Modulatory effects of R10 fraction of garlic (Allium sativum L.) on hormonal levels, T cell polarization, and fertility-related genes in mice model of polycystic ovarian syndrome.

Polycystic ovary syndrome (PCOS) is an inflammatory endocrine-metabolic disorder related to reproductive system characterized by polycystic ovarian morphology, androgen excess, and chronic anovulation. Current treatments haven't been very successful in PCOS treatment and the problem still remains as a challenge. Therefore, new approaches should be applied to overcome the disease. Previous studies demonstrated immunomodulatory effects of R10 fraction of garlic in the treatment of inflammatory conditions such as cancer. Considering previous studies suggesting immunomodulatory therapy for PCOS, therapeutic effects of R10 fraction was evaluated in a mouse model of PCOS. To do so, PCOS was developed by intramuscular injection of estradiol valerate. Treatment with R10 fraction, isolated from garlic, was performed and the alterations in hormonal levels (estradiol, progesterone, and testosterone), T cell polarization markers (IFN-γ, IL-4, and IL-17), and expression of fertility-related genes (Gpx3 and Ptx3) were evaluated. The results showed that hormonal levels were elevated in PCOS model comparing to normal animals but were markedly modulated after treatment with R10 fraction. Moreover, a severe disturbance in T cell polarization with a significant reduction of fertility-related genes expression were detected in PCOS-induced ovaries. Treatment with R10 fraction also represented modulatory effects on T cell polarization by increasing IL-4 and decreasing IL-17 and IFN-γ levels. Accordingly, fertility-related genes were also modulated following treatment with R10 fraction in PCOS. Our study elucidated that R10 fraction of garlic possess immunomodulatory effects alleviating PCOS symptoms. This approach could be adjusted to give rise the optimum therapeutic results and considered as a candidate therapeutic approach for PCOS.

Nutrient use efficiency (NUE) of wheat (Triticum aestivum L.) as affected by NPK fertilization.

Nutrient use efficiency is crucial for increasing crop yield and quality while reducing fertilizer inputs and minimizing environmental damage. The experiments were carried out in silty clay loam soil of Lalitpur, Nepal, to examine how different amounts of nitrogen (N), phosphorus (P), and potassium (K) influenced crop performance and nutrient efficiency indices in wheat during 2019/20 and 2020/21. The field experiment comprised three factorial randomized complete block designs that were replicated three times. N levels (100, 125, 150 N kg ha-1), P levels (25, 50, 75 P2O5 kg ha-1), and K levels (25, 50, 75 K2O kg ha-1) were three factors evaluated, with a total of 27 treatment combinations. Grain yields were significantly increased by N and K levels and were optimum @ 125 kg N ha-1 and @ 50 kg K2O ha-1 with grain yields of 6.33 t ha-1 and 6.30 t ha-1, respectively. Nutrient levels influenced statistically partial factor productivity, internal efficiency, partial nutrient budget, recovery efficiency, agronomic efficiency, and physiological efficiency of NPK for wheat. Nutrient efficiency was found to be higher at lower doses of their respective nutrients. Higher P and K fertilizer rates enhanced wheat N efficiencies, and the case was relevant for P and K efficiencies as well. Wheat was more responsive to N and K fertilizer, and a lower rate of P application reduced N and K fertilizer efficiency. This study recommends to use N @ 125 kg ha-1, P2O5 @ 25 kg ha-1 and K2O @ 50 kg ha-1 as an optimum rate for efficient nutrient management in wheat in mid-hills of Nepal.

Melatonin and Myo-Inositol: Supporting Reproduction from the Oocyte to Birth.

Human pregnancy is a sequence of events finely tuned by several molecular interactions that come with a new birth. The precise interlocking of these events affecting the reproductive system guarantees safe embryo formation and fetal development. In this scenario, melatonin and myo-inositol seem to be pivotal not only in the physiology of the reproduction process, but also in the promotion of positive gestational outcomes. Evidence demonstrates that melatonin, beyond the role of circadian rhythm management, is a key controller of human reproductive functions. Similarly, as the most representative member of the inositol's family, myo-inositol is essential in ensuring correct advancing of reproductive cellular events. The molecular crosstalk mediated by these two species is directly regulated by their availability in the human body. To date, biological implications of unbalanced amounts of melatonin and myo-inositol in each pregnancy step are growing the idea that these molecules actively contribute to reduce negative outcomes and improve the fertilization rate. Clinical data suggest that melatonin and myo-inositol may constitute an optimal dietary supplementation to sustain safe human gestation and a new potential way to prevent pregnancy-associated pathologies.

