Dubai, United Arab Emirates
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Nagy P.,Emirates Industries for Camel Milk and Products | Skidmore J.A.,Camel Reproduction Center | Juhasz J.,Emirates Industries for Camel Milk and Products
Animal Reproduction Science | Year: 2013

Despite their production potential and ability to survive on marginal resources in extreme conditions, dromedaries have not been exploited as an important food source. Camels have not been specifically selected for milk production, and genetic improvement has been negligible. High individual variation in milk production both within the population and within breeds provides a good base for selection and genetic progress. In this paper, we discuss the possibilities and constraints of selective breeding for milk production in camels, and include a summary of the use of embryo transfer at the world's first camel dairy farm. Embryo transfer is an integral part of the breeding strategy at the camel dairy farm because it increases selection intensity and decreases the generation interval. Using high milk-producing camels as donors and low producing camels as recipients, 146 embryos were recovered (6.1±1.0 embryos/donor; range: 0-18). Embryos were transferred non-surgically into 111 recipients (83 single and 28 twin embryo transfers). Pregnancy rate at 21 days and 5 months was 55% (61/111) and 45% (50/111), respectively. Finally, a total of 46 recipients delivered a live calf. These results document the utility of embryo transfer using high milk producing dromedaries as donors. © 2012 Elsevier B.V.

Wani N.A.,Camel Reproduction Center | Wernery U.,Central Veterinary Research Laboratory
Reproduction in Domestic Animals | Year: 2010

The present experiment was aimed to compare the effect of different protein supplementation sources, foetal calf serum (FCS), oestrous dromedary serum (EDS) and BSA, in experiment 1, and the effect of different concentrations of epidermal growth factor (EGF), in experiment 2, on in vitro nuclear maturation of the dromedary oocytes. Cumulus oocyte complexes (COCs) were harvested from the ovaries collected from a local slaughterhouse by aspirating the visible follicles in PBS supplemented with 5% FCS. Pooled COCs were randomly distributed to 4-well culture plates containing 500 μl of the maturation medium and cultured at 38.5°C in an atmosphere of 5% CO 2 in air for 32-36 h. The basic maturation medium consisted of TCM-199 supplemented with 0.1 mg/ml L-glutamine, 0.8 mg/ml sodium bicarbonate, 0.25 mg/ml pyruvate, 50 μg/ml gentamicin, 10 μg/ml bFSH, 10 μg/ml bLH and 1 μg/ml estradiol. In experiment 1, this medium was supplemented with 10% FCS, 10% EDS or 0.4% BSA, whereas in experiment 2, it was supplemented with 0.4% BSA and 0, 10, 20 or 50 ng/ml of EGF. The oocytes were fixed, stained with 1% aceto-orcein stain and their nuclear status was evaluated. Oocytes were classified as germinal vesicle, diakinesis, metaphase-I, anaphase-I (A-I), metaphase-II (M-II) and those with degenerated, fragmented, scattered, activated or without visible chromatin as others. There was no difference (p > 0.05) observed in the proportion of oocytes reaching M-II stage between the media supplemented with FCS (71.5 ± 4.8), EDS (72.8 ± 2.9) and BSA (72.7 ± 6.2). In experiment 2, a higher proportion (p < 0.05) of oocytes reached M-II stage when the medium was supplemented with 20 ng/ml of EGF (81.4 ± 3.2) when compared with the media supplemented with 10 ng/ml (66.9 ± 4.1) and control (67.2 ± 7.1) groups. It may be concluded that the maturation media for dromedary camel oocytes can be supplemented with any of the three protein sources, i.e. FCS, EDS and BSA without any significant differences on the maturation rates. Also, a supplementation of 20 ng/ml of EGF in the maturation medium seems to be optimal and improves the nuclear maturation of dromedary camel oocytes. © 2010 Blackwell Verlag GmbH.

