Proteomic and antimicrobial peptide analyses of Buffalo colostrum and mature Milk whey: A comparative study.

Buffalo colostrum is the initial mammary secretion after parturition, consisting of nutritional and bioactive components. In this study, we conducted a proteomic analysis of buffalo colostrum whey to identify bioactive proteins and peptides. A total of 107 differentially expressed proteins (DEPs) were identified in buffalo colostrum whey compared to those in mature milk. Gene Ontology analysis revealed that DEPs were primarily associated with immune response and tissue development. KEGG pathway enrichment suggested that colostrum actively enhances nascent immunity involved in interleukin and interferon signaling pathways. Furthermore, candidate antimicrobial peptides (AMPs) of whey protein hydrolysates from buffalo colostrum were characterized, which exhibits broad-spectrum activity against gram-positive and gram-negative pathogens. Overall, this study improves our understanding of protein variations in buffalo lactation, and contributes to the development of AMPs from buffalo colostrum.

Dietary naringin supplementation on hepatic yolk precursors formation and antioxidant capacity of Three-Yellow breeder hens during the late laying period.

In this study, the effects of naringin on hepatic yolk precursors formation and antioxidant capacity of Three-Yellow breeder hens during late laying period were evaluated. A total of 480 (54-wk-old) Three-Yellow breeder hens were randomly assigned to 4 groups (6 replicates of 20 hens): nonsupplemented control diet (C), and control diet supplemented with 0.1%, 0.2%, and 0.4% of naringin (N1, N2, and N3), respectively. Results showed that dietary supplemented with 0.1%, 0.2%, and 0.4% of naringin for 8 wk promoted the cell proliferation and attenuated the excessive fat accumulation in the liver. Compared with C group, increased concentrations of triglyceride (TG), total cholesterol (T-CHO), high-density lipoprotein cholesterol (HDL-C), and very low-density lipoprotein (VLDL), and decreased contents of low-density lipoprotein cholesterol (LDL-C) were detected in liver, serum and ovarian tissues (P < 0.05). After 8 wk of feeding with naringin (0.1%, 0.2%, and 0.4%), serum estrogen (E2) level, expression levels of proteins and genes of estrogen receptors (ERs) increased significantly (P < 0.05). Meanwhile, naringin treatment regulated expression of genes related to yolk precursors formation (P < 0.05). Furthermore, dietary naringin addition increased the antioxidants, decreased the oxidation products, and up-regulated transcription levels of antioxidant genes in liver tissues (P < 0.05). These results indicated that dietary supplemented with naringin could improve hepatic yolk precursors formation and hepatic antioxidant capacity of Three-Yellow breeder hens during the late laying period. Doses of 0.2% and 0.4% are more effective than dose of 0.1%.

Dietary naringin supplementation on laying performance and antioxidant capacity of Three-Yellow breeder hens during the late laying period.

In this study, the effects of 3 graded dietary levels (0.1%, 0.2%, and 0.4%) of naringin were studied in Three-Yellow breeder hens during the late laying period (55-62 wk). A total of 480 Three-Yellow breeder hens (54-wk-old) were randomly divided into 4 groups (6 replicates of 20 hens): basal diet group (C), and basal diets supplemented with 0.1%, 0.2%, and 0.4% of naringin (N1, N2, and N3), respectively. Results showed that dietary supplementation with 0.1%, 0.2%, and 0.4% of naringin for 8 wk increased the laying rate and egg mass, enhanced egg yolk color, and decreased the feed egg ratio (P < 0.05). Meanwhile, compared with hens in C group, there were more preovulatory follicles and higher ovarian index as well as an enhanced ovarian somatic cell proliferation in hens of N2 and N3 groups (P < 0.05). With 0.2% and 0.4% naringin, glutathione concentration, the activity of catalase and total superoxide dismutase, and the total antioxidant capacity of ovarian tissues and serum increased (P < 0.05), while the contents of malondialdehyde and hydrogen peroxide decreased (P < 0.05). Moreover, compared to C group, the transcription levels of antioxidant genes in ovarian tissues increased in hens from N2 and N3 groups (P < 0.05). In conclusion, supplementation with 0.2% and 0.4% naringin both could improve the laying rate, ovarian and serum antioxidant capacity of Three-Yellow breeder hens during the late laying period.

