Songklanakarin Journal of Science and Technology SJST R1 Vajdi Hokmabad

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1 The effect of Controlled Internal Drug Release device (CIDR) and progesterone sponge on short-term estrus synchronization in Zandi ewes during the breeding season Journal: Songklanakarin Journal of Science and Technology Manuscript ID SJST-0-0.R Manuscript Type: Original Article Date Submitted by the Author: -Apr-0 Complete List of Authors: Roshani, Behzad; Miyaneh Branch, Islamic Azad University, Miyaneh, Iran., Department of Animal Breeding Vajdi Hokmabad, Reza; Miyaneh Branch, Islamic Azad University, Miyaneh, Iran., Department of Veterinary Mohebalipour, Naser; Miyaneh Branch, Islamic Azad University, Miyaneh, Iran., Department of Agriculture Keyword: CIDR, Intra vaginal sponge, sheep, Estrus Synchronization, breeding season

2 Page of The effect of Controlled Internal Drug Release device (CIDR) and progesterone sponge on short-term estrus synchronization in Zandi ewes during the breeding season Behzad Roshani, Reza Vajdi Hokmabad *, Naser Mohebalipour Department of Animal Breeding, Miyaneh Branch, Islamic Azad University, Miyaneh, Iran Department of Veterinary, Miyaneh Branch, Islamic Azad University, Miyaneh, Iran. Department of Agriculture, Miyaneh Branch, Islamic Azad University, Miyaneh, Iran. * corresponding author: address: r.vajdi@m-iau.ac.ir, Cell phone: +00, Fax:+00

3 Page of Original Article The effect of Controlled Internal Drug Release device (CIDR) and progesterone sponge on short-term estrus synchronization in Zandi ewes during the breeding season Abstract The aim of this study was to investigate the effect of Controlled Internal Drug Release device (CIDR) and progesterone sponge on the short-term estrus synchronization in Zandi ewes during the breeding season. So 0 Zandi ewes were selected and divided into groups at the beginning of breeding season. The first group was the control group. The second group received: CIDR insertion and. ml of GnRH (day 0),. ml of natural prostaglandin (PGF α, day ) and. mg of PMSG (Pregnant Mare Serum Gonadotropin) with the removal of the CIDR (day ). For the third group, the same treatment was used, but medroxyprogesterone acetate-impregnated sponge (MAP) was inserted instead of CIDR. All of the ewes were mated naturally. There were significant differences in estrus, fertility, and pregnancy rates between the second group and the control group (P<0.0). It is recommended to use CIDR due to relative improvement of some reproductive parameters. Keywords: CIDR, Intra vaginal sponge, sheep, Estrus Synchronization, breeding season.. Introduction Estrus synchronization allows us to manipulate time of parturition so that we can have a better time of delivery in a year. Better management of the market, easy access to

4 Page of feed, and lower amount of allocated work and costs are the other advantages (Whitley & Jackson, 00). The most common method of estrus synchronization in ewe is the use of intravaginal devices (with synthetic progesterone or progestagen such as Fluro Gestone Acetate (FGA) or medroxyprogesterone acetate (Fukui, et al., )). The intravaginal sponges containing MAP or FGA are usually inserted into the vagina for 0- days and combine with a dose of ecg (equine chorionic gonadotropin) about hours before the removing of progesterone sponge (Zeleke et al., 00). In addition, Prostaglandin F α (PGF α ) can be injected hours before or at the time of removal of sponge (Beck et al., ; Dogan & Nur, 00; Ali, 00). Traditional treatments for estrus synchronization in ewe and goats consist of a long progestagen insertion associated with an i.m. injection of ecg given at the end of the treatment was developed over 0 years ago and is still recommended (Menchaca et al., 00). Due to the negative effect of long-term progesterone treatment on the subsequent fertility of ewes (Vinoles et al., 00), a functional alternative method, the short-term treatment, has recently been proposed for estrus synchronization in ewes (Ali, 00; Husein et al., 00). Short-term periods of treatment with progesterone sponges (- days) have been used successfully for synchronization of estrus in ewes within or out of the breeding season (Beck et al., ; Vinoles et al., 00; Ataman & Akoz, 00). In 00, the efficacy of using intravaginal CIDR for estrus synchronization has also been approved by the FDA (Food and Drug Administration of America) and now it is used for estrus induction in anestrus ewes for to days (Jackson et al., 0). In addition, a dose of GnRH has widely been used in cattle to

