Mating Fertility Research Articles

Sperm Storage and Use Following Multiple Insemination in Aedes albopictus: Encouraging Insights for the Sterile Insect Technique.

The key to success in the application of the sterile insect technique (SIT) relies on the ability of released, sterile males to outcompete their fertile wild male counterparts to mate with wild females. However, many insect species exhibit multiple-mating behavior, which can be a way for females to select paternity for their progeny. This study aims to recognize the consequences of potential double-matings during an SIT program and to detect any evidence of sperm selection favoring sperm from fertile mates. This report provides a descriptive analysis of the storage and use of sperm by female Aedes albopictus. Stable isotopes were used to mark the sperm of fertile and sterile males. Mated females were allowed to oviposit before dissecting the spermathecae to link the presence of each type of sperm to the sterility of the eggs laid. It was found that sperm in females inseminated by both males was distributed in the three spermathecae with no obvious pattern, mostly mixed but also separately, and no evidence of any mechanism for sperm selection, sperm precedence, or sperm competition in Ae. albopictus females could be found. The fact that only a few double-mated females were double-inseminated and could also produce semi-sterile eggs, together with the finding that the sperm of sterile males appeared to be no less viable than that of fertile males, is an encouraging outcome for SIT approaches.

A retrospective study on whelping seasonality of the Cirneco dell’Etna dog in Sicily, Italy

ABSTRACT Previous to this study, there is no data relating to the reproductive physiology of the Cirneco dell’Etna breed. The birth date of 141 bitches and 104 sires registered at the National Agency of the Italian Kennel Club (ENCI) from all Sicilian breeders, and data related their fertile matings, which produced 266 whelps over a 14-year period, (2009–2022) were collected. Total whelping produced 1069 pups. The mean bitch age at first whelping was 39.13 ± 21.7 months and the mean litter size was 3.84 ± 2.08. The birth period of the bitches and sires was significantly influenced by the month of the year (p < 0.01), as was the distribution of the first (p < 0.05) and subsequent whelping periods (p < 0.01). Whelping distributions show a significant difference between the highest peak in April (which roughly corresponds to a period of heat in February) and in July (which roughly corresponds to a period of heat in May) and the lowest peak in March. No influence of month on litter size and sex ratio was found. Available data suggests that some ancient breeds such as the Cirneco dell’Etna may show seasonal patterns as a vestige of the stricter seasonality of its closest relative the wolf.

Effect of Age at Breeding on Female African Giant Rat (Cricetomys gambianus) Fertility in Captivity

This study aimed to evaluate the influence of age at breeding one female African giant rat (FAGR) fertility. Twenty-four nulliparous FAGR of 6, 8, or 10 months old were allocated to one of the three groups according to their age. They were housed individually and fed ad libitum. After five days of accommodation, a male was transferred into a female cage, and daily vaginal smears were performed to evidence mating for a period not exceeding 30 days. Reproductive parameters were then evaluated. Results showed a significant reduction (P&lt;0.05) in pre-coitus cohabitation time in 8 (6.75±4.23days) and 10 (5.13±1.25days) months old females as compared to those aged six months (22.25±1.75days). There was an increase in mating rates, fertile mating, fertility, and stillbirth in 8 and 10 months old FAGR. The litter size significantly increased (P&lt;0.05) in 8 and 10 months old compared to 6 months old FAGR. Preimplantation embryo losses rates were 60.00±39.44, 66.66±34.96, and 35.00±21.19% in 6, 8, and 10 months old FAGR, respectively. In this study, fertility increased with age, so it was better in the FAGR older than six months.

A Novel Genetic Sexing Strain of Anastrepha ludens for Cost-Effective Sterile Insect Technique Applications: Improved Genetic Stability and Rearing Efficiency.

