Bull Vet Inst Pulawy 50, 97-100, 2006 INDUCTION OF MULTIPLE BIRTHS IN AKKARAMAN CROSS-BRED SHEEP SYNCHRONIZED WITH SHORT DURATION AND DIFFERENT DOSES OF PROGESTERONE TREATMENT COMBINED WITH PMSG OUTSIDE THE BREEDING SEASON MEHMET AKÖZ, BÜLENT BÜLBÜL1, MEHMET BOZKURT ATAMAN2 AND SÜLEYMAN DERE3 Department of Gynaecology and Obstetrics, 2 Department of Reproduction and Artificial Insemination, 3 Department of Animal Science, Faculty of Veterinary Medicine, University of Selcuk, 42075, Konya, Turkey 1 Department of Artificial Insemination and Biotechnology, Bahri Dagdas International Agricultural Research Institute, 42020 Karatay-Konya, Turkey e-mail:email@example.com Received for publication January 14, 2005. Abstract gonadotrophin (eCG), follicle stimulating hormone (FSH) (17, 20), pregnant mare serum gonadotrophin Determination of the optimal PMSG dose for inducing (PMSG) (7, 23), human chorionic gonadotrophin (hCG) increased the prolificacy in Akkaraman cross-bred ewes and mixed gonadotrophin preparations (13). Another synchronized with the different doses of fluorogestone acetate technique is actively immunizing ewes against (FGA) outside the breeding season was the aim of this study. androstendione (10), testosterone (3) and oestron (25). A total of 90 non-lactating ewes were randomly divided into The aim of this study was the determination of the two groups. Vaginal sponges containing 30 mg (group FGA1) and 40 mg (group FGA2) of fluorogestone acetate were optimal dose of PMSG for inducing increased inserted into the vagina of the ewes. The sponges were prolificacy in Akkaraman cross-bred ewes synchronized withdrawn on day 7 and 300 IU of PMSG (FGA1A, FGA2A), with different doses of fluorogestne acetate (FGA) 500 IU of PMSG (FGA1B, FGA2B) and 700 IU of PMSG outside the breeding season. (FGA1C, FGA2C) were injected intramuscularly in the named above subgroups of FGA1 and FGA2. Oestrus response, pregnancy, lambing, and multiple birth rates were 100%, Material and Methods 93.3%, 78.6% and 18.8% in the group FGA1A, 93.3%, 92.8%, 76.9% and 40% in the group FGA1B, 100%, 100%, 86.7% and 69.2% in the group FGA1C, 93.3%, 92.8%, 76.9% and 20% in A total of 90 non-lactating ewes (Merinos x the FGA2A, 92.8%, 100%, 84.6% and 36.4% in the group Akkaraman crossbred, F1) aging 18-24 months and 6 FGA2B, 100% 93.3%, 85.7%, 66.7% in the group FGA2C, healthy rams (Merino) aging 2-3 years with a known respectively. Multiple birth rates in the groups FGA1C and fertility were used. The study was carried out between FGA2C were significantly higher than in the other groups. In May 1 and 31, which is the period accepted as outside conclusion, the application of 700 IU PMSG was rather more breeding season in Konya, Turkey. effective than injections of 300 IU and 500 IU in ewes being The ewes were kept indoors at night and had access outside the breeding season. to grazing outdoors for most of the day. Indoors, the ewes were fed barley, wheat bran, and wheat straw Key words: Akkaraman ewe, fluorogestone, supplemented with vitamins. Water and mineral licks PMSG, oestrus synchronization, non-breeding season. were available ad libitum. At first, the ewes were randomly divided into two groups. Vaginal sponges (Chrono-gest, grey sponges, Methods to improve reproduction in ewes often aim Intervet, Turkey) containing 30 mg of FGA were to increase the proportion of ewes having multiple inserted into the vagina of ewes in the first group ovulations, and thereby increase lambing percentage (FGA1) (n=45) and vaginal sponges (Chrono-gest, white (24). Ovulation rates are increased by injection of sponges, Intervet, Turkey) containing 