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Artificial insemination using trypsin-treated sperm in the silkworm, Bombyx mori.

A new method was developed for artificial insemination of the silkworm. Virgin females artificially inseminated by this method oviposited fertilized eggs at a rate almost similar to that of females mated with males. Using inactive sperm collected from the seminal vesicles, we confirmed the previous finding by Omura, S., 1936a, Artificial insemination of Bombyx mori, Journal of Faculty of Agriculture, Hokkaido Imperial University 38, 135-150, and Omura, S., 1938, Studies of the reproductive system of the male of Bombyx mori II, Post-testicular organs and post-testicular behaviour of the spermatozoa, Journal of Faculty of Agriculture, Hokkaido Imperial University 40, 129-170, that the sperm of B. mori requires the secretion of the glandula prostatica for activation. Sperm also could be activated by trypsin. At an optimal concentration, 0.3 &mgr;g trypsin/ml in 50% semen solution, the fertilization rate as well as the number of eggs oviposited was almost equivalent to that obtained in normally mated moths. These results may contribute not only to basic studies on fertilization and reproduction in Lepidoptera but also to the development of long-term preservation of genetic resources by using cryopreserved sperm of B. mori and other Lepidoptera insects.

Journal Article↗

Initial and cyclic recruitment of ovarian follicles.

Mammalian ovaries consist of follicles as basic functional units. The total number of ovarian follicles is determined early in life, and the depletion of this pool leads to reproductive senescence. Each follicle develops to either ovulate or, more likely, to undergo degeneration. The dynamics of ovarian follicle development have interested endocrinologists and developmental biologists for many years. With the advent of assisted reproductive techniques in humans, the possibility of regulating follicle development in vivo and in vitro has gained clinical relevance. In this review, we focus upon key branching points during the development of ovarian follicles as well as factors involved in determining the eventual destiny of individual follicles. We discuss inconsistencies in the literature regarding the definitions of follicle recruitment and selection and propose to name the two major steps of follicle development as initial and cyclic recruitment, respectively. Because some of these disparities have arisen due to differences in the animal systems studied, we also compare the development of the ovarian follicles of both humans and rats. We also review the status of knowledge of several puzzling clinical issues that may provide important clues toward unlocking the mechanisms of follicle development.

Aging↗

Reproductive physician-scientists for the twenty-first century.

As we enter a new century, departments of obstetrics and gynecology in American medical schools face a number of challenges. A primary concern is the relative dearth of physician-scientists to explore basic mechanisms of cell function and relate these to the diseases of women. The question arises, what can we do to revitalize the spirit of investigation in many of our academic departments? This report reviews three programs designed to develop academic investigators: the Reproductive Scientist Development Program, the American Gynecological and Obstetrical Society/American Association of Obstetricians and Gynecologists Foundation Fellowship Program, and the Women's Reproductive Health Research Career Development Centers. On the basis of a decade of experience with the first two of these programs, the prospects for the third are promising. Clearly, the opportunities for obstetrician-gynecologist basic and clinical scientists are considerable. The question remains: Will leaders in the specialty work together to meet the challenge?

Education, Medical↗

Oocyte freezing: here to stay?

Oocyte freezing is an established technology but, in contrast to embryo freezing, it has very limited application in clinical IVF programmes. Is there a chance that oocyte freezing will become an integrated routine in assisted reproductive technology? The delicate cytological architecture of the oocyte with a cold-sensitive spindle and a hardening zona have made the frozen oocyte 'unwanted' in assisted reproductive technology. Nevertheless, empirical improvements in freezing protocols and the use of ICSI for fertilization have led to an increasing number of live births. This mitigates against a simple ban on oocyte freezing. While efficiency of oocyte freezing can certainly be further improved by basic research, it is clear that there are humanitarian reasons for considering oocyte freezing as a future fully utilized assisted reproductive technology. The storage of the female genome as a particulate entity can provide an alternative in case of moral, ethical, legal or religious concerns about embryo freezing. Oocyte freezing can also offer hope for oocyte donation and preservation of fertility for women facing ovarian loss. The message is one of cautious optimism when looking for a place for oocyte freezing in routine assisted reproductive technology.

Animals↗

Census of genes expressed in porcine embryos and reproductive tissues by mining an expressed sequence tag database based on human genes.