5 Page of manipulate patterns of follicular development in the ovaries (Macmillan et al., 00). Administration of GnRH causes ovulation of the dominant follicles in the atresian phase and induces a new wave of follicular growth within to days after treatment in every stage of the reproductive cycle in cattle (Twagiramungu et al., ). An alternative method is the use of simultaneous injection of GnRH and PGF α, that has also been used in sheep during the breeding season (Beck et al., ). The effective treatment of ewes in the synchronization of estrus by using the combination of GnRH and PGF α has been reported in the study conducted by Ataman et al. (00). The aim of this study was to investigate the effect of Controlled Internal Drug Release device (CIDR) and progesterone sponge on the short-term estrus synchronization in Zandi ewes during the breeding season.. Materials and Methods Islamic Azad University animal health and breeding department approved all protocols and procedures used in this study. 0 heads of - year old Zandi ewes with the average weight of - kg were selected. All were physically and reproductively healthy and pastured with mixed grass. The experiment was carried out in the countryside of Baladeh city situated in Mazandaran province (/0 northern & /0 eastern) of Iran in early September of 0 (the beginning of breeding season in this region). The ewes fed from pasturage freely and salt and water were available for them. All rams were separate from the herd. After determining the age of ewes and numbering, the ewes were moved to drylot and divided into groups of 0 heads. The first group was the control group that received no treatment. On day zero, the second group received (CIDR treatment) CIDR insertion (each CIDR contains 00 mg of

6 Page of natural progesterone, Pfizer Animal Health, New Zealand),. ml of GnRH (each ml contains 0.00 I.U. of buserelin acetate, Rooyan Darou, Iran, I.M.); on the sixth day,. ml of PGF α (Vetalyse, mg/ml Dinoprost tromethamine, Aburaihan Co., Iran) intramuscularly; and on the seventh day, with the removal of the CIDR, 00 I.U. of PMSG (Pregnant Mare Serum Gonadotropin) (Folligon, 00 I.U./ml, Intervet, Boxmeer and Holland) was injected intramuscularly to each of the ewes. The same treatment with the second group was used for the third group, but instead of CIDR, medroxyprogesterone acetate-impregnated sponges (each sponge contains 0 mg of medroxyprogesterone acetate) (Esponjavet 0 mg, Hipra-lab, Spain) were inserted intravaginally in ewes. The treatment used for the three groups has been presented in Table and fig.. After removal of the CIDR and sponge on the seventh day, rams were entered into the three groups of ewes for mating. One - year-old ram (clinically healthy with good reproductive history in last breeding season) was entered for every heads of ewes for weeks and for all groups. Data were collected through controlling the herd at delivery time. The number of lambs born per ewe (single or twin), their sex and birth weight were recorded. The estrus rate (Number of ewes showing estrus/total number of ewes in each group 00), pregnancy rate (Number of pregnant ewes/number of mated ewes in each group 00), fertility rate (Number of ewes lambing/number of mated ewes in each group 00), litter size (Number of lambs born in each group/total number of mated ewes in each group 00), multiple birth rate (Number of multiple lambing/total lambing in each group 00) were calculated in the three groups and the results were compared with each other.

7 Page of For the statistical analysis, the mean squares of reproductive parameters were compared with each other between the three groups, at first. Then, the coefficient of variation (CV) of the reproductive parameters were compared individually in each of the groups. Analysis of variance of reproductive parameters including the estrus rate, pregnancy rate, fertility rate, litter size and multiple birth rates in ewes were done using SPSS (ver.0) and Dunnett and Duncan methods. The comparison of the means was performed using Chi-square test at the probability level of %.. Results and Discussion As presented in Table, there were significant differences (P<0.0) between the CIDR group and the control group in estrus rate, pregnancy rate, and fertility rate but there were no significant differences (P>0.0) between the CIDR group and the sponge group in reproductive parameters. Furthermore, no significant differences (P>0.0) were observed between the sponge group and the control group in terms of reproductive parameters. Moreover, no significant differences (P>0.0) were found between the treatment groups and the control group in litter size, average birth weight and multiple birth rates. Based on the results of variance analysis (Table ), the highest variations are related to estrus rate, pregnancy rate, and fertility rate. Various sources of exogenous progesterone (CIDR and sponge) are used for inducing estrus and estrus synchronization in seven days. This pattern of estrus synchronization was used in a study conducted by Jackson et al. (0). They used CIDR and PG injection, only CIDR insertion for days and tested reproductive parameters between two treatment groups and control group. They found no significant differences (P 0.) between the treatments in lambing rate and multiple birth rates and CIDR treated

