1. 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
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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 ( ). 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 Sperm abnormalities and cigarette smoking. Lancet 1,
Goverde, H.J.M., Dekker, H.S.,Janssen, H.J.G., Bastiaans B.A., Rolland, R., Zielhuis, G.A 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 Cigarette smoking and sperm quality. Andrologia
Sepaniak, S., Forges, T., Gerard, H., Foliguet, B., Bene,M-C., Monnier-Barbarino, P 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,
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
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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