Amelioration of sperm fertilizability, thyroid activity, oxidative stress, and inflammatory cytokines in rabbit bucks treated with phytogenic extracts.

This study investigated the beneficial effect of phytogenic extracts on semen quality, reproductive hormones, thyroid activity, immunity, hepatic antioxidant activity, and fertility in rabbit bucks. We divided 70 bucks into seven groups (10 in each). Group 1 was fed a basal diet (control); groups 2, 3, and 4 were fed the control diet with 30, 60, and 90 mg/kg of turmeric, respectively; and groups 5, 6, and 7 were fed the control diet with 50, 75, and 100 mg/kg of garlic extract, respectively, for 8 weeks. Rectal and skin temperatures decreased, while follicle-stimulating hormone, luteinizing hormone, triiodothyronine, thyroxine, testosterone, immunoglobulin M, tumor necrosis factor-alpha, and interleukin-6 in blood serum and glutathione peroxidase in the liver increased in all groups (p < .05). Garlic extract (100 mg/kg diet) increased adenosine triphosphate and glutathione in the liver tissues. All treatments significantly increased net semen volume, percentages of progressive motility, livability, curled tail, and intact acrosomes of spermatozoa, sperm cell concentration, and outputs of total and motile spermatozoa, while significantly decreased percentage of sperm abnormality. In conclusion, dietary supplementation of turmeric or garlic extract can be used as a suitable tool for enhancing the hepatic antioxidant activity, immunity, and semen quality in rabbit bucks.

The effect of Myo-inositol on fertility rates in poor ovarian responder in women undergoing assisted reproductive technique: a randomized clinical trial.

BACKGROUND: Poor ovarian response to gonadotropin is a significant challenge in assisted reproductive technique (ART) and affect 9-24% of ART cycles. This study aimed to evaluate the effect of Myo-inositol on fertility rates in poor ovarian responder women undergoing assisted reproductive technique. METHODS: This study is a double-blinded randomized controlled study that involved 60 poor ovarian responders included in an ICSI program and divided into two groups; intervention group: 30 patients who have been assuming Inofolic (4 g myo-inositol + 400 µg folic acid) for the before the enrollment day; control group: 30 patients assuming folic acid (400 µg) for the same period. Controlled ovarian stimulation was performed in the same manner in the two groups. The main outcomeswere the assessment of oocytes retrievednumber and quality, ovarian sensitivity index,required dose of Gonadotropinsunits × 1000), fertilization rate, biochemical, and clinical pregnancy rate. RESULT: There is no significant difference in clinical characteristics between study groups. The number of oocytes retrieved, number of MII oocytes, number of embryos transferred, chemical, and clinical pregnancy were higher in the intervention group. However, they are not statistically significant in comparison to the control group. The ovarian sensitivity index and fertilization rate were significantly higher in the intervention group than the control group (P > 0.05). The required dose of gonadotropin significantly lower in the intervention group than the control group. CONCLUSION: Our results suggest that the supplementation myo-inositol in poor ovarian responders significantly improved the ART outcomes such as fertilization rate gonadotropin, ovarian sensitivity index (OSI) and significantly reduced the required unities of gonadotropin. Additionally, more extensive randomized controlled studies are needed. TRIAL REGISTRATION: Iranian Registry of Clinical Trials, IRCT20180515039668N1 , retrospectively registered since 2020-03-16.

Impacts of Petroleum Fuels on Fertilization and Development of the Antarctic Sea Urchin Sterechinus neumayeri.

Antarctic marine environments are at risk from petroleum fuel spills as shipping activities in the Southern Ocean increase. Knowledge of the sensitivity of Antarctic species to fuels under environmentally realistic exposure conditions is lacking. We determined the toxicity of 3 fuels, Special Antarctic Blend diesel (SAB), marine gas oil (MGO), and intermediate fuel oil (IFO 180) to a common Antarctic sea urchin, Sterechinus neumayeri. Sensitivity was estimated for early developmental stages from fertilization to the early 4-arm pluteus in toxicity tests of up to 24 d duration. The effects of the water accommodated fractions (WAFs) of fuels were investigated under different exposure scenarios to determine the relative sensitivity of stages and of different exposure regimes. Sensitivity to fuel WAFs increased through development. Both MGO and IFO 180 were more toxic than SAB, with median effect concentration values for the most sensitive pluteus stage of 3.5, 6.5, and 252 µg/L total hydrocarbon content, respectively. Exposure to a single pulse during fertilization and early embryonic development showed toxicity patterns similar to those observed from continuous exposure. The results show that exposure to fuel WAFs during critical early life stages affects the subsequent viability of larvae, with consequent implications for reproductive success. The sensitivity estimates for S. neumayeri that we generated can be utilized in risk assessments for the management of Antarctic marine ecosystems. Environ Toxicol Chem 2020;39:2527-2539. © 2020 SETAC.