A total of 98,898 expressed sequence tags (ESTs) derived from embryos and reproductive tissues in pigs were identified in the GenBank "est_others" database. Pig embryos were collected at 11, 12, 13, 14, 15, 20, 30, and 45 days after gestation. The reproductive tissues were sampled from testis, ovary, endometrium, hypothalamus, anterior pituitary, uterus, and placenta. A gene-oriented approach was developed to annotate these porcine EST sequences to census the genes expressed from these sources. Of the 33 308 mRNA sequences from the human genes used as references (data accessed on 1 November 2002), 9410 had the porcine EST homologs expressed in embryos and 11 795 had the EST homologs expressed in reproductive tissues. The entire genome contributes at least 28.3% of its genes to embryo development and 35.4% of its genes to reproduction. Using the EST entry numbers as indicators of gene expression, we determined that the gene expression patterns differ significantly between embryos and reproductive tissues in pigs. The basic active genes were identified for each source, but most of them are not coexpressed abundantly. Few genes were expressed on the Y chromosome (P < 0.01), but they may represent counterparts of the double-dose genes that remain active in an inactivated X chromosome in females but are needed for proper development and growth. The census provides a panel of transcripts in a broad sense that can be used as targets to study the mechanisms involved in embryo development and reproduction in pigs and other mammals, including humans.

Animals↗

Molecular mechanisms underlying pig oocyte maturation and fertilization.

Since the pig is not only an important farm animal, but also a model animal for biomedical applications, the development of reproductive technologies in this species has been very important. In vitro oocyte maturation and fertilization (IVM-IVF) are basic techniques for a number of oocyte- or embryo-related technologies. The practical aspects for pig oocyte IVM-IVF have been reviewed, while the molecular mechanisms underlying oocyte meiotic maturation and fertilization have not been well summarized, although accumulating data have been obtained in recent one decade. This review will focus on what is known about the molecular mechanisms of porcine oocyte maturation and fertilization such as first meiosis resumption, meiotic spindle assembly, second meiosis metaphase (MII) arrest during oocyte maturation, sperm-egg recognition and fusion, sperm acrosome reaction, second meiosis resumption, sperm chromatin decondensation, and pronucleus formation during fertilization, as well as the establishment of polyspermy block.

Acrosome Reaction↗

The bovine dominant ovarian follicle.

Central roles in reproductive biology (i.e., growth and development of the oocyte, steroidogenesis, and ovulation) are played by the ovarian dominant follicle (DF). The DF is different from other follicles because it can escape atresia (the fate of all other follicles), and if exposed to the LH surge, its cells will differentiate into the corpus luteum. The DF was originally studied by looking at the surface of ovary through a surgical approach. Current studies employ a less-invasive ultrasound technique to track the growth and development of the DF. Recruitment and selection, the processes that give rise to the DF, and dominance, the physiological state of the mature DF, are important areas of basic research. Results of these basic studies are easily translated into real-world problems in farm animal reproduction. Superovulation, for example, overrides the selection mechanism and increases the number of ovulations. Understanding the factors that affect the size of the recruited pool should increase success rates (i.e., number of collected embryos) for superovulation. In most animals, the DF is short-lived, existing for long enough to allow for the final maturation of the oocyte. Some DF become atretic because they mature during the luteal phase and are never exposed to the LH surge. For other DF, the LH surge redirects the DF toward its ultimate demise (i.e., luteinization, ovulation, and differentiation into the corpus luteum). The DF is managed pharmacologically within protocols for timed AI. When timed AI fails, there may be abnormal corpus luteum development and early embryonic loss; outcomes that are secondary to inadequate follicular cell maturation and incomplete oocyte capacitation in the DF. Future work on the DF will clarify its underlying biological functions so that a variety of needs in farm animal reproduction can be efficiently managed.

Journal Article↗

Fertility awareness and natural family planning.