40 mg of FGA gonadotrophins (22) such as human menopausal were inserted into the vagina of ewes in the second gonadotrophin (hMG) (6), equine chorionic group (FGA2) (n=45). The sponges were withdrawn on 98 day 7. The ewes, following the withdrawal of the Results sponges, were divided into three subgroups in each of FGA groups. Each subgroup consisted of 15 ewes. In the One of the ewes in the group FGA2B lost its subgroups of FGA1 and FGA2, 300 IU (FGA1A, vaginal sponge during the treatment and data analysis FGA2A), 500 IU (FGA1B, FGA2B), and 700 IU did not include this ewe. Oestrus response, pregnancy, (FGA1C, FGA2C) of PMSG (Folligon, Intervet, lambing, and multiple birth rates in the subgroups were Turkey) were injected intramuscularly to all ewes. summarized in the Table 1. After the injections of PMSG, the oestrus cycle of There were no statistically significant differences each ewes was followed twice at a 12-h interval using among the groups FGA1A, FGA1B, FGA1C, FGA2A, teaser rams. After the detection of oestrus, the ewes FGA2B, and FGA2C for oestrus, pregnancy, and were hand-mated. Pregnancy in ewes was determined lambing rates (P>0.05). But multiple birth rates in the using the real-time B-mode ultrasound (Scanner 480 groups FGA1C and FGA2C were significantly higher Vet, Pie Data Medical, Maastrich, Netherlands) with the than in the other groups (P<0.05). There were no 5 MHz linear-array transrectal probe on day 30 significant differences among the groups FGA1A, following the mating. The number of multiple embryos FGA1B, FGA2A, and FGA2B for multiple birth rates of each ewe was determined using the transrectal (P>0.05). ultrasonography on days 30 and 36 following the mating, as described by Schrick and Inskeep (21). The following traits were evaluated for each of the Discussion treated groups: - oestrus response (number of ewes showing Increasing rate of reproduction in sheep offers the oestrus/total ewes treated in each group x 100); best opportunity to increase the efficiency of lamb meat - pregnancy rate (number of pregnant ewes/number production (15). Methods improving the reproduction of ewes showing oestrus and mated in each often aim to increase the proportion of ewes having twin group x100); ovulations, and thereby increase multiple birth - lambing rate (number of ewes lambing/number of percentage (24). pregnant ewes in each group x 100); Gonadotrophins are used to induce superovulation - multiple birth rates (number of multiple in sheep (6). These compounds are administered over lambing/total lambing in each group). the last 2 - 3 d before progestagen sponge removal or The data were evaluated with chi-square analysis to injection of prostaglandin to synchronize oestrus (9). compare the oestrus response, pregnancy, PMSG, when injected immediately after the removal of lambing the progestagen sponge, may produce an increase in the and multiple birth rates among the groups at the 5% rate of ovulation (16, 19). and 1% level of significance (11). Table 1 Oestrus response, pregnancy, lambing, and multiple birth rates Oestrus response Pregnancy rate Lambing rate Multiple birth rate (%) (%) (%) (%) FGA1A 15/15 14/15 11/14 2/11 (300 IU PMSG) (100) (93.3) (78.6) (18.2) b (n=15) FGA1B 14/15 13/14 10/13 4/10 FGA1 (500 IU PMSG) (93.3) (92.8) (76.9) (40) b (n=15) FGA1C 15/15 15/15 13/15 9/13 (700 IU PMSG) (100) (100) (86.6) (69.2) a (n=15) FGA2A 14/15 13/14 10/13 2/10 (300 IU PMSG) (93.3) (92.8) (76.9) (20) b (n=15) FGA2B 13/14 13/13 11/13 4/11 FGA2 (500 IU PMSG) (92.8) (100) (84.6) (36.4) b (n=15) FGA2C 15/15 14/15 12/14 8/12 (700 IU PMSG) (100) (93.3) (85.7) (66.7) a (n=15) Different superscripts in columns differ significantly; a,b: P<0.05 99 In our study, the doses of 300 IU (18.2% and 20%) IU and 500 IU of PMSG in Akkaraman cross-bred ewes and 500 IU of PMSG (40% and 36.4 %) compared to in the breeding season. 