Males Of Proven Fertility Research Articles

Allium ampeloprasum var. Porrum (Alliaceae) Improves Metabolic and Reproductive Disorders Associated with Polycystic Ovary Syndrome in Wistar Rats.

To provide scientific evidence of the efficacy of Allium ampeloprasum against female infertility, the effects of the aqueous extract of the said plant (AE) were evaluated in rats with letrozole-induced polycystic ovary syndrome (PCOS). AE was administered orally to PCOS rats at doses of 192, 384, and 768 mg/kg. The positive control was co-treated with clomiphene citrate (1 mg/kg) and metformin (200 mg/kg). Normal and negative controls received distilled water. The vaginal contents of rats were examined daily under a microscope before (7 days) and during treatment. Treatments were administered orally for 15 days, and then, 6 rats from each group were sacrificed for biochemical and histological analyses. The remaining rats were mated with males of proven fertility for 5 days. The daily examination of vaginal smears allowed the evaluation of fertility index. After parturition, additional fertility parameters were determined. Results showed that in PCOS rats, AE decreased body weight (p < 0.001), abdominal fat weight (p < 0.001), serum levels of LH (p < 0.001), testosterone (p < 0.001), total cholesterol (p < 0.05), and LDL cholesterol (p < 0.01). HDL cholesterol increased and atherogenic indices decreased (p < 0.001). The number of Graafian follicles and corpora lutea increased, while cystic (p < 0.001) and atretic (p < 0.05) follicles decreased. AE also decreased oxidative stress in the ovaries, restored the estrous cycle, induced uterine epithelial cell hypertrophy, and improved fertility. These effects were attributed to phenols, flavonoids, terpenoids, and anthocyanins present in AE. The overall results justify the traditional use of A. ampeloprasum against female infertility and suggest its potential use as a dietary supplement for PCOS patients.

Abortifacient activity of &lt;em&gt;Aegle marmelos&lt;/em&gt; and &lt;em&gt;Laurus nobilis&lt;/em&gt; leaf extracts

Rapid elevation of population in India is the one of the major problems, which directly influence the economy of country and may lead to poverty. Government implemented number of family planning programs through the various surgical operations (tubectomy and laproscopy) and oral contraceptives. Usage of oral contraceptive pills may lead serious complications, and may induce congenital abnormalities. The primary goal of this research is to assess the abortifacient activity in rat models of two historically used medicinal plants, Laurus nobilis L. and Aegle marmelos (L.) Corr. Restructure the paragraph as the study included 18 female wistar rats (150-200 g) and six male wistar rats (male wistar rats were used only for copulation). Female rats in proestrous phase were isolated and allowed to mate with males of proven fertility using the mass mating technique in a 3:1 ratio for an overnight. Control animals received an equivalent volume of the dosing vehicle (1% tween 80) orally. Aqueous extract of Laurus nobilis (AQLN) leaves and Ethanolic Extract of Aegle Marmelos leaves (EEAM) at doses of 175 mg/kg and 250 mg/ kg of were orally administrated daily for 10 days from day 0 of pregnancy to day 9. On day 20th of pregnancy, all the animals were sacrificed under euthanasia and the uterine horns were isolated, later they were examined for number of abortifacient sites and deformities of fetuses. The number of live fetuses in animals treated with EEAM at two doses was substantially lower in Group-4 at 175mg/kg (2.63 + 0.36) and Group-5 at 250mg/kg (1.87 + 0.40) compared to the vehicle control group (p 0.05, p 0.01). The survival ratio decreased considerably from 52.2% to 28.8% as the dose increased. Similarly, the abortion rate was higher in group 5 compared to Group-4. AQLN demonstrated to have 100% abortifacient efficacy at 250mg/kg, while EEAM has 83.3%.

Gestational Sympathetic Stress Programs the Fertility of Offspring: A Rat Multi-Generation Study.

The exposure to sympathetic stress during the entire period of gestation (4 °C/3 h/day) strongly affects the postnatal reproductive performance of the first generation of female offspring and their fertility capacity. The aim of this work was to determine whether this exposure to sympathetic stress affects the reproductive capacity of the next three generations of female offspring as adults. Adult female Sprague–Dawley rats were mated with males of proven fertility. We studied the reproductive capacity of the second, third, and fourth generations of female offspring (the percentage of pregnancy and the number and weight of female offspring). The estrus cycle activity of the progenies was studied, and a morphological analysis of the ovaries was carried out to study the follicular population. The second generation had a lower number of pups per litter and a 20% decrease in fertile capacity. The estrus cycle activity of the third generation decreased even more, and they had a 50% decrease in their fertile capacity, and their ovaries presented polycystic morphology. The fourth generation however, recovered their reproductive capacity but not the amount of newborns pups. Most probably, the chronic intrauterine exposure to the sympathetic stress programs the female gonads to be stressed in a stressful environment; since the fourth generation was the first born with no direct exposure to stress during development, it opens studies on intrauterine factors affecting early follicular development.

