Validation of a spectrophometric method used for the measurement of spermatozoa concentration in rabbit semen

Abstract

ABSTRACT:In rabbits, the number of inseminated spermatozoa plays an important role in the fertility rate obtained after artificial insemination. However, methods for the correct determination of spermatozoa number are scarce. The aim of this study was to develop and validate a spectrophotometric method for the measurement of spermatozoa concentration corrected by the seminal particle/spermatozoa ratio. For this purpose 360 ejaculates were collected from 36 rabbit bucks. The semen samples were divided into three groups based on their particle/spermatozoa ratio subjectively determined by microscopic viewing. The spermatozoa concentration of each semen sample was evaluated by spectrophotometer and these results were compared with those obtained by using a haemotocytometer. The results of the standard spectrophotometric method were correlated with the results of the haemotocytometer count of spermatozoa (r=0.76; P<0.01). When the optical density was corrected by considering the P/S ratio, the correlation values greatly improved (r=0.94; P<0.01). The simplicity, rapidity and low cost of this method make it suitable for the routine measurement of spermatozoa concentration of heterospermic pools in rabbit artificial insemination centres.

Effect of Age, and Vitamin E and Selenium Administration on Semen Characteristics of New Zealand White Rabbit Bucks

“…Percentage of live/dead spermatozoa was estimated by counting the number present in 100 sperm in different fields. Sperm Cell Concentration was assessed spectrophotometrically according to Castellini et al (2007).Sperm membrane integrity percent (SMI %) was assessed according to Hypo-Osmotic Swelling Testby observing the response of sperm cell to hypotonic medium (Jeyendran et al, 1984). The hypo-osmotic solution (150 m Osm/l) was prepared by dissolving 7.35g sodium citrate and 13.51g fructose in 100 ml of distilled water.…”

Andrology laboratory review: Evaluation of sperm concentration

“…The correlation coefficients of duplicate readings increased from 0.75 to 0.97 when calibration lines specifically constructed for semen containing the estimated ratio of particles were used. In addition, the correlation coefficients between the results obtained with the spectrophotometer and results obtained with the hemocytometer increased from 0.76 to 0.94 [65]. These observations indicate that accurate sperm concentration estimates can be obtained using spectrophotometers when good quality assurance process are in place to calibrate and periodically validated the equipment.…”

Sexual behaviour response, testicular development and semen quality of New Zealand white rabbit bucks as influenced by dietary chitosan

“…A haemocytometer was used to determine sperm cells concentration by counting the spermatozoa in five large squares. Then the total number was multiplied by 50 according to Castellini et al (2007).…”