The reproduction of bopyrid isopod parasites is thought to occur immediately following host ecdysis, but direct observations supporting this hypothesis are limited. The aim of this study was to describe the reproductive behaviour of the bopyrid isopod Bopyrus crangorum relative to host ecdysis based on video recordings. Several hours after host ecdysis, biphasic moulting of female parasites was observed. The cuticle of the posterior body was shed before that of the anterior body at an interval of 1 h. Two hours after female moulting, the male repeatedly moved from its initial position between the female pleopods and stopped at the anterior end of the fifth oostegite, immediately above the gonopore. To our knowledge, this repeated visiting behaviour by males has not been previously observed in bopyrid isopods. Oviposition through the female gonopore occurred 33 min later. The male-removal experiment showed that females with their males removed after visits to the gonopore oviposited eggs, whereas females with their males removed before visits did not. We propose that repeated visits by males to the gonopore are attempts to inseminate the female. We hypothesised that sperm are released onto the external surface of each gonopore and that the eggs are fertilised as they pass through the opening, which would explain the synchronous development of fertilised eggs inside the marsupium. The present study provides new information on the life history of bopyrid isopods, which allows for a better understanding of the host–parasite relationship.

Expression of estrus (EST) near the time of fixed-time artificial insemination (FTAI) increases pregnancy success in beef females. This outcome has been associated with improved pregnancy establishment and maintenance, although research is still warranted to validate this theory. Hence, this experiment compared ovarian, uterine and conceptus factors associated with pregnancy establishment in Bos indicus beef cows according to estrous expression during a FTAI protocol. One hundred lactating multiparous Nelore cows received a 2 mg injection of estradiol benzoate and an intravaginal progesterone (P4) releasing device on day −11, a 12.5 mg injection of prostaglandin F2α on day −4, P4 device removal in addition to 0.6 mg injection of estradiol cypionate and 300 IU injection of equine chorionic gonadotropin on day −2, and FTAI on day 0. An estrous detection patch was attached to the tailhead of each cow on day −2, and estrous expression was defined as removal of >50% of the rub-off coating from the patch at FTAI. Overall, 39 cows expressed EST, 55 did not express EST (NOEST), and six cows lost their patch and were discarded from the experiment. Ovarian ultrasonography was performed at FTAI, and on days 7 and 15 of the experiment. Blood samples were also collected on days 7 and 15. Only cows without a corpus luteum (CL) on day 0, and with a CL on days 7 and 15 remained in the experiment (EST, n=36; NOEST, n=48). On day 15, cows were randomly selected within each group (EST, n=29; NOEST, n=30) for conceptus collection via transcervical flushing, followed by endometrial biopsy in the uterine horn ipsilateral to the CL. Within cows not assigned to conceptus collection, blood samples were collected for whole blood RNA extraction (day 20) and pregnancy status was verified by transrectal ultrasonography (day 30). Diameter of dominant follicle on day 0 and plasma P4 concentrations on day 7 were greater (P⩽0.02) in EST v. NOEST cows. Conceptus length and messenger RNA (mRNA) expression of prostaglandin E synthase and interferon-tau were greater (P⩽0.04) in EST v. NOEST cows. Moreover, EST cows diagnosed as pregnant on day 30 had greater (P<0.01) blood mRNA expression of myxovirus resistance 2 on day 20 compared with NOEST. In summary, estrous expression near the time of FTAI enhanced pregnancy establishment factors in B. indicus cows, including conceptus development and mRNA expression of interferon-tau.

The coffee berry borer, Hypothenemus hampei (Ferrari), can survive in residual coffee berries during the inter-harvest period, while new fructification only appears 2–3 months after the last harvest. The dispersal of colonizing females is an adaptation that enables the life cycle of the species to go ahead whenever his flight aptitude allows. This paper focuses on accurately determining the rate of inseminated females ready to reproduce when emerging from residuals berries to colonize new ones, which constitutes a characteristic of the live cycle far from common in Curculionidae. We dissected females caught in traps baited with a mixture of alcohols during the inter-harvest season, females from infested residual berries collected from branches, and virgin females obtained from pupae reared individually in the laboratory. After microscopic preparation with Giemsa stain, spermathecae were observed to identify the physiological status of each specimen. Out of the females found in the traps, 98.4% displayed recent and abundant insemination and 1.6% sporadic insemination. In contrast, in residual berries, most of females were recently inseminated (84.5%), followed by virgin females (10.5%) and older inseminated females (5%). In addition, the flight tests of the virgin females were negative. These results indicate that all colonizing females were inseminated, ready for flying and oviposition, females inside residual berries showed different physiological status, and virgin females could not migrate since they could not flight. The large number of inseminated females inside the residual berries, and the capacity of migrating females to colonize and reproduce, suggest that it is necessary to control residual berries and use traps to stop the dispersal and reproduction of this pest.