Effect of hCG administration on Day 7 post-mating on accessory corpus luteum development and progesterone concentration in llamas

A high embryo loss rate has been reported in llamas. As strategies that lead to an increase in plasma progesterone (P4) concentration might improve fertility, the aim of the present study was to evaluate if the administration of hCG on Day 7 post-mating is useful to develop an accessory corpus luteum (CL), increasing plasma P4 concentration. Twenty (n = 20) female llamas, ranging between 5 and 10 years of age and four (n = 4) males of proven fertility, ranging between 8 and 10 years of age were included in the study. Accessory CL developed in all treated llamas after hCG administration and plasma P4 concentration was significantly greater in treated than in control females (P˂0.0001). The diameter and vascularization of the original CL were not affected by treatment in pregnant llamas. However, in treated non-pregnant llamas, corpus luteum diameter was greater than in the control group from Day 14 post-mating until the end of the study (P˂0.001). In treated llamas, the accessory CL was detected throughout the study in pregnant and non-pregnant females, but its vascularization started to decrease around Day 16 post-mating in non-pregnant animals (P˂0.05). In conclusion, hCG treatment on Day 7 post mating was useful to induce an accessory CL and increase plasma P4 concentration in llamas. Thus, this treatment could be considered as a useful strategy to improve pregnancy rates in llama herds.

TARGETED DELETION OF MITOFUSIN 1 AND MITOSUFIN 2 CAUSES FEMALE INFERTILITY AND LOSS OF FOLLICULAR RESERVE

Mitofusin 1 (MFN1) and Mitofusin 2 (MFN2) are transmembrane GTPases that regulate mitochondrial fusion and are required for the maintenance of cellular homeostasis. The aim of the study was to determine the role of mitofusins in female reproductive competence and senescence using a mouse model with oocyte-specific deletion of Mfn1 and Mfn2. Mice with oocyte-specific double deletion of Mfn1 and Mfn2 were generated by mating Mfn1 flox/flox or Mfn2 flox/flox with Zp3-Cre Mfn1 and Mfn2 males. In all experiments, adult (8-weeks-old) Mfn1-/-/Mfn2-/- female mice were compared to wild-type (WT). To assess fertility, female mice from each group (n=5) were mated with adult WT males of proven fertility for 12 weeks. The ability to generate oocytes (germinal vesicle [GV] and metaphase II [MII]) was assessed after injection with PMSG (10IU) or PMSG and hCG (10IU). Unfertilized MII oocytes were collected 14 hours after hCG injection. Follicle development was assessed in ovaries from Mfn1-/-/Mfn2-/- and WT mice after fixation, paraffin embedding, and sectioning, followed by hematoxylin and eosin (H&E) staining. ATP levels were determined by bioluminescent assay. Mitochondrial DNA (mtDNA) copy number was measured in individual oocytes by cloning of mitochondria specific gene (Cox3) as a standard, followed by quantitative real-time PCR (qPCR). Expression levels of mitochondrial respiratory chain genes was evaluated with qRT-PCR. Student’s t-test and Chi-Square analysis were applied. Mature female Mfn1-/-/Mfn2-/- were infertile compared to WT females (0 ± 0 vs ± 5.62 ± 1.64 pups per litter; p < 0.0001). Mfn1-/-/Mfn2-/- mice showed a significant decrease in GV oocyte number (12.75 ± 1.70 vs 40.25 ± 3.5; p < 0.0001) and generated no MII oocytes (0 ± 0 vs 31.01 ± 3.5; p < 0.0001) in comparison to WT. Eight-week-old Mfn1-/-/Mfn2-/- mice ovaries had similar number of primordial, primary, and secondary follicles compared to WT. However, the number of early antral and antral follicles numbers was significantly decreased (17.2 ± 10.2 vs 55.3 ± 9.50; p < 0.03, and 2.3 ± 1.5 vs 16.3 ± 5.8; p < 0.04). At 6 months, Mfn1-/-/Mfn2-/- mice had a lower number of primordial, early antral, and antral follicles compared to WT. At 12 months, all follicle stages were found to be significantly lower in Mfn1-/-/Mfn2-/- mice. Decreased ATP production (0.62 ± 0.1 vs 1.23 ± 0.12; p = 0.002) and mtDNA copy number (446033 ± 53978 vs 706672 ± 72470; p = 0.003) were also observed in Mfn1-/-/Mfn2-/-oocytes compared to WT. Similarly, the expression of mitochondrial chain respiratory genes Cox1, Sdhb, Uqcrc2 and Atp5a1 were decreased in Mfn1-/-/Mfn2-/- oocytes compared to WT. Our findings demonstrate that lack of MFN1 and MFN2 results in infertility and accelerated loss of follicular reserve consistent with a diminished ovarian reserve (DOR) phenotype.

