Evaluation of a long-acting recombinant bovine FSH for multiple ovulation and embryo transfer in dromedary camels.

The present study was conducted to test a new super-agonist recombinant bovine FSH (rbFSH) to induce superovulation (SOV) in dromedary camels. In experiment I, a single IM injection of 40, 60, 80, 100, or 120 µg rbFSH was administered (4 donors per group) to determine the effective dose resulting in acceptable multiple ovulation and embryo yield. Administration of 40 µg was ineffective, while 100 and 120 µg were associated with increased numbers of developed follicles, corpora lutea, and recovered embryos compared to administration of 60 and 80 µg. In experiment II, donors were divided into treatment groups to compare rbFSH with two conventional protocols for SOV. Donors received a single dose of 2000 IU eCG in combination with 400 mg porcine follicle-stimulating hormone (pFSH; Folltropin-V®; Group 1, n = 29) or 500 µg of pFSH with 100 µg of pLH (Stimufol®; Group 2, n = 16). Group 3 (n = 19) received a single dose of 100 µg rbFSH. No difference was found in the size and number of follicles per donor. Response time, ovulation rate, and the number of corpora lutea and recovered embryos per donor were similar in all groups. The number of medium-sized and transparent embryos decreased while the number of small-sized and semi-transparent embryos increased in Group 3 (rbFSH) compared to the other two groups. The pregnancy rate of the recipients at 10 days post-ET, at two months of gestation, and the rate of early pregnancy loss (EPL) did not differ among the groups. In conclusion, a single IM administration of 100 µg rbFSH induces a successful superovulation in dromedary camels and has the advantage of reducing stress associated with multiple FSH administration of the conventional protocols.

Comparison between trans-vaginal and recto-vaginal techniques for transferring embryos in the dromedary camels.

The present study was conducted in two breeding seasons to compare the effect of two non-surgical techniques for embryo deposition intrauterine, trans-vaginal (TV) versus recto-vaginal (RV) techniques, on the pregnancy rate and early pregnancy loss (EPL) in dromedary camels. Embryos were collected from 70 donors and transferred to 210 recipients by TV (n = 256 transfers) or RV technique (n = 186 transfers). Pregnancy diagnosis was conducted on Day 10 after embryo transfer (ET) by using the progesterone-ELISA test and by trans-rectal ultrasonography at Day 60 of gestation. EPL was calculated as the recipients that were diagnosed pregnant on Day 10 post-ET and lost their pregnancy between Days 20-60 of their gestation. Using the RV technique in ET of a single embryo showed higher pregnancy rates at Day 19, especially with the embryos of folded, semi-transparent shapes, or those collected after superovulation with the recovery of >4 embryos per flush. While, the pregnancy rates at 60 days showed increases after ET with the RV technique of single, folded, transparent, and semi-transparent, medium-size embryos and/or those collected after superovulation with any number of the recovered embryos than those transferred by the TV technique. The rate of EPL was increased when the TV technique was used for ET of single, spherical, folded, semi-transparent, medium-sized embryos and those collected without or with the superovulation and recovery of >4 embryos per flush. In conclusion, using the RV technique to deposit the embryos intrauterine improves the pregnancy rate and reduces EPL compared to the TV technique.

Factors affecting outcomes of embryo transfer in dromedary camels: A retrospective study.

