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A very large majority of farm animals express seasonal variations in their production traits, thus inducing seasonal availability of fresh derived animal products (meat, milk, cheese and eggs). This pattern is in part the consequence of the farmer's objective to market his products in the most economically favourable period. It may also be imposed by the season-dependent access to feed resources, as in ruminants, or by the specific requirements derived from adaptation to environmental conditions such as water temperature in fish. But seasonal variations in animal products are also the consequence of constraints resulting from the occurrence of a more or less marked seasonal reproductive season in most farm animal species including fish, poultry and mammals. Like their wild counterparts, at mid and high latitudes, most farm animals normally give birth at the end of winter-early spring, the most favourable period for the progeny to survive and thus promote the next generation. As a consequence, most species show seasonal variations in their ovulation frequency (mammals and fish: presence or absence of ovulation; birds: variations or suppression of laying rates), spermatogenic activity (from moderate to complete absence of sperm production), gamete quality (variations in fertilisation rates and embryo survival), and also sexual behaviour. Among species of interest for animal production, fishes and birds are generally considered as more directly sensitive to external factors (mainly temperature in fish, photoperiod in birds). In all species, it is therefore advisable that artificial photoperiodic treatments consisting of extra-light during natural short days (in chickens, turkeys, guinea fowl, sheep and goats) or melatonin during long days (in goats, sheep) be extensively used to either adjust the breeding season to animal producer needs and/or to completely overcome seasonal variations of sperm production in artificial insemination centres (mammals) and breeder flock operations (poultry, fish farming). Pure light treatments (without melatonin), especially when applied in open barns, could be considered as non invasive ones which fully respect animal welfare. [ABSTRACT FROM AUTHOR]

Elephants have the longest pregnancy of all mammals, with an average gestation of around 660 days, so their embryonic and foetal development have always been of special interest. Hitherto, it has only been possible to estimate foetal ages from theoretical calculations based on foetal mass. The recent development of sophisticated ultrasound procedures for elephants has now made it possible to monitor the growth and development of foetuses of known gestational age conceived in captivity from natural matings or artificial insemination. We have studied the early stages of pregnancy in 10 captive Asian and 9 African elephants by transrectal ultrasound. Measurements of foetal crown–rump lengths have provided the first accurate growth curves, which differ significantly from the previous theoretical estimates based on the cube root of foetal mass. We have used these to age 22 African elephant foetuses collected during culling operations. Pregnancy can be first recognized ultrasonographically by day 50, the presumptive yolk sac by about day 75 and the zonary placenta by about day 85. The trunk is first recognizable by days 85–90 and is distinct by day 104, while the first heartbeats are evident from around day 80. By combining ultrasonography and morphology, we have been able to produce the first reliable criteria for estimating gestational age and ontological development of Asian and African elephant foetuses during the first third of gestation. [ABSTRACT FROM AUTHOR]

To apply otolith microstructure to examination of age and growth of juvenile chum salmon Oncorhynchus keta inhabiting coastal waters, formation of otolith increments was investigated for juveniles reared in a seawater aquarium and in net pens. In all otoliths examined, a distinctive check was formed at the time of sea entry of the fish. The deposition of otolith increments after the check was daily for rearing both in the aquarium (57 days) and in the net pens (26 days). Check formation associated with sea entry was also observed in otoliths of juvenile salmon collected 1 km off the coast of Shari, Hokkaido, Japan. Transmitted light observation of otoliths of those fish revealed a transition in otolith increment appearance from dark to light. Otolith Sr:Ca ratio remarkably changed from a low to a high level, coinciding with the transition in otolith appearance. It is suggested that the transition was associated with individual sea entry. This study demonstrated that the check and/or transition associated with sea entry are applicable to a benchmark for otolith increment counts of juvenile chum salmon inhabiting coastal waters. [ABSTRACT FROM AUTHOR]

Abstract: Artificial insemination (AI) was performed in spider monkeys; these primates are vulnerable to extinction and usually do not reproduce spontaneously in captivity. Uterine cycles were followed by daily assessment of vaginal cytology, and corroborated a posteriori by concentrations of 17-β estradiol and progesterone, measured by radioimmunoassay (RIA), in fecal samples collected once daily. Five females between 13 to 27 years old were inseminated intravaginally (with fresh semen) twice each during the periovulatory phase (Days 9–12 of the menstrual cycle; Day 0, first day of menstrual bleeding), from September to the first 3 weeks of November (most fertile months). Transcervical AI was not useful in this primate because the liquid portion of the semen completely solidified instead of liquefying as in other primates. Pregnancies were apparently achieved in 5 of 14 attempts. One female became pregnant after the first round of inseminations, delivered a healthy infant, was inseminated and got pregnant again (subsequently aborted). One female aborted, apparently due to an intramural uterine leiomyoma. Another two females stopped menstruating for a few months, then restarted menstruating (these females may have been pregnant and aborted). In conclusion, in spider monkeys: (1) captivity-induced stress did not inhibit reproduction; (2) fecal steroid hormones were useful to assess cyclicity; (3) the semen coagulum, which apparently is a tightly packed and large reservoir of spermatozoa, must not be discarded but used in AI; (4) old female spider monkeys did not have cessation of reproductive function. [Copyright &y& Elsevier]