Effect of addition of l-carnitine to cryopreservation extender on rabbit post-thaw semen parameters, antioxidant capacity, mitochondrial function, apoptosis and ultrastructure changes.

l-Carnitine (LC) is considered to be a natural antioxidant agent that could be used to improve the efficiency of reproduction. However, the precise machinery of the effect of LC supplementation on frozen-thawed rabbit sperm has not been evaluated. Thus, the aim of this study was to evaluate the effect of the addition of LC to a freezing medium on parameters and ultrastructure changes in frozen-thawed rabbit sperm. Rabbit bucks (7 months of age) were involved, and semen was collected using the artificial vagina method. Pooled rabbit semen was cryopreserved in a tris yolk fructose extender without any supplement (LC0, control group) or with LC at levels of 1, 2 or 4 mM (LC1, LC2 and LC4, respectively). The samples were then loaded into 0.25-ml straws and frozen over liquid nitrogen vapours before being plunged into the liquid nitrogen and stored at -196°C until evaluation. Data showed that the addition of LC significantly increased sperm motility, viability and membrane function, while sperm abnormalities decreased (p < .001). Sperm-like apoptosis (early, late and necrosis spermatozoa) was lower in the LC4 group compared with the other groups. l-Carnitine addition significantly enhanced the total antioxidant capacity and superoxide dismutase and glutathione peroxide activities and significantly reduced the protein carbonyl and malondialdehyde levels compared with the control group. Moreover, electron microscopy images demonstrated that LC addition (2 or 4 mM) preserved the acrosome and plasma membrane and protected the ultrastructure integrity of the cryopreserved spermatozoa in relation to the control group. Spermatozoa treated with LC exhibited higher mitochondria membrane potential (MMP) values compared with the control group. We conclude that the addition of LC (4 mM) to the freezing extender enhanced the quality, increased the antioxidant capabilities, preserved the ultrastructure integrity and reduced lipid and protein peroxidation as well as increased MMP activity of frozen-thawed rabbit sperm.

Differences in egg yolk precursor formation of Guangxi Ma chickens with dissimilar laying rate at the same or various ages.

Egg yolk formation and deposition are quite vital for oocyte maturation and egg production in hens. There are great differences in egg production and number of preovulatory follicles among individuals in Guangxi Ma chickens at the same or various ages, however, the causes of this phenomenon remain unclear. This study aimed to elucidate the differences in yolk precursors formation among Guangxi Ma hens at the same or various ages with dissimilar egg laying rate via comparing the synthesis and transportation of yolk precursors, serum estradiol (E2) levels, as well as the antioxidant capacity of the liver and serum from hens of 32-week-old (32W), 50-week-old with higher laying rate (50WH), 50-week-old with lower laying rate (50WL), 72-week-old with higher laying rate (72WH), and 72-week-old with lower laying rate (72WL). The results showed that the contents of triglyceride (TG), total cholesterol (T-CHO), high density lipoprotein cholesterol (HDL-C) and very low density lipoprotein cholesterol (VLDL-C) in the liver, serum and ovarian stroma, serum E2 levels, expression levels of proteins and genes related to yolk precursors formation, and the antioxidant capacity of liver and serum decreased significantly during aging process. TG, HDL-C and VLDL-C contents in serum and ovarian stroma, as well as HDL-C and VLDL-C levels in liver tissues of hens in 50WL and 72WL were significantly lower compared to hens in 50WH and 72WH, respectively. Meanwhile, expression levels of estrogen receptor ɑ and transcription levels of genes related to lipid transportation in liver tissues, and the antioxidant capacity of livers were remarkably lower in hens in 50WL and 72WL than those in 50WH and 72WH, respectively. However, no significant difference was detected in liver TG levels, serum E2 levels, genes related to fatty acids synthesis and serum antioxidant capacity between hens in 50WL and 50WH. In conclusion, the age-related decline of egg production in Guangxi Ma chicken maybe related to the age-related decreases in the synthesis and transportation of liver yolk precursor as well as the antioxidant capacity of liver and hens. And the non-age related decline of egg production may mainly attribute to the decreases in yolk precursors transportation ability and hepatic antioxidant capacity.