Nicotinamide Mononucleotide Supplementation Reverses the Declining Quality of Maternally Aged Oocytes.

Advanced maternal age is highly associated with a decline in oocyte quality, but effective approaches to improve it have still not been fully determined. Here, we report that in vivo supplementation of nicotinamide mononucleotide (NMN) efficaciously improves the quality of oocytes from naturally aged mice by recovering nicotinamide adenine dinucleotide (NAD+) levels. NMN supplementation not only increases ovulation of aged oocytes but also enhances their meiotic competency and fertilization ability by maintaining the normal spindle/chromosome structure and the dynamics of the cortical granule component ovastacin. Moreover, single-cell transcriptome analysis shows that the beneficial effect of NMN on aged oocytes is mediated by restoration of mitochondrial function, eliminating the accumulated ROS to suppress apoptosis. Collectively, our data reveal that NMN supplementation is a feasible approach to protect oocytes from advanced maternal age-related deterioration, contributing to the improvement of reproductive outcome of aged women and assisted reproductive technology.

Maternal conjugated linoleic acid alters hepatic lipid metabolism via the AMPK signaling pathway in chick embryos.

The effects of maternal conjugated linoleic acid (CLA) on embryonic development and hepatic lipid metabolism were investigated in chick embryos. A total of 180 Arbor Acres female broiler breeders (36 wk old) were randomly divided into the following 3 dietary treatment groups: a basic diet (control), a basic diet containing 0.5% CLA (CLA1), and a basic diet containing 1.0% CLA (CLA2). The females were fed for 8 wk, and the eggs from each group were collected and hatched during the last 2 wk. The results showed that the addition of dietary CLA increased the broken egg rate and reduced the fertilization rate and the egg hatchability (P < 0.05). CLA enrichment decreased the polyunsaturated and monounsaturated fatty acids and increased the saturated fatty acids in the yolk sac (P < 0.05). The yolk sac weight, body weight, and body length had a linear decrease with CLA supplementation (P < 0.05). In the developing chick embryo (at E14) and newly hatched chick (D0), the serum triglyceride concentration decreased with maternal CLA supplementation and was accompanied by a reduction in subcutaneous adipose tissue deposition. In addition, maternal CLA supplementation mediated the hepatic lipid metabolism by decreasing the mRNA expression of sterol regulatory element-binding proteins-1c (SREBP-1c), fatty acid synthase and acetyl-CoA carboxylase, and increasing the mRNA expression of adenosine 5'-monophosphate-activated protein kinase α (AMPKα), peroxisome proliferator-activated receptors α (PPARα), liver fatty acid-binding protein, adipose triglyceride lipase and carnitine palmitoyltransferase in embryonic chick livers (P < 0.05). A drop in SREBP-1c protein expression and an increase in the protein expression of p-AMPKα and PPARα were also observed in the liver of chick embryo (P < 0.05). In conclusion, maternal CLA supplementation regulated the fatty acid composition in the yolk sac, and mediated embryonic chick development and hepatic lipometabolism, and these effects may be related to the AMPK pathway. These findings suggest the potential ability of maternal CLA supplementation to reduce fat deposition in chick embryos.

The extracellular calcium-sensing receptor promotes porcine egg activation via calcium/calmodulin-dependent protein kinase II.