In this study, the embryonic development of the eggs in the Mastacembelus mastacembelus (Bank & Solender, 1794) was examined. At the same time, possibilities of artificial breeding through artificial insemination were investigated. Artificial insemination was achieved by mixing the eggs of the mature female and sperm of the mature male samples caught with gill nets (22 × 22) in Ataturk Dam Lake in Turkey. To this end, first in a Petri dish (100 × 20), the testes were cut into small pieces with a lancet and the mixture of sperm–testes-tissue was obtained. The fertilization rate of the eggs was found to be 80%. The diameter of the eggs ranged from 2.015to 1.147 mm. The perivitelline space formed 0.5 h after insemination. The first cleavage occurred at the animal pool 4 h after insemination. The oil droplets had fused to a single droplet 19 h after insemination. The blastoderm became an embryonic shield 30 h after insemination. The blastoderm covered almost half the egg 40 h after insemination and embryonic body was formed. The blastoderm covered almost the whole egg 50 h after insemination. Some somites were discernible 59 h after insemination. The embryonic body reached two-third of the circumference of the egg 70 h after insemination. The tail bud began to separate from the yolk 77 h after insemination. A newly hatched larva was observed at 85 h after insemination. [ABSTRACT FROM AUTHOR]

The structure of the seminal receptacles and associated accessory pouches in the reproductive tracts of four species of Australian ovoviviparous peripatopsids is described to identify features that may relate to their inferred mode of insemination. Euperipatoides rowelli is a dermal inseminator whereas Phallocephale tallagandensis, Ruhbergia bifalcata and Cephalofovea clandestina, species with head structures in males, are considered to be vaginal inseminators. Contrary to our expectations, E. rowelli, R. bifalcata and C. clandestina have very similar seminal receptacles and display two types of accessory pouches: funnel accessory pouches that open to the haemocoel and spherical accessory pouches in which destruction of excess sperm occurs. On the other hand, Ph. tallagandensis differs in the form of its accessory pouches. Although these pouches have no direct opening into the haemocoel, sperm were observed in the act of directly penetrating their walls to gain entry from the haemocoel. Hence, despite differences in the form of their accessory pouches, all species display features that provide the potential for entry of sperm into the female reproductive tract from the haemocoel. This finding has caused us to reconsider our initial assumptions of vaginal and dermal insemination as mutually exclusive reproductive strategies. Indeed, we propose that species of peripatopsids with head structures in males that were considered to be obligate vaginal inseminators may in fact be facultative, with vaginal insemination of sexually immature females when their uterus is short and free of developing embryos followed by dermal insemination to maintain the level of sperm in storage in the seminal receptacles. This study has also provided morphological evidence for mechanisms that may facilitate female control of paternity. [ABSTRACT FROM AUTHOR]

Contents Over the past decade, there has been increasing interest in the application of reproductive technology to the conservation and management of natural populations of deer. The application of assisted reproduction technologies within natural population of deer is in its infancy. However, its future potential is enormous, particularly in relation to genetic management or conservation. This paper reviews the present state of such technologies for a wild subspecies of red deer, the Iberian red deer ( Cervus elaphus hispanicus), by discussing the major components of oestrous synchronization, semen collection/cryopreservation and insemination techniques. In addition, findings made during the course of studies on natural populations have enormous potential for the understanding of novel reproductive mechanism that may not be uncovered by livestock or human studies. A summary of these results are also reviewed here. [ABSTRACT FROM AUTHOR]

Nuptial feeding is an important part of mating in several arthropods including some spiders. A male can contribute via the nuptial gift to his own offspring (paternal investment) and/or the nuptial gift may allow him to better transfer sperm or to make the female more willing to copulate (mating effort). From the female's perspective, however, weighing a male's quality according to the presence of a gift should coincide with her mating status, because male rejection by the virgin female may decrease her fitness by increasing the likelihood of remaining unmated. Therefore, female mate choice would depend on her mating status and virgin females are expected to be less choosy than mated females. I examined this hypothesis in the nursery web spider, Pisaura mirabilis , in which females are polyandrous and mate both with males with and without a gift. I found no support for my hypothesis; instead the presence of a gift was crucial in accomplishing successful copulation regardless of female mating status. Interestingly, sexual size dimorphism was found to be a significant predictor for duration of copulation. [ABSTRACT FROM AUTHOR]

Two species of loach, the mud loach ( Misgurnus mizolepis Günther) and the cyprinid loach ( M. anguillicaudatus Cantor), are commercially important in Korea both for food and ceremonial purposes. The mud loach has superior potential for aquaculture in terms of growth, whereas the cyprinid loach has a more desirable body shape and colour. This study was conducted to produce reciprocal diploid and triploid hybrids and to evaluate their potential benefits in culture by examining growth performance, morphometrics and gonad development. Reciprocal diploid and triploid hybridization was performed by artificial insemination without or with the induction of triploidy. The successful formation of diploid or triploid karyogamy was verified by flow cytometric analysis. Body weights of induced reciprocal diploid hybrids were intermediate between those of the two parental species, i.e. hybrids were heavier than cyprinid loach but lighter than mud loach. In contrast, the growth performance of triploid hybrids was similar to that of their corresponding maternal parents. Diploid hybrids showed intermediate morphometric traits between the two species; however, the body proportions of triploid hybrids were more similar to those of the maternal species than the paternal species. Histological analyses indicated that reciprocal diploid hybrids of both sexes were able to reach maturity, as evidenced by the presence of mature oocytes or spermatozoa in the gonad tissues. However, triploid hybrids showed stringent sterility at the gonadic level; the sizes of ovaries and testes were much smaller, and gonad development was abnormal and significantly retarded. [ABSTRACT FROM AUTHOR]

The authors detail an experiment establishing the impact of insemination times on the fertilization rate and hatch rate of eggs of obscure puffer fish, Takifugu obscurus. Fertilization rates in highly matured ova are low. The authors conclude that insemination must be done within 4 hours of ovulation for better results.

