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In December 2009, 8 female Himalayan tahrs (Hemitragus jemlahicus) were synchronized by inserting CIDR devices for 13 or 15 days. One day before removal of the CIDRs, a combination of pregnant mare serum gonadotropin (PMSG) 400 IU and hCG 200 IU (PG600®, Intervet, Boxmeer, the Netherlands) was intramuscularly injected into each animal and PGF2αwas injected at the time of CIDR withdrawal. Semen was collected by electrical stimulation, diluted, cooled to 5°C for 2h, and maintained in a refrigerator before performing AI twice. Pregnancy diagnosis was conducted by Doppler ultrasonography on March 2, 2010. Four female Himalayan tahrs were impregnated, showing a fetus present in their uteri. In the first experiment in which CIDRs were inserted into the vagina of 4 Himalayan tahrs for 13 days, 3 of the tahrs were transcervically inseminated with fresh semen, whereas 1 was inseminated with frozen–thawed semen 33h after CIDR removal. As a second insemination experiment, 46h after CIDR removal, all 4 tahrs were inseminated with diluted (1:3) fresh semen. While the tahr that was later inseminated with frozen–thawed semen did not become pregnant, the other 3 Himalayan tahrs became pregnant. In the second experiment, CIDR devices were inserted into the vaginas of 4 Himalayan tahrs for 15 days. Extended (1:7)-chilled semen was used for transcervical insemination 42 and 49h after CIDR withdrawal. When performing the second insemination, it was difficult or impossible to penetrate cervical canals. Thus, only 1 tahr became pregnant. These findings show 13 days of CIDR insertion, the dose of PMSG 400 IU and hCG 200 IU, and the use of PGF2αas beneficial for inducing oestrus synchronization, and prove that extended-chilled semen may also be used for artificial breeding of Himalayan tahrs. This is the first report of successful pregnancy induced by artificial insemination of fresh or extended-chilled semen in oestrus-synchronized Himalayan tahrs. [ABSTRACT FROM AUTHOR]

Artificial insemination (AI) using sex-selected sperm of bottlenose dolphins is currently used for the reproductive and social management of captive populations, but distance of males to the sorting facility represents a limitation of the procedure. Sorting and recryopreservation of previously frozen–thawed (FSF) sperm would facilitate the global application of this technology. Although a calf has been produced using FSF sperm (O’Brien et al.2009 Theriogenology 71, 98–107), a comprehensive examination of the in vitroquality of such samples is needed. The objective was to compare the in vitroquality of nonsorted (CNTR) and sorted (FSF) dolphin sperm before and after recryopreservation using straw (STR) and directional freezing (DF) methods. At all assessment intervals, sperm were evaluated for 1) motility parameters with computer-assisted sperm analysis (CASA); 2) plasma membrane integrity (viability) and acrosome integrity using propidium iodide/fluorescein isothiocyanate-labeled peanut agglutinin (PI/FITC-PNA) staining and 3) DNA denaturation using the sperm chromatin structure assay (SCSA). Semen from 3 ejaculates×3 males was cryopreserved by DF. After thawing, samples were divided into CNTR and FSF. The CNTR sperm were recryopreserved using STR and DF methods with assessments performed after the first thaw (PT1) and before recryopreservation (PF2). The FSF sperm were prepared for sorting using a density gradient centrifugation (DGC) method, stained with Hoechst 33342, sorted (SX MoFlo®, Dako, Fort Collins, CO, USA), then recryopreserved using STR and DF methods. The FSF sperm were assessed post-PT1, post-DGC, post-stain, post-sort, and at PF2. After the second thaw (PT2), CNTR and FSF samples were diluted (1:0.1, vol/vol) with Androhep Enduraguard™ (AE; Minitube of America, Verona, WI, USA), incubated at room temperature, and assessed at 0, 6, 12, 18, and 24h PT2. The PT1 samples retained high proportions of their PF1 total motility (TM) and progressive motility (PM) (mean±SD; 87.9±7.3% and 92.2±5.9%, respectively). The FSF sperm had improved (ANOVA; P<0.05) motility (TM, PM, VAP, VCL, VSL) and viability at PF2 compared with PF1. The FSF sperm recryopreserved using DF had higher (P<0.05) motility over the 24-h post-thaw incubation period compared with STR. The CNTR sperm DNA fragmentation remained unchanged throughout the process. The DNA fragmentation of FSF samples increased after staining (P<0.05), then decreased during the PT2 incubation period, stabilising at lower values (P<0.05) than CNTR from 6 to 24h PT2. This unusual pattern indicates a possible interaction between Hoechst 33342 and acridine orange. After recryopreservation, the viability of FSF sperm was higher (P<0.05) than that of CNTR sperm. Results indicate that bottlenose dolphin sperm undergoing cryopreservation, sorting, and recryopreservation are of adequate quality for use in AI. [ABSTRACT FROM AUTHOR]