Extracellular calcium is required for intracellular Ca2+ oscillations needed for egg activation, but the regulatory mechanism is still poorly understood. The present study was designed to demonstrate the function of calcium-sensing receptor (CASR), which could recognize extracellular calcium as first messenger, during porcine egg activation. CASR expression was markedly upregulated following egg activation. Functionally, the addition of CASR agonist NPS R-568 significantly enhanced pronuclear formation rate, while supplementation of CASR antagonist NPS2390 compromised egg activation. There was no change in NPS R-568 group compared with control group when the egg activation was performed without extracellular calcium addition. The addition of NPS2390 precluded the activation-dependent [Ca2+ ]i rise. When egg activation was conducted in intracellular Ca2+ chelator BAPTA-AM and NPS R-568 containing medium, CASR function was abolished. Meanwhile, CASR activation increased the level of the [Ca2+ ]i effector p-CAMKII, and the presence of KN-93, an inhibitor of CAMKII, significantly reduced the CASR-mediated increasement of pronuclear formation rate. Furthermore, the increase of CASR expression following activation was reversed by inhibiting CAMKII activity, supporting a positive feedback loop between CAMKII and CASR. Altogether, these findings provide a new pathway of egg activation about CASR, as the extracellular Ca2+ effector, promotes egg activation via its downstream effector and upstream regulator CAMKII.

Vitamin C protects carboplatin-exposed oocytes from meiotic failure.

CBP (carboplatin) is a second-generation chemotherapeutic drug of platinum compound commonly applied in the treatment of sarcomas and germ cell tumours. Although it is developed to replace cisplatin, which has been proven to have a variety of side effects during cancer treatment, CBP still exhibits a certain degree of toxicity including neurotoxicity, nephrotoxicity, hematotoxicity and myelosuppression. However, the underlying mechanisms regarding how CBP influences the female reproductive system especially oocyte quality have not yet been fully determined. Here, we report that CBP exposure led to the oocyte meiotic defects by impairing the dynamics of the meiotic apparatus, leading to a remarkably aberrant spindle organisation, actin polymerisation and mitochondrial integrity. Additionally, CBP exposure caused compromised sperm binding and fertilisation potential of oocytes by due to an abnormal distribution of cortical granules and its component ovastacin. More importantly, we demonstrated that vitamin C supplementation prevented meiotic failure induced by CBP exposure and inhibited the increase in ROS levels, DNA damage accumulation and apoptotic incidence. Taken together, our findings demonstrate the toxic effects of CBP exposure on oocyte development and provide a potential effective way to improve the quality of CBP-exposed oocytes in vitro.

Screening of angiogenesis inhibitors using a 3D vascular microfluidic chip to achieve contraception.

Angiogenesis plays a vital role in the process of embryo implantation, as it improves endometrial receptivity and guides embryo implantation, thus creating a favorable environment for subsequent development of the embryo. Hence, a theory of achieving contraception by inhibiting angiogenesis was put forward. Here, we screened the drugs inhibiting angiogenesis using cell scratch wound assay and a 3D biomimetic vascular microfluidic chip, then observed the effect of them on contraception by injecting these drugs into fertilized mice and observing if the embryos were implanted. We preliminarily verify the feasibility of contraception by inhibiting angiogenesis and gives a new direction in the development of contraceptive pills.

Blood metabolite concentration, milk yield, resumption of ovarian activity and conception in grazing dual purpose cows supplemented with concentrate during the post-partum period.

Cattle grazing on natural pasture face seasonal variation in pasture availability and nutritive value. Improving the nutrition of cows by providing supplementary feeds during periods of pasture or nutrient deficit is necessary for improved productivity. This study was conducted to determine the effect of feed supplementation during the post-partum period on the metabolic status and reproductive performance of 40 grazing Sanga and Friesian × Sanga cows. Twenty out of a total of 40 cows (10 Sanga and 10 Friesian × Sanga cows) were supplemented with 2.5 kg of concentrate per day for 16 weeks after calving. Parameters measured include daily milk yield, concentration of blood metabolites, resumption of ovarian activity and conception. Supplemented cows had higher partial milk yield than their non-supplemented counterparts (2.07 vs. 1.60 kg/day; P < 0.001). Sanga cows had lower milk yield than the Friesian × Sanga crossbreds (1.61 vs. 2.05 kg/day; P < 0.01). Supplemented cows had higher mean total protein (86.7 vs. 81.3 g/L; P = 0.007) and globulin (53.0 vs. 47.7; P = 0.014) concentrations than non-supplemented cows. Sanga cows had higher glucose (P = 0.027), total protein (P < 0.001) and globulin (P < 0.001) but lower triglyceride (P = 0.003) and progesterone (P = 0.023) concentrations than Friesian × Sanga cows. Supplemented cows had less proportion not cycling (20 vs. 55%; P = 0.022) and less days open (95.8 vs. 106 days; P = 0.032) than non-supplemented cows. The Friesian × Sanga cows had higher progesterone concentrations at first progesterone rise (3.34 vs. 1.32 ng/mL; P = 0.032) and shorter interval from calving to conception (96.7 vs. 106 days; P = 0.042). The results from this study indicate the beneficial effects of feed supplementation in terms of increased milk yield, better metabolic status and reproductive performance.