Effect of Bisphenol A on the Testis of Offspring of White Albino Rats during Pregnancy and Lactation: (Light and Electron Microscopic Study)

Introduction: The synthetic industrial and biomedical chemicals suspected to threat the health of the living organisms by disrupting their endocrine systems, referred to as endocrine disrupting chemicals (EDCs). Objective: To study the effects of Bisphenol A (BPA) on the natal and postnatal testis by the light and electron microscope to make sure if it is harmful or not on the development of male reproductive organs in albino rats. Matrial and Methods: Chemical used: Bisphenol A. The daily single oral dose was (0.3 mg/kg BW./day). Experimental animals: Adult female rats in proestrus cycle were caged overnight with adult males of proven fertility (1 male / 2 females / cage) and produced neonatal male rats.The offspring rats were divided into five main groups: Group I (Control offspring group), Group II (Experimental low dose treated offspring group with Bisphenol A (0.3 mg/kg BW./day) for two weeks, Group III (Experimental high dose treated offspring group; 12 mg/kg b.w/day), Group IV (First recovery group ), Group V (Second recovery group), Results: Chronic Bisphenol A administration produced testicular toxicity leading to marked serious histological changes of the testis including the spermatogenic cells, spermatids, Sertoli cells and Leydig cells. These testicular changes increased with the high dose intake of BPA indicating its cumulative toxic effects that caused hypospermatogenesis, leading to infertility. Conclusion: Bisphenol A administration causes testicular toxicity. So, these serious complications on testicular structure should be considered when buying any canned foods or drinks, because it is used nowadays on a large scale allover the world.

Association between a marker of sperm DNA damage and sperm indices in infertile males in Benin City, Nigeria: A cross-sectional study.

Background Studies have shown oxidative DNA damage is associated with male infertility.ObjectiveThis study determines the levels of 8-hydroxy-2'-deoxyguanosine (8-OHdG) and some markers of oxidative stress in seminal fluid of males investigated for infertility and men of proven fertility in Benin City, Nigeria.Materials and MethodsSemen samples produced by self or assisted masturbation were analyzed by microscopic technique according to the World Health Organization guidelines. Thereafter, samples were centrifuged and seminal fluid plasma separated and stored at -20°C prior to assay for 8-OHdG and oxidative stress biomarkers. Based on the sperm concentration/count, the overall samples were grouped into the following categories: normospermia (n = 20), oligozoospermia (n = 30), and azoospermia (n = 20). The control group comprised of 30 age-matched males of proven fertility. The seminal fluid 8-OHdG, total antioxidant status, superoxide dismutase and malondialdehyde (MDA) were assayed through ELISA and spectrophotometric methods, respectively.ResultsSeminal plasma level of 8-OHdG and MDA were significantly higher (p = 0.01) in infertile subjects than controls. The mean levels of 8-OHdG and MDA in infertile subjects were higher in azoospermia than oligospermia than normospermia and so, was least in the normospermia. Conversely, the mean levels of total antioxidant status and superoxide dismutase were significantly lower (p = 0.01) in infertile than fertile the control male subjects with levels higher in normospermia than oligospermia and least in azoospermia. Moreover, the seminal 8-OHdG correlated negatively with sperm count (r = -0.359, p = 0.01), percent motility (r = -0.388, p = 0.04), and percent morphology (r = -0.327, p = 0.02).ConclusionThe assessment of sperm DNA damage in addition to routine seminal fluid analysis may play an important role in specific diagnosis and management of male infertility.