Wani N.A.,Camel Reproduction Center | Wernery U.,Camel Reproduction Center | Hassan F.A.,Camel Reproduction Center | Wernery R.,Camel Reproduction Center | Skidmore J.A.,Camel Reproduction Center
Biology of reproduction | Year: 2010

In this study, we demonstrate the use of somatic cell nuclear transfer to produce the first cloned camelid, a dromedary camel (Camelus dromedarius) belonging to the family Camelidae. Donor karyoplasts were obtained from adult skin fibroblasts, cumulus cells, or fetal fibroblasts, and in vivo-matured oocytes, obtained from preovulatory follicles of superstimulated female camels by transvaginal ultrasound guided ovum pick-up, were used as cytoplasts. Reconstructed embryos were cultured in vitro for 7 days up to the hatching/hatched blastocyst stage before they were transferred to synchronized recipients on Day 6 after ovulation. Pregnancies were achieved from the embryos reconstructed from all cell types, and a healthy calf, named Injaz, was born from the pregnancy by an embryo reconstructed with cumulus cells. Genotype analyses, using 25 dromedary camel microsatellite markers, confirmed that the cloned calf was derived from the donor cell line and the ovarian tissue. In conclusion, the present study reports, for the first time, establishment of pregnancies and birth of the first cloned camelid, a dromedary camel (C. dromedarius), by use of somatic cell nuclear transfer. This has opened doors for the amelioration and preservation of genetically valuable animals like high milk producers, racing champions, and males of high genetic merit in camelids. We also demonstrated, for the first time, that adult and fetal fibroblasts can be cultured, expanded, and frozen without losing their ability to support the development of nuclear transfer embryos, a technology that may potentially be used to modify fibroblast genome by homologous recombination so as to generate genetically altered cloned animals.

Skidmore J.A.,Camel Reproduction Center
Society of Reproduction and Fertility supplement | Year: 2010

The reproductive efficiency of camels is low under natural pastural conditions and so the use of artifical insemination and embryo transfer are becoming increasingly important to improve their breeding potential. Methods to control their reproductive cycle are therefore essential. This review describes characteristics of the ovarian follicular wave pattern in camels and exogenous hormonal control of ovulation. It also summarizes the difficulties involved with artifical insemination and analyzing the highly gelatinous semen, and reports on the latest methods used to try and reduce the viscosity and liquefy camel semen. In addition an account is given of different hormonal and physical methods used to synchronise follicular waves, and various hormone treatments used to broaden the availability of ovulated, asynchronous and non-ovulated recipients are discussed.

Skidmore J.A.,Camel Reproduction Center | Morton K.M.,Camel Reproduction Center | Billah M.,Camel Reproduction Center
Animal Reproduction Science | Year: 2013

Artificial insemination (AI) is an important technique in all domestic species to ensure rapid genetic progress. The use of AI has been reported in camelids although insemination trials are rare. This could be because of the difficulties involved in collecting as well as handling the semen due to the gelatinous nature of the seminal plasma. In addition, as all camelids are induced ovulators, the females need to be induced to ovulate before being inseminated.This paper discusses the different methods for collection of camel semen and describes how the semen concentration and morphology are analyzed. It also examines the use of different buffers for liquid storage of fresh and chilled semen, the ideal number of live sperm to inseminate and whether pregnancy rates are improved if the animal is inseminated at the tip of the uterine horn verses in the uterine body. Various methods to induce ovulation in the female camels are also described as well as the timing of insemination in relation to ovulation. Results show that collection of semen is best achieved using an artificial vagina, and the highest pregnancy rates are obtained if a minimum of 150×106 live spermatozoa (diluted in Green Buffer, lactose (11%), or I.N.R.A. 96) are inseminated into the body of the uterus 24h after the GnRH injection, given to the female camel to induce ovulation.Deep freezing of camel semen is proving to be a great challenge but the use of various freezing protocols, different diluents and different packaging methods (straws verses pellets) will be discussed. Preliminary results indicate that Green and Clear Buffer for Camel Semen is the best diluent to use for freezing dromedary semen and that freezing in pellets rather than straws result in higher post-thaw motility. Preservation of semen by deep-freezing is very important in camelids as it prevents the need to transport animals between farms and it extends the reproductive life span of the male, therefore further work needs to be carried out to improve the fertility of frozen/thawed camel spermatozoa. © 2012 Elsevier B.V.