Simple SummaryTephritid flies, including the Mexican fruit fly Anastrepha ludens, are key agricultural pests responsible for billions of dollars of damage each year due to the female flies which lay eggs and develop maggots in the fruits of hundreds of species of economically-important plants. Integrated pest management practices such as the sterile insect technique were developed which rely on the mass rearing and release of millions of sterile males of the same pest species in order to suppress the pest reproductive capacity. The presence of females in early bisexual strains often caused high rearing and release costs plus damage to crops by oviposition. Therefore, using classical genetic tools, genetic sexing strains were developed for some fruit flies in order to produce male-only colonies which enhanced the technique efficacy, cost-efficiency and reduced undesirable side-effects such as fruit damage by the mass reared females. In this paper the process for the development and characterization of a new genetic sexing strain (GSS) for A. ludens is documented, using the low-dose irradiation and line selection techniques. The new GUA10 GSS, when compared with its predecessor Tapachula-7, promises to further increase the sterile insect technique performance given its higher quality, yield and genetic stability.Anastrepha ludens (Loew) is one of the most destructive insect pests damaging several fruits of economic importance. The sterile insect technique (SIT) is used under an area-wide integrated pest management approach, to suppress these pest populations. Mass rearing facilities were initially established to produce sterile males of bi-sexual strains in support of SIT. The first genetic sexing strain (GSS) for A. ludens, Tapachula-7, based on pupal color dimorphism, was a key development since the release of males-only significantly increases the SIT efficiency. In this study, we document the development of a novel pupal color-based GSS. Twelve radiation-induced translocation lines were assessed as potential GSS in terms of recombination rates and rearing efficiency at a small scale. The best one, GUA10, was cytogenetically characterized: it was shown to carry a single translocation between the Y chromosome and chromosome 2, which is known to carry the black pupae marker. This GSS was further evaluated at medium and large scales regarding its genetic stability, productivity and quality versus Tapachula-7. GUA10 presented better genetic stability, fecundity, fertility, production efficiency, flying ability, and male mating, clear indicators that GUA10 GSS can significantly improve the efficacy and cost-effectiveness of SIT applications against this pest species.

Social context and noise affect within and between male song adjustments in a common passerine

AbstractAcross populations, animals that inhabit areas with high anthropogenic noise produce vocalizations that differ from those inhabiting less noisy environments. Such patterns may be due to individuals rapidly adjusting their songs in response to changing noise, but individual variation is seldom explored. Therefore, we tested the hypothesis that male house wrens (Troglodytes aedon) immediately adjust their songs according to changing noise and that social context further modifies responses. We recorded songs, quantified noise, and defined social context within pairs as female fertile status and between males as number of conspecific neighbors. We used a reaction-norm approach to compare song trait intercepts (between-male effects) and slopes (within-male effects) as a function of noise. Individuals immediately adjusted song duration in response to changing noise. How they achieved adjustments varied: some sang shorter and others longer songs with greater noise, and individuals varied in the extent to which they adjusted song duration. Variation in song duration could be affected by competition as between-male noise levels interacted with number of neighbors to affect syllable duration. Neither within- nor between-male noise effects were detected for frequency traits. Rather, males with fertile mates sang lower-frequency songs and increased peak frequency with more neighbors. Among males, social context but not noise affected song frequency, whereas temporal structure varied between and within individuals depending on noise and social factors. Not all males adjusted signals the same way in response to noise, and selection could favor different levels of variation according to noise.

Deslorelin subcutaneous implants in Oryx dammah males for reproductive control

The aim of this study was to assess the effects of the deslorelin subcutaneous implant as a temporary contraceptive method in the Oryx dammah male. For this purpose, deslorelin at different doses, i.e. 14.1 mg and 9.4 mg, was subcutaneously implanted in three males (Phase 1) and one male (Phase 2) adult Oryx dammah, respectively. Quantitative behavior evaluation and androgen concentrations in feces and plasma were assessed before and after implant application. Fecal androgen concentrations observed in treated males were compared with those measured in one orchiectomized male and two females. Fecal androgen concentrations increased up to 15 days after the implant application, then progressively decreased, reaching the basal level at day150 in Phase 1. In Phase 2, levels remained high until day 60 and returned to basal level on day 120. Plasma testosterone concentration was higher on the day of implant application than three months later, but with variable ranges among males. A general increase of activity levels and hierarchical changes were observed after treatment, in accordance with hormonal variations. Despite males cohabiting with two fertile females during the observation period, no births were recorded. However, between the end of Phase 1 and the beginning of Phase 2, i.e. about 10–11 months after the first deslorelin implant, a fertile mating occurred leading to the birth of a calf. Therefore, we can hypothesize a contraceptive effect up to 10 months after the implant. Testicular histology performed on one male at the end of the Phase 2 showed no spermatogenetic activity.Our results suggest that deslorelin implant can be used to temporarily control reproduction in the Oryx dammah male. Behavior and fecal androgen measurements were useful and repeatable, non-invasive methods to monitor response.

