The experiment was conducted at the research laboratory under the Department of Animal Nutrition, Genetics & Breeding, Sher-e-Bangla Agricultural Univeristy, Dhaka during July 2013 to July 2014.
a. Preparation of the laboratory: Before starting the experiment, all the necessary electrical power driven or digital equipment were properly installed and checked for good condition.
b. Preparation for Collection of ovaries: Physiological saline of 0.9% NaCl was prepared for washing the ovaries. The solution was warmed at 25oC to 30oC and kept in a thermos box to maintain the temperature during transporting the ovaries form slaughterhouse to the laboratory. Dulbecco's Phosphate Buffered Saline (D-PBS) solution was also prepared by adding one pack of PBS salt (Sigma Chemical Co., USA) in one liter of distilled water. Then it was sterilized in autoclave and stored in a refrigerator for further use.
c. Processing and Trimming of ovaries:Ovaries from goats on unknown reproductive history were collected from local slaughterhouse.
d. Evaluation of ovary: After collection and trimming ovaries were evaluated on the basis of the following parameters.
# Measurement of weight, length and width: Individually right, left, with-CL and without-CL group ovaries were weighed in g in a digital balance and the weight was recorded in tabular form. The length and width in cm of right, left, with-CL and without-CL group ovaries measured with the help of a digital slide calipers.
# COunting of follicle on the surface of the ovary: There are numerous follicles on the surface of both ovaries. The number of visible follicles on the surface of different category of ovaries were counted and recorded.
#COCs aspiration and grading: The ovaries were washed 2-3 times in saline solution at 30oC. They were then placed in a beaker and kept in a water bath at 30oC. One ovary was picked up in hand. The 10 ml syringe was loaded with PBS (1.0-1.5ml), and the needle (19G) was put in the ovary parenchyma near the vesicular follicles (2 to 6 mm diameter) and all 2 to 6 mm diameter follicles were aspirated near the point at the same time. After aspirating the follicles from one ovary, the aspirated follicular materials were transferred slowly into a 90-mm Petridis, avoiding damage to the cumulus cells and the COCs were searched and graded under microscope at low magnification. The COCs were then classified into 4 grades as described by Khandoker et al. (2001). Briefly, grade A: oocytes completely surrounded by cumulus cells; grade B: oocytes partially surrounded by cumulus cells; grade C: oocytes not surrounded by cumulus cells and grade D: degeneration observed both in oocytes and cumulus cells, where grade A and B will be considered as normal COCs and grade C and D as abnormal.
e. Statistical Analysis: The statistical analysis was done by SPSS (Statistical Package for Social Science, SPSS inc. 1999, Microsoft corporation 1998).