To develop an appropriate cryopreservation protocol for spermatophores of black tiger shrimp, Penaeus monodon, three cryoprotectants (dimethyl sulphoxide (DMSO), methanol (MeOH) and ethylene glycol (EG)) at two concentrations (5% and 10%) were examined. Artificial implantation of spermatophores was also carried out to assess the fertilizing ability of fresh and post-thaw spermatophores. Spermatophores were collected during consecutive regenerations (15-day intervals) and assessed for qualitative and quantitative changes and also for fertilizing ability by implantation. The mean fertilization rate for artificial insemination using post-thaw spermatophore was 79.9±3.7%, lower than the fertilization rates observed for artificial implantation using fresh spermatophore and natural mating. Mean hatch rates for fresh spermatophore, frozen-thawed spermatophore and natural mating were 88.8±0.6%, 87.8±0.4% and 88.3±0.5%, respectively; and there was no difference among the three groups. The mean fertilization rate of spermatophores collected during the first stripping was higher (90.6±0.6) than during the second stripping (85.7±2.6), but the mean hatch rate was not different between the two strippings. The highest mean sperm viability (79.7±0.4%) was obtained from DMSO (5%), with no survival observed in the 10% MeOH treatment. Spermatophore weight, total sperm count and percentage of abnormal sperm were not different between spermatophores collected at the first and second stripping. This is the first study to report high fertilization and hatch rates from cryopreserved spermatophore using artificial implantation of spermatophore before spawning. [ABSTRACT FROM AUTHOR]

Typhlodromus phialatus and T. exhilaratus are morphologically close species. Their differentiation is based on the shape of the insemination apparatus and on idiosomal setae length. However, the setae length values are often intermediate between these two species and do not allow accurate identification. Furthermore, the handful of differences in insemination apparatus shape are also questionable as a means of differentiation. Synonymy between these species has thus been questioned. Three experiments were carried out. Idiosomal seta length measurements, molecular tests and cross experiments were conducted for three populations, identified as T. exhilaratus and T. phialatus according to the shape of their insemination apparatus. The results show that the variation range of seta lengths is great and that these criteria do not allow accurate separation of these populations into two species. However, molecular tests show a species-level nucleotide differentiation between them. Cross experiments confirm this result, showing complete reproduction incompatibility between the mites bearing different insemination apparatus shapes. Therefore, T. exhilaratus and T. phialatus could be considered to be two valid species and the insemination apparatus could be considered as a pertinent diagnostic criterion at the specific level. [ABSTRACT FROM AUTHOR]

Productivity and reproduction of herd have influence on dairy cattle herd profitability. Reproduction traits are dependent from genetic and non genetic factors. For improvement of breed, aspiring to increase productivity of dairy cows, the bulls having large amount of Holstein genes are used. The aim of the study - to test the influence of Holstein genes in bulls' genotype on the reproduction traits of their daughters: age at the first calving, days open, calving interval, number of services from calving to conception and calvings results. The information of a pedigree, calvings, inseminations, and part of genes of improving breed in a genotype of bulls of black and white cattle in Lithuania was used. In Lithuania, to improve dairy cattle productivity, bulls having large amount of Holstein genes are used. As a result of such selection work we have productive cows, which have days open, number of services from calving to conception and calving interval larger than cows, which have not been improved with Holstein breed genes and characterized by lower productivity. In order to avoid reproduction problems it is very important to choose the optimal time for insemination after calving, taking into account productivity of cows. [ABSTRACT FROM AUTHOR]

The eel has long been esteemed as an important food fish in the world, especially in Japan, and has been used as an experimental fish for many fields of fish physiology. However, the decreases in eel resources have been a serious concern in recent years. The catches of glass eels as seedlings for aquaculture have shown a long-term decrease in both Europe and East Asia. To increase eel resources, the development of techniques for artificial induction of maturation and spawning and rearing their larvae have been eagerly desired. Recent progress of reproductive physiology of fish, especially mechanisms of oocyte maturation and ovulation in female and of spermatozoa maturation in male, facilitate to establish techniques for hormonal induction of maturation and spawning in sexually immature eels. With persistent effort to development of rearing techniques of larvae, we have first succeeded to produce glass eel. These applied techniques are may contribute to understand the basic reproductive physiology of the eel. [ABSTRACT FROM AUTHOR]

Studies the reproductive cycle of the bottlenose dolphin. Characteristics of the estrous cycle; Artificial insemination using frozen-thawed spermatozoa; Excretory dynamics of urinary luteinizing hormone and ovarian steroid metabolites; Effect of a synthetic progesterone analog on reproductive hormone excretion.

Artificial insemination with frozen-thawed buck semen shows variable results which depend on many factors related to semen quality and the cryopreservation processing. We conducted experiments based on a new freezing method, directional freezing, of large volumes (8 ml). In the first experiment semen from three Saanen bucks, ages 1–2-years-old and genetically selected for milk improvement, was frozen individually. Two to three-years-old Saanen females (n = 164) were synchronized with controlled internal drug release (CIDR), pregnant mare serum gonadotrophin (PMSG) and prostaglandin. Double cervical inseminations were performed with frozen-thawed semen and fresh semen as control. In the second experiment we used pooled, washed frozen semen to examine the effect of washed seminal plasma.The motility after washing was 80–90% and after thawing was 55–65% for all bucks. The sperm concentration increased with the collections and the advance into the breeding season from 1.9 × 109 to 4.4 × 109 cell/ml average. Two inseminations were carried out at 8 h intervals. The first insemination was performed at 32 h after CIDR withdrawal with fresh and frozen-thawed semen. Pregnancy rates were assessed by ultrasonography conducted 40 and 90 days post-insemination (from three bucks). Results were 58, 67, 50% with fresh semen, and for frozen semen were 33, 37 and 53%; these results were significantly different in one of the three bucks (P < 0.005). In the second experiment with pooled, washed semen the pregnancy rate was 41.6%, which compared with the average results of the frozen semen in the first experiment 38.9% no significant difference was found. We conclude that freezing buck semen in large volumes (8 ml) is possible. Cryobanking of buck semen will facilitate a genetic breeding program in goats and preservation of biodiversity. Washed semen did not improve the fertility of the semen when Andromed bull extender is used. [Copyright &y& Elsevier]

