"Comparison of two closed vitrification methods for vitrifying dromedary camel (Camelus dromedarius) embryos".

A total of 184 dromedary camel embryos were vitrified using a novel vitrification kit specifically developed for camel embryos. These embryos were vitrified using a 3-step process by exposing them to vitrification solutions (VS) containing 20% foetal calf serum (FCS) with (+) or without (-) the addition of bovine serum albumin (BSA). Embryos were then further divided into two groups (

Individual male dependent improvement in post-thaw dromedary camel sperm quality after addition of catalase.

Cryopreservation is stressful to sperm cells inducing an increase in the production of reactive oxygen species and subsequently reducing post-thaw sperm quality. With the present study, there was evaluation of the protective effects of two antioxidants, epigallocatechin (1 mM) and catalase (500 IU/ml), added at thawing, as well as inter-individual variation on quality of cryopreserved dromedary camel spermatozoa. Semen was collected from six males and sperm, selected using single layer centrifugation, were cryopreserved. Post-thaw sperm quality was evaluated by assessing motility variables, viability and acrosome integrity then sperm were co-incubated with or without antioxidant (control) and further assessed at 1.5 and 3 h of the incubation period. Oxidative damage was measured colorimetrically for malondialdehyde production at 3 h of the incubation period. With the use of epigallocatechin there were not promising results, however, with use of catalase there were greater total and progressive motility, and values for some kinematic variables (P<0.05) at both incubation time points, although there were some differences among males. There was no overall effect of antioxidant based on production of malonaldehyde. The capacity of thawed sperm to fertilize, with and without addition of catalase at thawing, was studied using artificial insemination (n = 10 per treatment) with no differences between treatments (10% for both). It is concluded that catalase supplementations to semen extender prolong sperm survival, however, there is no improvement of in vivo fertilization as a result of this supplementation. There was an obvious male effect, necessitating further studies to understand the mechanisms of action of catalase.

Near-Newtonian Blood Behavior - Is It Good to Be a Camel?

From a certain level of exercise-intensity onward, hematocrit increases in horses, which brings more oxygen carriers into the bloodstream. Camels, however, when used in competitive racing could be even in need of iron supplementation and blood transfusions due to a severe reduction of their available hematocrit compared to their resting hematocrit. Since the extrinsic and intrinsic mechanical properties of camel erythrocytes (RBC) are so different compared to RBCs of other mammals, the question arises whether this observation might be a response to endurance exercise aiming at keeping the RBC count low. Rheometry indicated dromedary camel blood to behave almost Newtonian, which is unique amongst mammals. Shear thinning did increase with the hematocrit, but remained marginal compared to horses. As a result, camel whole blood viscosity (WBV) exceeded horse WBV at high shear rates, an effect, which was significantly augmented when the packed cell volume (PCV) was increased. Therefore, in camels any infusion of RBCs into the bloodstream can increase the cardiac work and the energy input into the endothelium more effectively, which should generate vascular remodeling in the long term. Yielding, however, was completely absent in camel blood, confirming low cohesion between its components at quasi-static flow. Camel blood remained a viscous liquid without a threshold even at unphysiologically high PCVs. This can help to washout lactate when camels start to dehydrate and might contribute to the sustained working ability of these animals. The subtle pseudoplastic behavior and the high viscosity contrast across the RBC membrane point to weak coupling between blood flow and red cell behavior. We predict that RBCs flow as separate entities and can show various types of motion, which can lead to friction instead of being collectively aligned to the flow direction. In comparison to horses, this behavior will become relevant at higher RBC counts in front of flow obstacles and possibly cause vascular remodeling if the PCV rises during strenuous exercise, a matter that should be avoided.

The use of some assisted reproductive technologies in old world camelids.

The use of camels for racing, milking and as show animals is growing in popularity, thus there is increased enthusiasm to breed more of the genetically superior animals. This review highlights recent developments in assisted reproductive techniques in camels, such as embryo transfer and artificial insemination, to ensure more rapid genetic progress. This paper discusses the difficulties involved in handling the semen due to its high viscosity and ways to reduce it. It also examines methods for short term liquid storage of fresh semen with and without the use of antioxidants to reduce oxidative stress. The widespread use of AI in camels is hindered by the lack of a reliable method for deep freezing and long term storage but various freezing protocols, cryoprotectants and freezing and thawing methods are discussed as well as different insemination techniques. Embryo transfer requires the donor to be superovulated and the recipients synchronized. This review discusses different protocols used for superovulation of donor animals and the problems involved. It also examines various methods to synchronize recipients, or how to make best use of non-synchronized or non ovulated recipients. Cryopreservation of embryos would greatly improve the wider use of ET and spread of genetics worldwide so methods for slow cooling and new methods of vitrification with promising results are discussed.