During the fall 2008 breeding season in Louisiana, 2 synchronization protocols for fixed-timed artificial insemination (FTAI) in White-tailed deer were assessed. The objective was to determine if GnRH at FTAI improved pregnancy rates in White-tailed deer. White-tailed does (n= 35) with a mean body weight of 65.8 kg (range: 54.0 to 79.8 kg) and average age of 5.9 years (range: 1.5 to 9.5 years) were stratified by weight, age, and last fawning date into 2 groups. Treatment and control synchronization protocols were randomly assigned to each group. Does were synchronized with a CIDR-G device for 14 d and were then subjected to either FTAI 60 hpost-CIDR removal (control, n= 17) or 100 μg (i.m) injection of GnRH at FTAI 60 h post-CIDR removal (treated, n= 18). At insemination (AI), insemination pipette placement score (IP; 0 = at cervical os to 3 = within uterine body), mucous scores (clear/cloudy, viscous/nonviscous, or no secretions), vulva assessments (hyperemic/nonhyperemic and swollen/not swollen), and sperm progressive motility were recorded. Does were inseminated with frozen-thawed sperm (5 × 107progressively motile cells pre-freeze) from 2 fertile bucks stratified across each treatment group. Starting 28 d following AI, intact bucks (ITB) were introduced into both groups for natural mating. Pregnancy was determined via transrectal ultrasonography 73 or 80 d postinsemination. Does, confirmed pregnant via ultrasonography, had fawns within the reported gestation range of 187 to 222 days. Placentomes were visualized and measured in AI pregnancies (range: 32.7 to 56.2 mm in length), whereas pregnancies derived from ITB presented no identifiable placentomes. However, crown-rump measurements were obtainable from ITB pregnancies (range: 13.4 to 21.7 mm). Five does were not included in the final analyses as they were either lost to predators or removed because of illness. IP (1.4 ± 0.24 v.1.3 ± 0.30; P= 0.671), mucous classifications (3.0 ± 0.35 v.2.5 ± 0.39; P= 0.311), vulva assessments (2.1 ± 0.29, 1.7 ± 0.23; P= 0.223), and sperm motility (1.6 ± 0.16, 1.7 ± 0.22; P= 0.829) were not different for pregnant and nonpregnant AI does, respectively. Treatment did not affect AI pregnancy rates (53 v.27%) or fecundity rates (1.6 v.1.3 offspring/doe) for the GnRH treated and control groups, respectively. Addition of GnRH to a 14-day estrus synchronization protocol did not result in significantly higher pregnancy rates compared with controls. More studies are needed to determine the effect of GnRH on White-tailed pregnancies following FTAI protocols. We have demonstrated that differentiating pregnancies derived from AI and ITB could be accomplished by utilizing transrectal ultrasonography as early as 73 d postinsemination in White-tailed does. [ABSTRACT FROM AUTHOR]

