• Ahmed Alaa Safwat Teaching Veterinary Hospital, Zagazig University, Faculty of veterinary medicine, Zagazig, Egypt, Corresponding author, E-mail:
  • Abd El-salam Eidaroos Department of Theriogenology, Faculty of veterinary medicine, Zagazig University, Zagazig, Egypt
  • Hany Abdalla Department of Theriogenology, Faculty of veterinary medicine, Zagazig University, Zagazig, Egypt
  • Asmaa A. Abdallah Department of Theriogenology, Faculty of veterinary medicine, Zagazig University, Zagazig, Egypt



bitch, estrus-induction, FSH-LH, vaginal cytology, progesterone, estrogen


The current study was designed to evaluate the changes in vaginal cytology and serum progesterone and estrogen concentration in anestrum bitches treated with a combination of FSH and LH. Seven healthy Mongrel anestrum bitches were treated with daily administration of 75 I.U. FSH plus 75 I.U LH for consecutive nine days and another three bitches were kept as a control without any treatment. Treated and control bitches were examined for the percentages of different vaginal-cytology cells, serum concentration of estrogen and progesterone, and signs of proestrus and estrus in a scheduled timeline. Five of the seven treated bitches (71.4%) showed a gradual decrease in the percentage of parabasal cells, a gradual increase in the percentage of cornified and RBCs, a rapid decrease in the estrogen concentration, and an increase in the progesterone concentration. These five bitches showed signs of proestrus and four of them showed male acceptance and were conceived. These five bitches had high estrogen concentrations before initiation of the treatment. The other two bitches had a low estrogen concentration before the initiation of the treatment and after treatment, they showed a gradual increase in the estrogen concentration, but they did not show characteristic changes in vaginal cytology or progesterone concentration. Control bitches had high estrogen concentrations before treatment, but they did not show characteristic changes in vaginal cytology, estrogen and progesterone concentrations, or signs of proestrus up to one month after the end of the treatment. In conclusion, treating anestrum bitches with a mixture of FSH and LH induced characteristic changes in vaginal cytology and serum estrogen and progesterone concentrations in the majority of bitches. These changes were associated with the induction of fertile estrus in more than half of the treated bitches, and this percentage increased to about 80% in bitches having initial high estrogen concentration.

Key words: bitch; estrus-induction; FSH-LH, vaginal cytology, progesterone; estrogen


● 1. Concannon PW. Reproductive cycles of the domestic bitch. Anim Reprod Sci 2011; 124: 200–10.

● 2. Concannon PW, Castracane VD, Temple M, Mon-tanez A. Endocrine control of ovarian function in dogs and other carnivores. Anim Reprod 2018; 6: 172–93.

● 3. Okkens AC, Bevers MM, Dieleman SJ, Van Haaften B, Van Sluijs FJ. Fertility problems in the bitch. Anim Re-prod Sci 1992; 28: 379–87.

● 4. Concannon PW. Endocrinologic control of normal canine ovarian function. Reprod Domest Anim 2009; 44: 3–15.

● 5. Hollinshead FK, Hanlon DW. Normal progesterone profiles during estrus in the bitch: A prospective analysis of 1420 estrous cycles. Theriogenology, 2019; 125: 37–42.

● 6. England GC, Russo M, Freeman SL. Follicular dy-namics, ovulation and conception rates in bitches. Reprod Domest Anim 2009; 44: 53–8.

● 7. McBride MW, Aughey E, O'Shaughnessy PJ, Jeffcoate IA. Ovarian function and FSH receptor characteristics dur-ing canine anoestrus. J Reprod Fertil 2001; (Suppl, 57): 3–10.

● 8. Kutzler MA. Estrus induction and synchronization in canids and felids. Theriogenology 2007; 68: 354–74.

● 9. Spattini G, Borghi V, Thuroczy J, Balogh L, Scar-amuzzi RJ, De Rensis F. Follicular development and plasma con-centrations of LH and prolactin in anestrous female dogs treated with the dopamine agonist cabergoline. Theri-ogenology 2007; 68: 826–33.