Association between variants in vitamin D-binding protein gene and vitamin D deficiency among pregnant women in china.

BACKGROUND: The prevalence of vitamin D deficiency and insufficiency is extremely high in pregnant women worldwide. However, the association between single nucleotide polymorphisms (SNPs) in vitamin D metabolic pathway genes and 25-hydroxyvitamin D (25(OH)D) concentration among Chinese pregnant women is seldom reported. The risk of adverse neonatal outcomes due to maternal vitamin D deficiency has not been well investigated. METHODS: A total of 815 pregnant women and 407 infants were enrolled in this study. Serum 25(OH)D concentration was detected. DNA was extracted from the maternal blood for genotyping genetic SNPs in vitamin D pathway. An XGBoost model was established based on SNPs combined with external variables. RESULTS: Mean serum 25(OH)D level was 15.67 ± 7.98 ng/mL among the pregnant women. Seventy-five percent of pregnant women had 25(OH)D deficiency in China. SNPs of GC (rs17467825, rs4588, rs2282679, rs2298850, and rs1155563) were significantly associated with maternal 25(OH)D concentration. The influence of variants of rs17467825, rs4588, rs2282679, and rs2298850 on maternal 25(OH)D might be modified by vitamin D supplementation and sunshine exposure. An XGBoost model was established for monitoring 25(OH)D status in pregnant women and provided clinical advice to reduce the risk of 25(OH)D deficiency. Mothers with 25(OH)D deficiency hinted a risk for macrosomia. CONCLUSION: A high prevalence of vitamin D deficiency in China has been confirmed. A clinical model was established to guide pregnant women to supplement vitamin D according to genotype. Furthermore, we suggest the effect of maternal vitamin D status on the risk of macrosomia.

Maturation of buffalo oocytes in vitro with acetyl-L-carnitine improves cryotolerance due to changes in mitochondrial function and the membrane lipid profile.

The effects of acetyl-l-carnitine (ALC) supplementation during IVM on subsequently vitrified buffalo oocytes were evaluated, followed by determination of the mitochondrial DNA copy number, measurement of mitochondrial membrane potential (MMP) and identification of the lipid profile of oocyte membranes as markers of oocyte quality after vitrification. Supplementation with ALC during IVM significantly improved the rates of oocyte cleavage and morula and blastocyst formation, and increased MMP after vitrification compared with unsupplemented vitrified oocytes (P<0.05). Using a bidirectional orthogonal projection to latent structures discriminant analysis based on positive ion matrix-assisted laser desorption ionisation time-of-flight mass spectrometry data, five phospholipid ions (m/z 728.7 (phosphatidylcholine (PC) 32:3), 746.9 (PC 32:5), 760.6 (PC 34:1), 768.8 (PC P-36:3) and 782.6 (PC 36:4); P<0.05) were identified as significantly more abundant in fresh oocytes than in unsupplemented vitrified oocytes. Meanwhile, three phospholipid ions (m/z 734.6 (PC 32:0), 760.6 (PC 34:1), and 782.6 (PC 36:4); P<0.05) were more abundant in ALC-supplemented vitrified oocytes than in unsupplemented vitrified oocytes. Therefore, supplementation with ALC during IVM may improve buffalo oocyte quality after vitrification by enhancing mitochondrial function and altering the phospholipid composition of vitrified oocyte membranes.