Information about fertility awareness helps to fulfil the broader definition of the services many family planning clinics offer. Although information about natural family planning is requested by a small number of clients seeking family planning advice, many more clients benefit from information about fertility awareness. Fertility awareness is far more than just basic reproductive anatomy and physiology; fertility awareness involves understanding basic information about fertility and reproduction, being able to apply it to oneself, and being able to discuss it with a partner or with a health professional. Fertility awareness is fundamental to understanding and making informed decisions about reproductive health and sexual health. If clients have a better understanding of fertility awareness, they are in a stronger position to make informed decisions about how they wish to manage their reproductive and sexual health, for example: (1) Fertility awareness information is used to help couples to plan pregnancies as well as to avoid them. This can be helpful to couples who are having difficulty conceiving, for the timing of intercourse or for the timing of some of the sub-fertility investigations. (2) The information is also useful when helping couples to understand how each method of family planning works--how the family planning method interrupts normal fertility, how the method will fail if not used correctly, and how fertility returns when the method is discontinued. (3) Women who are fully breastfeeding value the knowledge about reduced fertility, as do women during the perimenopausal years who value being given clear information about their declining fertility. (4) When counselling couples about the importance of avoiding sexually transmitted diseases it is important they understand sexually transmitted diseases may damage their fertility. (5) Couples who choose only to use a barrier method during the time they think the woman is fertile are a group who do not readily identify themselves to family planning providers. These couples often do not have adequate information about fertility awareness. Advances in technology and the understanding of ovulation, ovum and sperm survival have confirmed that the guidelines used to teach fertility awareness and natural family planning effectively identify the fertile phase of the menstrual cycle. Serial ultrasound studies on the ovaries during the menstrual cycle have confirmed the accuracy of the hormonal assays in pinpointing the likely time of ovulation. Ultrasound studies have also shown that subjective observations of the alterations in cervical mucus and the basal body temperature rise are accurate indicators of the fertile phase. Research on the chances of conception on each day of the menstrual cycle, using hormonal assays to estimate the time of ovulation, was carried out in 1994 by Weinberg and Wilcox. Their results showed that the timing of sexual intercourse, in relation to ovulation, strongly influences the chance of conception. Conception only occurred during a 6-day interval that ended on the estimated day of ovulation. The chances of conception fell to zero 24 hours after ovulation. Several different methods of natural family planning are taught; some methods depend on only using one of the indicators of fertility, others are based on two or more indicators. The main indicators of fertility are: observing the cervical mucus, recording the basal body temperature, palpating the cervix and a calculation based on the cycle length. Research studies performed using a combination of the indicators of fertility show that the failure rate using a combination is less than most of the studies which use a single indicator. In each case the method failure is far lower than the user failure. (ABSTRACT TRUNCATED)

Body Temperature↗

Seasonal effects on embryo transfer results in Brahman cows.

To determine factors affecting embryo donor reproductive performance in Bos indicus cows, 1,841 embryo collection records of 813 Brahman donor cows were analyzed using least-squares methods for unequal subclass numbers. The basic model included the main effects of either season or month, technician, and year of collection. Season and year did not affect (P>.10) the variance of number of ova collected. The most skillful technician collected 2.8 more ova per flush compared with the least skillful. Number of transferable embryos recovered per donor cow was affected by season (P<.06). Maximal recovery of transferable embryos occurred during the fall season (4.2) with a lower number (2.9) recovered during the winter season. Pregnancy rate in the recipients followed a seasonal pattern with lower (31.8%) pregnancy rates during the winter and higher (41.0%) ones in the fall. There was a trend toward higher pregnancy rates during February through October and lower rates December and January. Number of blastocysts recovered per donor was affected by season (P<.08), and year (P<.01). Number of blastocysts recovered per donor was lower (2.0) during the winter season than during the fall season (3.1). These data establish the concept of seasonal effects on embryo donor reproductive performance in Brahman cows.

Journal Article↗

Reproduction in captive common marmosets (Callithrix jacchus).

Though sexual maturation may begin at around one year of age, first successful reproduction of the common marmoset (Callithrix jacchus) is likely to be later, and it is generally recommended that animals not be mated before 1.5 years of age. The average gestation period is estimated to be 143 to 144 days. A crown-rump length measurement taken by use of ultrasonography during the linear, rapid, prenatal growth phase (between approx. days 60 and 95) can be compared against standard growth curves to estimate delivery date to within 3 to 4 days, on average. Marmosets produce more young per delivery than does any other anthropoid primate, and have more variation in litter size. Many long-established colonies report that triplets are the most common litter size, and there is documented association between higher maternal body weight and higher ovulation numbers. Higher litter sizes generally do not generate higher numbers of viable young. Marmosets are unusual among primates in having a postpartum ovulation that typically results in conception and successful delivery; reported median inter-birth intervals range from 154 to 162 days. However, pregnancy losses are quite common; one study of a large breeding colony indicated 50 percent loss between conception and term delivery. The average life span for breeding females is around six years; the range of reported average lifetime number of litters for a breeding pair is 3.45 to 4.0. Our purpose is to provide an overview of reproduction in the common marmoset, including basic reproductive life history, lactation and weaning, social housing requirements, and common problems encountered in the captive breeding of this species. A brief comparison between marmoset and tamarin reproduction also will be provided.