Sexual selection for low male voice pitch among Amazonian forager-horticulturists

Abstract Pitch is the most perceptually salient feature of the voice, yet it is approximately five standard deviations lower in men than in women, a degree of sexual dimorphism exceeding that of all extant nonhuman apes. Evidence from Western samples suggests that low-frequency vocalizations may have augmented male mating success ancestrally by intimidating competitors and/or attracting mates. However, data are lacking from small-scale societies. We therefore investigated sexual selection on male pitch (measured by fundamental frequency, fo) in a population of Bolivian forager-horticulturists, the Tsimane. We found that experimentally lowering fo in audio clips of men speaking increased perceptions of fighting ability but did not affect perceptions of prestige and decreased their attractiveness to women. Further, men with lower speaking fo reported higher numbers of offspring, and this was mediated by the reproductive rates of men's wives, suggesting that men with lower fo achieved higher reproductive success by having access to more fertile mates. These results thus provide new evidence that men's fo has been shaped by intrasexual competition.

Reproduction in female South American domestic camelids

Alpacas and llamas are induced ovulators. They show marked reproductive seasonality in the Andean region, but under Northern Hemisphere conditions of feeding and management, they are non-seasonal breeders. Puberty is attained when they reach 50% of adult body weight. When they are not exposed to a male, females show successive waves of follicular maturation and atresia. Growth, maintenance and regression of a follicle each require an average of 4 and 6 days in alpacas and llamas, respectively. After sterile mating, progesterone concentrations in blood were increased from day 5, reached maximum concentrations on day 7-8, and declined rapidly at 9-10 days after mating. A fertile mating results in formation of a corpus luteum that remains functional throughout gestation. The duration of gestation is 340-346 days. Almost all fetuses were found to occupy the left uterine horn, even though ovulation occurs from both ovaries with equal frequency. Several methods of pregnancy diagnosis have been described. Mating is recommended within 15-20 days after parturition to obtain good fertility rates and one offspring per year. The factors that contribute to high rates of embryonic mortality are unknown. Reproductive technologies, such as AI, superovulation, embryo transfer and IVF, have not been used very extensively in these species but can be successfully applied.

112 Comparative study between slow freezing and vitrification on the survival rate of cryopreserved alpaca embryos post-transfer

The objective of this study was to compare the effectiveness of cryopreserving in vivo-produced alpaca embryos by slow freezing v. vitrification. The embryos were produced from 9 female alpacas at Fundo Mallkini, Puno, Peru, located at 4300m elevation. The donor alpacas were synchronized by induction to ovulate with an injection of gonadotropin-releasing hormone (0.0084mg of buserelin acetate) and natural mating with vasectomized males to male receptive donors (day of ovulation induction was considered Day 0). On Day 2, the donors were injected 700IU of eCG. On Day 7, the donors received an injection of prostaglandin F2α (0.25mg of cloprostenol) and were mated on Day 8 by fertile males (2 matings 12h apart: 0600 and 1800h). The embryos were collected at 5.5 days after fertile mating and were graded as per IETS recommendations; most of the embryos were already expanded and hatched blastocysts. Embryos were washed and maintained in holding medium (1L PBS+1g Glucose+36mg sodium pyruvate+0.4% BSA+50mg kanamycin monosulfate) at 23°C for up to 1h and distributed into 2 groups for either slow freezing for direct transfer (n=14 embryos) or vitrification (n=10 embryos). Slow freezing consisted of transfer into freezing medium (9mL of 1.5M ethylene glycol+1mL of 1.0M sucrose prepared in holding media) at 23°C, placing in 0.25-mL straws and subjected to freezing at a rate of −0.5°C/ minute to −35°C and then plunging into LN. Vitrification followed a procedure described for camel embryos whereby embryos were exposed to solutions containing increasing amounts of glycerol and ethylene glycol for fixed periods and were then loaded into an open pull straw and plunged directly into LN for storage. The cryopreserved embryos were transferred into adult alpacas at the Community of Suitucancha, Junin, Peru (1500km from the farm where the embryos were collected and cryopreserved, 4200m elevation). Embryos in the slow-freezing group were thawed in warm water at 37°C for 30s and loaded directly into the embryo transfer gun for direct transfer into 7 alpaca recipients (2 embryos per recipient). Vitrified embryos were warmed by removing the open pull straw from the LN and transferring the embryos to 2 warming solutions at 37°C with decreasing levels of vitricants and containing 0.5M galactose with a final incubation at room temperature in holding media and then transferred into 5 alpaca recipients (2 embryos per recipient). The embryos were transferred into synchronized recipients by transcervical nonsurgical method. Pregnancy diagnosis was made by transrectal ultrasound examination at 45 days post-transfer. The pregnancy rates in the slow-freezing and vitrification groups, respectively, were 2/7 (29%) and 0/5 (0%); the difference was not significant (P&amp;gt;0.05) based on Fisher’s exact test. Twin pregnancies were not detected. We consider the result with slow freezing very promising, as in previous trials we had less than 18% pregnancies. More trials with larger number of embryos per cryopreservation method are being programmed.