Donor sperm production in heterologous recipients by testis germ cell transplantation in the dromedary camel.

The object of this study was to investigate if testis germ cell transplantation (TGCT) into a heterologous recipient would result in donor-origin spermatogenesis in the dromedary camel. First, we investigated a workable protocol for TGCT in camels, including donor cell isolation, enrichment by density gradient centrifugation (Percoll and Bovicoll), rete testis injection and microsatellite detection of donor and recipient genotypes. Second, the effects of three doses of Dolichos biflorus agglutinin (DBA), a glycoprotein that specifically binds to gonocytes or Type A spermatogonia, on testis germ cell depletion were investigated by direct injection into the rete testis of a male camel. Seven recipients were prepared with DBA treatment, two males were castrated at 4 weeks for depletion assessment and the remaining five received donor cells 4-6 weeks after treatment. On average, ~17 million cells were isolated per gram of testis tissue, with 19.5±1.9% DBA-positive (DBA+) cells. Percoll centrifugation yielded a 1.5-fold increase in DBA+ cells while Bovicoll centrifugation produced a 2.5-fold increase from the input cells of 18.6±2.1% DBA+ cells. Semen was collected from the recipients 13-20 weeks after transfer and the presence of donor DNA in the samples was determined using microsatellite markers. In two of the five recipients, all semen samples were shown to be positive for donor-derived cells. These results demonstrate for the first time that: (1) heterologous testicular germ cell transplantation in camels is feasible and the recipients are able to produce spermatozoa of donor origin and (2) DBA can be used effectively to deplete endogenous stem cells.

Effect of different freezing rates and thawing temperatures on cryosurvival of dromedary camel spermatozoa.

The objective of this study was to evaluate the effect of different freezing rates and thawing temperatures on the post-thaw quality of camel spermatozoa. Ten ejaculates from five male camels were frozen at five different freezing rates, achieved by placing the straws at specific heights above the surface of liquid nitrogen for different lengths of time (4 cm for 15 min; 1 cm for 15 min; 7 cm for 15 min; 7 cm for 5 min + 4 cm for 3 min; 4 cm for 5 min + 1 cm for 3 min) followed by storage in liquid nitrogen. Two thawing temperatures (37° for 30 s and 60 °C for 10 s) were subsequently tested. Post-thawing, the samples were evaluated for total and progressive motility, kinematics, membrane and acrosome integrity, and membrane functionality (hypoosmotic swelling test) at zero and 1 h post thawing. Total and progressive motility were significantly higher for the fastest freezing rate (at 1 cm) at 0 h (p < 0.01 for both), as were VCL (p < 0.01), VSL (p < 0.05) and STR (p < 0.05). Freezing at 4 cm produced the lowest values of STR compared to other treatments (p < 0.05). At 1 h, no differences in total motility were observed between freezing at 4 cm and 1 cm, both being significantly better than freezing rate 7 cm + 4 cm (p < 0.01). For progressive motility and VSL, only freezing at 1 cm was superior to the 7 cm + 4 cm combination (p < 0.001 and p < 0.05 respectively). Membrane integrity at 1 h was higher for freezing at 7 cm than at 1 cm (p < 0.01). For thawing temperatures, total motility and progressive motility at 0 h and 1 h (p < 0.001), and acrosome integrity at 1 h (p < 0.01) were higher for 60 °C thawing temperature than 37 °C. The kinematics VCL (p < 0.001), VSL and STR (p < 0.01), and VAP (p < 0.05) showed higher values for 60 °C thawing temperature than 37 °C at 0 h. After 1 h, higher values for VSL, VCL and VAP (p < 0.05) were observed for 60 °C than for 37 °C. In conclusion, a fast freezing rate would probably be beneficial for camel semen, and thawing should be conducted at 60 °C.