Acute and Reproductive Toxicity of the Aqueous Extract of the Dry Seeds of &amp;lt;i&amp;gt;Aframomum daniellii &amp;lt;/i&amp;gt; on the Female Wistar Rat Strain

This work was designed to investigate the acute and reproductive toxicity activity of the aqueous extract of the dry seeds of Aframomum daniellii on the female rats. The acute toxicity of the aqueous extract of Aframomum daniellii (A. daniellii) was evaluated with 6 female rats which were divided into 2 groups (1 Test group and the Control group) of 3 female rats each. The control group received distilled water (10 mL/kg/po) and the test group received a single dose of extract of A. daniellii at the dose of 2000 mg/kg. The reproductive toxicity was evaluated using 45 adult female rats which were divided into 5 groups. Group I, received distilled water (1 mL/100 g/po, neutral control); group II, received Clomiphene citrate (600 μg/kg/po, positive control); Groups III, IV and V (trials) received aqueous extract of A. daniellii at doses of 100, 200 and 400 mg/kg/po respectively. The animals were treated daily for 14 days. From the 6th day of treatment, the rats were mated with males of proven fertility for 8 days. On day 22, after laparotomy and delivery, the number of implantation sites, corpora lutea, resorption sites and pups were recorded. Concerning the acute toxicity, it was observed that, after the single dose of 2000 mg/kg administration of the aqueous extract of the dry seeds of A. daniellii, no deaths were recorded. Concerning the reproductive toxicity, no implantation and gestation were observed when compared to the control. However, the aqueous extract of A. daniellii caused a significant (p A daniellii is weakly toxic, but could negatively affect some reproductive parameters.

Birth of a Live Cria After Transfer of a Vitrified-Warmed Alpaca (Vicugna pacos) Preimplantation Embryo.

The alpaca (Vicugna pacos) is an important species for the production of fiber and food. Genetic improvement programs for alpacas have been hindered, however, by the lack of field-practical techniques for artificial insemination and embryo transfer. In particular, successful techniques for the cryopreservation of alpaca preimplantation embryos have not been reported previously. The objective of this study was to develop a field-practical and efficacious technique for cryopreservation of alpaca preimplantation embryos using a modification of a vitrification protocol originally devised for horses and adapted for dromedary camels. Four naturally cycling non-superovulated Huacaya females serving as embryo donors were mated to males of proven fertility. Donors received 30 μg of gonadorelin at the time of breeding, and embryos were non-surgically recovered 7 days after mating. Recovered embryos (n = 4) were placed individually through a series of three vitrification solutions at 20°C (VS1: 1.4 M glycerol; VS2: 1.4 M glycerol + 3.6 M ethylene glycol; VS3: 3.4 M glycerol + 4.6 M ethylene glycol) before loading into an open-pulled straw (OPS) and plunging directly into liquid nitrogen for storage. At warming, each individual embryo was sequentially placed through warming solutions (WS1: 0.5 M galactose at 37°C; WS2: 0.25 M galactose at 20°C), and warmed embryos were incubated at 37°C in 5% CO2 in humidified air for 20–22 h in 1 ml Syngro® holding medium supplemented with 10% (v/v) alpaca serum to perform an initial in vitro assessment of post-warming viability. Embryos whose diameter increased during culture (n = 2) were transferred individually into synchronous recipients, whereas embryos that did not grow (n = 2) were transferred together into a single recipient to perform an in vivo assessment of post-warming viability. Initial pregnancy detection was performed ultrasonographically 29 days post-transfer when fetal heartbeat could be detected, and one of three recipients was pregnant (25% embryo survival rate). On November 13, 2019, the one pregnant recipient delivered what is believed to be the world's first cria produced from a vitrified-warmed alpaca embryo.

The leaf aqueous extract of Myrianthus arboreus P. Beauv. (Cecropiaceae) improved letrozole-induced polycystic ovarian syndrome associated conditions and infertility in female Wistar rats

BackgroundMyrianthus arboreus P. Beauv. (Cecropiaceae) is a medicinal plant used to treat female infertility. The aqueous extract of M. arboreus leaves was found to improve the fertility of healthy female Wistar rats. In the present study, we proposed evaluating the effects of such an extract on an animal model of infertility caused by polycystic ovary syndrome (PCOS), in order to bring scientific evidence to the curative action of this plant against female infertility.MethodsFollowing a 21-day administration (gavage) of letrozole (1 mg/kg), animals with PCOS, indicated by overweight and an estrous cycle blocked in the diestrus phase, were co-treated with letrozole (1 mg/kg) and the aqueous extract of M. arboreus leaves at doses of 20, 110 and 200 mg/kg. The positive control received clomiphene citrate (1 mg/kg) and metformin (200 mg/kg). The negative control received distilled water. Each group of animals was made up of 10 female rats. Vaginal smear was examined 7 days before and during co-treatments. Co-treatments were orally administered for 30 consecutive days and 5 animals per group were sacrificed thereafter for biochemical and histological analyses. The 5 remaining animals in each group were crossbred with males of proven fertility for 5 consecutive days. The daily examination of vaginal smears allowed evaluating fertility index. Following parturition, gestation rate was calculated.ResultsThe aqueous extract of M. arboreus leaves reversed letrozole effects by decreasing body weight, abdominal fat accumulation, and serum levels of LH and testosterone (p < 0.001). Ovarian dynamic was improved and the number of tertiary, Graafian follicles (p < 0.001) and corpus luteum increased while that of cystic (p < 0.001) and atretic follicles (p < 0.01) decreased. These effects were associated with increased serum levels of estradiol, decreased ovarian oxidative stress, the resumption of the estrous cycle, the hypertrophy of uterine epithelial cells and increased fertility index and gestation rate.ConclusionsThese results justify at least in part, the traditional use of M. arboreus against female infertility and suggest that this plant could be a promising alternative treatment to improve symptoms associated with different PCOS phenotypes.