Animals↗

[A model of resource reallocation during physiological adaptation in Mediterranean fruit fly females].

Analysis of experimental data on longevity and fertility of females of Mediterranean fruit fly Ceratitis capitata demonstrates that flies lay fewer eggs on the average as the average life-span increases. At the same time, the values of individual life-span and the number of eggs laid are positively correlated. To explain these results, a resource model describing the allocation and reallocation of resources between two basic functions, reproduction and maintenance, was proposed. The reallocation of resources, is triggered by changes in environmental conditions such as diet changes, and the parameters of reallocation depend on the current environmental conditions. Modeling demonstrated that the results of experiments are greatly influenced by population selection and heterogeneity.

Adaptation, Physiological↗

Modelling the prevalence of Echinococcus and Taenia species in small ruminants of different ages in northern Jordan.

A base-line survey of Echinococcus granulosus, Taenia hydatigena and T ovis were undertaken in order to investigate the transmission dynamics of these parasites in northern Jordan. Intensity of E. granulosus infection, in sheep, increased with age in a linear fashion whilst the asymptotic prevalence was one. This implied that E. granulosus is in an endemic steady state with no evidence of protective immunity in the intermediate host. The mean number of cysts increased by 1.66 per year with approximately 0.320 infections per year, each infection consisting of 598 eggs to produce 5.2 cysts. The basic reproduction ratio (R0) was estimated to be 1.5-1.8. A similar pattern was suggested with E. granulosus in goats but the infection pressure appeared to be lower with only 0.128 cysts per year. Although infection in goats appeared to be endemic there was some evidence of departure from the model which might indicate that the model needs adjusting for this species. In the case of T. hydatigena the host age-intensity helminth distribution indicated that this parasite was hyperendemic in both sheep and goats, implying regulation by intermediate host immunity. Consequently, R0 was determined from asymptotic prevalence curves for T hydatigena and was calculated to be 4.0 and 3.1 in sheep and goats, respectively. The lower R0 in goats, together with the higher asymptotic age-intensity and age-prevalences, indicates that goats acquire immunity more slowly to T hydatigena in comparison to sheep. Taenia ovis was not detected in any animals.

Animals↗

Modelling the effect of surveillance programmes on spread of bovine herpesvirus 1 between certified cattle herds.

For the eradication of an infectious agent, like bovine herpesvirus 1 (BHV-1), surveillance and certification can be used to reduce the transmission between herds. The goal of surveillance is that a certified herd that becomes infected is detected timely so that infection of several other certified herds is prevented. What counts is whether the reproduction ratio R, i.e. the average number of certified herds infected by one infected certified herd can be kept below 1. To support policy makers in making decisions about the minimal demands for a surveillance programme in an eradication campaign of BHV-1 in cattle, two mathematical models were investigated. With these models, the basic reproduction ratio between herds was calculated. The surveillance programmes were characterised with sample size, sampling frequency, test sensitivity, herd size, vaccination status, and contacts between herds. When R between herds is below 1, then the surveillance programme is sufficiently good to prevent spread of infection, provided that R is estimated well. In the model based on bulk milk testing sample size was replaced by a threshold at which bulk milk can be found positive. The R between herds was mainly influenced by the vaccination status, sampling frequency, and contacts between herds. Herd size moderately affected the outcome. Test sensitivity and sample size, however, were of minor importance. If herds of 50 cows became free of BHV-1 without vaccination, then spread of infection between herds might be prevented when animals within herds are sampled once a year (milk or blood samples). This frequency needs to be intensified, being twice a year, for larger herds and/or herds with extensive contacts with other herds. When bulk milk is sampled instead, sampling should be done at least every 5 months and more intensively, being each month, with larger herd sizes and more contacts between herds.

Animals↗

Lonely hearts or sex in the city? Density-dependent effects in mating systems.

