The inhibition of sperm cAMP and ATP levels, using an FDA-approved medication, may impair sperm motility and, consequently, fertilization, thus paving the way for the development of a male contraceptive. The objective of this study was to define the potential impact of dipyridamole (Persantine®) on sperm functions. Spermatozoa collected from mature Swiss albino mice were incubated for up to 7 h with dipyridamole concentrations of 0, 2.5, or 25 μg/mL. Computer-assisted sperm motion analysis showed that dipyridamole inhibited sperm motility and progressive motility (P < 0.003), as well as average path velocity, straight-line velocity, curvilinear velocity, and amplitude of lateral head displacement (P < 0.01), in a dose- and time-dependent manner. Sperm levels of cAMP and ATP showed similar patterns of decline (P < 0.02). Dipyridamole also inhibited sperm capacity to produce 2-4 cell embryos and blastocysts (P < 0.0001). Dipyridamole inhibited sperm motility, fertilization, and levels of ATP and cAMP at 2.5 μg/mL concentration, which is similar to those concentrations observed in pharmacokinetic studies of men taking dipyridamole therapeutic doses of 150-200 mg once or twice daily. These results suggest that dipyridamole at therapeutic doses could have contraceptive or adverse effects in men if the medication produces similar inhibitory effect on human sperm functions. Currently, however, there have been no reports of contraceptive or adverse effects in men taking dipyridamole, although it is possible that most men are past the normal reproductive age when taking this antiplatelet medication.

This study compared the effect of a 7-day estradiol- and a 7-day GnRH-based FTAI protocol versus a presynchronized GnRH-based FTAI protocol (7 & 7 GnRH) on pregnancy per AI (P/AI) in multiparous postpartum Angus cows. Cows were blocked according to presence/absence of a CL, body condition score, and days postpartum and assigned randomly to one of three treatments: I) 7-day EB (n = 368), estradiol benzoate (EB) and intravaginal progesterone-(P4)-releasing device (IVPD) on Day -10, prostaglandin F2α (PG), equine chorionic gonadotropin (eCG) and estradiol cypionate (ECP) at IVPD removal on Day -3 with FTAI done on Day -1 (54 ± 2 h after IVPD removal); II) 7-day GnRH (n = 367), GnRH at IVPD insertion on Day -10, PG and eCG at IVPD removal on Day -3 with GnRH and FTAI on Day 0 (66 ± 2 h after IVPD removal); and III) 7 & 7 GnRH (n = 361), PG at IVPD insertion on Day -17, GnRH on Day -10, IVPD removal, PG and eCG on Day -3 with GnRH and FTAI on Day 0 (66 ± 2 h after IVPD removal). Preovulatory follicle diameter, corpus luteum (CL) presence, CL area, and P4 concentration were determined at IVPD removal, as well as estrus expression at FTAI and P/AI between 35 and 40 days after FTAI. Plasma P4 concentrations at IVPD removal were greater (P < 0.05) in cows subjected to the 7-day GnRH or 7 & 7 GnRH protocols and a greater number of cows (P < 0.05) had at least one CL at IVPD removal compared to cows subjected to the 7-day EB protocol. However, the 7 & 7 GnRH protocol resulted in more cows with two CLs at IVPD removal compared to the other two protocols. Moreover, cows synchronized with the 7 & 7 GnRH protocol had larger preovulatory follicles (P < 0.05) and larger CL area at IVPD removal (P < 0.05), and a greater rate of estrus expression before FTAI (P < 0.05) than cows synchronized with the 7-day EB, whereas those variables were intermediate in cows synchronized with the 7-day GnRH. Pregnancy per AI was greater (P < 0.05) in cows treated with the 7 & 7 GnRH (63.5 %) treatment compared to the 7-day EB (51.4 %) and the 7-day GnRH (51.3 %) protocols. In summary, the 7 & 7 GnRH protocol enhanced circulating P4 before IVPD removal, increased estrus expression rate before FTAI, and improved P/AI in postpartum Bos taurus grazing beef cows.