Small Ruminants: Prepubertal Oocyte Donors.

In vitro embryo production using oocytes of prepubertal females is named JIVET (juvenile in vitro embryo technologies). The aim of the JIVET is to increase genetic gain by shortening the generation interval in breeding programs. In this chapter we describe the methodology currently used in our laboratory for producing in vitro embryos from prepubertal sheep (90-120days old) and goats (30-45days old).Briefly, we obtain cumulus-oocyte complexes (COCs) from slaughtered female ovaries. These COCs are in vitro matured in TCM199 with 10% (v/v) fetal bovine serum (FBS) at 38.5°C in a humidified air atmosphere with 5% CO2 for 24h. The next day we collect ejaculates from males of proven fertility and select the most motile spermatozoa by density gradient. Following coincubation with mature COCs, presumptive zygotes and embryos are cultured in SOF for 8days at 38.5°C, 90% N2, 5% CO2, 5% O2, and 100% humidity to obtain blastocysts.

58 Transcervical transfer of blastocysts reveals detrimental effect on implantation rate in di (2-ethylhexyl) phthalate-exposed mice

Transcervical transfer is widely used in assisted reproduction techniques, such as cloning and the production of transgenic animals. Using this technique, the number of transferred blastocysts and implanted embryos may be quantified, representing a useful method for assessing the effect of xenobiotics on quality and functionality of the blastocyst. Di (2-ethylhexyl) phthalate (DEHP) is one of the most widely used plasticizers worldwide that causes several detrimental effects, such as decreased embryo quality and a low implantation rate when laboratory animals are treated with very high doses, ranging from 3000 to 3500 µg·kg−1d−1. The aim of this research was to evaluate the effect of environmentally relevant doses of DEHP on blastocyst functionality and ability to implant in an ex vivo study. The CD-1 mice were exposed subchronically (1 month) to 0, 20, 200, and 2000 µg·kg−1d−1 of DEHP via deposition in the mouth, and then they were subjected to a transcervical transfer procedure. The donor female mice were superovulated with pregnant mare serum gonadotropin (5IU) IP, followed by hCG (5 UI) IP 48h later. They were then mated with males of proven fertility. The recipient females (unexposed mice) had the same superovulation protocol as donors, but were mated with vasectomized males. Blastocysts were obtained 96h post-hCG by flushing the oviduct with Gardner-blastocyst medium and were classified according to morphological characteristics. Only expanded blastocysts were transferred transcervically under general anaesthesia protocol using isoflurane (induction: 3-5% and maintenance 2-2.5%). Four days after transcervical transfer, recipient females were injected with 200µL of Chicago Blue 1% IV, killed by cervical dislocation 2h later, following which implantation sites were counted and the implantation rate was calculated with the following formula (implanted blastocysts×100/transferred blastocysts). A decreased implantation rate was evident in 2000 µg·kg−1d−1 DEHP-treated females compared with controls (DEHP 2000: 11.67%±9.64v. control: 71.67%±6.0; P&amp;lt;0.05 according to 1-way ANOVA analysis followed by the Bonferroni post-hoc test). Taken together, these data suggest that the dose of 2000 µg·kg−1d−1 DEPH impairs embryo implantation. We confirm that transcervical transfer is a useful technique in toxicological studies to assess detrimental effects on implantation.