8 Page of ewes had less days to estrus and lambing (P 0.0) than control ewes (Jacson, et al. 0). Furthermore, this pattern of estrus synchronization was applied in the study carried out by Wheaton et al. (). They have reported that the interval between the transitional phase and estrus in ewes treated with CIDR lasted for days after CIDR withdrawal on average but in the control group that received no treatment, this interval lasted for days on average (Wheaton et al.,). However, Titi et al. (00) found that there was no difference between the control group and the CIDR group (treated for days) regarding the average lambing days. In fact, the days of lambing were similar for all groups and as the same as current study they found no significant differences (P>0.0) between the treatment groups and the control group in estrus rate. But they reported increases in fertility rate in the ewes treated with CIDR for days in combination with GnRH and PGF α which is in agree with the present study. This might be probably due to the issue that the ewes did not have synchronous follicular developments or active corpus luteum. Furthermore, the ewes may have shown a false estrus or a complete estrus cycle at the time of PGF α injection. In agree with the present study (short-term treatment with exogenous progestrone), Karaca et al. (00) used progestagen sponges in combination with GnRH, PGF α, and ecg for synchronization of estrus with two methods of short-term and long-term treatments. With the exception of fertility rate, there were no significant differences (P>0.0) between the other measured reproductive parameters in their study. The results showed higher fertility rate in short-term treatment ( days) than in long-term treatment ( days) with progestagen at the beginning of mating season (P<0.0). Vinoles et al. (00) concluded that the lower pregnancy rate observed after long-term progestagen treatment was related to a slower follicular turnover that promoted the ovulation of

9 Page 0 of persistent dominant follicles; and short-term treatment resulted in a higher pregnancy rate probably due to the ovulation of newly recruited growing follicles. Further, Barrett et al. (00) have declared that the injection of 00 IU of ecg after days of progestagen treatment had limited effects on the dynamics of ovarian follicular waves. In the present study, although a high dose of PMSG (. mg) was used in short-term treatment with progestagen ( days), no significant differences were observed in multiple birth rate and litter size. This is probably due to the difference in the protocol of using PGF α and GnRH dosage in Zandi breed witch naturally have lower multiple birth rate. However Menchaca and Rubianes (00) reported the lysis of the corpus luteum with the injection of PGF α that increased estrus rate and pregnancy percentage during the treatment with progestagen in the breeding season of sheep. Husein and Kridli (00) have reported that seasonal anestrus ewes treated with GnRH- PGF α showed increase in estrus responses and pregnancy rates. Also, the use of GnRH in current study improved estrus rate, pregnancy rate, and fertility rate. The disagreement observed among the various studies may be due to the increase in the number of follicles, follicular synchronization, and starting a new recruitment of follicular growth for ovulation at the time of starting a treatment protocol.. Conclusions Based on the results, it can be concluded that both CIDR and sponge can be used for estrus synchronization; but due to the ease of CIDR insertion and its fewer side effects and also improving the estrus, pregnancy, and fertility rates in comparison with the sponge, it is recommended to use CIDR for short-term treatment in the breeding season for estrus synchronization.

10 Page of References Ali, A. 00. Effect of time of ecg administration on follicular response and reproductive performance of FGA-treated Ossimi ewes. Small Ruminant Research.,. Ataman M.B. and Akoz M. 00. GnRH-PGF a and PGF a- PGF a synchronization in Akkaraman cross-bred sheep in the breeding season. Bulletin of the Veterinary Institute in Pulawy Barrett, D.M.W., Bartlewski, P.M., Batista-Arteaga, M., Symington, A. and Rawlings, N. C. 00. Ultrasound and endocrine evaluation of the ovarian response to a single dose of 00IU of ecg following a -d treatment with progestagen-releasing intravaginal sponges in the breeding and non-breeding seasons in ewes. Theriogenology., -. Beck, N.F.G., Davies, B. and Williams, S.P.. Oestrus synchronization in ewes: the effect of combining a prostaglandin analogue with a -d progestagen treatment. Animal Production., 0 0. Beck N.F.G. Jones M. Davies B. Peters A.R. and Williams S.P.. Oestrus synchronization in ewes the effects of combining a prostaglandin analogue with a GnRH agonist (boserelin). Animal Reproduction Science. -. Dogan, I. and Nur, Z. 00. Different estrous induction methods during the nonbreeding season in Kivircik ewes. Journal of Veterinary Medicine.,.