The present study aimed to use a comprehensive approach for evaluating 12 factors related to embryo quality (shape, size and transparency), donors (age, ET type, number of recovered embryos and day of uterine flushing), recipients (age), males (age and individual variations) and environment (season and year) which could affect the outcome of ET in terms of pregnancy (PR) and pregnancy losses in dromedary camels. During three breeding seasons, 116 donor females were mated repeatedly at 12- to 14-day intervals by fertile male camels (n = 33) without stimulation of the ovaries (WSPO). Superovulation (SPO ET) regimen was applied for each donor female twice or thrice per season. In the occasions of applying superovulation regimen, donor females having an ovulatory follicle were mated instead of GnRH administration and superovulation regimen was applied 4 days post-mating (MIX ET). The uteri of all donor females were flushed at Day 8 or 9 post-mating, and a total of 2,095 embryos were recovered and transferred individually to 924 recipient females. Pregnancy diagnoses were conducted at Day 10 after ET (Days 18-19 of gestation) by using progesterone assay and by transrectal ultrasonography (TRU) at Days 30 and 60 of gestation. By using logistic regression analysis, transparency of embryos and age of recipient females had significant effects on PR at Days 18-19 (p < .01), 30 (p < .01) and 60 (p < .01; p < .05, respectively) of gestation. The shape of embryos had significant effects on the PR at Days 30 (p < .05) and 60 (p < .01) of gestation. Type of ET and the breeding season (year) had significant (p < .05) effects on the PR at Day 30, while day of flushing had the same effect on PR at Day 60. Regarding the pregnancy losses, transparency and shape of the embryo, type of ET, breeding season had significant (p < .05) effect on the late embryonic mortalities (LEM) and shape and season of year had significant (p < .01 and p < .05, respectively) effect on LEM/early foetal mortalities (EFM). Regarding male individual factor, there was a tendency for a significant (p = .055) effect of male camels on the PR at Days 18-19 and rate of LEM. In conclusion, transferring a spherical, transparent or a large-sized embryo (>750 µm) into recipient females ageing between 8 and 11 y could greatly improve the PR from Days 18 to 60 of gestation. Also, embryo recovered from donor females with Mix ET type or embryos sired by certain male camel or at Day 8 post-mating of the donor could improve the 2-month PR. In addition, transferring a transparent or spherical-shaped embryo or embryos recovered from donor females with SPO or Mix ET could reduce the pregnancy losses during the first 2 months of pregnancy.

Factors affecting pregnancy rates and pregnancy losses after embryo transfer in dromedary camels.

The present study was conducted to estimate incidence of late embryonic and early fetal mortalities (LEM/EFM) after embryo transfer (ET) and to investigate factors that could affect proportions of LEM/EFM in dromedary camels. The uteri of 180 donors were flushed at Day 9 post-mating and 1309 ETs were conducted in 886 recipient females in two breeding seasons (2014-2015 and 2015-2016). Pregnancy diagnosis was conducted at Day 10 after ET (Day 19 of gestation) by using the progesterone-ELISA test and by transrectal ultrasonography at Day 60 of gestation. Pregnancy rates at Days 19 and 60 and proportions of LEM/EFM were 54.5%, 34.1%, and 37.5%, respectively. Using logistic regression analysis for evaluations, shape of embryos had an effect (P < 0.01) on pregnancy rates at Days 19 and 60. Also, number of recovered embryos per uterine flushing and breeding season (year) had an effect (P < 0.05) on pregnancy rates at Day 19 and 60, respectively. Regarding rate of LEM and EFM, size of embryos and year of ET had effects (P < 0.01 and P < 0.05, respectively) on the values for these variables. In conclusion, there is a marked occurrence of LEM/EFM in recipient females between Days 20 and 60 of gestation in dromedary camels. The pregnancy rate after ET might be affected by the shape and number of recovered embryos and the year. In addition, the rate of LEM/EFM after ET might be affected by the year and size of the embryos.

Passive immunization against inhibin increases testicular blood flow in male goats.

The present study investigated whether or not passive immunization against inhibin modulates testicular blood flow in goats. Male Shiba goats were injected with either 10 ml of inhibin antiserum (INH group; n = 5) or 10 ml of normal castrated goat serum (NGS group; n = 4). Concentrations of FSH, LH, testosterone (T), and estradiol (E2) in the plasma were measured by radioimmunoassay. Blood flow within the supratesticular (STA) and marginal testicular arteries (MTA) were measured by color pulsed-Doppler ultrasonography, and Doppler indices (resistive index; RI and pulsatility index; PI) were recorded. Results revealed significant increases in concentrations of FSH and E2 in the INH group compared to those in the NGS group (P < 0.05). Animals in the INH group had greater (P < 0.05) FSH concentrations than those in the NGS group in the period between 60 h and 144 h after treatment than at any other time. Estradiol concentrations were greater (P < 0.05) in the INH group than in the NGS group at 6 h (12.15 ± 2.09 pg/ml vs 5.49 ± 1.17 pg/mL), 12 h (8.27 ± 1.29 pg/mL vs 3.05 ± 0.38 pg/mL), and 36 h (9.35 ± 1.31 pg/mL vs 5.09 ± 0.46 pg/mL) after treatment than at any other time. Concentrations of LH and T did not significantly change between the two groups. Goats in the INH group had lesser (P < 0.05) RI of the STA than those in the NGS group and RI values were lesser at 24 h (0.37 ± 0.031 vs 0.49 ± 0.004) and 120 h (0.38 ± 0.028 vs 0.55 ± 0.048) after treatment than at any other time. Furthermore, values of RI and PI of the MTA were significantly lesser (P < 0.05) in the INH group compared to those in the control group at 48 h (RI of MTA: 0.21 ± 0.014 vs 0.37 ± 0.039; PI of MTA: 0.24 ± 0.016 vs 0.46 ± 0.058) after treatment than at any other time. In conclusion, passive immunization against inhibin has a stimulatory effect on testicular blood flow in goats by inducing decreases in the RI values of the STA and MTA.