Objective: Evaluate the White-tailed Deer (WTD) in vitro embryo production (IVP) and oocytes vitrified with Trehalose (TH) or Sucrose (SC). Design/methodology/approach: Total vitrified oocytes were placed into two different groups: TH (n=60) and SC (n=61). Samples were selected and analyzed for viability evaluation TH (n=5) and SC (n=5), nuclear status (NS) TH (n=4) and SC (n=5), Germinal Vesicle (GV), Metaphase I, or not evaluable (NE) after warming. In vitro maturation (IVM) was conducted for 36 h in supplemented TCM-199 medium. Immediately afterwards, oocyte NS was evaluated (n=88) [(GV, MI=immature), (MII=mature)]. In vitro fertilization (IVF) was performed in supplemented TALP medium for 24 h using frozen WTD semen (3x106 sperm/mL), NS was classified [Fertilized (F), Not fertilized (NF), or NE]. Results: After warming, viability for the TH group (n=5) was 60% versus 40% for SC group (n=5), however, oocytes in both groups were immature (GV and MI stage). For IVM, NS evaluations of the TH group (n=38) revealed no maturation versus 2% in the SC group (n=50) (MII stage=matured). IVF evaluations for the TH group (n=10) revealed no fertilization compared to 20% in the SC group (n=5). A statistical difference (p>0.05) was not found between the TH and SC groups. Limitations on study/implications: White-tailed Deer in vitro embryo production is not well documented. Findings/conclusions: Future research with a larger number of WTD oocytes is needed for further evaluation of oocyte vitrification IVP techniques as a model for endangered cervids.

Fixed-timed artificial insemination (FTAI) protocols for beef cattle in South America are primarily based on estradiol esters and intravaginal progesterone-releasing devices (IVPD). The objective of this study was to determine the optimal gonadotropin-releasing hormone (GnRH)-based protocol as an alternative to the use of estrogen-based protocols in grazing Bos taurus suckling beef cows. All cows received an IVPD on the day of protocol initiation and prostaglandin F2α (PG) plus equine chorionic gonadotropin (eCG) treatments at the time of IVPD removal. In Experiment 1, cows (n = 235) were randomly assigned to one of four treatments: (i) 7-day estradiol = 2 mg of estradiol benzoate (EB) at IVPD insertion on Day 9 and 1 mg of estradiol cypionate (ECP) at IVPD removal on Day 2; (ii) 7-day GnRH = 10 µg of GnRH at IVPD insertion on Day 10, IVPD removal on Day 3 and GnRH at FTAI; (iii) 7 & 7 estradiol = PG at IVPD insertion on Day 16, EB on Day 9 and ECP at IVPD removal on Day 2; (iv) 7 & 7 GnRH = PG at IVPD insertion on Day 17, GnRH on Day 10, IVPD removal on Day 3 and GnRH at FTAI. In Experiment 2, cows (n = 462) were randomly assigned to one of four treatments: (i) 6-day estradiol = EB at IVPD insertion on Day 9, IVPD removal on Day 3 and GnRH at FTAI; (ii) 7-day estradiol; (iii) 7-day GnRH; (iv) 7 & 7 GnRH. In Experiment 1, plasma progesterone concentrations and percentage of cows with a corpus luteum (CL) at IVPD removal, and pregnancy per AI (P/AI) were greater for cows subjected to GnRH-based protocols compared with cows subjected to estrogen-based protocols (p < 0.01). In Experiment 2, cows subjected to the 7 & 7 GnRH protocol had the greatest P/AI (p < 0.01). In summary, GnRH-based FTAI protocols resulted in similar or greater P/AI compared to estrogen-based FTAI protocols in grazing postpartum Bos taurus suckled beef cows. The greatest P/AI was attained with the 7 & 7 GnRH protocol.