Effect of emergence time on the reproductive parameters of mass-reared Anastrepha ludens (Diptera: Tephritidae) flies

Abstract In the mass rearing of fruit flies, the adult emergence time is gradual, due to factors such as variation in food consumption in the larval phase, the female oviposition behavior and environmental conditions, among others. Here, we investigate the effect of emergence time on sex ratio, fecundity, fertility, body size and mating performance in mass-reared Anastrepha ludens (Diptera: Tephritidae) flies. The results of the study indicated that the emergence of the flies is gradual and lasts approximately 96 h, but the highest proportion of emerged flies was observed at 48 h. The flies emerging at 24 and 48 h showed higher fecundity than those emerging at 72 and 96 h, but there were no significant differences in the sex ratio, fertility and male mating performance. Body size was significant, the flies that emerged at 24, 48 and 72 h were bigger than those emerged at 96 h. The discussion focuses on the effect of emergence time on the rearing processes and release of the Mexican fruit fly.

Links between parental life histories of wild salmon and the telomere lengths of their offspring.

The importance of parental contributions to offspring development and subsequent performance is self-evident at a genomic level; however, parents can also affect offspring fitness by indirect genetic and environmental routes. The life history strategy that an individual adopts will be influenced by both genes and environment; and this may have important consequences for offspring. Recent research has linked telomere dynamics (i.e., telomere length and loss) in early life to future viability and longevity. Moreover, a number of studies have reported a heritable component to telomere length across a range of vertebrates, although the effects of other parental contribution pathways have been far less studied. Using wild Atlantic salmon with different parental life histories in an experimental split-brood invitro fertilization mating design and rearing the resulting families under standardized conditions, we show that there can be significant links between parental life history and offspring telomere length (studied at the embryo and fry stage). Maternal life history traits, in particular egg size, were most strongly related to offspring telomere length at the embryonic stage, but then became weaker through development. In contrast, paternal life history traits, such as the father's growth rate in early life, had a greater association in the later stages of offspring development. However, offspring telomere length was not significantly related to either maternal or paternal age at reproduction, nor to paternal sperm telomere length. This study demonstrates both the complexity and the importance of parental factors that can influence telomere length in early life.

Breeding in the fat-tailed dunnart following ovarian suppression with the gonadotrophin-releasing hormone agonist Lucrin® Depot.

Lucrin Depot (AbbVie), a 1-month microsphere gonadotrophin-releasing hormone (GnRH) agonist preparation, was investigated as a potential agent to synchronise cycling in the fat-tailed dunnart (Sminthopsis crassicaudata). Forty-eight randomly selected females were treated with 5 or 10mgkg-1 Lucrin Depot (n=24 per dose). Eighteen females per treatment had their reproductive activity scored at 4, 8, 12 and 16 weeks using two ovarian (Graafian follicle and corpus luteum status) and two reproductive tract (uterine and vaginal muscularity and vascularity) parameters that formed a reproductive activity score. Six females per treatment were paired with a male at 4 weeks. Fertility was assessed between 8 and 16 weeks by pouch check, and thereafter by dissection. The effects of the 5 and 10mgkg-1 doses were statistically equivalent. Females showed suppression at 4-8 weeks, an increase in reproductive activity at 8-12 weeks and all were cycling normally at 16 weeks. Six pouch young were born at 12 weeks to two females treated with the 5mgkg-1 dose. Nine embryos were recovered at 16 weeks from two females treated with the 10mgkg-1 dose. In conclusion, Lucrin Depot can suppress breeding, and fertile mating can occur in subsequent cycles in the dunnart. There is potential for Lucrin Depot to be used as an assisted breeding tool, but it may need to be combined with ovarian stimulation treatment to achieve practical levels of synchronisation in the fat-tailed dunnart.