Preservation of dromedary camel embryos at 4 °C for up to 5 days: Factors affecting the pregnancy and pregnancy loss rates.

The aims of the present study were to evaluate the effect of cooling of the dromedary camel embryos on the pregnancy and pregnancy loss rates, and to investigate the factors which might affect the outcomes of the transfer of cooled embryos. After the donors (n = 56) had been super-ovulated and mated, they were flushed at Day 8 or 9 post-mating. Of 487 collected embryos, 110 were refrigerated at 4°C for up to 5 days in holding medium (HM), flushing medium supplemented with 10% fetal calf serum (FM + FCS) or TCM199 supplemented with 50% FCS and HEPES (TCM + FCS + HEPES). Both fresh (n = 377) and cooled embryos were transferred individually into synchronized recipients. Pregnancy diagnoses were carried out at Days 18-19, 30 and 60 post-mating of the donors. Transferring of fresh embryos into the recipients resulted in significantly higher pregnancy rates at Days 18-19 (53.1% vs. 38.2%, P < 0.01), Day 30 (46.4% vs. 31.8%, P < 0.01) and Day 60 (42.4% vs. 26.4%, P < 0.005) compared with those of cooled embryos, respectively. Pregnancy rates after transferring cooled embryos progressively decreased with the prolongation of the storage period. A significant difference in the pregnancy rate (56% vs. 13%, respectively, P < 0.05) was recorded only at Days 18-19 between cooled embryos held for one day and those held for 5 days. The pregnancy rates at Days 18-19, Day 30 and Day 60 were non-significantly higher when TCM + HEPES and FCS medium used for cooling of embryos compared to those of FM + FCS or HM medium. Cooling of spherical embryos resulted in significantly higher pregnancy rates at Days 30 (45.6% vs. 17.0%, respectively, P < 0.005) and 60 (42.1 vs. 9.4%, respectively, P < 0.005) and a significantly lower pregnancy loss rate (11.1% vs. 66.6%, respectively, P < 0.005) compared to those resulting from cooling of folded embryos. Neither the size of embryo nor the day of flushing had a significant effect on the pregnancy and pregnancy loss rates after the transfer of cooled embryos. In conclusion, pregnancy could be obtained after the transfer of camel embryos refrigerated for up to 5 days. In addition, higher pregnancy rates could be obtained if only spherical embryos are selected for cooling.

Monitoring of embryonic and fetal losses in different breeds of goats using real-time B-mode ultrasonography.

Compared to cattle and sheep, few studies had been undertaken to evaluate the incidence of embryonic and fetal losses (EFL) in goats. The objectives of the present study were to characterize the timing of EFL and to identify the factors that are associated with EFL in goats such as breed, age, parity, method of estrous synchronization, and breeding. Moreover, this study aimed to ensure whether a relationship existed between serum progesterone (P4) and EFL. Goats (n = 151) of different breeds (70 Zaraiebi, 42 Damascus, and 39 Cross goats [Baladi × Damascus]) were evaluated by ultrasonography to monitor EFL during different stages of gestation (D20-23, D26-29, D33-36, D40-45, and D47-54 after breeding). Blood samples were collected at D7, D20, and at each ultrasonographic scanning to clarify changes of serum P4 levels concurrently with EFL. Results revealed that 45 of 109 goats (41.28%) were exposed to EFL. A higher EFL % was observed between D20 to 23 and D47 to 54 (19.61%) compared with D47 to 54 to birth (11.76%). Moreover, a higher EFL % was observed in Zaraiebi goats compared with others. Age and goat parity had no significant effect on the EFL % in all goats. A high EFL % were observed in goats synchronized by P4 sponge, as well as artificially inseminated goats compared with goats with spontaneous estrus, and bred by natural mating, respectively. Serum P4 at D7 or D20 after breeding showed nonsignificant difference between normal pregnant goats and goats that experienced EFL. Unlike goats that experienced partial EFL, goats that experienced total EFL between D20 to 23 and D26 to 29 showed an abrupt P4 reduction (85.06%; P < 0.01) suggesting the probability of endocrine disruption of the CL. However, goats that were exposed to total EFL between D26 and 29 to D33 to 36 showed a low P4 reduction (24.90%; P < 0.05), which might be considered as an effect rather than a cause of EFL. In conclusion, different factors such as breed, estrous synchronization, breeding, and stage of pregnancy may be involved in EFL in goats. Therefore, improvement of the goat management in the early stage of pregnancy is important to decrease EFL % in goats. Although the P4 did not show any significant difference between normal pregnancy goats and goats that experienced EFL, CL disruption should be taken into the consideration, at least, in goats exposed to total embryonic losses.