Temperature dependency of whole blood viscosity and red cell properties in desert ungulates: Studies on scimitar-horned oryx and dromedary camel.

Background: The dromedary camel and the oryx antelope are exposed to excessive heat and solar radiation in their desert habitat. Desertification of areas with by now little rainfall may occur eventually. Well-adapted large animal species show us what is needed to survive in scorching regions. Methods: Four scimitar-horned oryx antelopes (Oryx dammah), 10 camels (Camelus dromedarius), nine South African Merino sheep, and 17 Nguni cows were tested for RBC aggregation, RBC elongation, and plasma viscosity. The temperature dependency of blood viscosity was tested in 10 camels and compared to human reference values. Results: Unlike sheep, Nguni cow, and dromedary camel, oryx RBCs aggregate in native plasma (M0:5.2 (3.3/6.7); M1:18.1 (16.7/27.9); Myrenne MA1). Elongation indices of oryx RBCs were intermediate to low (EImax: 22.6 (19.2/25.3); SS1/2 3.67 (2.52/4.95); Rheodyn SSD). Camel RBCs did not display the typical SS/EI curve by rotational ektacytometry. In-vitro blood viscosity (Physica MCR302) was lower in camels than in human blood at equal hematocrit. A decrease of temperature had only little effect on camel blood. At 10 s−1, blood viscosity in camel increased from 2.18mPa*s (2.01/2.37) at 42◦C to 4.39mPa*s (4.22/4.51) at 12◦C. In human blood, viscosity ranged from 8.21mPa*s (6.95/8.25) at 37◦C to 15.52mPa*s (14.25/16.03) at 12◦C. At 1000 s−1, blood viscosity in camel ranged from 2.00mPa*s (1.95/2.04) at 42◦C to 3.98mPa*s (3.88/4.08) at 12◦C. In human blood, viscosity ranged from 5.35mPa*s (4.96/5.87) at 37◦C to 11.24mPa*s (10.06/11.17) at 12◦C. Conclusions: Desert ungulates may need RBC membranes, which are fortified to withstand changes in osmolality during dehydration-rehydration cycles. This reduces RBC deformability. Dromedary camel blood does not undergo stark changes in viscosity with changes in temperature. Therefore, blood fluidity could be rather maintained during the day and night cycle. This should reduce the need of the vascularity to rhythmically adapt to changing shear forces when camels experience heterothermy.

An update on semen collection, preservation and artificial insemination in the dromedary camel (Camelus dromedarius).

Artificial insemination (AI) in domestic animals is an important tool to maximise the use of genetically superior males and thereby insure rapid genetic progress. However, the application of AI in camelids has been hindered by the difficulties involved in collecting, as well as handling the semen due to the viscous nature of the seminal plasma. This review describes the challenges of semen collection and discusses the role of seminal plasma as well as the reasons for the viscosity and how to liquefy it so that ejaculates can be more accurately evaluated. It also reports on the use of various extenders used for liquid storage of fresh and chilled semen and how pregnancy rates are affected by numbers of spermatozoa inseminated, site of insemination and timing of insemination in relation to GnRH injection given to induce ovulation. In addition, this paper reviews the latest research in cryopreservation of camel semen and addresses the various problems involved and possible improvements that can be made so that pregnancy rates can be increased with frozen semen.

Colloid centrifugation of fresh semen improves post-thaw quality of cryopreserved dromedary camel spermatozoa.