Timing of fertile period for successful pregnancy in American Bully dogs

Determination of the timing of the estrus cycle is essential for fertile mating. There are physiological variations among breeds, between bitches, and between cycles of the same bitch. If serial monitoring and many tools are applied, the exact moment of ovulation could be pin-pointed. However, it leads to time and costly difficulties. Progesterone concentrations during estrus cycles follow a specific pattern, and it is largely used in timing of fertile period. Although it has similar pattern in general, it is likely that breed-specific differences exist. The aim of this study was to investigate the way of timing the fertile period for successful pregnancy in American Bully dogs based on vaginal cytology and progesterone assay with minimized cost. To identify the empirical relations among reproductive characteristics, we performed statistical analyses on data from proestrus-to-estrus 27 American Bully dogs referred for 7 months. We found the significant correlations between the cyclic changes of vaginal cytology and progesterone assay. The relationship of serum progesterone concentrations with the days from vaginal discharge onset was analyzed through linear regression assay. In conclusion, we addressed two standards in the timing of fertile period with a minimal number of progesterone assays in the breeding management of American Bully dogs.

Manipulation of reproductive seasonality using melatonin implantation in Anglo-Nubian does treated with controlled internal drug release and equine chorionic gonadotropin during the nonbreeding season

The objective of this study was to compare the efficiency of hormonal treatments on ovarian activity and reproductive performance in anestrous Anglo-Nubian does during the nonbreeding season (February to May). A total of 48 multiparous does were divided into 2 groups (24 lactating does and 24 dry does). In each group, animals were allocated randomly into 2 equal subgroups (12 does each). In the first subgroup, does received a single 18-mg melatonin implant for 42 d followed by a controlled internal drug release (CIDR) device for 19 d in conjunction with 500 IU of equine chorionic gonadotropin (eCG) i.m. on the day of CIDR device removal. The second subgroup received CIDR combined with eCG in parallel with the first subgroup. Melatonin implantation induced a luteotrophic effect, expressed as an increasing number of corpora lutea, increased serum progesterone concentration, and reduced estradiol concentration. Regardless of treatment, dry does showed greater value of progesterone concentration. With the advancement of day of treatment, number of total follicles, small follicles, and medium follicles tended to increase to the greatest values at the day of CIDR device insertion. Furthermore, at day of mating, the numbers of large follicles reached the greatest value, which was associated with the lowest value of the number of corpora lutea. At day of mating, serum progesterone concentration achieved the lowest value, which increased until d 56 of pregnancy. The estradiol:progesterone ratio showed the opposite trend. The detrimental effect of reproductive seasonality, expressed as cessation of estrus behavior and fertile mating during the nonbreeding season, was successfully alleviated by the CIDR-eCG protocol. Furthermore, melatonin implantation in conjunction with the CIDR-eCG protocol enhanced conception rate and fecundity at d 28 of pregnancy and prolificacy at d 56 of pregnancy compared with does that were not implanted. Interestingly, does that failed to conceive did not come to heat again. In conclusion, the beneficial effect of melatonin implantation in conjunction with the CIDR-eCG protocol on the luteotrophic effect was reflected in the increasing number of corpora lutea, increasing progesterone concentration, and decreasing estradiol concentration. Furthermore, conception rate, prolificacy, and fecundity were improved compared with does that were not implanted during the nonbreeding season.