Comparison of a commercial bovine pregnancy-associated glycoprotein ELISA test and a pregnancy-associated glycoprotein radiomimmunoassay test for early pregnancy diagnosis in dairy cattle.

The present study aimed to compare the accuracy of a commercial PAG-ELISA test (Bovine Preg Test 29) and bovine pregnancy-associated glycoprotein radioimmunoassay (PAG-RIA) for diagnosing pregnancy at Day 28 after insemination in dairy cows. Transrectal ultrasonography (TRUS) was performed in 100 Holstein-Friesian cows at Day 28 after artificial insemination (AI; Day 0) to diagnose pregnancy. After TRUS examination, blood sample was collected from the coccygeal vessels of each cow to measure the concentrations of bPAGs by PAG-RIA test and Bovine Preg Test 29. Milk samples were collected at Days 0, 21 and 28 for measurement of progesterone (P4) by ELISA test. The cows were re-examined by TRUS at Day 42 to confirm the pregnancy diagnoses. The actual gold standard was based on TRUS outcomes at Day 28 that agreed with the outcomes of PAG-RIA test or PAG-ELISA test. If the outcomes of TRUS at Day 28 and PAG-RIA test and PAG-ELISA test did not agree, the gold standard was based on the outcome of TRUS at Day 42. Out of 100 inseminated cows, 41 were confirmed pregnant at Day 28 after AI. Based on the actual gold standard, the sensitivity of TRUS, PAG-ELISA and PAG-RIA tests for diagnosing pregnant cows at Day 28 were 92.7%, 90.2% and 100%, while the specificity of the three tests for diagnosing non-pregnant cows were 91.5%, 98.3% and 94.4%, respectively. The overall accuracy of the three tests were 92%, 95% and 97%, respectively. The degree of agreement (Kappa±S.E.) between PAG-RIA and PAG-ELISA test was 0.90 ±0.04. The degrees of agreement between PAG-RIA and PAG-ELISA and TRUS at Day 28 were 0.80±0.05 and 0.76±0.06, respectively. In conclusion, the commercial PAG-ELISA test is a highly accurate method for diagnosing early pregnancy in dairy cows on Day 28 after AI and may be used as an alternative method to the TRUS and the PAG-RIA test.

Effect of a single injection of gonadotropin-releasing hormone (GnRH) and human chorionic gonadotropin (hCG) on testicular blood flow measured by color doppler ultrasonography in male Shiba goats.

Although color Doppler ultrasonography has been used to evaluate testicular blood flow in many species, very little has been done in goat. Eight male Shiba goats were exposed to a single intramuscular injection of either gonadotropin-releasing hormone (GnRH group; 1 µg/kg BW) or human chorionic gonadotropin (hCG group; 25 IU/kg BW). Plasma testosterone (T), estradiol (E2) and inhibin (INH) were measured just before (0 hr) and at different intervals post injection by radioimmunoassay. Testis volume (TV) and Doppler indices, such as resistive index (RI) and pulsatility index (PI) of the supratesticular artery, were measured by B-mode and color Doppler ultrasonography, respectively. The results indicated an increase in testicular blood flow in both groups, as RI and PI decreased significantly (P<0.05), but this increase was significant higher and earlier in hCG group (1 hr) than in the GnRH group (2 hr). A high correlation was found for RI and PI with both T (RI, r= -0.862; PI, r= -0.707) and INH in the GnRH group (RI, r=0.661; PI, r=0.701). However, a significant (P<0.05) correlation was found between E2 and both RI (r= -0.610) and PI (r= -0.763) in hCG group. In addition, TV significantly increased and was highly correlated with RI in both groups (GnRH, r= -0.718; hCG, r= -0.779). In conclusion, hCG and GnRH may improve testicular blood flow and TV in Shiba goats.

