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Carvacrol (C 10 H 14 O), an efficient phenolic antioxidant substance for several cell types, may become a useful antioxidant for female germ cells and embryo culture. This study investigates the effects of carvacrol supplementation on bovine oocytes in in vitro maturation (IVM) and embryo production. In total, 1222 cumulus-oocyte complexes were cultured in TCM-199 + alone (control treatment) or supplemented with carvacrol at the concentrations of 3 µM (Carv-3), 12.5 µM (Carv-12.5), or 25 µM (Carv-25). After IVM, the oocytes were subjected to in vitro fertilization and embryo production, and the spent medium post-IVM was used for evaluating the levels of reactive oxygen species and the antioxidant capacity (2,2-diphenyl-1-picryl-hydrazyl-hydrate and 2,2'-azinobis-3-ethyl-benzothiozoline-6-sulphonic acid quantification). A greater ( P < 0.05) antioxidant potential was observed in the spent medium of all carvacrol-treated groups compared with the control medium. Moreover, the addition of carvacrol to the maturation medium did not affect ( P > 0.05) blastocyst production on days 7 and 10 of culture; however, the total number of cells per blastocyst was reduced ( P < 0.05) in two carvacrol-treated groups (Carv-3 and Carv-25). In conclusion, carvacrol demonstrated a high antioxidant capacity in the spent medium after oocyte maturation; however, although embryo production was not affected, in general, carvacrol addition to IVM medium reduced the total number of cells per blastocyst. Therefore, due to the high antioxidant capacity of carvacrol, new experiments are warranted to investigate the beneficial effects of lower concentrations of carvacrol on embryo production in cattle and other species.

This study investigates the impact of eugenol (EU) supplementation on bovine oocyte in vitro maturation (IVM) and antioxidant capacity, as well as in vitro embryo production and quality after conventional in vitro fertilization (IVF). A total of 1077 cumulus oocyte complexes were cultured in TCM-199 + without EU supplementation (control treatment) or supplemented with EU at the concentrations of 10 μM (EU-10), 20 μM (EU-20), or 40 μM (EU-40). After IVM, the oocytes were subjected to IVF and embryo culture. The addition of EU at 40 μM to the IVM medium improved (P < 0.05) the antioxidant capacity and cleavage rate when compared to the control treatment. Moreover, a positive correlation (r = 0.61, P < 0.03) was observed between cleavage rate and EU concentration. The addition of EU at concentrations of 10 and 20 μM decreased (P < 0.05) the calreticulin (CALR) levels in expanded blastocysts when compared to the control treatment and EU-40 treatment. However, the EU-10 and EU-20 treatments had a greater (P < 0.05) mean total cell number (TCN) per expanded blastocyst when compared to the control treatment and EU-40 treatment. In conclusion, the addition of EU to the enriched culture medium during IVM of bovine oocytes improved the antioxidant capacity of the spent medium, as well as the cleavage rate and embryonic quality (i.e., TCN/expanded blastocyst), and reduced the endoplasmic reticulum stress (i.e., CALR levels) in the embryos. Thus, we recommend enriching the IVM medium with 10 μM EU for in vitro bovine embryo production., (Copyright © 2022 Elsevier Inc. All rights reserved.)

Dicliptera verticillata is a medicinal plant traditionally used in western Cameroon to cure female infertility. This experiment was designed to assess the effects of the aqueous extract of Dicliptera verticillata (AEDv) on fertility and gestation in female rats. Oral increasing doses of AEDv were administered to immature female rats over 20 d. After this time, some animals were mated with fertile males and some fertility parameters were assayed; the other animals were euthanized for preliminary toxicity parameters analysis. The effects of AEDv on the different stages of gestation were assayed on selected animals previously controlled for estrous cycle regularity and mated. AEDv led to an increase in serum, uterine and ovarian proteins as well as in ovarian and uterine weights (P < 0.05) in immature female rats. Hepatic proteins significantly decreased (P < 0.01) in high dose-treated animals (50 and 100 mg/kg) compared with controls. The number of implantation sites and the fertility rate were significantly lower (P < 0.05), while the antifertility activity increased significantly (P < 0.05) in treated rats compared with controls. When administered from the 1st to the 5th day of pregnancy, AEDv led to a decrease of more than 60% in the implantation rate in high dose-treated rats (50, 100, and 400 mg/kg). From the 6th to the 9th day, the implantation, gestation rates and the number of fetuses decreased significantly in all treated groups. From the 11th to the 20th day, a 50% resorption and decrease in gestation rate were reported in 50 mg/kg dose-treated animals. AEDv possesses weak contraceptive and abortifacient effects during pregnancy.

