigs fed BSC and FBSC did not exhibit significant differences in their performance, but the intestinal morphology of the FBSC group had improved intestinal morphology, especially the villi height. In conclusion, both fresh and fermented banana stems can be recommended in a low protein diet as feed for crossbred pigs in an improved production system. This is relevant for raising pigs in mountainous areas, as it has the potential to reduce feed cost and maintain production performance

Association of a non-synonymous SNP of IL17RA gene with litter size traits in Large White and Landrace pigs

Interleukin-17 receptor A (IL17RA) is one of the cytokine receptors of the pro-inflammatory interleukin-17 (IL17) cytokine family. The IL17 and IL17RA genes are involved in inflammatory and immune responses as well as reproductive process of mammals. The purposes of this study were to examine polymorphisms in the porcine IL17RA gene and to assess its effects on litter size traits in Large White and Landrace pigs. Three non-synonymous single nucleotide polymorphisms (SNPs) in the porcine IL17RA gene were verified. The porcine IL17RA c.785C>T (p.Ala262Val) was found to be segregating in the Large White and Landrace pigs. No polymorphisms in the coding region of the porcine IL17RA gene at the two non-synonymous SNPs loci of c.997G>A (p.Val333Ile) and c.1962T>G (p.Asp654Glu) were found. The porcine IL17RA c.785C>T polymorphism was significantly associated with the total number born (TNB) and the number born alive (NBA) in Large White pigs (PT was significantly associated with the TNB, NBA, total birth weight (TBW), and total weaning weight of piglets at 21 days (TWW) in Landrace pigs (P<0.05). These results supported the importance of the porcine IL17RA gene in the litter size traits of pigs. Thus, the porcine IL17RA could be used as a potential candidate gene for improving litter size traits in pig breeding

Wound healing effect of supercritical carbon dioxide Datura metel l. leaves extracts : an in vitro study of anti-inflammation, cell migration, MMP-2 inhibition, and the modulation of the sonic hedgehog pathway in human fibroblasts

Datura metel L. (thorn apple) has been used in Thai folk wisdom for wound care. In this study, we chose supercritical carbon dioxide extraction (scCO2) to develop crude extraction from the leaves of the thorn apple. The phytochemical profiles were observed using liquid chromatography-quadrupole time-of-flight mass spectrometry (LC-QTOF-MS). The biological activities of D. metel were performed through antioxidant assays, anti-inflammation based on the Griess reaction, the migration assay, the expression of matrix metalloproteinase-2 (MMP-2), and regulatory genes in fibroblasts. Dm1 and Dm2 extracts were obtained from scCO2 procedures at different pressures of 300 and 500 bar, respectively. Bioactive compounds, including farnesyl acetone, schisanhenol B, and loliolide, were identified in both extracts. The antioxidant properties of both D. metel extracts were comparable to those of l-ascorbic acid in hydrogen peroxide-induced fibroblasts with no significant difference. Additionally, Dm1 and Dm2 significantly inhibited the nitrite production levels of 1.23 ± 0.19 and 1.52 ± 0.05 μM, respectively, against the lipopolysaccharide-treated group (3.82 ± 0.39 μM). Interestingly, Dm1 obviously demonstrated the percentage of wound closure with 58.46 ± 7.61 and 82.62 ± 6.66% after 36 and 48 h of treatment, which were comparable to the commercial deproteinized dialysate from the calf blood extract. Moreover, both extracts were comparable to l-ascorbic acid treatment in their ability to suppress the expression of MMP-2: an enzyme that breaks down collagen. The gene expressions of SHH, SMO, and GLI1 that control the sonic hedgehog pathway were also clearly upregulated by Dm1. Consequently, the scCO2 technique could be applied in D. metel extraction and contribute to potentially effective wound closure

Antioxidant activities of plant extracts and essential oil-cyclodextrin complexes and their effect on lipid accumulation in porcine adipocytes in vitro