Accuracy of B-mode ultrasonography for diagnosing pregnancy and determination of fetal numbers in different breeds of goats.

A total of 150 does of different breeds were used to evaluate the accuracy of transrectal (TRU) and transabdominal ultrasonography (TAU) for pregnancy diagnosis and fetal number estimation and to study the effect of breed and age of goats on the accuracy of ultrasonography. All does were scanned by TRU at days 17-22, 24-29 and 31-36 and by TAU at days 39-44 and 46-51 after breeding. The sensitivity (Se) of TRU for detecting pregnant does increased significantly (81.6% vs. 97.7%; P<0.0005) from days 17-22 to 24-29. There was a significant (P<0.05) effect of breed of goats on the specificity (Sp) of TAU for diagnosing non-pregnancy between days 39 and 51. In addition, there was a significant (P<0.01) effect of breed of goats on the Se and Sp of TRU for determination of does carrying multiples and singles, respectively, between days 24 and 36. The sensitivity of TAU for diagnosing pregnant does tended to be significantly (P=0.06) higher in young does (1.5-2 yr) than in older does (>2-9 yr) at days 39-44. The accuracy of TRU for determination of does carrying singles was significantly (P<0.05) higher in young does than that in older ones at days 24-29. In conclusions, TRU could be accurately used for early pregnancy diagnosis and fetal number estimation at days 24-29 and days 31-36, respectively, with accuracy similar to that of TAU at days 39-51. Furthermore, breed and age of does might influence the accuracy of ultrasonography for pregnancy diagnosis and fetal numbers estimation.

Relationship of progesterone, bovine pregnancy-associated glycoprotein-1 and nitric oxide with late embryonic and early fetal mortalities in dairy cows.

The aim of the present study was to determine the relationship of progesterone (P4), bovine pregnancy-associated glycoprotein-1 (bPAG-1) and nitric oxide (NO) levels with late embryonic (LEM; day 28 to day 42) and early fetal mortalities (EFM; > day 42 to day 56) in dairy cows. Transrectal ultrasonography (6-8 MHz) was performed in 100 Holstein-Friesian cows at days 28, 42 and 56 after artificial insemination (AI; day 0) to diagnose pregnancy and to monitor the fate of the embryo. After ultrasound scanning of each cow, a milk sample was collected for assessment of P4 by an ELISA test and a blood sample was collected for assessment of bPAG-1, by using a double-antibody radioimmunoassay, and serum NO metabolites (nitrate + nitrite). Based on ultrasonographic examinations and bPAG-1-RIA, 41 of 100 inseminated cows were confirmed pregnant at day 28 after AI. Nine cows suffered of LEM, and 6 cows suffered of EFM and the overall pregnancy loss rate was 36.6% (15/41) between days 28 and 56 of pregnancy. By logistic regression analysis, there were no significant relationships between the level of P4 and bPAG-1 at day 28 after AI and the occurrence of LEM and EFM. Also, there were no significant relationships between the levels of P4 and bPAG-1 at day 42 and the occurrence of EFM. On the other hand, a significant relationship (P<0.05) was found between NO level at day 28 and the occurrence of LEM. In conclusion, measurement of the serum NO concentration at day 28 of pregnancy might help to predict the outcome of pregnancy by day 42 in dairy cows but further studies are needed to confirm this.

Accuracy of transrectal palpation for early pregnancy diagnosis in Egyptian buffaloes.

The present study was undertaken to evaluate the accuracy of transrectal palpation (TRP) for diagnosing early pregnancy in buffaloes and the false diagnoses of the TRP test by using the pregnancy-associated glycoprotein radioimmunoassay (PAG-RIA) test. Pregnancy was diagnosed in 168 buffalo-cows once by TRP and PAG-RIA test between days 31 and 55 after breeding. The sensitivity of TRP for detecting pregnant buffalo-cows was 37.5% at days 31-35, increased to 93.8% at days 46-50 and reached 100% at days 51-55 (P < 0.01). All cases of false negative diagnoses (n = 10) had PAG concentration higher than the threshold (≥1.8 ng/mL) for diagnosing pregnancy. The specificity of TRP for detecting non-pregnant buffalo cows ranged between 90.9%, and 100% between days 31 and 55. All cases of false positive diagnoses (n = 5) made by TRP had PAG concentrations lower than the threshold for diagnosing pregnancy. It could be concluded that TRP is an accurate method for diagnosing pregnant and non-pregnant buffalo cows from day 46 after breeding.

