A study on the differences in general parameters and DNA integrity of spermatozoa of smokers and non-smokers Diganta Dey1, Abhijit Banerjee1, Sugat Sanyal1, Parbati Banerjee1, Kankan Biswas1, Dhrubajyoti Biswas1, S.M. Bhattacharya2, Satadal Das1 and Ratnamala Ray1 1Dept. of Pathology, Ashok Laboratory – Clinical Testing Centre, Kolkata; 2Ramkrishna Mission Seva Pratisthan, Kolkata This poster template is from http://www.swarthmore.edu/NatSci/cpurrin1/posteradvice.htm. It is free, free, free for non-commercial use. But if you really like it, I’m always thrilled to get postcards from wherever you happen to be presenting your poster. Have fun. Sincerely, Colin Purrington, Department of Biology, Swarthmore College, Swarthmore, PA 19081, USA. Email: cpurrin1@swarthmore.edu Conclusions The outcome of this study suggested that cigarette smoking may exert a detrimental effect on nuclear integrity of individual spermatozoa and overall sperm count when consumed extensively (>20 cigarette per day). Thus this should be taken into account during the treatment of infertile Indian patients. Introduction A retrospective analysis was conducted to evaluate the effects of cigarette smoking on some standard semen parameters such as sperm count, motility, hypo-osmotic swelling and morphology. Additionally the consequence of cigarette smoking on DNA integrity of spermatozoa was analyzed. Results Table: 1 Comparison between conventional semen parameter and DNA integrity among Non-smoker and Smoker groups. No significance difference was obtained. Materials and methods 181 study subjects of mean age 32.8±2.3 years were selected from the men who visited the Ashok Laboratory for sperm function test in last two years (2008-2009). All the subjects were instructed to collect semen samples by masturbation after three-day period abstinence. Detailed medical and smoking history was taken with the help of a questionnaire. On this basis 92 subjects who had never smoked was considered as Non-Smoker, 66 subjects who had smoked cigarettes from more than one year prior to the test and still consuming cigarettes at average rate of 8.0±2.95 per day, was regarded as Smoker and 23 persons who had smoked >20 cigarette per day was considered as Heavy Smoker. Fourteen, ten and three subjects, who had azoospermia, from Non-Smoker, Smoker and Heavy Smoker group respectively, was excluded from this study. The standard semen parameters were measured microscopically using differential staining. The DNA integrity of spermatozoa was analyzed and scored using the comet assay along with associated scoring software. The standard semen parameters and sperm DNA integrity of smoker (n=56) and Heavy Smoker (n=20) group was compared against Non-Smoker (n=78) group by Student t-test and p<0.05 was considered statistically significant. Figure:6 Variation of sperm normal morphology (%) among different Smoker groups Figure:7 Papanicolaou staining for the determination of normal sperm morphology Table: 2 Comparison between conventional semen parameter and DNA integrity among Non-smoker and Heavy-smoker groups. Sperm count and DNA integrity were significantly reduced in Heavy smoker in compare to Non-smoker group. Figure:8 Variation of sperm Head DNA Integrity (%) among different smoker groups Literature cited Evans, H.J., Fletcher, J., Torrance, M., Hargreave, T.B. 1981. Sperm abnormalities and cigarette smoking. Lancet 1, 627 - 629 Goverde, H.J.M., Dekker, H.S.,Janssen, H.J.G., Bastiaans B.A., Rolland, R., Zielhuis, G.A.1995. Semen quality and frequency of smoking and alcohol consumption – An explorative study, Int J Fertil 40(3), 135 – 138. Holzeki, G., Gall, H., Hermann, J. 1991. Cigarette smoking and sperm quality. Andrologia 23. 141 Sepaniak, S., Forges, T., Gerard, H., Foliguet, B., Bene,M-C., Monnier-Barbarino, P. 2006. The influence of cigarette smoking on human sperm quality and DNA fragmentation, Toxicology 213, 54 – 60 Vine, M.F. 1996, Smoking and male reproduction: a review. Int. J. Androl. 19, 323 - 337 Figure:9 Computer attached Fluorescence Microscope and the image analysis software Figure:2 Variation of sperm count (million/ml) among different Smoker groups Figure:3 Variation of sperm motility (%) among different smoker groups Figure:10 SYBR Green stained spermatozoa after electrophoresis Acknowledgments We are grateful to the entire unit of Ashok Laboratory; with special thanks to Dr. Ratnamala Ray, Dr. Abhijit Banerjee, Dr. Parbati Banerjee & Dr. S.M. Bhattacharya. For further information Please contact diganta_dey2006@yahoo.com More information on related projects can be obtained at www.cometassayindia.org Figure 1. Frequency of Non-smoker, smoker and Heavy smoker among the study subjects Figure:11 Data analysis by TriTek comet software Figure:4 Variation of sperm rapid progressive motility (%) among different Smoker groups Figure:5 Variation of Hypo-osmotic swelling (%) among different Smoker groups