Inbreeding effects on in vitro embryo production traits in Guzerá cattle

Inbreeding has been associated with the impairment of reproductive performance in many cattle breeds. Although the usage of reproductive biotechnologies has been increasing in bovine populations, not much attention has been given to the impact of inbreeding over cow’s performance on artificial reproduction. The objective of this study was to estimate the impact of inbreeding on in vitro embryo production in a Guzerá breed population. The inbreeding coefficient (F), calculated as half of the co-ancestry of the individual’s parents, was used as an estimate of inbreeding. The inbreeding coefficients of the donor, sire (used on in vitro fertilization) and of the embryos were included, separately, in the proposed models either as classificatory or continuous variables (linear and quadratic effects). The percentage of non-inbred individuals (or embryos) and mean F of donors, embryos and sires were 29.38%; 35.76%; 42.86% and 1.98±2.68; 1.32±3.13; 2.08±2.79, respectively. Two different models were considered, one for oocyte production traits and other for embryo production traits. The increase of F of the donor significantly (P<0.05) impaired the number of viable oocytes (NOV), number of grade I oocytes (NGI) and number of cleaved embryos (NCLV). Moreover, the donor’s F influenced the percentage of grade I oocytes (PGI), percentage of viable embryos (PEMB) and percentage of cleaved embryos that turned into embryos (PCXE). No significant (P>0.05) effects were observed for the sire (father of the embryos) inbreeding coefficient over the traits analysed. Embryo’s F influenced (P<0.05) the number of viable embryos (NEMB), percentage of viable embryos (PEMB) and percentage of cleaved embryos that turn into embryos (PCXE). Results suggested that an increase in the inbreeding coefficient might impair the embryos ability to survive through challenges imposed by the in vitro environment. Submitting highly inbred Guzerá female donors to in vitro embryo production may, in the long-term, have negative implications on the number of embryos obtained per cow and increase the relative costs of the improvement programmes based on this technology. High levels of inbreeding should be avoided when selecting Guzerá female donors and planning in vitro fertilization mating.

Semen evaluation and fertility assessment in a purebred dog breeding facility

Semen quality in dogs has not been assessed in a longitudinal study that includes endpoints of female fertility and pregnancy. Although use of artificial insemination with chilled semen is increasingly used in canine reproduction, the resultant level of predictability and odds of fertile matings for dogs is still not fully understood. This research provides, for the first time, comprehensive semen evaluation in a large population of dogs in which fertility has been tracked. Duplicate ejaculates were obtained from 39 Labrador retriever males of the Guide Dogs for the Blind (San Rafael, CA, USA) breeding program. Sperm endpoints were determined in fresh semen and extended chilled semen at 48 hour after collection. Evaluation included total and progressive motility, average path velocity, morphology, membrane lipid peroxidation, presence of sperm reactive oxygen species, sperm chromatin structure, and mitochondrial DNA copy number. Male age ranged from 1 to 10 years and were grouped as young (Y; 1–3 years, n = 21), middle aged (M; 4–6 years, n = 13), and senior (S; 7 years or greater, n = 5) for analysis. The effects of age and sperm state (fresh vs. chilled) on the above sperm endpoints were determined using a linear mixed effects model. Semen endpoint values for all parameters were established for this group of fertile males. Progressive motility was only lower in the senior male chilled samples compared to all other groups, fresh and chilled (P < 0.05). Velocity decreased with increasing age and was lower overall in chilled samples (P < 0.05). Percent morphologically normal sperm was lower in senior dogs compared with the other age groups (P < 0.05). The presence of reactive oxygen species was lower in chilled samples compared with fresh (P < 0.05). For sperm chromatin structure, the senior-aged group had a higher %COMPαt than the middle-aged group (P < 0.05). Bayesian analysis determined that no differences were seen in total motility, membrane lipid peroxidation, and mitochondrial DNA copy number, with regard to conception rate or average litter size between age groups or between fresh and chilled samples. We observed no effects from semen quality on fertility or fecundity regardless of age, despite the differences found in semen quality. The use of advanced laboratory tests to evaluate sperm parameters beyond the standard motility, morphology, and concentration will open investigation to more specific and sensitive fertility tests in canine reproduction.

Long-term contraception in a small implant: A review of Suprelorin (deslorelin) studies in cats.