Effect of &lt;i&gt;in utero&lt;/i&gt; exposure to &lt;i&gt;n&lt;/i&gt;-hexane extract of &lt;i&gt;Ricinus communis&lt;/i&gt; var. minor (RICOM 1013J) on postnatal foetal weight changes in Sprague-Dawley albino rats

In traditional medicine practice among the Bassa people of central Nigeria, the seeds of Ricinus communis (Var. minor) are widely used as anticonceptive agents. This study purposes to evaluate the effect of fetal in-utero exposure to extract of Ricinus communis (var. minor) on postnatal growth in rats. Eighteen (18) adult female rats in proestrous phase of the estrous cycle were divided into 3 groups and allowed to mate with males of proven fertility using a male: female ratio 1:3. Group 1 rats were injected subcutaneously on day 2 of pregnancy with 0.2ml of olive oil and served as control, while group 2 animals received 20mg/kg of the extract on day 2 of conception. The rats in group 3 were similarly administered 20mg/kg of the extract subcutaneously on days 2 and 5 of pregnancy. At birth, body weights of pups were taken on days 0, 7, 14 and 28. The results showed that in utero exposure to Ricinus communis var. minor (RICOM-1013-J) significantly decreased postpartum birth weight (p &lt; 0.05) in the first week of life only, while Days 14 and 28 were normal compared to control. The study showed that in utero exposure to therapeutic doses of Ricinus communis var. minor do not have may have no adverse or deleterious effects on foetal development and postpartum growth in rats.Keywords: Ricinus communis; In-utero; n-Hexane; Postnatal; Foetal; Weight changes

Rabbit semen fertility after heterospermic insemination with spermatozoa of different capacitation time

The influence of mixing heterogeneous sperm populations in rabbit semen fertility was studied. Eighty female rabbits which were divided in 4 groups (1,2,3 and 4) in = 20.) and 3 males of proven fertility were employed. The animals of group 1,2 and 3 were inseminated with semen from male A j, A2 and A3, respectively, while those of group 4 were inseminated with a mixture containing equal number of progressively motile spermatozoa from each of the above mentioned males ( A 1 + 2 + 3 ) (heterospermic insemination). Animals of each group were divided into 4 subgroups according to the insemination time (15,10, 5 and 0 hours prior the expected ovulation). In each group, differences were observed (P&lt;0.05) between subgroups at the percentage of animals that delivered, indicating differences between males at the time and the duration of sperm capacitation in vivo. Based on the percentage of the animals that delivered and the litter size, semen fertility was improved after the application of heterospermic insemination, at all insemination times. This improvement was attributed to the extension of the time during which capacitated spermatozoa were present in the female genital tract due to the mixture of heterogeneous sperm populations of different capacitation time.

Effect of the Aqueous Extract of &amp;lt;i&amp;gt;Schumanniophyton magnificum&amp;lt;/i&amp;gt; Harms on Sexual Maturation and Fertility of Immature (K. schum) Female Rat

The aim of the study was to bring scientific evidence of the curative action of the stem bark aqueous extract of Schumanniophyton magnificum (ASMa) on the sexual maturation and fertility of the immature female Wistar rat. Forty immature female rats were randomized and divided into 4 groups of ten animals each and orally treated with the ASMa at doses of 0 (distilled water), 200 mg/kg, 400 mg/kg and 800 mg/kg /BW/day for 30 consecutive days. Body weight and food intake were recorded throughout the experimental period. The precocity of the puberty onset in treated animals was evaluated through the determination of their age at vaginal opening. At the end of the experimental period, 5 animals in each group were sacrificed and blood samples were collected for hormonal assay. Their ovaries and uteri were removed, blotted, weighted and prepared for biochemical analysis. The remaining rats (5 per group) were crossed with males of proven fertility. After laparotomy (ten days after mating) and delivery, fertility and gestational parameters were recorded. It was noticed that, body weight gain increased significantly at all doses although there was no significant difference in food intake. The sexual maturation of treated animal was reduced to 5 days when compared to control. This was associated with the simultaneous elevation of FSH and LH (p mg/kg. However, the ovarian cholesterol significantly decreased (p mg/kg and 800 mg/kg. The animal treated at doses of 400 mg/kg and 800 mg/kg exhibited an early fertilization (2 - 3 days) when compared to the control one (9 - 14 days). The number of implantation site significantly increased (p mg/kg and 800 mg/kg) after laparotomy as well as the number of alive fetuses after delivery and gestational rate (80% and 100%) respectively. These results provide the strong evidence on the induction of the onset puberty and gonadotropin synthesis, the improvement of the ovarian folliculogenesis and the fertility effect of ASMa in immature rats.