Fat deposition is important for pig production because excessive fat decreases the economic value of meat production. Plant extracts have been used as feed additive in pig diets to improve oxidative stability of meat. However, only a few studies had investigated the effect of plant extracts on lipid accumulation of porcine adipocytes. Therefore, this study examined antioxidative potential of plant extracts and essential oil-cyclodextrin complexes and their effect on lipid accumulation in porcine adipocytes. Porcine preadipocytes were isolated from dorsal subcutaneous adipose tissue of a 5-month-old female pig. Preadipocytes were differentiated in the presence of extracts, then lipid accumulation was determined using oil red O staining. Results showed that green tea and grape seed extracts had the highest total phenolic and total flavonoid contents and also possessed the highest antioxidant activities. Among essential oil-cyclodextrin complexes, thyme oil had the highest antioxidant activities by all assays. With a regard to the lipid accumulation, all of plant extracts and essential oil-cyclodextrin complexes decreased lipid accumulation in adipocytes from 32.61 to 91.71% compared with the control. Green tea extract, thyme and galangal oil products decreased lipid accumulation about 60% lower than that of the control. In conclusion, green tea and grape seed extracts, and thyme oil powder illustrated high antioxidant potentials, while lipid accumulation in porcine adipocytes was greatly reduced by green tea extract, thyme and galangal oil products. The research finding is beneficial in the development of natural feed additives to improve oxidative stability and reduce excessive subcutaneous fat in pig production

Nutritional values and nutrient digestibility of ground perilla cake (Perilla frutescens) in growing pig diets

This study determined the nutritional values of ground perilla cake (GPC) and the potential for dietary supplementation in growing pigs based on in vitro ileal digestibility (IVID) and apparent total tract digestibility (ATTD). The IVID evaluated at four dietary supplementation levels: 0, 5, 10, and 20%. The ATTD was measured by using twenty-four grower crossbred pigs. Pigs were randomly assigned to three dietary supplementation levels (0, 5, and 10%). From these analytical results, GPC raw material contained crude protein (CP) content (31.54%). That total essential amino acid (EAA) was 138.34 mg/g, mainly leucine (28.87 mg/g), and contained notably limiting amino acids for pigs, such as lysine (19.52 mg/g) and methionine (10.94 mg/g). The ether extract content (EE) was 10.52%, and the major free fatty acid (FFA) was linolenic acid (C18:3n3; 55.97%) and the fat-soluble vitamins included γ-tocopherol (367.25 μg/100g). In addition, GPC contained minerals such as phosphorus (1.02%), potassium (0.83%), calcium (0.46%), and iron. However, crude fiber (CF) had a notably high content (24.43%). Increasing GPC levels in pig diets reduced the IVID of dry matter (DM), EE, and CF (P<0.05), especially the 20% GPC supplement. The IVID of the CP did not differ among the groups. Furthermore, the results for the ATTD of the CP and EE in the 5% GPC supplement group were significantly better than that of the other groups (P˂0.05). We found the potential of GPC as an alternative protein source. Moreover, it contained high energy and polyunsaturated fatty acid

Effect of porcine IL-6 polymorphism on litter size traits in commercial pig breeds

This study aimed to verify the polymorphisms in the porcine IL-6 gene and to elucidate its effects on litter size traits in Large White and Landrace sows. Four single nucleotide polymorphisms (SNPs) of the porcine IL-6 gene (g.91506415A>G, g.91507983A>G, g.91508173C>T, and g.91508716C>T) were genotyped using the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. There was no polymorphism observed on the three SNPs (g.91506415A>G, g.91507983A>G, and g.91508716C>T) of the porcine IL-6 gene. The porcine IL-6 g.91508173C>T polymorphism was found to be segregating in Large White and Landrace sows. The porcine IL-6 g.91508173C>T polymorphism was significantly associated with the total number born (TNB) and the number of piglets weaned alive (NWA) traits in Large White sows (PT polymorphism was significantly associated with the TNB, number born alive (NBA), and NWA traits in Landrace sows (PT polymorphism was associated with litter size traits. These findings confirmed the importance of the IL-6 gene as a candidate gene for litter size traits in pigs

Association of genetic markers with sex determination in Thai red tilapia

The objectives of this study were to verify the polymorphism on sex-linked marker loci and to assess their associations with phenotypic sex characteristics in red tilapia. Four sex-linked genetic markers of Amh, SCAR4, SCAR5, and Oni3161 were genotyped in the Thai red tilapia population. The Amh marker was significantly associated with the phenotypic sex of red tilapia with an accuracy of 46.2%. No significant association of SCAR4, SCAR5, and Oni3161 marker polymorphisms with phenotypic sex characteristics was observed in this study. However, the combinations of these two, three, and four markers were increasingly associated with phenotypic sex characteristics for red tilapia with an accuracy of 62.8, 68.4, and 72.4%, respectively. These results indicate that these combined genetic markers were associated with the phenotypic sex of red tilapias. These findings confirmed the importance of these genetic markers as candidate markers for sex determination in the Thai red tilapia population