Crichton E.G.,Camel Reproduction Center | Pukazhenthi B.S.,Smithsonian Conservation Biology Institute | Billah M.,Camel Reproduction Center | Skidmore J.A.,Camel Reproduction Center
Theriogenology | Year: 2015

The cryopreservation of dromedary camel (. Camelus dromedarius) sperm has proved challenging with little success reported. The routine application of artificial insemination with frozen semen would assist the flow of valuable genetic material nationally and internationally. The current study sought to examine the effects of cholesterol (cholesterol-loaded cyclodextrin [CLC]) preloading on camel sperm cryosurvival. Ejaculates (n=3 males; 3 ejaculates per male) were collected using an artificial vagina during the breeding season and extended in HEPES-buffered Tyrode's albumin lactate pyruvate (TALP) and allowed to liquefy in the presence of papain (0.1mg/mL) before removal of the seminal plasma by centrifugation. Sperm pellets were resuspended (120 million/mL) in fresh TALP and incubated (15minutes; 37°C) with 0, 1.5, or 4.5mg CLC/mL. Sperm suspensions were then centrifuged and reconstituted in INRA-96 containing 20% (v:v) egg yolk and 2.5% (v:v) methylformamide, loaded in 0.5-mL plastic straws, sealed, and cooled for 20minutes at 4°C. Straws were frozen over liquid nitrogen (4cm above liquid; 15minutes), plunged, and stored. Sperm motility, forward progressive status, and acrosomal integrity were recorded at 0 and 3hours after thawing and compared with these same parameters before freezing. Aliquots also were stained with chlortetracycline hydrochloride to assess spontaneous sperm capacitation status before freezing and post-thaw. Pretreatment with CLC (1.5and 4.5mg/mL) enhanced cryosurvival. Post-thaw sperm motility was highest (P<0.05) in 1.5mg CLC/mL immediately after thawing (44%) and after 3hours incubation at room temperature (34%). Highest post-thaw sperm progressive status was also achieved in the presence of 1.5 CLC. Greater proportions of spermatozoa retained acrosomal membrane integrity when cryopreserved in the presence of CLC, but there was no difference between 1.5 and 4.5 CLC. Although thawed spermatozoa underwent spontaneous capacitation during invitro incubation, cryopreservation and CLC treatment exerted no effect. In summary, dromedary camel sperm benefit from exposure to CLC before cryopreservation; this may facilitate the routine collection and storage of sperm from this species. © 2015 Elsevier Inc.

Morton K.M.,Camel Reproduction Center | Billah M.,Camel Reproduction Center | Skidmore J.A.,Camel Reproduction Center
Reproduction in Domestic Animals | Year: 2011