The present study evaluated the effect of the aqueous extract from leaves of E. speciosa on some physiological and biochemical parameters of reproduction and the onset of puberty in pregnant mare serum gonadotropin (PMSG)-primed immature female rats. High pressure liquid chromatography (HPLC) was used to quantify the phenolic compounds in the methanol/methylene chloride (1:1) extract, the ethanolic and ethyl acetate fractions and the aqueous residue of E. speciosa. E. speciosa (0, 8, 32 or 64 mg/kg) were administered for 15 days to 24 non-PMSG-primed and 24 primed rats with 0.01 IU of PMSG. At the end of the treatment period, animal were sacrificed and their body, ovarian, uterine weight, ovarian protein or cholesterol level, as well as data on puberty onset were recorded. Of the 16 polyphenolic compounds quantitatively revealed in the extracts and fractions of E. speciosa after HPLC analysis, quercetin, rutin, apigenin and eugenol were the most abundant. Non-primed rats showed a significant increase (P < 0.05) in the uterine relative weight at the dose of 8 mg/kg when compared with the other treatments. The uterine proteins and the ovarian cholesterol (P < 0.05), respectively, showed a reduction at doses of 64 mg/kg and 32 mg/kg in non-primed rats. However in PMSG-primed rats, a significant decrease (P < 0.05) was observed in ovarian cholesterol at 64 mg/kg. In conclusion, E. speciosa potentializes the PMSG-inducing effect on folliculogenesis in PMSG-primed rats.

Background: Senecio biafrae is a medicinal plant widely used in traditional medicine to cure female infertility. Some effects have been pharmacologically demonstrated on immature female rats but in vivo and in vitro investigations are still necessary for determining its mechanism of action. The aim of the present study was to evaluate the estrogenic and FSH-like effects of the plant extracts and fractions on some fertility parameters in immature female rats and on in vitro survival and growth of swine preantral follicles., Methods: 21-23 days old female Wistar rats orally received extracts and fractions of S. biafrae at 0, 8 and 64 mg/kg doses over 20 days. The LH, FSH, estradiol and progesterone serum levels were evaluated as well as the ovarian cholesterol, uterus and ovaries masses and proteins. The numbers of follicles at different developmental stages were recorded in ovarian cortexes after histology. Slices of swine ovarian cortexes were cultured along 1 or 7 days in alpha-minimum essential medium (α-MEM) and fixed for morphological analysis of preantral follicles. The fresh control, cultured control (CIV control) and different Senecio biafrae-treated ovarian fragments were analyzed for preantral follicles development. Treatments that showed the best follicle growth in culture were submitted to AgNOR test. The aqueous and MeOH/CH 2 Cl 2 extracts as well as the ethyl acetate and hexane fractions of S. biafrae were submitted to the HPLC for analysis of polyphenolic secondary metabolites., Results: Ovarian and uterine proteins were significantly high (p < 0.01) in animals treated with the two dosages of ethyl acetate and n-butanol fractions. The same result was recorded with uterine proteins in animals treated with the hexane fraction. The FSH level significantly dropped with all ethanolic extract doses and with the 64 mg/kg dosage of the methanol/methylene chloride (MeOH/CH 2 Cl 2 ) extract while LH was reduced (p < 0.01) in almost all the treated groups. Estradiol level was significantly increased (p < 0.001) in the three groups receiving the extracts, but reduced (p < 0.001) in the three groups receiving the fractions of the plant. The progesterone level increased with almost all the treated groups. Primary and secondary follicles augmented (p < 0.01) in MeOH/CH 2 Cl 2 extract and n-butanol fraction while tertiary follicles increased with the same extract and the ethyl acetate fraction (p < 0.05). Treatments with aqueous and ethanolic extracts as well as ethyl acetate fraction led to a significant increase (p < 0.05) in the number of morphologically normal follicles after 7 days of culture as compared to the CIV control. The number of AgNOR dots per follicle was significantly low (p < 0.05) in all cultured groups as compared to the fresh control, except the ethyl acetate 2.8 ng/ml dosage. The same observation was done with AgNOR dots per cell in the 2.8 ng/ml dosage aqueous extract-treated fragments. The phenolic compounds mainly encountered in the plant, independently of the extract or fraction are apigenin, eugenol and rutin., Conclusion: Extracts and fractions of S. biafrae have an important FSH-like effect which induces follicular survival and growth., Competing Interests: Declaration of competing interest The authors declare that there are no conflicts of interest., (Copyright © 2020 Elsevier B.V. All rights reserved.)