The objectives were to determine the efficacy of the HeatWatch™ (HW) electronic estrus detection system for monitoring behavioral estrus (including duration and intensity) in red deer hinds and to evaluate pregnancy rate to AI after detected estrus. Red deer hinds (Cervus elephus; n=50) were allocated into two treatment groups: AI following synchronization (CIDR/PMSG) and observed estrus (induced estrus group: IE; n=25) or AI following the detection of natural estrus (NE; n=25) without hormonal treatment. Hinds were fitted with two HeatWatch (HW) electronic estrus detection transmitters, one above the tail (bottom) and one between the tuber coxae of the pelvic girdle (top), and visual observations for mounting activity began with the aid of young sterile red deer stags (18 months old) fitted with marking harnesses. Hinds in both groups were inseminated (10–12 h after observed estrus) with frozen–thawed red deer semen using a transvaginal/cervical AI approach. Following a 26-day period of AI, hinds were placed with a mature fertile stag for an additional 30-day natural breeding period. Pregnancy diagnosis was performed 57 and 86 days after the start of AI. While the hinds were housed with the young stags, 82% were detected in estrus by visual appraisal of stag crayon marks, but only 32% of these were detected by HW. In contrast, in the hinds housed with the mature stag, 93% detected in estrus by crayon marks were also detected by HW. The top HW transmitter consistently recorded more mounts (P<0.05) than the bottom transmitter. The pregnancy rate was numerically better in IE versus NE hinds (42% versus 29%, P>0.10). In summary, there were no differences (P>0.10) in the intensity (number) or duration of mounts (detected by HW) during estrus in IE versus NE hinds, and HW was most effective in detecting estrus in the presence of a heavier, mature stag versus a younger stag. When used in combination with transvaginal AI, an overall first-service pregnancy rate of 36.6% was achieved with AI of frozen–thawed semen in red deer hinds following detected estrus. [Copyright &y& Elsevier]

This paper reports on cryopreservation of sperm from turbot (Scophthalmus maximus). The effects of various extenders, cryoprotectants and sperm–egg insemination ratios on motility score and/or fertilization capacity of post-thaw turbot spermatozoa were examined to optimize cryopreservation procedures. Post-thaw motility of frozen sperm obtained with extender TS-2 was higher than those achieved with extenders D-15 and modified plaice Ringer solution (MPRS). The most effective cryoprotectant was determined to be 10% dimethyl sulfoxide (DMSO). Fertilization of small egg batches (2 ml eggs) with frozen sperm resulted in average fertilization rate (FR) (70.1±8.9%) and hatching rates (HR) (46.8±5.2%) similar to the fertilization rates (74.7±8.0%) and hatching rates (47.5±6.8%) of fresh sperm. The minimal density of frozen sperm required to obtain satisfactory fertilization rate was determined to be 2000:1 (sperm/egg). Fertilization of larger egg batches (40 ml eggs) with sperm frozen in 1.8-ml cryovials provided similar fertilization rates (71.6±7.3%) to that of the fresh sperm (76.2±5.7%), whereas the hatching rates (34.7±3.6%) of eggs fertilized with frozen sperm were slightly lower than those of fresh sperm (41.1±3.7%). Our results demonstrated that cryopreservation technique for turbot sperm in 1.8-ml cryovial could be used for hatchery purposes. [Copyright &y& Elsevier]

The present work optimizes procedures for artificial insemination of European catfish, Silurus glanis L., with goals to increase collection of sperm and hatching rate. We address technical aspects of immobilizing solution (IS), activation solution (AS), process of insemination, activation of gametes and elimination of eggs stickiness with a practical solution for artificial propagation in hatcheries. Spermiation was stimulated and could be sustained over a period of 1 month after weekly carp pituitary (CP) injection of 5 mg kg-1 b.w. Males produced significantly the largest quantity of sperm, 0.12–0.13·109 spermatozoa kg-1 b.w., after third and fourth injections. The best IS for sperm was determined as solution containing 170–200 mM NaCl, 30 mM Tris–HCl, pH 7 and activation solution for activation of gametes containing 17 mM NaCl, 5 mM Tris–HCl, pH 7. The ratio of 1:2 rather than ratio of 1:1 in the control batch (volume of activation solution in milliliters/grams of eggs) increased the hatchability up to 68%. The highest hatching success of 82–88% from four individual females were found for sperm number of 800–80,000 spermatozoa egg-1, in comparison to 26% hatching at 80 spermatozoa egg-1. The best time for application of alcalase enzyme for elimination of egg stickiness was 3 min after gamete activation at concentration of 20 ml enzyme l-1 with exposure of 2 min. [Copyright &y& Elsevier]

Reports that sperm cryopreservation and artificial insemination are important management tools for giant panda breeding and the preservation of extant genetic diversity. Examination on the influence of freeze-thawing on sperm function and capacitation; Assessment of sperms from a number of giant pandas before and after rapid cryopreservation; Discovery on the ability of giant panda spermatozoa to undergo capacitation with or without chemical accelerators and withstand a rapid cryopreservation protocol.