Reproduction, Fertility and Development is an international journal publishing original research , review and comment in the fields of reproduction and developmental biology in humans, domestic animals and wildlife

Reproduction, Fertility and Development is an international journal publishing original research , review and comment in the fields of reproduction and developmental biology in humans, domestic animals and wildlife

Transvaginal ultrasound-guided oocyte retrieval (commonly called OPU) and in vitro embryo production (IVP) in cattle has shown significant progress in recent years, in part, as a result of a better understanding of the full potential of these tools by end users. The combination of OPU and IVP (OPU-IVP) has been successfully and widely commercially used worldwide. The main advantages are a greater number of embryos and pregnancies per unit of time, faster genetic progress due to donor quick turn around and more elite sires mating combinations, larger spectrum of female age (calves, prepuberal, heifer, cow) and condition (open, pregnant) from which to retrieve oocytes, a reduced number of sperm (even sexed) required to fertilize the oocytes, among other benefits. OPU-IVP requires significant less donor preparation in comparison to conventional embryo transfer (<50% of usual FSH injections needed) to the extent of no stimulating hormones (FSH) are necessary. Donor synchronization, stimulation, OPU technique, oocyte competence, embryo performance, and its impact on cryopreservation and pregnancy are discussed.

Reproduction, Fertility and Development is an international journal publishing original research , review and comment in the fields of reproduction and developmental biology in humans, domestic animals and wildlife

Objective: Evaluate the use of Ethylene Glycol (EG), Dimethyl Sulfoxide (DMSO), Sucrose and Fetal Bovine Serum (FBS) as cryoprotectants and their effect on the organization of chromosomes and the arrangement of microtubules, during the vitrification process in goat oocytes matured in vitro and in the development of preimplantation embryos produced in vitro. Design/methodology/approach: In vitro matured oocytes were divided into 3 groups (control group, cryoprotectant exposed group, vitrified group). A mixture of 15% EG, 15% DMSO, 0.4 M sucrose and 20% FBS was used for the vitrification using the Cryotop device. In vitro matured oocytes were warmed and afterwards each group was divided into two more groups. Both groups were subjected to immunofluorescence, the first group to observe the damage produced to the chromosomes and microtubules and the second group to observe the effect on the in vitro embryo development. Results: The combined use of 15% EG, 15% DMSO, 0.4 M Sucrose and 20% FBS during vitrification did not prevent cryoinjuries in goat oocytes and in vitro produced embryos, since embryo development was disrupted before the blastocyst stage by stopping cleavage at the morula stage. This disruption was associated with chromosome decondensation and the absence of a microtubule network, thereby hindering chromosomal segregation. Limitations on study/implications: The effect of conventional cryoprotectants on chromosomes and microtubules arrangement on vitrified goat oocytes and in vitro embryo production. Findings/conclusions: The combined use of 15% EG, 15% DMSO, 0.4 M sucrose and 20% FBS as vitrification cryoprotectants did not prevent cryoinjuries in caprine oocytes and did not improve caprine embryo development in vitro.

Reproduction, Fertility and Development is an international journal publishing original research , review and comment in the fields of reproduction and developmental biology in humans, domestic animals and wildlife

The purpose of this study was to compare the maturation, cellular division, and blastocyst production rates of water buffalo oocytes at 18, 21 and 24 hours of incubation. These oocytes were obtained through follicular puncture, assessed, and matured in BO-IVM medium at 38.5°C, and with a 5% of CO2, at 18 (n=31), 21 (n=34) and 24 hours (n=33). Fertilization was done in a BO-IVF medium at 38.5°C, and with a 5% of CO2, for 18 hours; the maturation rate was assessed a day after fertilization. The embryos were cultured in a BO-IVC medium, at 38.5°C, 5% CO2, 7% O2 and 88% N2, and the cell division rate and number of embryos were assessed at 5 and 7 days after fertilization, respectively. The maturation rate was comparable (P>0.05) among the three groups, while blastocyst production and cell division rates were higher in the group of oocytes matured for 18 hours (P<0.05) than in the 24-hour group. The blastocyst production and cellular division rates in oocytes that were matured for 21 hours were comparable (P>0.05) to the other groups. After 21 hours of maturation, oocytes started to degenerate, which resulted in lower cellular division and blastocyst production rates.