Efficacy of Ovsynch protocol in cyclic and acyclic Egyptian buffaloes in summer.

The aim of the present study was to evaluate the efficacy of an Ovsynch protocol in the cyclic and non-cyclic Egyptian buffaloes in the summer. The present study was carried out on 21 Egyptian pluriparous buffalo cows (11 cyclic and 10 acyclic) and 8 heifers (5 cyclic and 3 acyclic). All animals were administered 100 microg GnRH i.m. at Day 0, 500 microg of PGF2 proportional, variant i.m. at Day 7 and a second dose (100 microg) of GnRH i.m. at Day 9. Ovarian structures of the animals were recorded by means of transrectal ultrasonography (6-8 MHz linear-array transducer) daily from Day 0 (before treatment) to Day 9 (the second GnRH administration) and thereafter, at 12-h interval till ovulation or for a maximum of 48 h. After ultrasound examination, a blood sample was taken daily from each animal for progesterone analysis. The buffalo cows were inseminated at 16-20 h after 2nd GnRH administration. After the first administration of GnRH, 46% (5 of 11) of cyclic and 50% (5 of 10) of acyclic cows and 40% (2 of 5) of cyclic and 33% (1 of 3) of acyclic heifers responded by ovulation or luteinization of the follicles. The mean diameters (+/-S.E.M.) of the follicles in the responding cows and heifers were significantly higher than those in non-responding animals (9.7 +/- 0.4 mm vs. 6.7 +/- 0.6 mm, P < 0.0001; 11.1 +/- 1.5 mm vs. 7.3 +/- 0.7 mm, P < 0.05, respectively). After the second administration of GnRH, 81% (9 of 11) of cyclic and 60% (6 of 10) of acyclic cows and 80% (4 of 5) of cyclic and 33% (1 of 3) of acyclic heifers ovulated, respectively. Ovulations occurred earlier (P = 0.059) and over a wider range of time in the non-cyclic cows (26 +/- 4.8 h; range: 12-36 h) and in a heifer (12 h,) than in the cyclic cows (34.7 +/- 1.3 h; range 24-36 h) and heifers (33 +/- 3 h; range: 24-36 h). The conception rate was 18% (2 of 11) and 0% (0 of 10) in cyclic and non-cyclic cows, respectively. In conclusion, the Ovsynch protocol could be used effectively for synchronization of ovulation in cyclic cows and heifers in summer.

Estimation of gestational age in Egyptian native goats by ultrasonographic fetometry.

The main aim of the present study was to estimate the gestational age of Egyptian goats by B-mode ultrasound measurement of embryonic or fetal parts throughout pregnancy. Trans-rectal (TR) ultrasonography (7 MHz) was carried out on 15 pregnant Egyptian does at Day 10 post mating on alternate days until Day 25 and then once at 3-5-day intervals until Day 50. Trans-abdominal (TA) ultrasonography (3.5-5 MHz) was carried out on the same animals from Days 25 to 130 at 3-5-day intervals. After imaging the embryo or the fetus, the following parameters were measured: length of the embryo or fetus (CRL), heart rate (FHR), biparital diameter (BPD), trunk diameter (TD), placentome size (PS), umbilical cord diameter (UCD) and femur length (FL). The average of days at which the embryonic vesicle was first determined by TR and TA ultrasonography was 16.98+/-1.97 and 27.87+/-3.48, respectively. The embryo proper with a beating heart was first determined by TR and TA ultrasonography at an average of 22.36+/-2.66 and 30.36+/-4.75 days, respectively. All the fetal measures were significantly (P<0.0001) correlated with the gestational age. With the exception of fetal heart rate (R(2)=0.551), all the measured fetal structures were highly correlated (R(2)> or =90) with the gestational age. In conclusion, the age of embryo or fetus in Egyptian does can be estimated by ultrasound measuring the crown rump length, biparital diameter, trunk diameter, placentome size, umbilical cord diameter and femur length.

Effect of pretreatment with insulin on the response of buffaloes with inactive ovaries to gonadotrophin-releasing hormone agonist treatment in summer.