Viable F1 hybrids were obtained from crosses of female Macrobrachium nipponense and male Macrobrachium hainanense involving spermatophore transfer and artificial insemination. This represents the first successful known case of hybridization of two Macrobrachium species by means of artificial insemination. The hatching rate was over 90%. About 20–60% of newly hatched larvae metamorphosed to postlarvae. The morphological characteristics of the hybrids resembled a combination of features of both parents. Malate dehydrogenase (MDH) and esterase (EST) isozyme electrophoresis indicated parents and F1 hybrids showed co-dominant expression of the paternal and maternal alleles controlling the isozymes and confirmed the hybridization. [Copyright &y& Elsevier]

We developed a simple, easy method with a microplate to artificially incubate Japanese crayfish (Cambaroides japonicus) eggs for their cultivation. We prepared 6-, 12-, 24-, 48-, and 96-well microplates containing sterile water heated to 5, 10, 15, and 20 °C. Fourteen experimental groups for each water temperature were prepared for each of different water volumes (0.125–10 ml) in each well. One embryonic egg was placed in each well. Experiments were also conducted with water collected from the lake where the eggs were harvested from and held at 15 °C. In the microplates with sterile water, high proportions of eggs hatched (60–100%) at 15 °C in all volumes of water, although the proportions of hatching were low (0–20%) at 5, 10, and 20 °C. All eggs died in the experiments that used lake water. We conclude that the 96-well would be the most effective size to hatch crayfish eggs in, because of its convenience. This method using a microplate is simpler and easier compared with methods of previous studies to artificially incubate crayfish eggs, and therefore it might be useful to incubate eggs of other freshwater crayfish species. [Copyright &y& Elsevier]

The application of artificial insemination techniques as a way to overcome the lack of mating of the closed thelycum Farfantepenaeus paulensis under large scale conditions is reported. The performance of artificially inseminated females (49.4±11.6 g) maintained in conventional versus unisex maturation systems was compared. Groups of unilaterally eyestalk-ablated females were artificially inseminated with one compound spermatophore soon after molting. The percentage of fertilized spawns increased from 26% before the use of artificial insemination to 57% afterwards. The reproductive performance of inseminated females held in conventional or unisex maturation systems showed no significant differences. Artificial insemination was considered a practical approach to overcome the lack of mating of F. paulensis under large-scale conditions with acceptable reproductive performance. The results also indicate the possibility of holding females separately from males, which could mean significant improvements in maturation systems through the increase in the stocking density of spawning females. [Copyright &y& Elsevier]

The Asian elephant (Elephas maximus) population in the wild has been in decline for several decades and breeding in captivity has not been self-sustaining. The use of artificial insemination (AI) can help overcome many of the difficulties associated with breeding elephants in captivity; however, the ability to store semen for extended periods of time is critical to the successful application of AI to elephants. The objective of the present study was to assess the effects of four different semen extenders and the presence of egg yolk on the viability and motility of Asian elephant semen stored at 4 °C. High quality ejaculates (n=4) were collected from two Asian elephant bulls by rectal massage. Aliquots of each ejaculate were extended in four different diluents (Beltsville thawing solution (BTS); Tris–citric acid (TCA)/fructose-based; Beltsville F5 (BF5); dextrose-supplemented phosphate-buffered saline (PBS)) with or without egg yolk then cooled and stored at 4 °C. The percentages of viable (viability) and motile (motility) sperm were evaluated at 8, 24 and 48 h following collection. The addition of egg yolk significantly reduced the percentage loss in viability from initial collection to 48 h compared to extenders without egg yolk (17.0±8.2 versus 32.6±8.9 decline in percent viable sperm in the population, respectively; P<0.05). Extender and egg yolk affected (P<0.005) total motility and percent progressively motile sperm at all evaluation times during incubation. TCA + egg yolk maintained higher (P<0.05) levels of progressive motility compared to other extenders supplemented with egg yolk. These results indicate that Asian elephant semen extended in TCA diluent supplemented with egg yolk can maintain at least 50% viability and motility when stored at 4 °C for 48 h. [Copyright &y& Elsevier]

In many animals reproductive success is determined after insemination by the interaction of male and female processes. While sperm competition is reasonably well understood in some taxa, other processes, such as cryptic female choice and differential early embryo mortality resulting from genetic incompatibilities, are less well understood. The relative importance of these different factors contributing to reproductive success is difficult to assess. Here we control for male-mediated effects (which are often considerable and can mask more subtle processes) through the artificial insemination of known numbers of sperm in the domestic fowl to reveal that male reproductive success is nontransitive across females: the success of a particular male depends on the background against which his sperm compete for fertilization. Two potential processes could account for this effect: cryptic female choice (sperm choice) or differential early embryo mortality. Regardless of the mechanism, nontransitivity of male reproductive success has important evolutionary consequences, including the maintenance of variation in male fitness. [ABSTRACT FROM AUTHOR]

For decades, attempts to breed elephants using artificial insemination (AI) have failed despite considerable efforts and the use of various approaches. However, recent advances in equipment technology and endocrine-monitoring techniques have resulted in 12 elephants conceiving by AI within a 4-year period (1998–2002). The successful AI technique employs a unique endoscope-guided catheter and transrectal ultrasound to deliver semen into the anterior vagina or cervix, and uses the “double LH surge” (i.e., identifying the anovulatory LH (anLH) surge that predictably occurs 3 weeks before the ovulatory LH (ovLH) surge to time insemination. This study describes the 6-year collaboration between the National Zoological Park (NZP) and the Institute for Zoo Biology and Wildlife Research (IZW), Berlin, Germany, that led to the refinement of this AI technique and subsequent production of an Asian elephant calf. The NZP female was the first elephant to be inseminated using the new AI approach, and was the fifth to conceive. A total of six AI trials were conducted beginning in 1995, and conception occurred in 2000. Semen was collected by manual rectal stimulation from several bulls in North America. Sperm quality among the bulls was variable and was thus a limiting factor for AI. For the successful AI, semen quality was good to excellent (75–90% motile sperm), and sperm was deposited into the anterior vagina on the day before and the day of the ovLH surge. Based on transrectal ultrasound, ovulation occurred the day after the ovLH surge. Pregnancy was monitored by serum and urinary progestagen, and serum prolactin analyses in samples collected weekly. Fetal development was assessed at 12, 20, and 28 weeks of gestation using transrectal ultrasound. Elevated testosterone measured in the maternal circulation after 36 weeks of gestation reliably predicted the calf was a male. Parturition was induced by administration of 40 IU oxytocin 3 days after serum progestagens dropped to undetectable baseline levels. We conclude that AI has potential as a supplement to natural breeding, and will be invaluable for improving the genetic management of elephants, provided that problems associated with inadequate numbers of trained personnel and semen donors are resolved. Zoo Biol 23:45–63, 2004. © 2004 Wiley-Liss, Inc. [ABSTRACT FROM AUTHOR]