The reproductive physiology of the Pacific white-sided dolphin, Lagenorhynchus obliquidens, was characterized to facilitate the development of artificial insemination (AI) using cryopreserved spermatozoa. Specific objectives were to: 1) describe reproductive seasonality of the Pacific white sided dolphins; 2) describe urinary LH and ovarian steroid metabolites during the estrous cycle; 3) correlate LH and ovarian steroidal metabolite patterns to ultrasound-monitored follicular growth and ovulation; and 4) assess the efficacy of synchronizing estrus, sperm collection/cryopreservation, and intrauterine insemination. Ovulations (64%, n=37) and conceptions (83%, n=18) occurred from August to October. Peak mean serum testosterone (24 ng/ml), cross-sectional testicular area (41.6 cm²), and sperm concentration (144.3x107 sperm/ml) occurred in July, August, and September respectively. Spermatozoa were only found in ejaculates from July to October. Estrous cycles (n=22) were 31 d long and were comprised of a 10 d follicular and 21 d luteal phase. Ovulation occurred 31.2 h after the onset of the LH surge and 19.3 h after the LH peak. Follicular diameter and circumference within 12 h of ovulation were 1.52 and 4.66 cm respectively. Estrus synchronization attempts with altrenogest resulted in 17 (22%) ovulatory cycles with ovulation occurring 21 d post-altrenogest. Ten AI attempts using cryopreserved semen resulted in five pregnancies (50%). The mean gestation length was 356 days (range 348-367). These data provide new information on the Pacific white-sided dolphin's reproductive physiology and collectively enabled the first application of AI in this species. [ABSTRACT FROM AUTHOR]

Artificial insemination (AI) involves the accurate detection of oestrus, the ability to pharmacologically induce ovulation, or both. This is particularly important in flying-foxes (Pteropusspp.) that do not have an overt behavioural oestrus. Pregnant mare serum gonadotropin (PMSG) stimulates folliculogenesis and ovulation in the Little Red Flying-fox (P. scapulatus; Towers PA and Martin L 1985 Proc. Aust. Soc. Reprod. Biol. 17, 115). In this study, a dose rate of 15 IU PMSG was used. Our study investigated if 15 IU PMSG would sufficiently induce ovulation in the larger, Black Flying-fox (P. alecto) or if a higher dose (30 IU) would be necessary. Before the mating season, a single injection of PMSG (Folligon®, Intervet Australia Pty Ltd, Bendigo East, Australia) was administered i.m. to eight adult females at 15 IU (n= 4) or 30 IU (n= 4) on Day 0. On Day 4, semen was collected by electro-ejaculation from adult males and inseminated intravaginally into females under isoflurane (Forthane®, Abbott Australasia Pty Ltd, Sydney, Australia) anaesthesia. Ovaries and reproductive tracts were surgically removed on Day 6, fixed, serially sectioned and stained using Gomori''s Stain. Histological sections were examined for evidence of ovarian activity and the presence of ova and spermatozoa in the reproductive tracts. Preliminary observations showed evidence of ovulation in both groups in the form of at least one CL in either ovary and an ovum in the ipsilateral oviduct or uterine horn. Ovaries of females stimulated with 30 IU PMSG differed noticeably from those treated with 15 IU in containing multiple, large, collapsed luteinized follicles with retained oocytes. Vascularization and glandular hypertrophy of the endometrium was also more evident in the higher dose group. No spermatozoa were observed in any of the excised tracts. These results suggest that whereas both doses of PMSG induce ovulation, the administration of 30 IU PMSG may over-stimulate the ovaries. This, in turn, could lead to an unphysiological environment for successful fertilization and embryonic development. A dose level of 15 IU or slightly above may be sufficient for subsequent attempts to stimulate folliculogenesis and ovulation in P. alecto. The absence of spermatozoa suggests that the site of insemination, the number of spermatozoa inseminated, or both requires further investigation if AI is to be successfully implemented in these species. We conclude that the ovarian responses to PMSG indicate that pharmacological induction of ovulation can be successfully achieved and thereby utilized in AI programs of endangered Pteropusspecies. [ABSTRACT FROM AUTHOR]