Global oyster abundance has declined ~85 % over the past 200 years, primarily because of overharvesting (Beck, Brumbaugh, and Airoldi 2011; Kirby 2004). Healthy oyster reef systems benefit the environment in many ways, including water-quality improvement, shoreline protection, increased biological and habitat diversity, and carbon sequestration. To maintain these environmental benefits, reef-restoration efforts that produce healthy, sustainable oyster reefs are essential. To this end, the US Army Corps of Engineers (USACE) has been involved in reef-restoration projects in many locations, including extensive efforts in the Chesapeake Bay (Virginia, Maryland), coastal regions of New York and New Jersey, and the Gulf of Mexico. There are many benefits to creating and maintaining oyster reef systems that are well connected, for both oysters and other organisms within the reef and surrounding habitats. This technical note presents the current knowledge of benefits and costs to restore oyster-reef connectivity along the East and Gulf Coasts of North America. Connectivity of oyster reefs can refer to the physical location of reefs with respect to one another as well as to the dynamics of the genetic links within a metapopulation or to the extent to which larval transport and recruitment unite reef communities. For the purposes of this technical note, connectivity is defined as the spatial aggregation of reefs, though we address impacts of genetic and larval flow as well. Reef connectivity positively affects many ecosystem services and dynamics but can also have unintended consequences (that is, negative externalities). This technical note reviews the benefits and costs of increasing connectivity and presents a brief example of how trade-offs may occur between these potentially opposing ecological objectives. Here, we focus on the eastern oyster, Crassostrea virginica, which inhabits the East and Gulf Coasts of North America, though many of the concepts and principles discussed may apply to other oyster species as well.

Reproduction, Fertility and Development is an international journal publishing original research , review and comment in the fields of reproduction and developmental biology in humans, domestic animals and wildlife

Domestic cat invitro embryo production (IVEP) begins with IVM of oocytes to produce mature oocytes; that is, MII. The domestic cat (Felis catus) has been used as a model to carry out assisted reproductive technology (ART) research for application in wild feline species that may be threatened or endangered. The objective of this research was to evaluate oocyte maturation of domestic cats in different reproductive stages: (1) prepubertal, (2) oestrus, (3) pregnant, and (4) anoestrus. The present study was carried out at the Universidad Autónoma Metropolitana Unidad Xochimilco in Mexico City. Unless otherwise stated, all reagents used were from Sigma-Aldrich. The domestic cat ovaries were obtained from a veterinary clinic using salpingo-oophorectomy hysterectomy (OSH). Ovaries were classified as one of the following: (1) prepubertal (female cats under 6 months of age); (2) in oestrus (one or more 2-mm mature follicles); (3) pregnant (presence of fetuses with one or more corpora lutea; and (4) anoestric (ovaries without follicular activity). The ovaries were transported (<2h) in NaCl solution (0.157M) with ampicillin (10 000 IU mL−1), streptomycin (10 000 µg mL−1) and amphotericin (25µg mL−1) to the laboratory. The cumulus–oocyte complexes (COCs) were obtained by ovary microdissection with modified Tyrode’s medium supplemented with sodium lactate (10mM), HEPES (0.50mM) and polyvinyl alcohol (0.01%). COCs were washed twice with TCM-199 medium with Earle’s salts supplemented with bovine serum albumin (BSA, 3mg mL−1), cysteine (0.1mg mL−1), HEPES (1.4mg mL−1), sodium pyruvate (0.25mg mL−1), sodium lactate (0.6mg mL−1), L-glutamine (0.15mg mL−1) and gentamicin (0.055mg mL−1). The wash medium was also used for IVM, but supplemented with human menopausal hormone (Merional® IBSA; 4.5IU mL−1). Oocyte maturation was performed with TCM-199 medium supplemented with BSA, in an atmosphere of 38.5°C, 5% CO2, 95% air, and humidity at saturation for 48h. To evaluate IVM, 300μg mL−1 of hyaluronidase was used to remove the granulosa cells for 5min at 38°C. Next, the oocytes were fixed with paraformaldehyde (4%) for 15 min; washed with a mounting solution (Imacel, invitro); then, 1.5μg mL−1 of 4’,6 diamidino-2-phenylindole dihydrochloride (DAPI) was added. The stained oocytes were evaluated under a microscope (Eclipse E600, Nikon) equipped with a fluorescence lamp and a UV filter (excitation: 330–380nm). The Student’s t-test and the Chi-squared test (χ2) were used for statistical analyses (α=P<0.05). A total of 210 ovaries were obtained from 105 female cats: prepubertal (n=38), oestrus (n=25), pregnant (n=18), and anoestrus (n=24), with a total of 1405 oocytes recovered. The meiotic maturation between the different reproductive stages after 48h of culture was similar in prepubertal (48%), oestrus (46%), pregnant (43%), and anoestrus (45%) groups and did not show a significant difference (P>0.05). This study shows that the domestic cat reproductive stage does not significantly affect the production of mature oocytes for use in ART.