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The silkworm, Bombyx mori, has a dimorphic sperm system. The eupyrene sperm is the sperm to fertilize eggs and the apyrene sperm plays a crucial role for assisting fertilization. Heat-treated (33°C for 96h) Daizo (DH) males, one of the strains in the silkworm, produce only eupyrene sperm, while in triploid males only apyrene sperm are functional. Though both types of males are found to be sterile, double copulation of the two males with a single female greatly increases fertility. Here we examined the fertilizing ability of eupyrene and apyrene sperm by means of an artificial insemination technique previously established in B. mori. Neither the eupyrene sperm collected from DH males, nor the apyrene sperm from triploid males have the ability to fertilize eggs. Artificial insemination with the mixture of eupyrene and apyrene sperm leveled up the frequency of fertilized eggs to more than 80%. When cryopreserved DH sperm (eupyrene sperm) were subjected to the same experiment, more than 95% fertilized eggs were obtained. These results confirmed that apyrene sperm play an important and indispensable role in fertilization in B. mori. Separate collection of functional eupyrene sperm and functional apyrene sperm and success of fertilization by means of the artificial insemination technique are applicable for further studies to elucidate the function of apyrene sperm. J. Exp. Zool. 297A:196–200, 2003. © 2003 Wiley-Liss, Inc. [ABSTRACT FROM AUTHOR]

Eggs of the Japanese eel, Anguilla japonica, were obtained by hormonally induced maturation and ovulation, and then categorized as buoyant and non-buoyant types in seawater after insemination. The water content of the buoyant eggs, 93.1%, was significantly higher than that of the non-buoyant eggs, 87.5%. Furthermore, the buoyant eggs had 1.5 times more free amino acids (FAA), 573.5 μmol/g dry weight (DW), than the non-buoyant eggs, 385.8 μmol/g DW, while no significant difference in egg lipid content or protein content was found. Compared to other pelagic eggs reported in the literature, the FAA content of the buoyant eggs was relatively low, although the water content was similar. These results suggest that there is a close relation between the quality of Japanese eel eggs obtained by typical hormonal inducement and the FAA content of the eggs, but the roles of FAA in the acquisition of egg buoyancy and fulfilling the early nutritional requirements are comparatively minor. [Copyright &y& Elsevier]

The site of semen deposition (vaginal, cervical or uterine) and the time required for transvaginal artificial insemination (AI) using the Gourley Scope method of AI vs. a standard speculum-guided AI gun was assessed in three species of farmed deer (fallow, red and white-tailed deer). Pregnancy rates using these two transvaginal AI methods were also compared in a commercial herd of fallow does. The estrous cycles of fallow (n=53), red (n=27) and white-tailed (n=27) deer were synchronized using half of a Norgestomet ear implant inserted for 14 days. Following implant removal, does were artificially fix-time inseminated with frozen-thawed semen using either a 0.5 ml standard French-style speculum-guided AI gun or the Gourley Scope method of AI. The time-frame (i.e., duration in minutes) for the AI procedures performed on each animal and the location of semen deposition was recorded. To assess effectiveness of AI, transrectal ultrasonography was conducted in fallow deer 45 days post-AI for pregnancy diagnosis. In fallow does and red deer hinds, use of the Gourley Scope (96.3 and 100%, respectively) enabled a greater (P<0.05) percentage of intrauterine inseminations compared to the speculum-guided AI gun (73.1 and 42.9%, respectively). In contrast for white-tailed does, both the speculum-guided AI gun and the Gourley Scope resulted in a similar proportion of intrauterine inseminations (92.3 and 100%, respectively). The duration for performing the two transvaginal AI techniques were also similar in red and white-tailed deer. However in fallow does, AI with the aid of the Gourley Scope took longer (P<0.05) to conduct than the speculum-guided AI gun (6.6±0.72 and 4.3±0.34 min, respectively). The pregnancy rate using both methods of AI combined was overall 32.1% for fallow does. The total percentage of fallow does that conceived with the speculum-guided AI gun was 42.3% whereas the pregnancy rate using the Gourley method of AI was only 22.2% (P<0.05). Although both AI methods may be acceptable for use in farmed deer, the standard speculum-guided AI gun is simpler and produced more pregnancies compared to the Gourley Scope method in fallow deer, despite a greater proportion of intrauterine inseminations using the Gourley Scope. [Copyright &y& Elsevier]

Examines the sperm competition mechanisms in wild bird population. Efficacy of anticopulator; Determination on the frequency and timing of extra-pair inseminations; Fertilization probability caused by female behavior.

Presents information on the endangered species of the giant panda, while examining ways in which artificial insemination can used to protect the species. Where and when artificial insemination was applied; What has been identified as contributory to the failure of sperm-egg binding and fusion; What tests conducted showed; Analysis of the results of the tests.