Colloids have been successfully used in a number of species to improve sperm populations for IVF and for cryopreservation The usefulness of Single Layer Centrifugation (SLC) for freezing dromedary camel spermatozoa in two different extenders was evaluated by examining the motility, viability, acrosome status, DNA integrity, and ability of cryopreserved sperm to penetrate oocytes in vitro in a heterologus IVF system. Two ejaculates from each of five males were divided into four aliquots: two were processed by SLC (selected) while two were centrifuged without colloid (control). Pellets were cryopreserved in Green Buffer or INRA-96® containing 3% glycerol and evaluated at 0 and 1 h post thawed. The SLC improved post-thaw total and progressive motility at 0 (both P < 0.0001) and 1 (P < 0.001; P < 0.01, respectively) h, and STR (both P < 0.05) and BCF (both P < 0.001) at 0 h. Sperm viability and acrosome integrity (both P < 0.001) were improved at both time points. Sperm frozen in Green Buffer had greater total and progressive motilities at 0 (both P < 0.001) and 1 (both P < 0.001) h than INRA-96® samples. Spermatozoa in Green Buffer also had a greater VAP, VCL and VSL at 0 h and improved viability and acrosome integrity at 0 h (P < 0.05; P = 0.001, respectively) and 1 h (P < 0.05; P < 0.001, respectively). Viability of SLC spermatozoa was improved in Green Buffer at 1 h (P < 0.05). Oocyte penetration (P < 0.05) and pronuclear formation (P < 0.01) were greater with SLC-selected spermatozoa than non-selected spermatozoa, regardless of extender. No difference was observed between treatments or extenders in the mean number of spermatozoa per oocyte penetrated. The SLC spermatozoa had less (P < 0.01) DNA fragmentation compared to controls. The DNA fragmentation was moderately and negatively correlated with penetration (r = -0.4162; P = 0.02) and pronuclear formation (r = -0.3390; P < 0.01). In conclusion, colloid centrifugation of spermatozoa and cryopreservation in Green Buffer improves post thaw motility variables and IVF performance of dromedary camel spermatozoa.

Optimization of the cryopreservation of dromedary camel semen: Cryoprotectants and their concentration and equilibration times.

Research into an optimal cryoprotectant, its concentration and equilibration time underlies the successful cryopreservation of dromedary camel spermatozoa. This study assessed the cryo-efficiency of different cryoprotectants, their concentration and equilibration time and any interactions. In experiment 1, semen samples (n = 4 males; 2 ejaculates/male) were frozen using Green Buffer containing one of four cryoprotectants (3% glycerol, ethylene glycol, methyl formamide, dimethyl sulfoxide) and using 4 equilibration times (10 min, 0.5, 1 and 2 h). Glycerol and ethylene glycol provided the best motility recovery rates and different equilibration times were not significant for any cryoprotectant nor were any interactions noted. However different equilibration times were pertinent for improved kinematic parameters BCF and VSL. In experiment 2, glycerol and ethylene glycol were evaluated at 4 concentrations (1.5, 3, 6, 9%) with 0.5 h equilibration (n = 4 males, 3 ejaculates/male). Sperm motility recoveries, kinematics and acrosome status were assessed. Higher values for LIN and STR were found with ethylene glycol. At 0 and 1 h post thaw 3 and 6% of either cryoprotectant resulted in better motility values than 1.5%. Acrosome integrity was compromised at 9% cryoprotectant. There were interactions between cryoprotectant and concentration in total motility at 0 and 1 h. For glycerol, total motility recoveries were best at 3-9%; for ethylene glycol 1.5-6% were best at 0 h and 3-6% at 1 h. In conclusion, 3-6% glycerol or ethylene glycol offered the best cryoprotection for camel sperm while different equilibration times were not critical.

Evaluation of cholesterol- treated dromedary camel sperm function by heterologous IVF and AI.

Cholesterol (cholesterol-loaded cyclodextrins: CLC) treatment of dromedary camel sperm prior to freezing enhances cryosurvival. The present study first validated the efficacy of a heterologous zona-free goat oocyte assay (n=115 oocytes) to evaluate camel sperm function in vitro (Experiment 1: n=6 bulls), then examined the effects of CLC treatment (1.5mg/mL CLC; CLC+) versus no treatment (0 CLC) of fresh (Experiment 2: n=4 bulls) and frozen-thawed (Experiment 3: n=5 bulls) camel sperm to penetrate, de-condense and form pro-nuclei in in vitro-matured goat oocytes. Finally, the ability of fresh 0 CLC and CLC+ sperm to fertilize in vivo was studied by artificially inseminating super-ovulated females (n=7-9 per treatment) and examining embryo production (Experiment 4: n=4-5 bulls/treatment). Camel spermatozoa penetrated (60%) and formed pro-nuclei (33%) in goat oocytes demonstrating the utility of this heterologous system for assessing sperm function in vitro. For fresh spermatozoa, 0 CLC-treated sperm performed better than their CLC+ counterparts for all parameters measured (P<0.05). In contrast, cryopreservation resulted in a sharp decline in sperm-oocyte interaction in 0 CLC aliquots but remained unaltered in CLC+ aliquots demonstrating a protective effect of cholesterol treatment. There was no difference between treatments in the in vitro fertilizing ability of frozen-thawed sperm or in the numbers of embryos retrieved following AI with fresh 0 CLC or CLC+ sperm. We conclude that although CLC treatment of dromedary camel sperm improves sperm motility it fails to confer an advantage to them in terms of improved in vitro sperm-oocyte interaction or in vivo fertilization under the conditions tested.