In addition to gametes, mammalian internal fertilisation has required the evolution of assorted anatomical, physiological and biochemical devices to deal with intra- and inter-sexual conflict such as sperm competition and female cryptic choice respectively. The seminal coagulum of primates and other mammals is viewed as one of such devices. Among primates, the seminal coagulum characteristically occurs in multi-male and multi-female species, leading us to suppose that it intervenes in sperm competition. However, it can also provide cues to the female reproductive tract about male desired or undesired traits, and therefore deter or favour sperm survival and migration. The present work investigates whether the seminal coagulum of the black-handed spider monkey enhances sperm fertilisation chances by improving the female reproductive tract conditions, and if the female reproductive tract is 'blind' to semen or behaves selectively towards ejaculates of different males. A series of artificial inseminations were done in five females, using the ejaculates of three different males, one at a time, and measuring the presence of distinct types of sperm inside the uteri at 10, 30 and 60 min following the insemination. The presence of coagulum, menstrual phase, and male and female identity only affected fast, straight-moving sperm, with larger amounts of fast sperm appearing inside the uteri when ejaculates had seminal coagulum, as well as when in the periovulatory phase. There was great intra-uterine fast-sperm variation regarding which male's semen inseminated which female. The results provide evidence to account for sexual conflict in the spider monkey as well as a methodological approach to this kind of study. [ABSTRACT FROM AUTHOR]

The article reports on the intention of the Department of Conservation of New Zealand to examine the use of artificial insemination of kakapos during the breeding season in Codfish Island, New Zealand. It mentions that dominant male kakapos mate with more females than less dominant male. It means that the representation of genes in the gene pool of kakapo is distorted towards specific males. Therefore, the use of artificial insemination will increase the diversity of genes within the critically endangered species. Richard Henry has been chosen as the sperm donor.

Introduction: A 33-year-old infertile patient presented with two early miscarriages in a spontaneous pregnancy and a pregnancy achieved by intrauterine insemination, respectively. She subsequently had nine failed IVF cycles. Despite the fact that >10 oocytes were retrieved in each cycle, all were characterized by abnormal fertilization demonstrated by the presence of only one pronucleus (IPN) and nonextraction of the secondnd polar body when examined 9-20 h following either standard insemination or intracytoplasmic sperm injection (ICSI). About 50% of these embryos further cleaved by day 3 post fertilization. While embryo transfer was performed in all cycles, no pregnancy was achieved. The aim was to analyse the genetic composition of oocytes and embryos in this patient in order to provide counselling regarding prognosis and treatment options. Materials/Methods: In her last cycle, ICSI was performed for all 10 oocytes. All first polar bodies (PB) were biopsied and analysed by fluorescence in-situ hybridization (FISH) probes for chromosomes 13, 18, 21, X and Y. In addition, all cleaving embryos were subjected to blastomere biopsy, followed by FISH analysis, as above. Since none of the embryos were found to be euploid, they were further subjected to single cell PCR analysis using a panel of highly polymorphic markers from different chromosomes (6, 7, 17, 19, X and Y). Results: FISH analysis of the first PB demonstrated normal segregation of chromosomes in meiosis 1. FISH analysis of single blastomeres from five cleaving embryos demonstrated three diploid and two mosaic triploid embryos. However, haplotype analysis demonstrated that all embryos including the diploid ones, had two sets of maternal alleles. Conclusion: We conclude that the diploid embryos represent parthenogenetic activation of a diploid oocyte. Furthermore, since ICSI was performed, the triploid embryos must have resulted from fertilization of a diploid oocyte by a single spermatozoon. Thus the abnormality detected in this case results from failure to extrude the second PB. Although abnormality of the spermatozoa has not been ruled out, this most likely represents failure in the oocyte activation machinery leading to incompetence of the oocyte to complete meiosis II as well as to decondense the sperm nucleus into the male pronucleus. [ABSTRACT FROM AUTHOR]