The aim of the present study was to evaluate the effect of a pretreatment with insulin on the response of buffalo cows with inactive ovaries to gonadotrophin-releasing hormone agonist (GnRHa) treatment during hot summer months (July and August). Thirty-six Egyptian buffalo cows with inactive ovaries were randomly allocated into three groups: (1) group treated with GnRHa (G1, n = 16) in which each buffalo received an intramuscular injection of 250 mg of GnRHa (Day 0; gonadorelin, Fertagyl); (2) group treated with insulin before the GnRHa injection (G2, n = 8) in which each buffalo received a subcutaneous injection of biphasic insulin at a dose of 0.25 IU kg(-1) bodyweight once daily starting at Day -3 for 3 consecutive days, followed by an intramuscular injection of 250 mg of GnRHa on Day 0; and (3) the control group (G3, n = 12) in which each buffalo cow received an intramuscular injection of 2.5 mL of sterile saline on Day 0. The ovaries of all animals were examined by trans-rectal ultrasonography (5 MHz) on Days -7, -3 and 0 and continued thereafter at 4-day intervals until oestrus or the end of the experiment. On the same days that ultrasound examinations were performed blood samples were collected to measure the progesterone concentrations. Administration of insulin for 3 days before GnRHa injection (G2) significantly (P < 0.05) increased the diameter of the largest follicle from 6.85 +/- 0.64 to 12.4 +/- 0.88 mm. There was a significant (P < 0.01) increase in the oestrous induction rate in G2 compared with both G1 and G3 during the first 12 days after the treatment interval. It is concluded that pretreatment with insulin for 3 days before GnRHa injection increases the diameter of the dominant follicle and therefore the oestrous induction rate of acyclic buffaloes.

Comparison of accuracy of transabdominal ultrasonography, progesterone and pregnancy-associated glycoproteins tests for discrimination between single and multiple pregnancy in sheep.

The aim of the present study was to evaluate and, compare the accuracy of transabdominal ultrasonographic (US) and the progesterone (P4-RIA) and ovine pregnancy-associated glycoprotein (ovPAG-RIA) tests for the discrimination between single and multiple pregnancy in sheep. One hundred pregnant AwassixMerino ewes were scanned by transabdominal ultrasonography (3.5 MHz linear-array transducer) at Days 43-56 and 81 of these ewes were scanned at Days 76-87 of gestation. The ewes were scanned in dorsal recumbency at the bare area of the inguinal regions (without pre-scanning shaving of the ventral abdominal wall). After each scan, blood samples were withdrawn from the jugular vein to estimate the levels of P4 and ovPAG by radioimmunoassay. At lambing, 61 ewes gave birth to single lambs and 39 ewes gave birth to multiples. The sensitivity of the transabdominal US, the P4-RIA and the ovPAG-RIA tests for determining ewes carrying multiples was 54, 64.1 and 64.1% at Days 43-56. At Days 76-87 of gestation these accuracies were 60.0, 66.7 and 76.6% for the US, P4-RIA and PAG-RIA tests, respectively. The specificity of the transabdominal US, the P4-RIA and the ovPAG-RIA tests for determining ewes carrying singles, was 78.6, 60.7 and 62.3% at Days 43-56 and 78.4, 64.7 and 70.6% at Days 76-87 of gestation, respectively. It is concluded that the accuracy of transabdominal ultrasonographic (without pre-scanning shaving of the ventral abdominal wall), the P4- and the ovPAG-RIA tests for determination of the fetal numbers in AwassixMerino crossbred ewes is too low to be used in the field.

Accuracy of transrectal ultrasonography for determination of pregnancy in sheep: effect of fasting and handling of the animals.