Genotype of MTNR1A gene regulates the conception rate following melatonin treatment in water buffalo.

Buffaloes have tendency to show seasonal reproduction and remain in anestrus due to limited ovarian activity during summer. The seasonal reproductive behavior is ascribed the effect of melatonin related to photoperiod. Treating animals with melatonin could be a possible strategy to overcome the problem. The role of MTNR1A gene has not been fully explained in the buffalo. Therefore, we conducted a study on 114 buffalo heifers to detect the polymorphic site in MTNR1A gene and further treated them with melatonin implants to investigate the role of most frequent genotype following melatonin treatment on pregnancy. The present investigation is the first to investigate the association between melatonin treated different MTNR1A genotype buffalo and pregnancy. We confirmed SNP at position 72 in 812 bp fragment exon II of MTNR1A gene. RFLP of PCR products with Hpa I enzyme resulted in three genotypes: TT (812bp), CT (812, 743, 69bp) and CC (743, 69bp). Next, buffaloes of each genotype (TT, CC, CT; n = 28 for each) were treated with melatonin implants to compare the conception rate with their corresponding untreated control (n = 10 for each genotype). Melatonin concentrations were higher (P < 0.05) for the treatment groups of all genotypes compared to their respective untreated control from day 1-28. The pregnancy rate was significantly associated with the MTNR1A genotype. The conception rate was higher (P < 0.05) for TT genotype than for the other genotypes of buffaloes treated with melatonin. Furthermore, buffaloes of TT genotype treated with melatonin started exhibiting estrus activity soon from second week of melatonin treatment (14.1 ±â€¯2.1; range: 10-17 days) and were found to be 7.8 times more likely to become pregnant compared to other genotypes following melatonin treatment. In conclusion, TT genotype of MTNR1A gene is more sensitive to melatonin treatment that favours pregnancy in buffaloes during summer.

Effects of hypertonic dextrose and paraffin solution as non-antibiotic treatments of clinical endometritis on reproductive performance of high producing dairy cows.

The objective of this study was to compare the reproductive performance of cows affected by clinical endometritis (CE) following treatment with an intrauterine infusion of 50% dextrose solutions (DEX) and liquid paraffin (LP) as alternative therapies with routine treatments including PGF2α injection and intrauterine infusion of oxytetracycline (OTC). Moreover, the reproductive indices of cows treated with endometritis were compared with those of healthy ones. At postpartum reproductive tract examination (28-35 DIM) in one Iranian dairy farm, cows with CE without any selection were assigned to four groups: (a) OTC, a common treatment in Iran, was administered (5 g) to 396 cows, (b) PGF2 α (PG) was injected to 496 cows, (c) dextrose solution (DEX): intrauterine infusion of 50% dextrose solution (200 ml) was done in 427 cows, and (d) liquid paraffin (LP) was administered (100 ml) to 423 cows via intrauterine route. We further assigned 2,233 clinically healthy cows to the control group. The incidence of endometritis was 41.6% in this study. Based on the results of reproductive indices including median days to first AI, days open (DO), first service conception rate, conception rate in 2nd and 3rd services, conception rate in all three services, pregnancy rate < 100 days and pregnancy rate < 200 days, except for median days to first AI in other reproductive indexes, reproductive performance was significantly lower in LP group compared with the healthy cows and other treatment groups (p < 0.05). Except for the first service conception rate and proportion of cows pregnant before 100 days in milk (DIM), there existed no significant difference between the DEX group and the control as far as reproductive performance is concerned (p ≥ 0.05). The first service conception rate was recognizably lower in DEX group compared with OTC and PG (p < 0.05). In conclusion, the use of a non-antibiotic special solution of dextrose 50% is a good alternative to antibiotic agents concerning the treatment of CE in dairy cows.

Effects of Dietary L-arginine Supplementation from Conception to Post- Weaning in Piglets.