Optimum conditions for hydrostatic pressure treatment for duplication of chromosome set in gynogenetically activated fighting fish, Betta splendens (Regan), eggs were identified. Maximum survival of heterozygous gynogens was 50%, when 2.5-min-old eggs, after insemination with UV irradiated tilapia sperm, were pressure shocked at 7000 psi for 6 min. The frequency (21%) of homozygous gynogenetic fry was high, when the 34min (post-insemination) old eggs, which were inseminated with tilapia sperm, were pressure shocked for 5 min. Sex ratio of gynogenetic progeny suggested that the mechanism of sex determination in this fish is homogametic female and heterogametic male. [ABSTRACT FROM AUTHOR]

Triploidy was induced in the fighting fish, Betta splendens Regan, by varying all possible combinations of temperature (37–41°C), time after insemination (2–3 min) and shock duration (2–4 min). Heat shock at 39°C for 3 min duration initiated 2.5 min after insemination gave high frequencies of triploids (86%) as assessed from chromosome number and red blood cell nuclear volume. There was no significant difference in the growth rate of triploid and diploid fish. Gonadal development in both sexes was retarded in triploids at 5 months of age. Eggs fertilized with milt from triploids developed to gastrulation. Beyond gastrulation there was increasing mortality associated with abnormalities and none of them hatched. The display frequencies of air gulping, erection of operculum and fins, striking and biting, and undulating movements were fewer in triploids compared to diploids. It appears that triploids are less aggressive than diploids. The aggressive behaviour of fighting fish may be related Io their reproductive activity. [ABSTRACT FROM AUTHOR]

The sperm of tench, Tinea tinca L., is characterized by a milky colour and consistency, and is of very low density. After collecting the sperm, motion of spermatozoa was recorded even without water activation. A better motility value (value 4.36 on average) was observed in spermatozoa collected in immobilizing solution (collecting medium) and stored for 3h, when compared with spermatozoa without collecting medium. Average total and relative numbers of spermatozoa were 12.16 ⊗ 109 per male and 18.50 ⊗ 109 per kg of body weight, respectively. When testing the effect of activating solution in artificial propagation of tench, the highest fertilization rates (81.3 and 85% in two cases) were found for NaCl solution with an osmotic concentration of 34 or 69 mOsmol and for fresh water, respectively. The fertility rate was reduced significantly (P « 0.01) by any increase above 105 mOsmol in NaCl concentration in the activating solution. In the tests of optimal method of artificial fertilization, the highest hatching rate of sac fry (71.35%) was found in sperm collected into immobilizing solution. The application of immobilizing solution significantly increased the number of sac fry at the levels P«0.1 and P«0.01, if compared with intact sperm stored for 3 h and fresh sperm, respectively. [ABSTRACT FROM AUTHOR]

Most of the progress in female great ape reproduction has focused on monitoring ovarian and endocrine activity. Perhaps of more importance has been the gradually evolving interest and willingness to consider biotechnology as a potential, viable approach for enhancing reproductive performance. Artificial insemination (AI), in vitro fertilization (IVF), and embryo transfer offer an array of possibilities for combating infertility and understanding the fundamental differences and similarities among great ape species. Multidisciplinary efforts have assessed reproductive competence, from the simple (i.e., perineal tumescence and urinary occult blood in chimpanzees) to the complex (i.e., ovum recovery and IVF following exogenous hormone treatment in chimpanzees and gorillas). Hysterosal-pingography and laparoscopy have been used to determine uterotubal patency and to identify pathological conditions in gorillas. Assays for steroid metabolites in serial urine samples, to permit accurate assessments of ovarian cyclicity. have been developed and validated for all great apes species. Stimulation of follicular recruitment and maturation h4s been achieved following administration of clomiphene citrate (chimpanzees and gorillas) and human menopausal gonadotropin (gorillas), Clomiphene citrate and human chorionic gonadotropin have been used to regulate ovulation for the Al of chimpanzees. Pregnancies have resulted in chimpanzees and gorillas following Al using fresh and cryopreserved/thawed semen; however, conception rates vary. Embryos have been nonsurgically recovered from chimpanzees after timed matings and follicular oocytes have been recovered from chimpanzees and gorillas by transabdominal laparoscopy after exogenous hormone treatment. [ABSTRACT FROM AUTHOR]

Sound production during reproductive behaviour, dyadic encounters and distress situations was investigated in the callichthyid catfish Corydoras paleatus. Sounds were broad-band, pulsed, acoustic signals produced during abduction of the pectoral spines. Only males emitted trains of sounds during courting and trains of sounds of shorter duration during dyadic encounters. Several males, which are usually smaller than females, courted one gravid female without obvious cooperation or competition between them. During mating, one previously vocalizing male clasped the female's barbels with one pectoral spine and inseminated the eggs. The number of successful spawnings, days until spawning, and number of eggs laid was not related to the number of males (one, two or three) combined with one female. Males did not behave aggressively towards each other during courting or in dyadic encounters. In distress situations, when fish were hand held, both sexes and juveniles produced single sounds. The dominant frequency was negatively correlated with body size and the sound duration was positively correlated with relative length of pectoral spines (standardized to body length). This acoustical behaviour in C. paleatus differs considerably from Hoplosternum thoracatum, a representative of the callichthyine subfamily, in which vocalization was observed during territorial behaviour in males and aggressive behaviour in both sexes. This is the first report of a major difference in vocalizing behaviour within one teleost family. [ABSTRACT FROM AUTHOR]