Two synchronization protocols for fixed-timed AI (FTAI) in White-tail deer were evaluated over a 2-year period. InYear 1, White-tail does (n = 38), with a mean body weight of 55 kg and mean age of 2.4 years, were stratified by weight, age, and last fawning date randomly across 2 estrous cycle synchronization treatment groups. Does received either a CIDR-G device for 14 days (CIDR 14) and underwent AI 60 h post-CIDR removal, or received a CIDR-G device for 7 days (CIDR 7) with 1 mg of estradiol benzoate (i.m.) at CIDR insertion (Day 0), 25 mg of prostaglandin F2α (PGF2α; Dinoprost) on Day 6 and 1 mg of estradiol benzoate (i.m.) on Day 7 with AI occurring 52 h post-CIDR removal. In Year 2, White-tail does (n = 36), with a mean body weight of 60 kg and mean age of 3.9 years, were stratified as described for Year 1 and allotted to 2 treatments. Does received a CIDR-G device for 14 days and were inseminated 60 h post-CIDR removal (Treatment A) or were synchronized in the same way as does inTreatment A but also received 200 IU (i.m.) of eCG (Sigma, St. Louis, MO) at CIDR removal (Treatment B). All does received 1 mg of Domosedan (i.v.) before initiating the AI procedure. In both years, electroejaculated semen was collected from a single buck and frozen for AI. In both years, clean-up bucks were introduced into the pen with does for natural cover at no less than 14 days following AI. A gestation length of 195 7 days was used to determine whether the fawn(s) resulted from AI or from natural mating. In Year 1, FTAI pregnancy rates were not different between the CIDR 14 (56%) and CIDR 7 (24%) treatment groups. Likewise, pregnancy rates were not different between CIDR 14 FTAI (56%) and clean-up bucks (75%). However, the FTAI pregnancy rate was lower (P < 0.001) for the CIDR 7 treatment group (24%) compared with the clean-up bucks (100%). Treatments did not affect fecundity rates; however, those females pregnant from FTAI had lower (P < 0.001) fecundity rates compared with those females pregnant from natural cover (1.2 v. 1.9 fawns/doe). In Year 2, treatment did not affect FTAI pregnancy rates across treatments (33% for Treatment A and 55% Treatment B); however, fewer (P = 0.024) does in Treatment A were pregnant following FTAI (33%) compared with those pregnant from clean-up bucks (75%). There was no difference in FTAI fecundity rates across treatment groups (1.3 v. 1.7 for Treatments A and B, respectively) or between FTAI treatments and natural cover (1.3, 1.7, and 1.5 for Treatments A, B, and clean-up bucks, respectively). These results indicate that the use of a 14-day CIDR synchronization protocol with FTAI occurring 60 h after CIDR removal yields acceptable pregnancy rates of White-tail does following transcervical AI. [ABSTRACT FROM AUTHOR]

The purpose of this investigation was to develop a non-invasive technique to artificially inseminate snakes using the corn snake, Elaphe gutatta, as the model representative for this taxon. Semen was collected by first applying pressure to the lower abdomen in a continuous distal motion toward the cloaca to remove any feces or urates. The cloaca was then gently washed using phosphate-buffered saline, and a more localized pressure was applied to each side of the vent to evert the hemipenes and, subsequently, the ejaculate. The semen was collected using a sterile transfer pipette and placed into 70 to 90 L of medium (TL-HEPES solution; Cambrex Bio Science, Inc., Baltimore, MD, USA04–616F) in a sterile microcentrifuge tube, and then analyzed for overall motility, rate of forward progression (RFP, 0–5), and concentration. Based on a previously reported procedure, 10 females were inseminated with either fresh (n = 5) or cooled semen (n = 5; refrigerated for 3 days) one week after recovering from a hibernation period required to stimulate reproduction in this species. The overall sperm motility and concentration for females inseminated with fresh or cooled semen was 92%, 9.6 million sperm mL-1; and 85%, 6.1 million sperm mL-1, respectively. Immediately prior to insemination, the same method for expressing feces and urates in the males was applied to the females. The insemination dose (50 L semen per oviduct) was drawn into a 1-mL latex- and silicone-free tuberculin syringe (Norm-Ject; VWR, Batavia, IL, USA) that was connected to a feeding/dosing needle (EJAY International, Issaquah, WA, USA) with a ball tip to prevent any potential damage during the insemination. The tip of the needle was then moved around the inner tissue of the vent to relax the cloaca, and the insertion continued until resistance was found indicating the vicinity of the oviducts. The extended semen was carefully deposited on both sides, and then the needle was slowly withdrawn. The offspring were tested for parentage to verify the success of the insemination. Blood was collected from the dorsal aorta posterior to the cloaca and stored in 10 mM Tris at 4C. The DNA was extracted using a phenyl : chloroform : isoamyl alcohol (PCI) extraction method. Eight microsatellite loci were used for the paternity exclusion analysis: Eob1, Eob3, Eob10, Eob13, Eob16, Eob34, Eob366, and Eob373 (IDT, Coraville, IA, USA). All males and females in the collection were tested, and parental candidates were excluded if 2 or more allele mismatches occurred. From the total number of females inseminated, 3 females laid 51 eggs. Two females inseminated with cooled semen laid 36 eggs, of which 5 eggs were hatched, and the remaining were either unfertilized (n = 25) or non-viable (n = 6). All 5 hatched eggs were laid by one of the females. The third female inseminated with fresh semen laid 15 eggs, resulting in 5 hatching and 10 unfertilized eggs. The parentage test validated the AI a success as the alleles correlated between the adults and the offspring. In conclusion, artificial insemination was successful using both freshly collected and cooled (3 days) semen. Further studies are underway to improve the success rate in order to maximize the efficiency of this technology, and thus assist in the genetic preservation of endangered snake species. [ABSTRACT FROM AUTHOR]