Cholesterol addition aids the cryopreservation of dromedary camel (Camelus dromedarius) spermatozoa.

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 (0.1 mg/mL) before removal of the seminal plasma by centrifugation. Sperm pellets were resuspended (120 million/mL) in fresh TALP and incubated (15 minutes; 37 °C) with 0, 1.5, or 4.5 mg 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 20 minutes at 4 °C. Straws were frozen over liquid nitrogen (4 cm above liquid; 15 minutes), plunged, and stored. Sperm motility, forward progressive status, and acrosomal integrity were recorded at 0 and 3 hours 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 (1.5 and 4.5 mg/mL) enhanced cryosurvival. Post-thaw sperm motility was highest (P < 0.05) in 1.5 mg CLC/mL immediately after thawing (44%) and after 3 hours 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 in vitro 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.

Patterning and post-patterning modes of evolutionary digit loss in mammals.

A reduction in the number of digits has evolved many times in tetrapods, particularly in cursorial mammals that travel over deserts and plains, yet the underlying developmental mechanisms have remained elusive. Here we show that digit loss can occur both during early limb patterning and at later post-patterning stages of chondrogenesis. In the 'odd-toed' jerboa (Dipus sagitta) and horse and the 'even-toed' camel, extensive cell death sculpts the tissue around the remaining toes. In contrast, digit loss in the pig is orchestrated by earlier limb patterning mechanisms including downregulation of Ptch1 expression but no increase in cell death. Together these data demonstrate remarkable plasticity in the mechanisms of vertebrate limb evolution and shed light on the complexity of morphological convergence, particularly within the artiodactyl lineage.

Reproductive physiology in female Old World Camelids.

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.

Cross-species chromosome painting among camel, cattle, pig and human: further insights into the putative Cetartiodactyla ancestral karyotype.

The great karyotypic differences between camel, cattle and pig, three important domestic animals, have been a challenge for comparative cytogenetic studies based on conventional cytogenetic approaches. To construct a genome-wide comparative chromosome map among these artiodactyls, we made a set of chromosome painting probes from the dromedary camel (Camelus dromedarius) by flow sorting and degenerate oligonucleotide primed-PCR. The painting probes were first used to characterize the karyotypes of the dromedary camel (C. dromedarius), the Bactrian camel (C. bactrianus), the guanaco (Lama guanicoe), the alpaca (L. pacos) and dromedary x guanaco hybrid karyotypes (all with 2n = 74). These FISH experiments enabled the establishment of a high-resolution GTG-banded karyotype, together with chromosome nomenclature and idiogram for C. dromedarius, and revealed that these camelid species have almost identical karyotypes, with only slight variations in the amount and distribution patterns of heterochromatin. Further cross-species chromosome painting between camel, cattle, pig and human with painting probes from the camel and human led to the establishment of genome-wide comparative maps. Between human and camel, pig and camel, and cattle and camel 47, 53 and 53 autosomal conserved segments were detected, respectively. Integrated analysis with previously published comparative maps of human/pig/cattle enabled us to propose a Cetartiodactyla ancestral karyotype and to discuss the early karyotype evolution of Cetartiodactyla. Furthermore, these maps will facilitate the positional cloning of genes by aiding the cross-species transfer of mapping information.

Embryo transfer in the dromedary camel (Camelus dromedarius) using asynchronous, meclofenamic acid-treated recipients.