Two very basic ideas in sexual selection are heavily influenced by numbers of potential mates: the evolution of anisogamy, leading to sex role differentiation, and the frequency dependence of reproductive success that tends to equalize primary sex ratios. However, being explicit about the numbers of potential mates is not typical to most evolutionary theory of sexual selection. Here, we argue that this may prevent us from finding the appropriate ecological equilibria that determine the evolutionary endpoints of selection. We review both theoretical and empirical advances on how population density may influence aspects of mating systems such as intrasexual competition, female choice or resistance, and parental care. Density can have strong effects on selective pressures, whether or not there is phenotypic plasticity in individual strategies with respect to density. Mating skew may either increase or decrease with density, which may be aided or counteracted by changes in female behaviour. Switchpoints between alternative mating strategies can be density dependent, and mate encounter rates may influence mate choice (including mutual mate choice), multiple mating, female resistance to male mating attempts, mate searching, mate guarding, parental care, and the probability of divorce. Considering density-dependent selection may be essential for understanding how populations can persist at all despite sexual conflict, but simple models seem to fail to predict the diversity of observed responses in nature. This highlights the importance of considering the interaction between mating systems and population dynamics, and we strongly encourage further work in this area.

Animals↗

Fundaments of plant cybernetics.

A systemic approach is proposed for analyzing plants' physiological organization and cybernesis. To this end, the plant is inspected as a system, starting from the integration of crown and root systems, and its impact on a number of basic epigenetic events. The approach proves to be axiomatic and facilitates the definition of the principles behind the plant's autonomous control of growth and reproduction.

Cybernetics↗

Sperm motility parameters to evaluate the seminal quality of Boa constrictor occidentalis, a threatened snake species.

Semen quality analysis constitutes a powerful tool to evaluate the fertility potential of males in threatened species. The Argentine boa constrictor or lampalagua (Boa constrictor occidentalis) is a threatened snake species and has been included in Appendix I of CITES. The objective of this work is to characterize the sperm of B. c. occidentalis on the bases of dynamic parameters to improve this species conservation. Dynamic parameters were measured in sperm samples using videomicroscopy and image analysis software. The sperm population showed a high degree of heterogeneity in velocity parameter values and 95% of the cells showed a linear pattern of movement. Studies in other species indicate that the number of motile spermatozoa and their movement speed is directly correlated with fertilization success. This work will help to establish basic parameter values for the evaluation of the reproductive potential of populations of B. c. occidentalis and to resolve questions referred to its reproductive strategies.

Animals↗

Dynamics of Forest Insect Density: Bifurcation Approach

Six basic phase portraits differing in character and in the number of their equilibrium behaviour regimes were suggested in the classification of forest insect population dynamics. The portraits graphically illustrate six types of dynamic behaviour of numbers of the system "phytophage-entomophage": two types of stable equilibrium dynamics (stationary and oscillatory) and four types corresponding to outbreaks of mass reproduction-fixed, permanent, reversive and true outbreaks, respectively. In this paper we consider the single parameter model, with its parametric domains realizing a wide spectrum of phase dynamics, both basic and transitional. The changes in the phase portraits are accompanied by the bifurcations in the model. The results of these analyses are in agreement with the "stability principle of mobile ecological systems" and may supply its parameter substantiation at the model level. Copyright 1997 Academic Press Limited

Journal Article↗

A two-generation reproductive toxicity study of 2,4-dichlorophenol in rats.

A two-generation reproductive toxicity study was conducted with 2,4-dichlorophenol (2,4-DCP), an agent suspected of exerting endocrine disrupting effects. Wistar-Hannover rats, 24/sex/group, were given diet containing 2,4-DCP at dose levels of 0, 500, 2000 or 8000 ppm to examine the potential effects of the test substance on parental animals and their offspring over 2 successive generations. Neither clear systemic nor reproductive toxicity of 2,4-DCP was apparent in the 500 ppm group. In the 2000 ppm group, mean body weight gain and food consumption of females were lowered significantly during the treatment period. Effects on body weights and food consumption were more serious in the 8000 ppm group, both males and females being significantly affected. Reproductive effects of the test substance were also observed in the 2000 and 8000 ppm groups dose-dependently. Observations included significantly increased uterine weights of F1 and/or F2 female weanlings and reduced numbers of implantation sites and live births of F1 parental females. These results suggest that 2,4-DCP has weak reproductive toxicity, possibly based on endocrine activity. However, the basic mechanisms for apparent estrogenic effects of 2,4-DCP remain to be elucidated.

Administration, Oral↗