700 IU (69.2% and 66.7%) were found to be less effective to induce multiple births. This might be Acknowledgments: The authors would like to because the dose used in our study was not sufficient to thank Mr Şefik SAYANER for providing the stimulate additional follicular development or weak experimental animals. response of the breed used in this experiment. Regarding this, the use of 500 IU of PMSG at sponge withdrawal was reported to improve the fertility by about 35-70% References (2), but the response among different breeds can be varied (12). The multiple birth rates obtained in the 1. Ak K., İleri K.İ., Horoz H., Şenünver A., Alkan S., groups FGA1C (69.2%) and FGA2C (66.7%) were Rahimi H., Kaşıkçı G.: Kıvırcık koyunlarda mevsim içi similar to findings of Bekyürek (2). Romano et al. (19) östrus senkronizasyonu. İ Ü Vet Fak Derg 1994, 20, 261- reported that, PMSG at 250 IU did not increase the 266. fertility or prolificacy rate during the breeding season. 2. Bekyürek T.: Anöstrüs dönemindeki Tuj koyunlarında Similarly, 300 IU of PMSG did not sufficiently increase östrüsün uyarılması. Tr J Vet Anim Sci 1993, 18, 11-15. 3. Cognié Y., Scaramuzzi R.J. : Les techniques the multiple birth rates in our study. physiologiques d’accroissement de la fertilité de la Koyuncu et al. (14) synchronized the Kıvırcık ewes prolificité chez les ovins. Actes du 3ème Congres using pessaries containing progesterone and Mondial de Reproduction et Selection des Ovins et superovulated them with different doses of PMSG (500 Bovins à Viande, Ed. INRA, Paris, 1988, 2, 623-636. and 700 IU) during breeding season. Prolificacy rates 4. Çoyan K., Ataman M.B., Erdem H., Kaya A., Kasıkcı G.: were 158% and 196% in the 500 and 700 IU groups, Synchronization of estrus in cows using double PGF2 respectively. The authors demonstrated that progestagen alpha, GnRH - PG2 alpha and hCG- PGF2 alpha and PMSG treatment increased prolificacy significantly combination. Revue Méd Vét 2003, 154, 91-96. in Kıvırcık ewes. In this study, FGA + PMSG treatment 5. Esen F., Bozkurt T.: Akkaraman ırkı koyunlarda flushing ve östrüs senkronizasyonu uygulamasının dölverimi also provided sufficient synchronization and multiple üzerine etkisi. Tr J Vet Anim Sci 2001, 25, 365-368. birth rates. 6. Evans A.C.O.: Ovarian follicle growth and consequences Özbey and Tatlı (18) synchronized the Awassi ewes for fertility in sheep. Anim Reprod Sci 2003, 78, 289- for 14 d with sponges containing 40 mg of FGA and 306. superovulated by 500 IU of PMSG injection and oestrus 7. González-Reyna A., Márquez-Garciá E., Lizárraga-Tracy and twinning rates were 100% and 46%. These rates H., Martínez-González J.C.: Dose response effects of were higher than our ones obtained by using 500 IU of PMSG on ovulation rate and follicular development in PMSG. This difference may be explained by the Pelibuey ewes treated with Syncro-mate-B implants. influence of breed differences on response to the Small Rum Res 1999, 31, 149-155. 8. Gökçen H., Ünal E.F, Tümen H., Deligözoğlu F., Soylu treatment protocol. Also, Karagiannidis et al. (12) M.K., Çelik İ.: Kızgınlıkları değişik yöntemlerle reported that response to the different PMSG doses sinkronize edilerek tohumlanan Merinos koyunlarında among various breeds was different. dölverimi üzerinde araştırmalar. U Ü Vet Fak Derg 1992, In our study, results in groups FGA1C (100%, 11, 81-90. 