Comparative Investigation of Several Sperm Analysis Methods for Evaluation of Spermatotoxicity of Industrial Chemical: 2-Bromopropane as an Example.

Abstract

Reproductive toxicity of 2-bromopropane (2BP), a substitute for ozone layer-depleting chloro-fluorocarbon, was found among the workers in an electronics factory in Korea in 1995. Furthermore the importance of testicular toxicity has been realized since the problem of endocrine disruptors arose all over the world, but manual methods must rely on subjective assessment. Recently, computer-assisted sperm analysis (CASA) was proposed but this system requires vast investment. We then investigated the applicability of the MTT method with a microplate and sperm quality analyzer (SQA) as simple, rapid, and economic instrumental methods for the examination of sperm quality in rats, comparing it with the manual microscopic method and CASA. Epididymal fluid derived from male F344/N Slc (Fischer) rats intraperitoneally injected with 2BP in the dose range of 125-1,000 mg/kg/d twice a week (total 8 times) were examined by these methods as a model experiment. Sperm count measured by the manual method and CASA in the epididymal fluid, absorbance by the MTT method and sperm motility index value by the SQA method were significantly lower in the 2BP 1,000 mg/kg administered group than in the control group. This result suggests that the MTT method can detect oligospermia. With the microplate and microplate reader, the efficiency of detection becomes much better. Sperm analyses by the MTT method with the microplate reader and the SQA method are available for reproductive toxicity study in rats.

Influence of injection timing on severity of cadmium-induced testicular toxicity in mice

“…Fluorescence of sperm heads under ultra-violet beam was monitored and counted in RAT-IDENT mode automatically. Data were expressed as total number of sperm heads per tissue sample (Ohtani et al, 2004).…”

High sensitivity of testicular function to titanium nanoparticles

“…Fluorescence of sperm heads under an ultra-violet beam was monitored and counted in RAT-IDENT mode automatically. Data were expressed as total number of sperm heads per tissue sample (Ohtani et al, 2004).…”

The Benefits and Limitations of the Sperm Quality Analyzer IIB in Assessing Porcine Sperm Motility

“…The SQA-IIB and similar devices measuring only the scatter of light rather than tracking individual sperm, are thus best used to monitor the demise of total sperm motility. This has already been applied to detect spermatotoxicity of industrial chemicals when administered to laboratory rats [Ohtani et al 2004]. The use of human sperm survival studies to detect toxicity within in vitro fertilization (IVF) laboratories is well recognized [Critchlow et al 1989], and the SQA-IIB has also been shown to provide an automated option in measuring sperm motility as an end-point in this bioassay [Iemmolo et al 2005].…”