We evaluated the effects of the vitrification solution (i.e., ethylene glycol (EG) + dimethyl sulfoxide (DMSO) with or without propylene glycol (PG)) and of exposure time on the re-expansion and hatching rates of vitrified Bos indicus embryos. In vitro produced embryos (n = 1050) were divided into seven groups: control group (non-vitrified embryos) and six vitrification groups with different cryoprotectant concentrations and exposure times. After vitrification, embryos were cultured for determination of re-expansion and hatching rates. Vitrification with 25% DMSO +25% EG (exposure for 1 min and 20 s) resulted in the highest re-expansion (65.2%) and hatching (68.2%) rates. The lowest re-expansion and hatching rates were observed in vitrification with 12.5% DMSO + 25% EG + 12.5% PG with both tested exposure times (i.e., 3 min + 1 min and 1 min + 20 s). A combination of DMSO + EG is efficient to preserve blastocysts, especially following a short exposure time., (Copyright © 2018. Published by Elsevier Inc.)

The objective of this study was to assess the effects of bovine embryo vitrification by applying three different vitrification solutions containing ethylene glycol (EG) and dimethylsulphoxide (DMSO) at different concentrations (10, 20 or 25% each) combined with 1.0 M glucose or 1.0 M sucrose, on the in vitro hatching and expansion rates. Healthy oocytes were selected for in vitro maturation and fertilization from 200 bovine ovaries, and subsequently cultured up to the blastocyst stage (n = 800). Control (n = 200) and vitrified cells (n = 100 per treatment; 600 in total) were cultured for an extra 24 or 48 h to evaluate hatching and expansion, respectively. Vitrification significantly decreased embryonic re-expansion and hatching rates independently of the tested solution when compared with control embryos, but solutions with 25% EG + 25% DMSO resulted in the highest re-expansion (75%) and hatching (70%) rates, independently of the added sugar. The addition of sucrose resulted in higher rates of re-expanded and hatched embryos when compared with glucose addition. We concluded that the combination of 25% EG + 25% DMSO and 1.0 M sucrose allowed hatching and expansion of vitrified-warmed bovine embryos produced in vitro.

Ethnopharmacological Relevance: Senecio biafrae is a plant from the huge family of Asteraceae used in the African pharmacopoeia for the treatment of many ailments among which is infertility., Material and Methods: The aqueous extract, which was primarily subjected to polyphenol analysis, has been administered to immature female rats for 20 days at 8, 32, 64 and 128 mg/kg of body weight. The day following the treatment, the animals were sacrificed; their serum, ovary and uterus were retained respectively for reproductive hormones, ovarian and uterine proteins, and ovarian cholesterol assays., Results: Light body weight gain variation of treated animals was observed during the experimental period. A significant increase (p ˂ 0.05) in serum estradiol and proteins as well as in uterine weight (p ˂ 0.01) of all Senecio biafrae treated animals was noted. No significant variation was noticed in the ovarian weight and follicle numbers., Conclusion: The various biochemical and physiological parameters of fertility were significantly improved with the aqueous extract of Senecio biafrae, thus attesting some of its traditional usage., (Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.)

Ethnopharmacological Relevance: In Cameroon, most women use traditional medicine for the treatment of pregnancy and childbirth complaints. In order to identify some of the medicinal plants locally used to alleviate these complaints, an ethnobotanical survey was undertaken in five villages of Menoua Division (West-Cameroon)., Materials and Methods: Interviews were conducted through structured questionnaires among 24 traditional healers and 179 women living either in the town of Dschang or in 4 neighboring villages. After having recorded the interviewee personal information on issues related to medicinal plants utilization, a literature investigation on their therapeutic or pharmacological effects and phytochemical composition was conducted., Results: A total of 88 medicinal plants species used to treat 24 conditions occurring during or after pregnancy and belonging to 70 genera or 34 families were recorded. Maximum medicinal uses of plants are reported for the treatment of the following ailments: swelling of legs and ankles (23%), facilitation of delivery (22%), cleaning of the baby (12%). Most herbal remedies are prepared with the leaves (30%), leaves+stems (28%) and whole plant (23%) as maceration (76%). The majority of women who used medicinal plants were very satisfied (75 %) and it is reported that most of these plants are used in the treatment of women health conditions., Conclusion: Many herbal remedies used for the treatment of pregnant women׳s health conditions in Menoua division-West Cameroon have been revealed. It would therefore be judicious for our government and research institution to evaluate the therapeutic and toxicological potentials of these plants in order to valorize their use., (Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.)