Transcervical administration of polidocanol foam prevents pregnancy in female baboons

BackgroundOur objective was to conduct a pilot study to determine if transcervical administration of polidocanol foam (PF) with or without doxycycline or benzalkonium chloride (BZK) would prevent pregnancy in baboons. MethodsIn study phase 1, adult cycling baboons underwent a hysterosalpingogram to evaluate tubal patency prior to transcervical infusion of 20 mL of 5% PF followed by 1 mL of saline containing 100 mg doxycycline (5%/doxy; n=5), 3% PF plus doxycycline (3%/doxy; n=4), 3% PF with 0.01% BZK (3%/BZK; n=4) or no additional treatment (control; n=9). Immediately following treatment, animals received intramuscular depot medroxyprogesterone acetate (DMPA, 2 mg/kg) to suppress cyclicity during healing and were then socially housed with males of proven fertility. The primary outcome was pregnancy within six cycles of resumption of menses (efficacy phase 1). During study phase 2, PF-treated females from study phase 1 contributed additional cycles (6–8) of exposure (efficacy phase 2), and 5 control females who had recovered from medical abortion (after study phase 1 pregnancy) were subsequently treated with 5% PF (with DMPA) and exposed to breeding (efficacy phase 1; n=3 six cycles, n=2 five cycles). ResultsAll females resumed normal menstrual cycles and mating activity after DMPA. During efficacy phase 1, 7/9 (78%) control females became pregnant. In contrast, fewer pregnancies occurred in PF-treated females: 5% PF 0/5 (0%), 5%/doxy 1/5 (20%), 3%/doxy 1/4 (25%) and 3%/BZK 1/4 (25%). During efficacy phase 2, only one additional pregnancy occurred (3%/BZK). ConclusionsA single transcervical treatment with 5% PF prevented pregnancy in most baboons. Cotreatment with doxycycline or BZK did not improve results. ImplicationsTranscervical intrauterine administration of PF resulted in a high rate of tubal occlusion with prevention of pregnancy; refinements are needed to increase the contraceptive rate following a single treatment to near 100%.

A platform for evaluating sperm RNA biomarkers: dysplasia of the fibrous sheath—testing the concept

To investigate the potential use of correlative microarray-based transcript pairs as candidate markers for male fertility using dysplasia of the fibrous sheath (DFS) as an affected model. It is widely recognized that microarray technology may be limited by cost and that the quality of the transcript remains relatively unknown. To address these issues, we analyzed the stable transcript pairs by qPCR with a systematic primer design process. Experimental study. University. Men with proven fertility and men with a diagnosis of DFS. None. Primer sequences for six genes of interest were designed using Oligo7 and Primer3Plus. Primer specificity was initially assessed in silico by searching the ENSEMBL, University of California Santa Cruz, and National Center for Biotechnology Information databases for nontarget complementary sequences throughout the genome. The ability of transcript pairs to classify samples from males of proven fertility away from DFS was assessed. In conjunction with identifying four new stable transcript pairs, comparison of the DFS qPCR C(t) correlation coefficients revealed the disruption of four stable fertile sample transcript pairs. This suite of transcript pairs resolves DFS. The results show that with effectively designed primers, qPCR may provide an affordable molecular assay to assess male fertility status.

Improved Fertilization Success Using Laparoscopic Oviductal Artificial Insemination with Low Sperm Numbers in Domestic Cats.

Artificial insemination (AI) is a valuable tool for infusing genetic variation into stagnant populations, especially using frozen-thawed spermatozoa, and for propagating physically or behaviorally incompatible individuals. Within zoological institutions, AI has received growing interest for genetic management of endangered felids, provided that AI success can be optimized for applied usage. Traditionally, laparoscopic AI in cats involves treatment with equine chorionic gonadotropin (eCG) followed 80-85h later by human CG (hCG) to induce follicular development and ovulation, with subsequent bilateral sperm deposition into the uterine lumen. However, hCG, a large glycoprotein, may remain in circulation for several days post-injection, generating undesirable secondary ovulations. Uterine AI also requires relatively high numbers of spermatozoa to achieve sperm transport through the uterotubal junction and fertilization within the oviduct. Furthermore, sperm recovery from male cats frequently is poor, limiting the number of spermatozoa available for AI. Alterations in the AI protocol, using short-acting porcine luteinizing hormone (pLH) as the ovulatory signal and oviductal AI for sperm deposition, could improve fertilization success while requiring fewer spermatozoa. Our objectives in this study were to assess pregnancy and fertilization success in cats treated with one of two gonadotropin regimens (eCG/hCG vs. eCG/pLH) and inseminated laparoscopically at two sperm deposition sites, the uterus (UT) and the contralateral oviduct (OV). Sixteen female domestic cats were randomly assigned to either eCG (100 IU)/hCG (75 IU) or eCG/pLH (1000 IU) treatment groups. All 16 females ovulated following gonadotropin treatment and were inseminated with low sperm numbers (1x106 motile sperm/site; 5 µl volume) in one uterine horn and one contralateral oviduct using freshly collected semen from a different male for each site. Semen samples were obtained from two males of proven fertility via artificial vagina. Pregnant females were spayed at 20-21 days post-AI and recovered fetuses assessed for paternity using genetic analysis. Comparing gonadotropin regimens, similar numbers of females became pregnant following eCG/hCG (75%, 6/8) versus eCG/pLH (63%, 5/8). The number of corpora lutea (CL) at AI was similar between regimens, but hCG treatment increased the number of CLs at day 20 post-AI. Although hCG and pLH treatments produced similar numbers of normal fetuses, implantation abnormalities (e.g. empty gestational sacs, malformed placentae) were observed in the hCG, but not pLH, group. Overall, 11 (of 16; 69%) females became pregnant (5 cats from OV AI only, 2 from UT AI only, and 4 from both sites). In comparing insemination sites, more fetuses resulted from OV AI (36/49; 73%) than UT AI (13/49; 27%). To assess capacity for term pregnancies, three additional females were treated with eCG/pLH and inseminated in one oviduct and the contralateral uterine horn. All three females became pregnant (2-3 fetuses each) and healthy kittens were produced. In summary, laparoscopic oviductal AI with low sperm numbers in eCG/pLH-treated females resulted in high pregnancy and fertilization percentages in domestic cats. These findings suggest that this technique may have value for propagating endangered nondomestic cat species. Our recent success in using oviductal AI in eCG/pLH-treated ocelots (Leopardus pardalis) to produce a healthy ocelot kitten supports this cross-species applicability. (platform)