69.2%) and FGA2C (100%, 66.7%) for oestrus and 9. Haresign W.: Controlling reproduction in sheep. New twinning rates were similar to those obtained by Esen developments in sheep production. In: British Society of and Bozkurt (5) who treated the Akkaraman ewes for 14 Animal Production. Ed: CFR Slade and TLJ Lawrence, d with pessaries containing 40 mg of FGA and injection 1990, 14, 23-37. of 600 IU of PMSG at the time of pessary removal. 10. Haresign W.: Manipulating of reproduction in sheep. J Reprod Fert Suppl 1992, 45, 127-139. They reported that the oestrus and twinning rates in 11. Inal S.: Biyometri, S. U. Veteriner Fakültesi Basımevi, breeding season were 94% and 57%, respectively. Konya,Turkey, 1998, p. 12. Ak et al. (1) treated the Kıvırcık ewes for 14 d with 12. Karagiannidis A., Varsakeli S., Karatzas G., Brozos C.: sponges containing FGA and injected them with 500 IU Effect of time of artificial insemination on fertility of of PMSG and 500 IU of hCG at pessary removal. Their progestagen and PMSG treated indigeneous Greek ewes, oestrus and twinning rates were 100 and 90%. The rates during non-breeding season. Small Rum Res 2001, 39, were lower in the groups FGA1B and FGA2B than those 67-71. reported by Ak et al. (1) and this may be due to lack of 13. Knights M., Baptiste Q.S., Dixon A.B., Pate J.L., Marsh hCG injection in our experiments. Relevant to this, D.J., Inskeep E.K. Lewis P.E.: Effects of dosage of FSH, vehicle and time of treatment on ovulation rate and Çoyan et al. (4) reported that hCG was a good prolificacy in ewes during the anestrous season. Small exogenous source of LH to accelerate the ovulation. Rum Res 2003, 50, 1-9. In our study, oestrus and pregnancy rates in the 14. Koyuncu M., Uzun Ş.K., Şengül L.: Kıvırcık groups FGA1A-B and FGA2A-C agreed with those koyunlarında progesterone ve farklı dozda PMSG observed by Gökçen et al. (8), who superovulated the kullanımının kızgınlık denetimi ve döl verimini arttırma Merino ewes with vaginal sponge + 500 IU of PMSG olanakları. Tr J ve Anim Sci 2001, 25, 971-974. and received 92% oestrus and 85% pregnancy rates. 15. 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Noël B., Mandiki S.M.N., Perrad B., Bister J.L., Paquay C., Toutges M.J., Reynolds L.P., Redmer D.A.: R.: Terminal follicle growth, ovulation rate and hormonal Ovulation rate in ewes synchronized with syncro-mate-B secretion after melatonin pre-treatment prior to FGA- (SBM) and follicle stimulating hormone. Small Rum Res PMSG synchronisation in Suffolk ewes at the onset of 2003, 48, 1-8. the breeding season. Small Rum Res 1999, 32, 269-277. 23. Wani G.M., Buchoo B.A., Wani N.A.: Use of human 18. Özbey O., Tatlı P.: İvesi koyunlarda flushing ve gonadotrophin in superovulation of Southdown sheep. sinkronizasyon uygulamalarının döl verimi üzerine etkisi. Small Rum Res 1997, 25, 93-94. U Ü Vet Fak Derg 2001, 20, 109-116. 24. Wilkins J.F.: Method of stimulating ovulation rate in 19. Romano J.E., Rodas E., Ferreira A., Lago I., Benech A.: Merino ewes may affect conception but not embryo Effects of progestagen, PMSG and artificial insemination survival. Anim Reprod Sci 1997, 47, 31-42. time on fertility and prolificacy in Corriedale ewes. Small 25. Wilson P.A., Cox R.I., Wong M.S.F., Paul D.R.: Rum Res 1996, 23, 157-162. Improved prolificacy in Merino ewes immunized against 20. Rubianes E., Castro T., Kmaid S., Carbajal B., Benquet a combination of androgens and oestrogens. Proc Aust N., Pinczak A.: Superovulatory response to FSH Soc Reprod Biol 1986, 18, 41. treatments after different progesterone priming in ewes. Small Rum Res 1999, 33, 159-162.
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