The ability to cryopreserve epididymal sperm from mature postmortem bucks has long been of interest to both wildlife conservationists and deer ranchers. At present, there is loss of valuable genetics from the hunter harvest of trophy males. Increasing adult body weight and antler size of adult males would be of substantial economic value to the deer hunting industry. In this preliminary trial, 6 yearling pen-raised Whitetail does (47 to 58 kg), in good body condition, were isolated from all bucks prior to the onset of the fall breeding season. Females were synchronized for AI with a 14-day caprine CIDR and 200 IU of eCG (IM) at the time of CIDR removal. Does were timed AI 60 to 63 h after eCG with one 0.5-mL straw of frozen–thawed Whitetail sperm. All sperm used for AI were harvested from a single mature Whitetail buck that was hunter harvested during the previous hunting season. Within 3 h after death, the testes with scrotum were removed, enclosed in a plastic Ziploc bag, and then placed in a Styrofoam ice chest containing frozen cold packs. The ice chest was transported to the laboratory where sperm were extracted at 4C in the late evening (<12 h postmortem) by flushing the cauda epididymides with Triladyl one-step extender (Minitube, Verona, WI, USA) in a retrograde flow from a small incision made in the cauda. The sperm–Triladyl mixture was flushed from the cauda incision into a sterile 50-mL tube using a 10-mL plastic syringe modified by heating and then stretching the tip until small enough to thread into the vas deferens. The sperm plus extender was then held at 4 to 10C for 12 h and frozen at a concentration of <50 million/straw using a commercial bull freezing protocol (Genex Custom Collection Center, Baton Rouge, LA, USA). A random sample of straws was then thawed, resulting in an overall post-thaw motility of 60%. The remaining straws were left frozen in liquid nitrogen until the next breeding season. On the first of December, does (n = 6) were given 0.1 mL of Domosedan (Pfizer Animal Health, Groton, CT, USA) IV and inseminated transcervically using a modified caprine speculum. All does were handled in a custom-built deer barn, and AI was performed by one technician in a drop-bottom deer chute (Deer Handler; Delclayna, Alberta, Canada). At 48 days after AI, 3 of the 6 does (50%) were diagnosed pregnant by transrectal ultrasonography. All pregnant females gave birth, producing 5 offspring (1 male and 1 female singletons and a set of mixed sex triplets) that ranged from 1.9 to 4.3 kg and had a mean gestation length of 196 days (range = 190–203 days). In summary, results indicate that live offspring can be produced from epididymal sperm harvested from mature hunter-harvested Whitetail bucks. Further experiments are needed to optimize techniques and protocols for the harvesting and usage of these gametes. [ABSTRACT FROM AUTHOR]

The article reports on the latest massage technique in getting semen samples in birds introduced by Spanish avian insemination expert Juan Blanco to help the Kakapo Recovery Program in New Zealand. The nation's Department of Conservation will utilize the collected sperm to test several solutions that maintain the sperm alive and to analyze how fertile the frozen sperm after it is defrosted. According to DOC kakapo scientist Ron Moorhouse, the success of the semen-freezing experiments could pave the way for the preservation of the sperm of Richard Henry, an elderly bird which contributed a significant role in the genetic diversity population of kakapo.