11 Page of Fukui, Y., Ishikawa, D., Ishida, N., Okada, M., Itagaki, R. and Ogiso, T.. Comparison of fertility of estrous synchronized ewes with four different intravaginal devices during the breeding season. Journal of Reproduction and Development.,. Husein, M.Q. and Kridli, R.T. 00. Effect of progesterone prior to GnRH-PGF α treatment on induction of oestrus and pregnancy in anoestrus Awassia ewes. Reproduction in Domestic Animals.,. Husein, M.Q., Ababneh, M.M. and Abu-Ruman, D.S. 00. The effects of short or long term FGA treatment with or without ecg on reproductive performance of ewes bred out-of-season. American Journal of Animal and Veterinary Sciences. (),. Jackson C.G. Neville T.L. Mercadante V.R.G. Waters K.M. Lamb G.C. Dahlen C.R. and Redden R.R. 0. Efficacy of various five-day estrous synchronization. Small Ruminant Research. 0, Karaca F. Ataman M.B. and Coyan K. 00. Synchronization of estrus with short-and long- term progestagen treatments and the use of GnRH prior to short - term progestagen treatment in ewes. Small Ruminant Research., -. Menchaca A. and Rubianes E. 00. New treatments associated with timed artificial insemination in small ruminant. Reproduction. Fertility and Development. 0-.

12 Page of Menchaca, A., Miller, V., Salveraglio, V. and Rubianes, E. 00, Endocrine, luteal and follicular responses after the use of the Short-Term Protocol to synchronize ovulation in goats, Animal Reproduction Science. 0,. Macmillan, K.L., Segwagwe, B.V.E. and Pino, C.S. 00. Associations between the manipulation of patterns of follicular development and fertility in cattle. Journal of Animal Reproduction Science.,. Titi, H.H., Kridli, R.T. and Alnimer, M.A. 00. Estrus synchronization in sheep and goats using combinations of GnRH, progestagen and prostaglandin F α. Reproduction in Domestic Animals. (), -. Twagiramungu H. Guilbault L.A. and Dufour J.J.. Synchronization of ovarian follicular waves with a gonadotropin- releasing hormone agonist to increase the precision of estrus in cattle: a Review. Journal of Animal Reproduction Science. -. Vinoles, C., Forsberg, M., Banchero, G. and Rubianes, E. 00. Effect of longterm and short-term progestagen treatment on follicular development and pregnancy rate in cyclic ewes. Theriogenology., -00. Wheaton, J.E., Windels, H.F. and Johnston, L.J.. Accelerated lambing using exogenous progesterone and the ram effect. Journal of Animal Reproduction Science. 0, -. Whitley, N.C. and Jackson, D.J. 00. An update on estrus synchronization in goats: a minor species. Journal of Animal Science., 0.

13 Page of Zeleke, M., Greyling, J.P.C., Schwalbach, L.M.J., Muller, T. and Erasmus, J.A. 00. Effect of progestagen and PMSG on oestrous synchronization and fertility in Dorper ewes during the transition period. Small Ruminant Research.,.

14 Page of Table. Average age and weight and the treatment used for the groups of ewes Number Average Age Average Groups Treatment (Head) (year) Weight (kg) Control 0.. no treatment CIDR Day 0: insertion of CIDR and injection of. mg GnRH simultaneously + th day: injection of. mg PGF α + th day: removal of CIDR and injection of. mg PMSG, simultaneously Sponge 0.0. st day: insertion of sponge and injection of. mg GnRH simultaneously + th day: injection of. mg PGF α + th day: removal of sponge and injection of. mg PMSG, simultaneously Table. Assessment of changes in reproductive traits in the different experimental groups Parameters Control group Sponge group CIDR group Number of ewes (head) Male lamb (head) Female lamb (head) Total number of lambs (head) The average weight of male lambs (kg)... The average weight of female lambs (kg)... The total average weight of lambs (kg)... Number of delivery (birth) 0 Estrus rate (%) 0 b ab 00 a Number of twins Litter size (%).. 0 Pregnancy rate (%) 0 b ab 00 a Fertility rate (%) 0 b ab 00 a Multiple birth rates (%) a b represent a significant difference between the groups (P<0.0)

15 Page of Table. Analysis of Variance and Coefficient of Variation Sources of Variation Mean Squares Degree of Estrus Multiple Pregnancy Fertility Freedom Litter size rate birth rate rate rate Treatment. * 0.. *.* 0. Experimental Error..... Coefficient of Variation (%) * represent significant differences at % level CIDR insertion +. mg GnRH, i.m. CIDR group 0 Sponge group 0 Spong insertion +. mg GnRH, i.m.. mg PGF α, i.m.. mg PGF α, i.m. Fig. : Treatments used for CIDR and Sponge groups CIDR removal +. mg PMSG i.m. sponge removal +. mg PMSG i.m. Naturally mating Naturally mating Day Day