ex funebri charta

Bukowska D., Kempisty B., Antosik P., Jackowska M.,
Woźna M., Margarita Lianeri M., Jaśkowski J. M.

Association between the number and quality of bitch COC’s and selected donor factors

Since the first artificial insemination in the dog was applied, assisted reproductive technologies (ART) have undergone several modifications (3, 11, 12). The implementation of in vitro techniques led to the development of in vitro maturation (IVM) and in vitro embryo production (IVP) procedures (14, 17, 25). Since that time, dogs have been used for physiological and genetic studies of human diseases. One of the most important concerns in studies involving IVM and IVF of bitch oocytes is the quality of gametes (1, 4, 18). The classification of dog oocytes is based on the analysis of the cytoplasmic appearance and the presence or absence of cumulus cells. The number and quality of canine cumulus-oocyte complexes (COC) that are suitable for in vitro maturation and consequent embryo culture depend on several factors. Moreover, the method of COC collection is considered to be one of the most important factors. An average number of COCs collected from a bitch may range from 10 up to several dozen or even over 100 (3, 11, 22). On the other hand, regardless of the oocyte collection method, the number and morphology of collected COCs are influenced by some bitch-related factors such as: season, breed, age, size and weight of ovaries, presence or lack of functional structures on the ovarian surface, stage of oestrus cycle, and physiological or pathological condition of the uterus (2, 6, 16, 22). Previously published data on the importance of the above-mentioned factors for the oocyte number and quality are quite often contradictory. Pyometra is a pathological condition of the uterus, occurring predominantly in bitches under the influence of progesterone at metoestrus. However, the plasma progesterone concentration in bitches with pyometra is not different from those of pregnant or normally cycling females (15, 21, 23). The most frequently preferred treatment of this disease is ovariohysterectomy (24). Therefore, the collection and morphological assessment of COCs and IVF are applied to salvage the genetic potential of these females. The development of ART in dogs may be important in the future to salvage gametes from genetically and evolutionarily precious bitches of other canid species. The aim of the present study was to investigate whether selected factors related to oocyte donors influence the number and quality of collected COCs.
Material and methods.
Animals. Ovaries were collected from 84 sexually mature pure bred bitches and mongrels aged from 18 months to 13 years. The animals originated from a local dog shelter or were directed to our veterinary clinic (Department of Agricultural Veterinary, University of Agriculture, Poznan, Poland) due to various reasons. Ovaries, oviducts and uteri were surgically removed under general anesthesia induced with ketamine hydrochloride (KetalarTM, Sankyo Co., Ltd., Tokyo, Japan) administered i.m., following pre-anesthetic medication with atropine sulphate (Fuso Pharmaceutical Industries, Ltd., Osaka, Japan), and maintained by inhalation of halothane, nitrous oxide and oxygen. The surgery was performed at the Department of Agricultural Veterinary, University of Agriculture, Poland. Bitches were diagnosed with pyometra on the basis of clinical signs and a clinical examination, i.e. hemogram, cytology of vaginal smears, radiography, and ultrasonography. All bitches with pyometra showed polydipsia, complete lack of appetite or its reduction (anorexia), apathy, peripheral leukocytosis, enlarged abdominal integuments, and increased pulse and respiration rate; additionally, 75% of them developed purulent discharge from the reproductive organs. In all investigated bitches, abdominal ultrasound examination with a 5/7.5 MHz microprobe ...
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