A total of 40 (Day 7) embryos were recovered from the uteri of 10 superovulated camels. Recipient camels (n = 30) were prepared by injection with 20 (1/4)g of the gonadotrophin-releasing hormone analogue buserelin (i.v.) to induce ovulation and then treatment with 1 g meclofenamic acid (a prostaglandin synthetase inhibitor), orally, once on Day 7 and twice daily on Days 8 and 9 after ovulation and thereafter at a dose of 1 g day(-1) until 8 days after embryo transfer. Embryos were transferred into recipients on Day 8 (n = 10), Day 10 (n = 10) or Day 12 (n = 10) after ovulation and another 10 embryos were transferred into untreated recipients on Day 8 after ovulation as controls. In addition, serum samples from all recipient camels were recovered daily throughout the period of meclofenamic acid administration and for a further 7 days after treatment had ceased and were assayed for progesterone concentrations. Results showed that whereas only one of 10 of the control group of recipients (10%) was diagnosed pregnant, a total of eight of 10 Day 8 (80%), six of 10 Day 10 (60%) and seven of 10 Day 12 (70%) recipients were diagnosed pregnant by ultrasonography 12 days after the embryo had been transferred. Subsequently, however, four pregnancies were lost when the conceptus was aged between 22 and 60 days, but this is not considered above the early fetal mortality rate expected in camels after natural mating or after transferring camel embryos to untreated recipients. Serum progesterone concentrations remained elevated, above 2 ng mL(-1), throughout the period of meclofenamic acid administration in all recipient camels; thereafter, concentrations remained above 2 ng mL(-1) in pregnant animals, whereas in non-pregnant camels concentrations had declined to baseline values (<1 ng mL(-1)) within 3 days of the end of the treatment period. In conclusion, treatment of recipient camels with meclofenamic acid reduced the need for tightly timed synchrony between donor and recipient because pregnancies were established in recipients that had ovulated as much as 5 days ahead of the donor.

Investigation of factors affecting pregnancy rate after embryo transfer in the dromedary camel.

The uteri of 36 adult dromedary camels were flushed non-surgically three times each with 90-120 mL of embryo flushing medium 7 days after ovulation. A total of 242 embryos were recovered, of which 139 were transferred non-surgically to recipient camels that were either at different levels of synchrony with respect to the Day 7 donor (+1 to -3 days; n = 58), or were at Day 6 after ovulation, but received one of the following treatments: (i) none (controls, n = 15); (ii) 150 mg progesterone-in-oil injected intramuscularly once daily during Days 5-20 after ovulation inclusive (n = 16); (iii) 500 mg flunixin meglumine given intravenously 15 min before transfer of the embryo (n = 6); (iv) 20 microg of the gonadotrophin-releasing hormone (GnRH) analogue buserelin given on Day 5 after ovulation (n = 12); or (v) the embryo was cooled to 4 degrees C and held at this temperature in an insulated container for 24 h before being transferred (n = 32). Jugular vein blood samples, taken daily from all the recipient camels during Days 0-20 after ovulation, were assayed for progesterone concentration and closely timed serial samples taken from the camels receiving flunixin meglumine or GnRH were assayed for 13,14-dihydro-15-keto prostaglandin F2alpha (PGFM) or oestradiol concentrations. The pregnancy rate increased to a maximum of 67% when ovulation in the recipient was negatively synchronized to have occurred 1 day behind that in the donor, and it fell dramatically when the level of asynchrony between recipient and donor increased to +1 (9%) or -3 (10%) days. It was not improved by daily injections of progesterone (44%), flunixin meglumine given before transfer (16%), or GnRH given on Day 5 (33%). Of the 32 embryos that were cooled to 4 degrees C before being transferred to Day 6 recipients, 20 resulted in pregnancies (63%) to give a success rate similar to that attained with the control fresh embryos (67%). Serum progesterone concentrations in the recipients increased to a mean +/- SEM of 2.6 +/- 0.8 ng mL(-1) by Day 8 after ovulation and, in those that were pregnant, levels remained elevated at 3-5 ng mL(-1) for the remainder of the sampling period; in non-pregnant recipients the concentrations declined to <1 ng mL(-1) by Day 11. Plasma PGFM concentrations in the flunixin meglumine-treated camels remained low (40-90 pg mL(-1)) compared with those in the untreated control camels, in which peak values of around 180 pg mL(-1) were reached within 10 min after transfer after which a steady decline occurred until resting concentrations of 90-100 pg mL(-1) were reached by 110 min after transfer. Treatment with GnRH on Day 5 after ovulation produced a transitory increase in serum oestradiol-17beta concentrations for 24 h. However, from Day 8, oestradiol concentrations in both the GnRH-treated and the untreated camels increased steadily to reach 2.5-3.5 pg mL(-1) by Day 12.