In vitro differentiation of spermatogonial stem cells using testicular cells from Guangxi Bama mini-pig.

In this study, we attempted to establish a culture system for in vitro spermatogenesis from spermatogonial stem cells (SSCs) of Bama mini-pig. Dissociated testicular cells from 1-month-old pigs were co-cultured to mimic in vivo spermatogenesis. The testicular cells were seeded in minimum essential medium alpha (α-MEM) supplemented with Knockout serum replacement (KSR). Three-dimensional colonies formed after 10 days of culture. The colonies showed positive staining for SSC-associated markers such as UCHL1, PLZF, THY1, OCT4, Dolichos biflorus agglutinin, and alkaline phosphatase. Induction of SSCs was performed in α-MEM + KSR supplemented with retinoic acid, bone morphogenetic protein 4, activin A, follicle-stimulating hormone, or testosterone. The results showed that STRA8, DMC1, PRM1, and TNP1 were upregulated significantly in the colonies after induction compared to that in testis from 1-month-old pigs, while expression levels of those genes were significantly low compared to those in 2-month-old testis. However, upregulation of ACROSIN was not significant. Replacement of α-MEM and KSR with Iscove's modified Dulbecco's medium and fetal bovine serum did not upregulate expression of these genes significantly. These results indicate that SSCs of Bama mini-pig could undergo differentiation and develop to a post-meiotic stage in α-MEM supplemented with KSR and induction factors.

Treatment with acetyl-l-carnitine during in vitro maturation of buffalo oocytes improves oocyte quality and subsequent embryonic development.

Oocyte quality is one of the important factors in female fertility, in vitro maturation (IVM), and subsequent embryonic development. In the present study, we assessed whether acetyl-l-carnitine (ALC) supplementation during in vitro maturation of buffalo oocytes could improve oocyte quality and subsequent embryonic development. To determine the optimal level of ALC supplementation, we matured cumulus-oocyte complexes in maturation medium supplemented with 0, 2.5, and 5 mM ALC. The oocytes with a polar body were selected for parthenogenetic activation (PA) and in vitro fertilization (IVF). We found that oocytes matured in 2.5 mM ALC had significantly higher PA blastocyst rate (P < 0.05) and blastocyst cell number than those of unsupplemented oocytes (P < 0.05) and a significantly higher IVF blastocyst rate than that of oocytes matured in 5 mM ALC (P < 0.05). In all further experiments, we supplemented the maturation medium with 2.5 mM ALC. We then tested whether ALC supplementation could improve various markers of oocytes and cumulus cells. We compared cell proliferation; concentrations of reactive oxygen species (ROS), intracellular ATP, estradiol, and progesterone; mitochondrial distribution; mitochondrial DNA copy number (mtDNA); and expression levels of four genes encoding oocyte-derived factors (GDF9, BMP15) and steroid hormones (StAR, P450scc) between the supplemented and unsupplemented oocytes and cumulus cells. Cumulus cells matured with ALC supplementation were more prolific than those matured without ALC supplementation (P < 0.05). Oocytes treated with ALC had lower concentrations of intracellular ROS (P < 0.05) and a higher rate of diffuse mitochondrial distributions (P < 0.05) than those of untreated oocytes. Additionally, the mtDNA was higher in the ALC-treated oocytes (P < 0.05) and cumulus cells (P < 0.05) than that in the untreated cells. The ALC-treated maturation medium had a higher postmaturation concentration of estradiol than that of the untreated medium (P < 0.05). Finally, the gene expression levels of P450scc and GDF9 were greater in ALC-treated oocytes and cumulus cells than those in untreated cells (P < 0.05). Therefore, in buffalo, our results suggest that ALC affects mitochondrial function, regulates oocyte-derived paracrine factors, and increases the production of steroid hormones, leading to increased quality of matured oocytes and improved embryonic development in vitro.