● 10. Ohtaki T, Fujiwara H, Watanabe G, Ono M, Taya K, Tsumagari S. Changes in luteinizing hormone pulse fre-quency and prolactin levels in bitches in response to estrus induction by cabergoline-its cases where it is delayed to induce estrus. J Vet Med Sci 2020; 82: 1773–80.

● 11. Zöldág L, Fekete S, Csaky I, Bersenyi A. Fertile es-trus induced in bitches by bromocryptine, a dopamine agonist: a clinical trial. Theriogenology 2001; 55: 1657–66.

● 12. De Rensis F, Spattini G, Ballabio R, Scaramuzzi RJ. The effect of administering a dopamine agonist (Cabergo-line) on follicular and luteal development during pro-estrus and estrus in the female greyhound. Theriogenology 2006; 66: 887–95.

● 13. Cirit Ü, Bacinoglu S, Cangul IT, Kaya HH, Taş M, Ak K. The effects of a low dose of cabergoline on induc-tion of estrus and pregnancy rates in anestrous bitches. Anim Reprod Sci 2007; 101: 134–44.

● 14. Verstegen JP, Onclin K, Silva LD, Concannon PW. Effect of stage of anestrus on the induction of estrus by the dopamine agonist cabergoline in dogs. Theriogenology 1999; 51: 597–611.

● 15. Walter B, Otzdorff C, Brugger N, Braun J. Estrus induction in Beagle bitches with the GnRH-agonist implant containing 4.7 mg Deslorelin. Theriogenology 2011; 75: 1125–9.

● 16. Fontaine E, Mir F, Vannier F, Gérardin A, Albouy M, Navarro C, Fontbonne A. Induction of fertile oestrus in the bitch using Deslorelin, a GnRH agonist. Theriogenolo-gy 2011; 76: 1561–6.

● 17. Rezende RS, Eurides D, Barbosa CP, Lacerda MS, Sampaio RL, Gomes AL. Use of a GnRH synthetic analog (buserelin) for estrous induction in female dogs. Arquivo Brasileiro de Medicina Veterinária e Zootecnia 2018; 70: 656–60.

● 18. Wanke MM, Farina J, Loza MH, Rebuelto M, Concannon PW. Induction of estrus in bitches with normal and persistent anestrus using human menopausal gonado-tropin (hMG). Theriogenology 1997; 47: 935–42.

● 19. Arnold S, Arnold P, Concannon PW, Weilenmann R, Hubler M, Casal M, Fairburn A, Eggenberger E, Rüsch P. Effect of duration of PMSG treatment on induction of oestrus, pregnancy rates and the complications of hyper-oestrogenism in dogs. J Reprod Fertil 1989; (Suppl, 39): 115–22.

● 20. Stornelli MC, Mitacek MG, Giménez F, Bonaura MC, Dorna IV, de la Sota RL, Stornelli MA. Pharmacoki-netics of eCG and induction of fertile estrus in bitches using eCG followed by hCG. Theriogenology 2012; 78: 1056–64.

● 21. England GC, Allen WE. Repeatability of events dur-ing spontaneous and gonadotrophin-induced oestrus in bitches. Reproduction 1991; 93: 443–8.

● 22. Al-Hamedawi TM. Induction of fertile estrus in bitches using equine chorionic gonadotropine (ecg) and human chorionic gonadotropine (hcg). The Iraqi J Vet Med 2013; 37: 102–5.

● 23. Johnston SD, Root Kustritz MV, Olson PS. Canine and feline theriogenology. Saunders: 2001.

● 24. Manda S, Rani NL, Suresh K, Makkena S. Vaginal exfoliative cytology as a tool in diagnosing reproductive disor-ders in bitches. Intas Polivet 2004; 5: 354–6.

● 25. Team RC. R: A language and environment for statis-tical computing. R Foundation for Statistical Computing, Vi-enna, Austria. http://www. R-project. org/. 2013.