Contents: Artificial insemination (AI) is one of the most widely used reproductive technologies, and there is considerably interest in commercializing this technology in camels. Storage of semen extender frozen (at -20°C) is of considerable interest to scientists working with camels, as transportation of diluents at refrigeration temperature is not always possible given the hot, arid and remote conditions that dromedary camels exist in. Therefore, this study was conducted to compare the fertility of fresh camel semen, after dilution in fresh or frozen-thawed green buffer (GB), after AI into single and multiple ovulating female camels. No differences were observed in any sperm characteristics (motility, membrane integrity, acrosome integrity or morphology) when semen was diluted in fresh or frozen-thawed GB (p>0.05). Sperm motility was increased by dilution (fresh: 70.7±4.9% and frozen: 68.8±3.1%) compared with the motility of sperm in neat semen (35±2.85%; p<0.05), and sperm motility changed from oscillatory to forward progressive after dilution. Pregnancy rates were higher (p<0.05) for single ovulating camels inseminated with semen diluted in fresh (72.7%) compared with frozen-thawed GB (27.3%), and fertilization rates were also higher (p<0.05) for multiple ovulating camels inseminated with semen diluted in fresh (83.3%) compared with frozen-thawed GB (11.1%). These results clearly demonstrate the detrimental effect of freezing and thawing semen diluent on the fertility of fresh camel semen. However, further studies are required to elucidate the mechanism responsible for this reduction in fertility. Moreover, these results demonstrate that the fertility of fresh camel semen diluted in fresh GB is high enough to be considered commercially viable. © 2010 Blackwell Verlag GmbH.

Wani N.A.,Camel Reproduction Center | Skidmore J.A.,Camel Reproduction Center
Theriogenology | Year: 2010

In Experiment 1, studies were conducted to apply the transvaginal ultrasound guided ovum pick-up (OPU) technique in dromedary camels after their ovarian super-stimulation and in vivo oocyte maturation. In Experiment 2, the developmental potential of two commonly used oocyte types, i.e., in vivo matured oocytes collected by OPU and abattoir derived in vitro-matured oocytes was compared after their chemical activation. In Experiment 3, developmental competence of oocytes collected from super-stimulated camels by OPU, matured either in vivo or in vitro, was compared after their chemical activation. Mature female dromedary camels super-stimulated with a combination of eCG and pFSH were given an injection of 20 μg of the GnRH analogue, buserelin 24, 26, or 28 h before the scheduled OPU. For collection of cumulus oocyte complexes (COCs) the transducer was guided through the vulva into the cranial most portion of the vagina and 17-gauge, 55 cm single-lumen needle was placed in the needle guide of the ultrasound probe and advanced through the vaginal fornix and into the follicle. Follicular fluid was aspirated using a regulated vacuum pump into tubes containing embryo-flushing media. Aspirates were searched for COCs using a stereomicroscope, and they were then denuded of cumulus cells by hyaluronidase and repeated pipetting. The oocytes were classified as mature (with a visible polar body), immature (with no visible polar body), activated (with divided or fragmented ooplasm) and others (degenerated and abnormal).Overall an average of 12.12 7.9 COCs were aspirated per animal with an oocyte recovery rate from the aspirated follicles of about 77%. The majority (> 90%) of the collected COCs by OPU were with loose and expanded cumulus cells. The proportion of matured oocytes obtained at 28-29 h (91.2 4.1) and 26-27 h (82.1 3.4) were higher (P < 0.005) when compared with those obtained at 24-25 h (40.4 16.3) after GnRH administration. In Experiment 2, a higher proportion (P < 0.05) of in vivo matured oocytes cleaved (84.6 2.1 vs. 60.9 6.6) and developed to blastocyst stages (52.4 4.1 vs. 30.5 3.3) when compared with in vitro matured oocytes collected from slaughterhouse ovaries. In Experiment 3, no difference was observed between the developmental competences of oocytes, collected from super stimulated camels, matured in vitro with those collected after their in vivo maturation. In conclusion, about 80. -90% mature oocytes can be collected by ultrasound guided transvaginal ovum pick-up from super-stimulated dromedary camels 26. -28 h after GnRH administration. The developmental response, to chemical activation, of in vivo matured oocytes collected by ultrasound guided transvaginal OPU is better than in vitro matured oocytes obtained from slaughterhouse ovaries. However, no difference was observed in the developmental competence of oocytes collected by OPU whether they were matured in vivo or in vitro. © 2010 Elsevier Inc.

