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Abstracts. II International Symposium on Animal Biology of Reproduction, Nov. 19-22, 2008, São Paulo, SP, Brazil. Ovarian crypts and germinative epithelium in Pinnipeds A.S.D. Machado 1 , P.C. Papa 1 1 Sector of Anatomy, Department of Surgery, Faculty of Veterinary Medicine and Animal Sciences, University of São Paulo, São Paulo, Brazil. Introduction Ovarian crypts were described for many pinnipeds species, besides the bitch others species such as, queen, cow and woman. Their functions are not fully elucidated. Researches suggest that mesenchymal cells in the tunica albuginea are bipotent progenitors for both primitive granulosa and germ cells (1). South–American-fur-seal (Arctocephalus australis) reproduction is largely influenced by environmental conditions. The species presents a single annual estrus in early summer, generating only one offspring and shows a rapid corpus luteum involution afterwards. This research intends to elucidate the cellular composition of the ovarian crypts and to study its role in Arctocephalus australis ovarian physiology. Material and methods Ovaries of 13 Arctocephalus australis (5 adult and 8 young), with death by natural causes were collected in the reproductive colony of Cabo Polônio –Uruguay during 2006 and 2007 (license CITES 000735). The samples were dissected and fixed by infusion of 4% formalin and Karnovsky modified solution. Subsequently, 0.5 cm 3 fragments were obtained for optical and electronic microscopy analysis. Slides were submitted to hematoxylin/eosin, Masson’s trichrome, aldehyde/fuchsin and picrosirius staining protocols for microscopical characterization. Immunohistochemistry for Ki-67 nuclear antigen was made in order to determine whether and which ovarian crypt cells are able to proliferate. Two superior canine teeth of each animal were collected for age determination. Results and discussion Morphologically ovarian crypts are tubular structure constituted by collagen (Fig.1–Bar 50μm) and elastic fibers (Fig.2–Bar 20μm) distributed in the ovarian parenchyma directed to the tunica albuginea (Fig.3–Bar 50μm). Internally they are covered by germinative epithelium, which express great amount of Ki-67 antigen in epithelial cells (Fig.4–Bar 10μm). Scanning Electronic Microscopic (SEM) showed different sizes of cells and presence of small intra-crypt lumen (Fig.5–Bar 3μm). No important differences were found between the ages studied. Ki-67 staining points towards an involvement of ovarian crypts in ovarian cell proliferation, probably contributing to neooogenesis (round shaped cells containing prominent nuclei). References (1) Bukovsky A, Caudle MR, Svetlikova M, Upadhyaya NB. 2004. Reprod Biol Endocrinol, 2:20. Acknowlegments: CNPq, CAPES, PROFAUMA and DINARA/UY E-mail: mamiferomarinho@gmail.com Fig.1 Fig.2 Fig.3 Fig.4 Fig.5 Anim. Reprod., v.6, n.1, p.215, Jan./Mar. 2009 215
Abstracts. II International Symposium on Animal Biology of Reproduction, Nov. 19-22, 2008, São Paulo, SP, Brazil. Ovarian activity induction in puerperium and in post puerperium anestrus Holstein dairy cows by administration of organic phosphorus and vitamin B12 complex 216 A.Grebogi, F.P.Souza, L.E. Kozicki, J.C.S. Breda, A.B.M. Giacomeli Pontifical University Catholic of Paraná, Curitiba, PR, Brazil. Introduction The reproductive efficiency displayed by Brazilian cows is low, with intervals between deliveries nearby 20 months (3). Puerperal and metabolic diseases might be responsible for such lower fertility by lengthening the intervals between delivery and conception (7) as a result of minerals diet deficiency (8). However, animal response in respect to phosphorus (P) administration has been variable, showing to be positive (6) or with no effect (9). The aim of the present study was to induce the return of anestrus cows in lactation towards an ovarian cyclical behavior. Materials and Methods A total of 50 Black and White Dutch cows, with a milk production of 11000 liters/305 days were used. The cows were maintained on corn silage, barley residue, "tifton" hay, ration, soybean bran, ground corn, protected fat, sodium bicarbonate, mineral salt and calcareous. Determination of animals in anestrus was based on the following endpoints: the anestrus condition since the delivery and the absence of a dominant ovarian follicle in any of the ovaries established by means of rectal palpation. The treatment protocol was: at the first day, intramuscular (IM) injection of 4 g of organic phosphate added with vitamin B12, 3 g at the third and sixth day after respectively. All animals were treated and separated to one of the three following groups: G1 - 22 animals in anestrus post partum (pp) from 30 to 60 days; G2 - 23 animals in anestrus pp from 61 to 90 days; G3 - 5 animals in anestrus pp from 91 to 150 days. The response of the treatment was considered as the manifestation of visible estrus up to 23 days after injection. Results and Discussion The results in regard to the percentual of the animal responses by manifesting the estrus (59.0; 65.2 and 80.0 % in the G1, G2 and G3 groups respectively) did not demonstrate significance between the groups in spite of the statement of Carvalho et al. (2003) in regard to P as being the mineral of the breeding. The animals belonging to group G3 were the ones that produced the best answer to the treatment (P>0.05) most probably because were the ones that reached the lactation peak and being in the process of stabilization milk production according account from Frandson et al.(2005). Also, no significance has been found by comparing the groups in regard to the days obtained for the estrus response as a consequence of the treatment employed in the experiment. However, the administration of inorganic P may account for the shortening of the time in which those animals stayed in anestrus (11.4 ± 7.4; 14.8 ± 6.5 and 13.0 ± 9.7 days for the G1, G2 and G3 group, respectively). Better reproductive performance of milk cows was achieved by Lamothe et al. (1976) by the presence of P high levels in the uterine fluids during the estrus. Doyle et al.(1990) found out expressive improvement in the endometrium of cows after administration of P improving the reproduction cyclical characteristic. In conclusion, the IM administration of organic P conjoined with vitamin B12 did not disclose significant effects in regard to the studied reproductive parameters, even though it has been observed a response of the experimental animals to the estrus within two weeks. References (1) Carvalho FAN, Barbosa FA, Mcdowell LR. 2003. Nutrição de bovinos a pasto. Belo Horizonte: Papel Form Ed.; (2) Doyle JC, Huston JE, Thompson PV. 1990. Theriogenology, 34:21-31; (3) Ferreira AM. 1991. Manejo reprodutivo e sua importância na eficiência da atividade leiteira. Coronel Pacheco: Embrapa-CNPGL, 47p. (Documentos, 46); (4) Frandson RD, Leewilkw W, Sails AD. 2005. Anatomia e fisiologia dos animais de fazenda. 6ª ed. Rio de Janeiro: Guanabara Koogan. pp.408-409; (5) Lamothe P, Busquet D, Guay P. 1976. Theriogenology, 6:353-365; (6) Mcdowell LR, BaverB, Galdo E, Koger M, Loosli JK, Conrad JH. 1982.. J. Anim. Sc.,55:964-970; (7) Stevenson JS, Call EP. 1988. J. Dairy Sc.,71:2572-2583; (8) Underwood EJ. 1996. The mineral nutrition of livestock. Aberdeen: FAO/CAB; (9) Valle ER, Souza JC, Nunes SG. 1982. In: Anais da XIX Reunião Anual da Sociedade Brasileira de Zootecnia, 1982. Piracicaba, SP. Piracicaba : ESALQ. pp. 217-218. E-mail: Kozicki.l@pucpr.br Anim. Reprod., v.6, n.1, p.216, Jan./Mar. 2009
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