● 26. Jeffcoate IA. Endocrinology of anoestrous bitches. J Reprod Fertil 1993; (Suppl, 47): 69–76.

● 27. Tsuchida M, Sakurai D, Komura N, Nakagata N, Su-zuki H. Induction of oestrus by administering Inhibin antise-rum along with equine chorionic gonadotropin in anoestrous bitches. Reprod Domest Anim 2021; 56: 1398–405.

● 28. Verstegen J, Onclin K, Silva L, Concannon P. Ter-mination of obligate anoestrus and induction of fertile ovarian cycles in dogs by administration of purified pig LH. Reproduction 1997; 111: 35–40.

● 29. Takao Y, Honda T, Ueda M, Hattori N, Yamada S, Maeda M, Fujiwara H, Mori T, Wimalasena J. Immuno-histochemical localization of the LH/HCG receptor in human ovary: HCG enhances cell surface expression of LH/HCG receptor on luteinizing granulosa cells in vitro. Mol Hum Reprod 1997; 3: 569–78.

● 30. Saint-Dizier M, Jaffré N, Reynaud K, Remy B, Thoumire S, Chastant-Maillard S. Expression of follicle-stimulating hormone and luteinising hormone binding sites in the bitch ovary during the follicular phase. Reprod Fertil Dev 2008; 20: 925–34.

● 31. Hirata M, Tanihara F, Taniguchi M, Takagi M, Tera-zono T, Otoi T. Follicular development of canine ovaries stimulated by a combination treatment of eCG and hCG. Vet Med Sci 2018; 4: 333–40.

● 32. Jurczak A, Domosławska A, Bukowska B, Janowski T. Equine chorionic gonadotropin and human chorionic gonadotropin stimulation increase the number of luteinized follicles and the progesterone level compared with cabergo-line stimulation in anoestrus bitches. Reprod Domest Anim 2016; 51: 562–8.

● 33. Stornelli MC, García Mitacek MC, Praderio RG, Nunez Favre R, de la Sota RL, Stornelli MA. Prolactin, andros-tenedione and igf 1 serum concentrations during induced follicular growth by ecg administration in the bitch. Reprod Domest Anim 2016; 5 1: 130–4.

● 34. Nak D, Nak Y, Şimşek G. Comparison of the use of cabergoline and gonadotrophin to treat primary and sec-ondary anoestrus in bitches. Aust Vet J 2012; 90: 194–6.

● 35. Concannon PW, Temple M, Montanez A, Newton L. Effects of dose and duration of continuous GnRH-agonist treatment on induction of estrus in beagle dogs: competing and concurrent up-regulation and down-regulation of LH re-lease. Theriogenology 2006; 66: 1488–96.

● 36. Kaya D, Aslan S, Kaya S, Kuru M, Kacar C, Schäfer-Somi S. Clinical and endocrine short-term effects of gnrh ana-logue deslorelin in prepubertal bitches: does a “flare-up” occur? Kafkas Univ Vet Fak Derg 2013; 19:299–304

● 37. Kutzler MA, Wheeler R, Lamb S, Volkmann DH. Deslorelin implant administration beneath the vulvar muco-sa for the induction of synchronous estrus in bitches. In: Proceedings of Third EVSSAR European Congress 2002: 96.

● 38. Kusuma PS, Tainturier D. Comparison of induction of oestrus in dogs using metergoline, metergoline plus human chorionic gonadotrophin, or pregnant mares' serum gonadotrophin. J Reprod fertil 1993; (Suppl, 47): 363–70.

● 39. Nakao T, Aoto Y, Fukushima S, Moriyoshi M, Ka-wata K. Induction of estrus in bitches with exogenous gonado-tropins, and pregnancy rate and blood progesterone profiles. Nihon Juigaku Zasshi. The Japanese Journal of Veterinary Science 1985; 47: 17–24.




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Veterinary Medicine and The One Health Concept