Evaluation of potential embryo toxicity of albendazole sulphoxide in CF1 mice

Benzimidazole compounds are used in both humans and animals for controlling helminth parasites. Albendazole has teratogenic effects attributed to its active metabolite albendazole sulphoxide. The aim of this work was to evaluate the effect of the latter compound when administered to pregnant CF1 mice during the preimplantation period. Females were superovulated by intraperitoneal injection of 10 IU of eCG and 10 IU of hCG (48h later) and were paired with males of proven fertility. Albendazole sulphoxide (200 mg/kg) was orally administered by gavages at day 1, 2 or 3 of pregnancy; the control group received only the vehicle (carboxymethylcellulose). Females were killed by cervical dislocation at day 4 of pregnancy and embryos were flushed from uteri with Ham F10 media supplemented with bovine serum albumin (0.4%). Number of collected embryos per female, percentage of morphologically normal embryos, differentiation rate and number of cells per embryos were recorded. The variables were analyzed on a per litter basis by Kruskal-Wallis test. There was no effect of albendazole sulphoxide on parameters evaluated (P>0.05). We conclude that the preimplantation mouse embryo development was not significantly affected by albendazole sulphoxide.

Effects of Dietary Urea on timing of embryo cleavages and blood components in Mice

The objectives of the present study were to investigate the effects of dietary urea supplementation (1.0% and 3.0%) on oocytes quality, timing expected of embryo cleavages, offspring numbers and weights, blood components and rectal temperature in mice. Sixty of growing albino mice were classified into three groups; the control group was given basal control diet and the other two groups were fed on basal control diet supplemented with 1.0% and 3.0% urea. Body weights were recorded at the beginning and after 5 weeks. Thereafter, five female mice of each group were injected with 7.5 IU of pregnant mare serum gonadotropin (PMSG) for determination of oocyte quality after 48h of injection. The fifteen female mice of each group were injected with 7.5 IU of PMSG followed by 7.5 IU of human chorionic gonadotropin (hCG) after 48h and mated with males of proven fertility. Five mated females of each group were used for determination of embryo cleavages to four cell stage and the other five mated females were used for determination of embryo cleavages to eight cell stage upon 59-60 and 70 h of hCG injection, respectively. Rectal temperatures were recorded and blood samples were collected. The remaining five mated females of each group were left for parturition. The offspring number, litter size and male:female ratio were recorded. Hematocrit and hemoglobin concentrations were determined in blood whereas urea, total protein, albumin, glucose calcium and phosphorus concentrations were determined in plasma. The results indicated that offspring number and weight of litter size at birth were significantly (P<0.05) increased in the urea groups compared to control group. Percentage of good quality oocytes was high (70%) in control group compared to 3% urea group (60%). Dietary 3% urea was delayed cleavages to four-cell stage embryos at the expected time. Dietary urea was significantly (P<0.05) increased concentrations of hematocrit and hemoglobin in blood and urea, total protein, globulin, glucose, potassium and phosphorus in plasma. In conclusions, although 3% dietary urea decreased oocytes quality and timing expected of embryo cleavages to four cell stages, it increased significantly (P<0.05) offspring number and weight of litter size.