The domestic cat has been used as a model to carry out comparative research in assisted reproduction, to be applied in wild cats.The efficiency in domestic cat IVM concerning the reproductive status and/or cultivation times has previously been investigated; however, the studies were carried out separately.The objective of this research was to evaluate the maturation of oocytes of domestic cats of different reproductive stages using two different in vitro culture times.The ovaries were obtained by Ooforo-Salpingo-Hysterectomy of cats that were of the following groups: 1) prepubertal, 2) follicular, 3) pregnant or 4) in anestrus.Maturation was carried out with TCM199 medium supplemented with BSA for 24 h and 48 h.On average, 29 ± 25, 20 ± 15, 17 ± 9 and 17 ± 13 oocytes/cat were recovered from the prepubertal follicular, pregnant, and anestrus stages, respectively, but did not show a significant difference (P > 0.05).Also, meiotic maturation did not show a significant difference between the different reproductive stages at 24 h and 48 h, respectively (P > 0.05).However, in the prepubertal and follicular stages, greater oocyte maturation numbers were observed at 48 h compared to 24 h (P < 0.05).In contrast, the aforementioned result was not observed in the pregnant and anestrus stages (P > 0.05), indicating that the in vitro culture duration is an important factor during in vitro maturation of domestic cat oocytes.

This report assesses potential impacts to aquatic resources resulting from proposed navigation channel expansion activities within Mobile Bay, Alabama.This work was conducted for the U.S. Army Corps of Engineers (USACE) Mobile District, to support development of a supplemental Environmental Impact Statement.Changes in water quality and hydrodynamics were evaluated for potential impacts to benthic macroinvertebrates, wetlands, submerged aquatic vegetation, oysters, and fish.The assessment includes extensive characterization of baseline conditions, evaluation of estimated post-project conditions related to aquatic resource habitat (e.g., changes in salinity, dissolved oxygen).An analysis of potential impacts related to a 0.5-m sea level rise (SLR) scenario were also evaluated.Results suggest that no substantial impacts in aquatic resources within the study area are anticipated due to project implementation, as the area of greatest potential changes to environmental conditions are already adapted to natural shifts in salinity (and other factors), and to conditions resulting from the existing navigation channel.Although SLR has the potential to alter aquatic resource habitats with Mobile Bay, additional impacts related to project implementation remain negligible under the 0.5-m SLR scenario.