The article reports on the pregnancy Atlantic bottlenose dolphin at SeaWorld Adventure Park in San Diego, California. The pregnancy of the dolphin, named Sandy, was already predetermined. The dolphin was artificially inseminated with sorted sperm carrying only the X chromosome, which produces females. She gave birth to a healthy female dolphin, named Sofi. At SeaWorld, half the dolphins are males, and half females. To mimic the primarily female social groups in the wild, researchers there plan to make more female dolphins. By selecting for specific gender scientists may improve the populations of captive sea mammals and reduce the need to capture wild animals.

The article reports on measures undertaken by Australian marine scientists to save the endangered grey nurse shark from extinction. Australian marine scientists are planning radical measures to prevent the extinction of the shark. Under the plan, a seven gill shark was artificially inseminated and monitored in an effort to perfect the technique for use in grey nurse sharks. In the next 10 years, Nick Otway and other marine biologists from the New South Wales State Fisheries Department will attempt to develop artificial uteri in which the shark embryos can develop. Researchers have found that grey nurse sharks exhibit intrauterine cannabilism; once the pups are about 10cm long, they have developed a fully functional set of jaws and teeth and they then set about trying to eat each other. Environmental groups such as the Humane Society International and the Nature Conservation Council have pointed out that on average, one grey nurse is killed every month either by shark nets or commercial and recreational fishing. Therefore, there is absolutely no point breeding sharks in test tubes without securing a safe place to release them in the wild.

This article reports that due next August, it's the offspring of a white rhino couple at the Budapest Zoo that received a bit of help from artificial insemination expert Thomas Hildebrandt of the Institute for Zoo and Wildlife Research. Hildebrandt says attempts to breed rhinos in captivity have been largely unsuccessful, sharing close quarters is apparently a turnoff for the beasts, whose normal mating activities involve a fair dose of aggression. The team hopes to repeat the success with captive northern white rhinos, a handful of which still roam in the war-torn Democratic Republic of the Congo.

In vitro fertilization of follicular oocytes harvested from ovaries and matured in vitro was attempted for 55 minke whales (Balaenoptera acutorostrata) captured for Japanese research purposes in the Antarctic Ocean during the period from November 1995 to March 1996. In Experiment 1, effects of culture duration (96 h or 120 h) on maturation of follicular oocytes and addition of caffeine (5 mM) and/or heparin (100 mu g/ml) on sperm penetration and pronuclear formation were investigated. Spermatozoa recovered from the vasa deferentia of four mature males were diluted (5-fold) and frozen at -80 deg.C. The post-thawed and pooled spermatozoa were used for in vitro insemination. A higher (P < 0.05) proportion of the oocytes cultured for 120 h (34.2%of 260) progressed beyond the second metaphase stage than of the oocytes cultured for 96 h (26.0% of 262). For the matured oocytes, higher rates of penetration (P < 0.05) and pronuclear formation (P < 0.01) were obtained in the oocytes cultured for 120 h (55.1% and 40.4%) than in those cultured for 96 h (32.4 % and 20.6916). Addition of caffeine and heparin did not show a significant effect. In Experiment 2, follicular oocytes matured for 120 h and then inseminated were cultured to examine the subsequent development in two culture systems (with and without co-cultured cumulus cells). Of 448 inseminated oocytes, cleaved embryos (2--16 cells) were observed with (5.8%) and without (4.9%) co-cultured systems. No cleavage was observed in 54 ova without insemination. These results indicate that in vitro fertilization of minke whale in vitro matured follicular oocytes with cryopreserved spermatozoa is possible, yielding cleaved embryos." [ABSTRACT FROM AUTHOR]