This paper reports preliminary studies designed to evaluate hybridization through natural mating and artificial insemination between female Penaeus japonicus Bate, 1888 and male Penaeus kerathurus Forskäl, 1775. Although both species were able to mature in captivity, natural copulation was not achieved. Different techniques, including manual squeezing in the base of the fifth pair of pereiopods. electrical stimulation and dissection, were used for the artificial insemination experiments. However, only dissection gave spermatophores. Stained sperm showed no malformations and its viability was very high. Seven out of 10 inseminations (70%) produced eggs, but the development of embryos was abnormal. It is suggested that the high genetic distance between these species is responsible for these results. [ABSTRACT FROM AUTHOR]

This article proposes that serious consideration should now be given to the establishment of banks of genetic material from rare and endangered species. In the light of advances in the technology associated with semen assessment and preservation, the limitations that have so far discouraged the practice of semen freezing and banking can now be minimized and circumvented. The authors review these developments and suggest that semen preservation should be reconsidered as a practical means of preserving the genetic diversity currently contained within captive stocks of wild animals. [ABSTRACT FROM PUBLISHER]

The study was done to understand the preservation of Washed Spermatozoa of Mehsana Buck at Refrigeration temperature. The washed spermatozoa from 78 semen ejaculates, 26 each from 3 Mehsana bucks, were preserved at 4 ± 10C temperature in SM and TCFEY diluents up to 72 hours and the physical characteristics of spermatozoa were studied to assess the suitability of diluents. The individual sperm motility and live spermatozoa decreased significantly whereas abnormal spermatozoa increased significantly (P<0.05) at each 12 hrs interval of preservation in two diluents. The effect of diluents on spermatozoan characteristics was less pronounced in SM than TCFEY diluent. Among the interaction studies, stage of preservation affected the live spermatozoa and their morphology. The findings suggested superiority of SM diluent over TCFEY for preservation of Mehsana buck semen. [ABSTRACT FROM AUTHOR]

Abstract: Artificial inseminations using sperm masses were applied to intra-specific crosses of Penaeus (Litopenaeus) occidentalis, as validation of the technique. Reciprocal, inter-specific crosses of P. occidentalis and P. (Litopenaeus) stylirostris were performed; complementary, inseminations of both species were also accomplished with sperm masses of P. (Litopenaeus) vannamei. Intra-specific inseminations generated viable spawns with 54.2% of normal embryos; on the contrary, inter-specific spawns were all unfertile, with a high rate of eggs showing abnormal intracellular corpuscles. Based on recent discoveries about sperm maturation/capacitation, a new approach to study inter-specific sperm–egg compatibility in the sub-genus Litopenaeus is discussed. [ABSTRACT FROM AUTHOR]

The low efficiency of estrus detection leads to inappropriate time for AI insemination in swamp buffalo. The ovulation synchronization (Ovsynch ) protocol has successfully applied in bovine and riverine buffalo but not in swamp buffalo . Our previous studies demonstrated that Ovsynch can be applied with a 34.6% pregnancy rate (Chaik hun et al., 2009). The aim of this study was to observed ovulation time and changes in hormonal profiles during ovsynch of Thai swamp buffaloes . The experiments were carried out in twelve buffaloes, estrus induction was performed by PGF2? (D-13) - PGF2? (D- 2) - GnRH(D0 ) - PGF2? (D7) - GnRH (D9) (Presynch - Ovsynch) treatment. Blood samples were collected from jugular vein on days 0, 4, 7, 9 and every 4 h after second GnRH injection. Plasma progesterone and estradiol concentrations were determinated by radioimmunoassay technique. The ovarian changes observed by ultrasonography and behavior of estrus signs investigated by rectal palpation and visual observation. Ten buffaloes (83.3%) ovulated with the average ovulated dominant follicles was 1. 24 ± 0.2 mm. and the me an + SD o f ovulation time was 30 + 2.8 h . A low level of progesterone (0.36 ± 0.15 ng/ml.) was maintained during the period from day 9 to day 12. During the estrus , estradiol 17- level fluctuated in a pulse fashion with the average level was 3.84 ± 1.0 pg/ml. recorded at about 24 h after the onset of heat. From the rectal palpation show that 12 h after the highest (+3) tone of uterus was the basic criteria for indicated ovulation time in swamp buffalo. The results show high efficiency of ovsynch program for induce ovulation and indicate the interval between the last GnRH administration of the ovsynch protocol and AI should be 28-32 h in Thai swamp buffalo. [ABSTRACT FROM AUTHOR]

The article reports on the controversial issues faced by the tiger breeding facility, Hengdaohezi Feline Breeding Centre, in Harbin, China. With about 800 Siberian tigers in its collection, the facility is facing media attention due to violent videos taken from its tourism programs. The Convention on International Trade in Endangered Species of Wild Fauna and Flora (CITES) meeting, held in Hague, Netherlands on June 2007, likewise criticizes China's tiger-breeding program and threatens to reintroduce its 600 tigers into the wild. Meanwhile, scientists are also considering the genetic diversity of Siberian tigers through artificial insemination to avoid their extinction.

The aim of this study was to evaluate the various alternations between long and short days of sunlight in an attempt to optimise the production of semen collected for artificial insemination (AI) under field conditions. The results suggest that alternation between long days and short days allowed for the production of AI doses all year round in large scale under field conditions. The alternation 45 LD / 45 SD seems to be the better treatment to optimise the production of AI doses. Under these conditions the production of AI doses was not influenced by the rhythm of collection. More investigations are necessary to optimise the rhythm of weekly collection. [ABSTRACT FROM AUTHOR]

Examines the effect of dietary vitamin E supplementation on fertilising ability of pheasant semen following artificial insemination. Sperm storage enhancing effect of alpha-tocopherol; Age of the birds; Number of fertilized eggs after artificial insemination (AI); Percent egg fertility; Percent decline of egg fertility during second week following AI.