Rationale:Deslorelin (Suprelorin®; Virbac) is a gonadotropin-releasing hormone (GnRH) agonist licensed in select countries for the long-term suppression of fertility in adult male dogs and male ferrets. This article summarizes studies investigating the use of deslorelin implants for the long-term suppression of fertility in male and female domestic cats.Evidence base:Slow-release deslorelin implants have been shown to generate effective, safe and reversible long-term contraception in male and female cats. In pubertal cats, a 4.7 mg deslorelin implant suppressed steroid sex hormones for an average of approximately 20 months (range 15–25 months) in males and an average of approximately 24 months (range 16–37 months) in females. Reversibility has been demonstrated by fertile matings approximately 2 years post-treatment in both male and female adult cats. In prepubertal female cats of approximately 4 months of age, puberty was postponed to an average of approximately 10 months of age (range 6–15 months) by a 4.7 mg deslorelin implant.Challenges:The large variability in the duration of suppression of gonadal activity makes the definition of the optimal time for reimplantation quite challenging. In addition, the temporary stimulation phase occurring in the weeks following deslorelin implantation can induce in adult female cats a fertile estrus that needs to be managed to avoid unwanted pregnancy. Longer duration and larger scale controlled field studies implementing blinding, a negative control group and a carefully controlled randomization to each group are needed. Furthermore, the effects of repeated treatment need to be investigated. Finally, the effect of treatment on growth and bone quality of prepubertal cats needs to be assessed. However, the ease of use, long-lasting effects and reversibility of deslorelin implants are strong positive points supporting their use for controlling feline reproduction.

Molecular and cytogenetic evaluation for potential genotoxicity of hydrocortisone

ObjectiveTo assess the risk of hydrocortisone sodium succinate through different end points of genotoxicity. MethodsThe study examined the induction of chromosomal aberrations in bone marrow cells, morphological sperm abnormalities, the effect on dominant lethal gene and protein synthesis. Hydrocortisone was given intraperitoneally at three dose levels 26, 39 and 52 mg/kg body weight which was equivalent to the therapeutic doses in man. ResultsThe results showed that single dose treatment with different doses had no effect on chromosomal aberrations. The dose of 52 mg/kg body weight induced significant percentage of chromosomal aberrations in bone marrow cells after repeated treatment for 7 and 14 days. Significant effect of morphological sperm abnormalities was demonstrated only after treatment with the dose of 52 mg/kg body weight. For examining the dominant lethal mutation, male mice were injected with dose of 39 mg/kg body weight for 5 consecutive days. Mating between treated males and virgin untreated females were performed at different time intervals. The results showed that the percentage of fertile mating at 1–7 and 8–14 days reduced to 50% and 60% respectively compared with control group while no effect was recorded at 15–21 days. The percentage of dominant lethal mutation reached 0.32%, 4.4% and 0% in mating intervals respectively indicating pronounced effect of hydrocortisone at the interval 8–14 days which represented by the late spermatids. The results also showed that the repeated treatment with the dose of 52 mg/kg body weight inhibited protein synthesis which contributed to the cytotoxic effect of the drug. ConclusionsIt is concluded that long term treatment with large doses of hydrocortisone may have genotoxic effect.

Genetic relationships in bipolar species of the protist ciliate, Euplotes

Protists thrive in polar oceans, where they represent a major driving force for globally important biogeochemical cycles and a key food-web component. Their biogeography is frequently associated to bipolar patterns of distribution. Although conceptually well supported by apparently unrestricted migration rates, the experimental certification of these patterns copes with the protist paucity of morphological characters with taxonomic value and difficulties in applying conventional species concepts. We studied three marine species of the ciliate Euplotes, E. euryhalinus, E. nobilii, and E. petzi, for their bipolar distribution by comparing the SSU-rRNA gene sequences and mating interactions of Antarctic, Patagonian, and Arctic strains. Each species was analogously found not to carry significantly varied SSU-rRNA gene sequences, implying a common occurrence of trans-equatorial genetic mixing. However, mating analyses revealed significant inter-species differences. Scarce Antarctic × Arctic strain mating compatibility distinguished E. petzi from E. euryhalinus and E. nobilii, in which mating pairs between Antarctic and Arctic strains were successfully induced. Yet, E. nobilii was the only one of the two species to show cross-fertilizing and fertile mating pairs. Taking the biological concept of species as discriminatory, it was thus concluded that only E. nobilii warrants the definition of genuine bipolar species.