Functional morphology of the male gonopods, female holding systems and vulvae of Libinia spinosa are described using scanning electron microscopy and histological sections, establishing differences between immature and mature individuals. The first gonopod of mature males, larger than 35.1 mm of carapace width (CW), is long, slender, and ending in two wing-like processes; abdominal, characterized by the presence of a sub-terminal seminal canal, and sternal, closed and where the cuticle holds setae. In immature males, smaller than 29.9 mm CW, both processes are shorter, and the tip of the abdominal process closed. Six different types of setae are described for both gonopods. Internally, the gonopods have rosette glands in immature and mature individuals. The presence of secretions, which may be involved in the moulting process, was observed in immature males towards the external cuticle corresponding to the rosette glands. The rosette glands may be related to reproduction in mature males. Females presented a press-button holding system and a biconcave vulva. In immature females, smaller than 36.1 mm CW, the vulva aperture was closed by integument bulging to the outside. In mature females, larger than 42.6 mm CW, the locking system was almost fused to the cuticle, and the vulvae showed an open biconcave form with a more inner flexible membrane. Morphological characteristics shown by the gonopods of mature males develop synchronously with the acquisition of physiological maturity, not with morphometrical maturity. The mature holding systems and vulvae of females are developed synchronously with physiological and morphometrical maturity.

Considerable diversity has been found in the reproductive behaviour of benedeniine (capsalid) monogenean parasites. Mating has been observed in Benedenia sp. 1 from the gills of Lutjanus carponatatus; externally attached spermatophores are not involved and parasites indulge in mutual cross-insemination with intromission. In contrast, there is evidence of spermatophore involvement in Benedenia sp. 2 from the fins of Lethrinus miniatus; mating was not observed but an individual was found carrying an external ovoid spermatophore attached by a stalk lodged in the vagina. In specimens of Benedeniella macrocolpa and B. posterocolpa, in which the male reproductive system was functional and the female system not fully developed, the everted cirrus was seen to be lodged in the parasite's own uterus, with, in some individuals, the tip of the cirrus inside the ootype. This is the first time that the copulatory organ has been observed inside the female reproductive tract of the same individual, not just in benedeniines but in monogeneans in general, and is also the first demonstration that monogeneans are capable of self-insemination via the uterine route.

This study summarizes optimization of techniques for common carp artificial propagation including improvements of activation solution (AS), the process of insemination, and elimination of egg stickiness. The optimum gamete ratio for good fertilization and hatching rate ranged from 8490 to 23672 spermatozoa per egg, when dechlorinated tap water was used. Optimal ratio between eggs (weight in g) and AS (in ml) was defined as 1:1 to 1:2. Different concentrations of AS such as NaCl from 0 to 34 mM (0–68 mOsmol kg–1) did not change fertilization and hatching rates. An AS adopted for carp spermatozoa (45 mM NaCl, 5 mM KCl, 30 mM Tris–HCl, pH 8) was compared with other saline AS; only the 51 mM (102 mOsmol kg–1) NaCl solution decreased fertilization and hatching rate. The AS containing 20 mM Tris–HCl at pH 9 increased fertilization and hatching rates compared to dechlorinated tap water of pH 7 or to AS of pH 6 and 7. Adhesiveness from the eggs was successfully removed by incubation in Alcalase DX (PLN 04715) using two successive steps with different enzyme concentrations. The first step with an enzyme concentration of 2 ml l–1 was applied from 8 to 20 min after fertilization. Later in a second step, the best time for application of alcalase enzyme at a concentration of 20 ml l–1 was for 45 and 60 s at 20 min after fertilization leading to fertilization and hatching rates of 80–87%. The α-Chymotrypsin (EC 3.4.21.1. Merck) was also found effective for elimination of stickiness. Results withα-Chymotrypsin enzyme indicate that the response to success in elimination of stickiness is highly variable mainly due to differences in the environment, quality of water and carp strains.