Skidmore J.A.,Camel Reproduction Center
Animal Reproduction Science | Year: 2011

This review summarizes the basic reproductive physiology of dromedary and Bactrian camels. Camels are seasonal breeders with a relatively short breeding season during the cooler months. The onset of the breeding season can be influenced by local environmental factors such as temperature and pasture availability although decreased libido of the male as the environmental temperature increases is also a factor. Oestrous behaviour is highly variable in duration and intensity and is therefore unreliable for the detection of oestrus and difficult to relate to follicular activity in the ovaries. Camels are induced ovulators and thus normally only ovulate in response to mating. In the absence of mating, ovarian follicles tend to regress after a period of growth and maturity, whereas if male and females are kept together the female gets mated when the dominant follicle measures between 1.3 and 1.7. cm in diameter and the corpus luteum that develops has a lifespan of only 10-12 days. Peripheral concentrations of oestradiol increase with increasing follicle diameter until the follicle reaches 1.7. cm in diameter at which time they start to decrease even if the follicle continues to grow. The concentrations of progesterone remains low in non-mated animals but in mated camels it increases 3-4 days after ovulation (day of ovulation = Day 0) to reach maximum concentrations on Days 8-9 before decreasing rapidly on Days 10-11 in the non-pregnant animal. Ovulation can also be reliably induced using either Gonadotrophin Releasing Hormone (GnRH) or human Chorionic Gonadotrophin (hCG) but only when the follicle measures between 1.0 and 1.9. cm in diameter. Ovulation does not typically occur from follicles that grow beyond 2.0. cm in diameter but these follicles typically develop echogenic strands of fibrin as the follicle degenerates. The gestation period of camels is 13 months but the time of resumption of follicular activity following parturition is highly variable and influenced by nutritional status and lactation. Females that lose their offspring or have offspring which are weaned have a mature follicle develop within 10-12 days whilst in well-fed lactating females mature follicles do not develop until 30-60 days postpartum. © 2010 Elsevier B.V.

Skidmore J.A.,Camel Reproduction Center | Billah M.,Camel Reproduction Center
Reproduction, Fertility and Development | Year: 2011

The aim of the present study was to investigate the use of exogenous progesterone and equine chorionic gonadotrophin (eCG) in non-ovulated and ovulated, asynchronous dromedary camel recipients being prepared for an embryo transfer programme. The uteri of 12 mated donor camels were flushed non-surgically 7 days after ovulation and 42 embryos were recovered. In Experiment 1, 16 embryos were transferred non-surgically to recipients on Day 3 or 4 after ovulation (ov+3 and ov+4, respectively). Each recipient received a daily dose of 75mg, i.m., progesterone-in-oil from 2 days before embryo transfer until 6 days after ovulation. Thereafter, the progesterone dose was reduced to 50mg on Day 7 and finally to 25mgday-1 on Days 8 and 9. Nine of 16 recipients (56%; ov+3, n=4; ov+4, n=5) became pregnant compared with none of eight non-progesterone treated controls, into which embryos were transferred on Day 4 after ovulation. In Experiment 2, 18 non-ovulated recipients received 75mg, i.m., progesterone-in-oil daily from 3 days before until 12 days after non-surgical transfer of a Day 7 blastocyst, at which time pregnancy was diagnosed by ultrasonography. All pregnant recipients continued to receive 75mg progesterone-in-oil daily for a further 6 days, when each camel received 2000IU, i.m., eCG. Progesterone treatment was then reduced to 50mgday-1 and, when a follicle(s) ≥1.3cm in diameter were present in the ovaries, each animal received 20 μg buserelin to induce ovulation. Once the corpora lutea had developed, progesterone treatment was reduced to 25mgday-1 for a final 3 days. Fourteen of 18 recipients (78%) became pregnant and seven of these (50%) remained pregnant after eCG treatment. Of the seven pregnancies that were lost, two were lost before eCG treatment, two did not respond to eCG treatment and three responded to eCG treatment and ovulated, but lost their pregnancies 6-8 days after the last progesterone injection. © CSIRO 2011.

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