Restoration of ovarian function and natural fertility following the cryopreservation and autotransplantation of whole adult sheep ovaries.

STUDY QUESTIONIs it possible to restore ovarian function and natural fertility following the cryopreservation and autotransplantation of whole ovaries, complete with vascular pedicle, in adult females from a large monovulatory animal model species (i.e. sheep)?SUMMARY ANSWERFull (100%) restoration of acute ovarian function and high rates of natural fertility (pregnancy rate 64%; live birth rate 29%), with multiple live births, were obtained following whole ovary cryopreservation and autotransplantation (WOCP&TP) of adult sheep ovaries utilizing optimized cryopreservation and post-operative anti-coagulant regimes.WHAT IS KNOWN ALREADYFertility preservation by WOCP&TP requires successful cryopreservation of both the ovary and its vascular supply. Previous work has indicated detrimental effects of WOCP&TP on the ovarian follicle population. Recent experiments suggest that these deleterious effects can be attributed to an acute loss of vascular patency due to clot formation induced by damage to ovarian arterial endothelial cells.STUDY DESIGN, SIZE, DURATIONStudy 1 (2010–2011; N = 16) examined the effect of post-thaw perfusion of survival factors (angiogenic, antioxidant, anti-apoptotic; n = 7–8) and treatment with aspirin (pre-operative versus pre- and post-operative (n = 7–9)) on the restoration of ovarian function for 3 months after WOCP&TP. Study 2 (2011–2012; N = 16) examined the effect of cryoprotectant (CPA) perfusion time (10 versus 60 min; n = 16) and pre- and post-operative treatment with aspirin in combination with enoxaparine (Clexane®; n = 8) or eptifibatide (Integrilin®; n = 8) on ovarian function and fertility 11–23 months after WOCP&TP.PARTICIPANTS/MATERIALS, SETTING, METHODSBoth studies utilized mature, parous, Greyface ewes aged 3–6 years and weighing 50–75 kg. Restoration of ovarian function was monitored by bi-weekly blood sampling and display of behavioural oestrus. Blood samples were assayed for gonadotrophins, progesterone, anti-Müllerian Hormone and inhibin A. Fertility restoration in Study 2 was quantified by pregnancy rate after a 3 month fertile mating period and was confirmed by ultrasound, hormonal monitoring and live birth. Ovarian function was assessed at sacrifice by ovarian appearance and vascular patency (Doppler ultrasound) and by follicular histology.MAIN RESULTS AND THE ROLE OF CHANCEIn Study 1, survival factors were found to have no benefit, but the inclusion of pre-operative aspirin resulted in four ewes showing acute restoration of ovarian function within 3 weeks and a further six ewes showing partial restoration. The addition of post-operative aspirin alone had no clear benefit. In Study 2, combination of aspirin with additional post-operative anti-coagulants resulted in total acute restoration of ovarian function in 14/14 ewes within 3 weeks of WOCP&TP, with 9/14 ewes becoming pregnant and 4/14 giving birth to a total of seven normal lambs. There was no difference between anti-coagulants in terms of restoration of reproductive function and fertility. In contrast, the duration of CPA perfusion was highly significant with a 60 min perfusion resulting in ovaries of normal appearance and function with high rates of primordial follicle survival (70%) and an abundant blood supply, whereas ovaries perfused for 10 min had either resorbed completely and were vestigial (7/14) or were markedly smaller (P < 0.01). It is concluded that both the degree of CPA penetration and the maintenance of post-operative vascular patency are critical determinants of the success of WOCP&TP.LIMITATIONS, REASONS FOR CAUTIONBefore application of this technology to fertility preservation patients, it will be critical to optimize the CPA perfusion time for different sized human ovaries, determine the optimum period and level of anti-coagulant therapy, and confirm the normality of offspring derived from this procedure.WIDER IMPLICATIONS OF THE FINDINGSThis technology holds promise for the preservation of fertility in women. It could also potentially be applied to the cryopreservation of other reproductive or even major organs (kidneys) where there are considerable difficulties in storing donated tissue.STUDY FUNDING/COMPETING INTEREST(S)Funding was received from the Medical Research Council, University of Nottingham. The authors confirm that they have no conflict of interest in relation to this work.