The present study was performed to investigate the effect of previous fasting and lifting of the abdomen of the ewes during transrectal ultrasonographic scanning on the results of early pregnancy diagnosis. Ewes of four flocks (A, B, C and D; all Awassi x Merino ewes, n = 1247 ) aged 0.7-10 years were used in this study. These ewes were estrus synchronized and artificially inseminated. From 2 weeks later onwards, fertile rams were kept with the ewes of flocks A, B and C ( n=949 ) for natural breeding, while ewes of flock D ( n=298 ) were re-inseminated 17 days later. Transrectal ultrasonography (5 MHz) was carried out in ewes of flocks A, B and C on four separate occasions but only once in ewes of flock D. For final analysis, animals were divided over two groups: ewes of Group 1 ( n=949 scans) were scanned in a standing position within the milking parlor. Animals of Group 2 ( n=764 scans) were scanned by the same operator and with the same scanning technique, but these ewes were fasted for 12h prior to scanning and the abdominal wall was lifted, just in front of the udder during scanning. The sensitivity of the test for diagnosing pregnancy at Days 18-24, 25-30, 31-40 and 41-50 was 21.8, 32.3, 63.3 and 50% in Group 1, and 46, 92.5, 92.3 and 96.8% in Group 2, respectively. Only within Group 1, the sensitivity of the test was higher in young ewes (0.7-2 years) than in older ones (>2-10 years). Significant differences were observed at scan periods Days 18-24 and Days 41-50 of gestation. It is concluded that, fasting prior to scanning and lifting the abdomen during scanning significantly improve the accuracy of transrectal ultrasonographic pregnancy diagnosis in Awassi x Merino ewes.

Early pregnancy diagnosis in sheep by progesterone and pregnancy-associated glycoprotein tests.

The aim of this study was to compare the accuracy of the progesterone (P4) and pregnancy associated glycoprotein (PAG) tests for determination of early pregnancy in sheep. Estrus was synchronized in 182 Awassi x Merino ewes and blood samples were collected at Days 0 (day of the insemination), 18, 22, 29, 36, and 50 after artificial insemination (AI). Plasma P4 concentrations at Days 0 and 18 were determined by double antibody radioimmunoassay, while PAG concentrations at Days 22, 29, 36 and 50 were determined by a heterologous, double-antibody radioimmunoassay (RIA) using the bovine PAG 67 kDa subunit as tracer and standard and rabbit antiserum raised against a mixture of caprine 55 and 59 kDa PAG subunits as the first antibody. The discriminatory value for diagnosis of pregnancy by the P4 and the PAG-RIA tests was > or = 1 ng/ml. Based on lambing data, the accuracy for diagnosing pregnant (sensitivity) and non-pregnant ewes (specificity) and predictivity of both tests were calculated. The sensitivity, specificity, positive and negative predictive values for P4 and PAG tests were 100, 95.4, 81.6, and 100% at Day 18 (P4) and 93.5, 100, 100 and 98.7% at Day 22 (PAG), respectively. For diagnosis of non-pregnant ewes the PAG test had significantly higher specificity than the P4 test (P < 0.05). It is concluded that ovine pregnancy can be reliably diagnosed at Day 22 after AI by using a heterologous radioimmunoassay of PAG.

Evaluation of false transrectal ultrasonographic pregnancy diagnoses in sheep by measuring the plasma level of pregnancy-associated glycoproteins.

The present study was undertaken to investigate to what extent pregnancy diagnoses made by transrectal ultrasonography could be confirmed by measurements of plasma concentration of ovine pregnancy-associated glycoproteins (ovPAG). A total of 424 Awassi x Merino ewes were synchronized for estrus and examined by transrectal ultrasonography. In Experiment 1, the ewes (n = 156) were repeatedly scanned in a standing position on d 29, 36 and 50 of gestation. Similarly, the ewes (n = 268) in Experiment 2 were scanned on d 24, 29 and 34 of gestation, but these ewes were fasted for 12 h prior to the examination and the abdominal wall of each animal was lifted up by the hands of the assistant during the scanning. Blood samples were withdrawn after each transrectal ultrasonographic examination in both experiments. Ovine PAG concentrations were measured in plasma by a heterologous radioimmunoassay and the cut-off value for pregnancy was > or = 1 ng.mL-1. Based on the lambing performance, in Experiment 1, altogether 47 false negative and 38 false positive diagnoses were made by transrectal ultrasonography in 24 and 33 ewes, respectively between d 29 and 50 of gestation. In Experiment 2, altogether 8 false negative and 13 false positive diagnoses both were made in 7 ewes between d 24 and 34 of gestation. In both experiments, all ewes with false negative diagnoses had ovPAG concentrations higher than the threshold level for pregnancy diagnosis and all ewes with false positive diagnoses had ovPAG concentrations lower than the threshold of pregnancy. Furthermore, by the PAG-RIA test all lambed or aborted ewes (n = 63) were correctly diagnosed as pregnant and with three exceptions, all non-lambed ewes (n = 361) were correctly diagnosed as non-pregnant during the examined periods of both experiments.