Objective: To obtain a scientific basis of the use of plant-derived preparations by many rural people in Cameroon, for their primary health care needs in the treatment of diseases such as cancer., Methods: The antiproliferative effect of 11 plants methanol crude extracts on four cancer cells using sulforhodamine-B assay and their antioxidant activities using 1-diphenyl-2-picrylhydrazyl radical and nitric oxide radical scavenging ability were investigated. The Ekebergia senegalensis (E. senegalensis) and Protea elliotii (P. elliotii) extracts were selected based on their antioxidant and anticancer activities, and partition in hexane, dichloromethane, ethyl acetate, butanol and methanol was done. Each fraction was submitted to antioxidant and anticancer activities, and the effect of the dichloromethane fraction (the most antiproliferative fraction) on NCI-H460 cell cycle was determined by flow cytometry., Results: The most antiproliferative substances were found for the extracts from E. senegalensis, P. elliotii, Terminalia macroptera and Vitellaria paradoxa. Whereas the most antioxidant substances were found for the extracts from Cissus populnea, E. senegalensis, P. elliotii, Terminalia macroptera, Vitellaria paradoxa, and Gardenia aqualla. Dichloromethane fraction of P. elliotii was found to be highly antiproliferative to NCI-H460 cancer cells and showed S phase arrest cell cycle progression. Ethyl acetate n-butanol and methanol fractions showed quite strong antioxidant activity for both E. senegalensis and P. elliotii, as compared to that of gallic acid., Conclusions: Overall, the antiproliferative and antioxidant activities of some of the extracts lend some support to their use in the traditional medicine of Adamawa Region, Cameroon to treat cancer., (Copyright © 2014 Hainan Medical College. Published by Elsevier B.V. All rights reserved.)

Background: Senecio biafrae (Asteraceae) is a medicinal plant widely used by traditional healers in the western region of Cameroon for the treatment of female infertility. This experiment was designed to evaluate the effect of the aqueous extract from leaves and stems of S. biafrae (AESb) on the onset of puberty and some biochemical and physiological parameters of reproduction in immature Wistar female rats., Methods: Different doses of AESb were daily and orally administered to immature female rats (13 animals/group) for 30 days. At the end of the treatment period, six animal of each experimental group were sacrificed and their body, ovarian, uterus weight; uterine, ovarian protein or cholesterol level as well as data on puberty onset recorded. The remaining animals of each group were used for the fertility test and some gestational parameters recorded., Results: A linear increase in the growth rate of all animals was observed. The body weight gain in animals treated at the dose of 8 mg/kg of AESb significantly increased (p < 0.05) after 25 days of treatment while those receiving the doses of 32 and 64 mg/kg presented a significantly low body weight gain starting from the 19th day till the end of the treatment period. The ages (days) of animals at vaginal opening (VO) was significantly reduced (p < 0.05) in those treated with the doses of 32 (41.25 ± 0.51) and 64 mg/kg (41.42 ± 0.54) as compared to control animals (43.33 ± 0.73). AESb significantly increased (p < 0.05) the ovarian weight and the number of corpora lutea in animals treated with 8 mg/kg as well as the uterine weight and protein levels irrespective of the dose. No significant effect of the extract on various fertility and gestational parameters was registered., Conclusion: The overall results of the present study provide evidence on the puberty onset induction and ovarian folliculogenesis effect of AESb in immature female rat.

Ethnopharmacological Relevance: Infertility affects many women in Cameroon, with a rate of about 25%. This study aimed at collecting and documenting information on herbal remedies traditionally used for the treatment of women infertility in Baham subdivision (Western Region of Cameroon)., Materials and Methods: Fieldwork was undertaken as an ethnopharmacological survey involving thirty-two traditional medicine practitioners interviewed in 8 villages of the Baham sub-division. Personal information on interviewees as well as issues related to medicinal use of plants were recorded using structured questionnaires. A literature investigation on the therapeutic or pharmacological properties of recorded medicinal plants was further undertaken., Results: From this inventory, a total of 46 plant species belonging to 43 genera and 26 families have been registered. These plants are used in 32 recipes and prepared as maceration (43%) or decoction (40%) of only one plant (25%) or of the mixture of two (22%), three (28%), four (22%) or even seven (3%) medicinal plants. Globally, they are given orally during 30 days, at an average dosage of two glasses per day. The literature confirms the use of the majority of these plants for the treatment of the woman infertility and illnesses that are associated to it., Conclusion: This research shows that traditional healers of the Baham subdivision use various recipes of medicinal plants for the treatment of female infertility. The valorization of this potential could be important for the conservation of these plants and the improvement of women reproductive health., (Copyright © 2011 Elsevier Ireland Ltd. All rights reserved.)