The aim of this study was to explore how some reproductive methodologies may affect the sex ratio. We first confirmed the association between the maturation stage of bovine oocytes at the time of in vitro fertilisation (IVF) and the sex ratio of in vitro-derived embryos. Secondly, we studied whether the time of insemination, prior to or after ovulation, could alter the sex ratio in sheep. In the first experiment, bovine oocytes were matured in vitro for 16 h; then oocytes were either fertilised in vitro immediately after extrusion of the first polar body or IVF was delayed for 8 h. The proportion of cleaving embryos and their development to the 8-cell stage was enhanced with delayed insemination. Moreover, delaying IVF produced a male-to-female sex ratio of 1.67:1.00, which was significantly different from the expected 1:1 ratio (p < 0.05), whereas more female embryos were produced when oocytes were fertilised in vitro immediately after polar body extrusion (sex ratio of 1.00:0.67; p < 0.05). In the second experiment, 380 ewes were inseminated at different times before or after ovulation, producing 537 lambs. Significant differences in the sex ratio were obtained when we compared the sex of the offspring of ewes inseminated during the 5 h preceding ovulation (more females) with those inseminated during the 5 h after ovulation (more males). Our results suggest that the differential ability of X- or Y-bearing spermatozoa to fertilise oocytes depending either on time of insemination or oocyte maturation state, may be due, at least partially, to ‘intrinsic’ differences in the physiological activity of X- or Y-bearing spermatozoa before fertilisation.

Despite growing discussion of the potential benefits of openness in relation to children conceived by donated gametes, the majority of parents do not intend to tell a future child about his/her origins. As a result little is actually known about the experiences and concerns of families who do choose openness. This is a descriptive study of 83 DI Network members who did intend to or had told a child conceived, using donor insemination, about his/her origins. Quantitative and qualitative data focused on their concerns and experiences of telling. Health professionals face the challenge of providing the opportunity for discussion of these issues both before treatment and afterwards during childhood.

A sperm cryopreservation protocol adapted from turbot, was tested on sea bass using either 250-μL straws or 1.5-mL cryovials. A dilution to 1/3 in Mounib's extender and a cooling rate of −65 °C·min−1 allowed frozen sperm to recover an initial motility similar to that of fresh sperm at thawing; however, significant differences in motility (P < 0.001, n = 10 fish semen) were observed at further post-activation times, the motility decrease being faster in thawed sperm. At the experimental scale, triplicate inseminations of 2-mL aliquots (approximately 2 000 eggs) showed a significant fertility decay of thawed sperm compared to that of fresh sperm (P < 0.01, n = 12 fish semen) when a discriminating 35·103 spermatozoa to egg ratio was applied. When 70·103 and 200·103 spermatozoa per egg were provided in the same experimental conditions, no significant difference appeared between the fertilisation rates of fresh and thawed sperm. In order to validate the procedure for production or cryobank purpose, a scaled-up protocol was established. Two and 50 mL batches of eggs (approximately 2·103 and 50·103 eggs, respectively) were inseminated in triplicate using either fresh or thawed individual sperms of 5 males with 200·103 spermatozoa per egg. The mean fertility decreased by 23.5 % due to cryopreservation. This decline was explained by the loss of fertility of only one sperm, and only in large-volume conditions, probably due to the delay of use after thawing.

Studies were conducted during the dry period February to May 1986 in River Shelliekha, Bahr El Arab fly belt, Sudan, to obtain baseline data on catch and age compositions, reproductive status and trypanosome infection rates of trapped and fly round samples of Glossina morsitans submorsitans. The objective was to assess the tsetse situation before the establishment of trypanosomiasis treatment stations at the northern limits of the fly belt. Trapped tsetse included significantly higher proportions of teneral and non-teneral females and a lower male:female ratio. Insemination rates were over 98% in both samples. In any one group, the frequency of pregnancy with egg predominated, followed by the second, first and lastly the third instar larva. There were significant differences between the two groups of females in the proportions of nullipars and pregnancy with any one of the larval instars. Abortion was the predominant reproductive abnormality and no relationship could be found between abortion rate and the sampling method, age or trypanosome infection rate of females. Age compositions were similar in the two samples, save for age categories 0 and 1 for females and two and four for males. Flies were infected with Trypanosoma vivax and T. congolense only, though T. brucei infections could also be diagnosed in livestock.

The field application of a laparoscopic technique to permit intra-uterine insemination of ewes with frozen-thawed semen was examined in two trials, conducted over successive years, to (i) determine the optimum time of insemination relative to sponge removal/pregnant mares's serum gonadotropin (PMSG) injection and (ii) establish the relationship between semen dose and conception rate. Pooled semen was used in both trials, and each involved > 900 ewes in a number of commercial flocks.

Maximum conception rates were achieved when insemination was conducted between 54 h and 60 h after sponge removal/ PMSG injection. However, there was no significant difference in conception rate when motile sperm numbers were reduced from 52·2 × 106to 13·0 × 106 motile sperm per uterine horn.

The overall conception rates (pooled over flocks and treatment groups) were 56% and 58% for the two trials, with a wide degree of variation between flocks in both cases (45% to 79% for trial 1 and 45% to 69% for trial 2). However, there was evidence for consistent differences in conception rates between the six flocks involved in both years of the trials.