Weaned piglets experience sudden changes in their dietary patterns such as withdrawal from the easily digestible watery milk to a coarse cereal diet with both systemic and intestinal disruptions coupling with the expression of pro-inflammatory proteins which affects the immune system and the concentrations of haptoglobin including both positive and negative acute-phase proteins in the plasma. L-arginine is an important protein amino acid for piglets, but its inadequate synthesis is a nutritional problem for both sows and piglets. Recent studies indicated that dietary supplementation of L-arginine increased feed intake, uterine growth, placental growth and nutrient transport, maternal growth and health, embryonic survival, piglets birth weight, piglet's growth, and productivity, and decreased stillbirths. L-arginine is essential in several important pathways involved in the growth and development of piglets such as nitric oxide synthesis, energy metabolism, polyamine synthesis, cellular protein production and muscle accretion, and the synthesis of other functional amino acids. However, the underlying molecular mechanism in these key pathways remains largely unresolved. This review was conducted on the general hypothesis that L-arginine increased the growth and survival of post-weaning piglets. We discussed the effects of dietary L-arginine supplementation during gestation, parturition, lactation, weaning, and post-weaning in pigs as each of these stages influences the health and survival of sows and their progenies. Therefore, the aim of this review was to discuss through a logical approach the effects of L-arginine supplementation on piglet's growth and survival from conception to postweaning.

Supplementation of biotin to sperm preparation medium enhances fertilizing ability of spermatozoa and improves preimplantation embryo development.

PURPOSE: Motility of spermatozoa helps not only in planning the type of infertility treatment but also directly reflects the success rate in assisted reproductive technology (ART). Previously, biotin, a water-soluble vitamin, has been shown to increase the motility and longevity of cryopreserved human spermatozoa. The present study was designed to understand the molecular basis of the beneficial effects of presence of biotin in sperm wash medium on early embryo development. METHODS: The effect biotin supplementation to sperm wash medium on the sperm parameters were assessed in swim-up fraction of normozoospermic and asthenozoospermic ejaculates collected from infertile men. Fertilization and early embryo development was studied using Swiss albino mice. RESULTS: Even though both biotin and pentoxifylline (PTX) enhanced the motility of spermatozoa from normozoospermic and asthenozoospermic samples, biotin group exhibited higher in vitro survival. Using mouse model, we observed that presence of biotin or PTX in sperm wash medium improved the fertilization rate and blastocyst rate compared to control. Blastocysts from these groups had significantly higher total cell number (P < 0.01) and lower apoptotic index. In silico target prediction revealed that GTPase HRas (HRas), tyrosine-protein phosphatase nonreceptor type 1 (PTP1B), and glucokinase are the probable targets for biotin. Solution-state Nuclear Magnetic Resonance (NMR) studies confirmed that biotin interacts both with human HRas and PTP1B. CONCLUSION: Our results indicate that presence of biotin in sperm wash medium can improve the fertilization potential and preimplantation embryo development and can be considered as a safe alternate to PTX.

Effects of melatonin supplementation prior to Ovsynch protocol on ovarian activity and conception rates in anestrous Murrah buffalo heifers during out of breeding season.

Buffalo heifers have tendency to show anestrus during summer season. Melatonin has been used for correcting summer dependent anestrous via inducing resumption of ovarian activity. Therefore, the investigation was conducted to compare efficacy of melatonin for induction of estrus and conception rate with Ovsynch protocol in summer anestrous Murrah buffalo heifers. Thirty, summer anestrous Murrah buffalo heifers were selected and divided into two groups- treatment (n = 20; 12 melatonin implants) and control (n = 10; no treatment). On day 28 post-implant insertion, animals of both the groups were subjected to Ovsynch protocol. Blood sampling and ovarian ultrasonography were conducted to measure plasma melatonin, progesterone concentration and ovarian dynamics, respectively. No animal in either group showed estrus during first 28 days post-implant insertion. However, estrus induction rate was 100% after Ovsynch protocol in both groups. As compared to controls, treatment group exhibited higher (p < 0.05) plasma melatonin on days 1, 4, 8, 15, 22 and 28 post-melatonin, with highest concentration on day 4. The progesterone concentration increased (p < 0.05) on days 15 and 22 post-melatonin treatment. The treatment group had larger (p < 0.05) preovulatory follicle on day of AI, subsequently developed larger (p < 0.05) corpus luteum and higher plasma progesterone concentrations by day 12 post-AI as compared to the control group. The overall conception rate was 50 and 20% in treatment and control groups, respectively. In conclusion, melatonin treatment along with Ovsynch protocol improved the luteal profiles as well as the conception rate in buffalo heifers when compared with animals treated with Ovsynch protocol alone during summer season.

Phyto-oestrogens affect fertilisation and embryo development in vitro in sheep.

Phyto-oestrogens such as isoflavones are natural compounds that can profoundly affect reproductive function. In the present study, we tested whether including isoflavone compounds (genistein, biochanin A, formononetin) in the maturation medium would affect the outcomes for ovine oocytes in vitro. Each isoflavone compound was evaluated at five concentrations (0, 2.5, 5, 10, 25µgmL-1) and the entire protocol was repeated four times. Cumulus-oocyte complexes were randomly allocated to the treatments, then fertilised and cultured in vitro. Compared with control (0µgmL-1), the lower concentrations of isoflavone (2.5, 5 and 10µgmL-1) had no detectable effect on the rates of cleavage or embryo development, or on embryo total cell counts (TCC). However, the highest concentration (25µgmL-1) of all three isoflavones exerted a variety of effects (P<0.05): genistein decreased cleavage rate, blastocyst rate and blastocyst efficiency (blastocysts produced per 100 oocytes); biochanin A decreased cleavage rate and blastocyst efficiency; and formononetin decreased blastocyst rate and blastocyst efficiency. Biochanin A (25µgmL-1) reduced embryo TCC specifically at the hatched blastocyst stage (P<0.05). We conclude that the presence of isoflavones at 25µgmL-1 during IVM decreases the cleavage rate and inhibits blastocyst hatching.

A retrospective analysis of adverse effects of an in vivo fluoroquinolone antibiotic enrofloxacin treatment on oocyte quality in the common marmoset.

Here we report a retrospective analysis of negative effects of routine enrofloxacin treatment of recurrent diarrhea on the ovary and the developing oocytes of the common marmoset, a small New World primate. The most deleterious effect on oocytes was observed about two months post treatment suggesting that the enrofloxacin effect is on early growing follicles. Manifestations of toxicity included decreased numbers of growing follicles and recovered culturable oocytes, as well as signs of early atresia of granulosa cells. In addition, increased amounts of holed stroma after treatment strongly suggested increased death of the early growing follicles. Of the oocytes judged to be of adequate quality for culture, maturation rates were not affected but fertilization of in vitro matured MII oocytes and subsequent cleavage rates were severely reduced in the enrofloxacin treated animals. Further, the arrested oocytes, which failed to mature or fertilize, showed obvious meiotic spindle abnormalities.

Biological effects of polyphenol-rich extract and fractions from an oenological oak-derived tannin on in vitro swine sperm capacitation and fertilizing ability.

Although excessive ROS levels induce sperm damage, sperm capacitation is an oxidative event that requires low amounts of ROS. As the antioxidant activity of the ethanol extract (TRE) of a commercial oenological tannin (Quercus robur toasted oak wood, Tan'Activ R®) and its four fractions (FA, FB, FC, FD) has been recently reported, the present study was set up to investigate the biological effects of TRE and its fractions in an in vitro model of sperm capacitation and fertilization. Boar sperm capacitation or gamete coincubation were performed in presence of TRE or its fractions (0, 1, 10, 100 µg/ml). TRE at the concentration of 10 µg/ml (TRE10) stimulated sperm capacitation, as it increased (p < .001) the percentage of spermatozoa with tyrosine-phosphorylated protein positivity in the tail principal piece (B pattern) (67.0 ±â€¯10.6 vs. 48.6 ±â€¯9.0, mean ±â€¯SD for TRE10 vs. Ctr respectively). Moreover T10 significantly (p < .001) increased oocyte fertilization rate (91.9 ±â€¯4.0 vs. 69.0 ±â€¯14.8, TRE10 vs. Ctr respectively). An opposite effect of TRE at the concentration of 100 µg/ml (TRE100) on both sperm capacitation (B pattern cell percentage 33.3 ±â€¯29.2) and fertilizing ability (fertilization rate 4.9 ±â€¯8.3), associated with a higher sperm viability (66.9 ±â€¯9.3 vs. 35.4 ±â€¯10.8, TRE100 vs. Ctr respectively) (p < .001), was recorded. The potency of the TRE fractions seems to be highest in FB followed by FC, faint in FD and nearly absent in FA. Our results show that TRE and its fractions, in a different extent, exert a powerful biological effect in finely modulating capacitation and sperm fertilizing ability.