Papers

Effect of prenatal exposure to oestrogen on quality of semen: comparison of twins and singleton brothers

Lone Storgaard, managing director ^a, Jens Peter Bonde, professor ^a, Erik Ernst, associate professor ^b, Claus Yding Andersen, senior researcher ^d, Kirsten Ohm Kyvik, associate professor ^e, Jørn Olsen, professor ^c. 

^a Department of Occupational Medicine, Aarhus University Hospital, DK 8000 Aarhus, Denmark, ^b Laboratory for Reproduction, Institute of Anatomy, University of Aarhus, DK 8000 Aarhus, ^c Danish Epidemiology Science Centre, University of Aarhus, ^d Laboratory of Reproductive Biology, University Hospital of Copenhagen, DK 2100 Copenhagen, Denmark, ^e Danish Twin Registry, Institute of Community Health and Centre of Health and Social Policy, DK 5000 Odense, Denmark

Correspondence to: L Storgaard lstor{at}aaa.dk

The decline in sperm count and increase in disorders of the male reproductive tract may be due to high concentrations of prenatal exposure to oestrogens.^1-2 As the concentration of free oestrogens in plasma is much greater in twin pregnancies from the first weeks of gestation, and greater for dizygotic twins than monozygotic twins,^3-4 we studied sperm count in twins and singleton brothers to see if twin brothers have lower sperm counts and if the lowest values are for dizygotic twins.

	Participants, methods, and results

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From the population based Danish twin registry, we selected 250 monozygotic and 250 dizygotic pairs of twin brothers and from the Danish civil registration 500 pairs of singleton brothers. All the men were 20-45 years old and born in Denmark.

Of the 2000 men, 778 (38.9%) agreed to participate (40% of the singletons (396), 39% of the dizygotic twins (197), and 37% of the monozygotic twins(185)). Because of our estimates of suitable sample size, we stopped enrolment when we had collected semen samples from 105 singleton brothers, 104 monozygotic twins, and 107 dizygotic twins.

The men produced semen by masturbation and we analysed it in less than 1.5 hours in a mobile laboratory at the participants' home (n=113) or at a stationary laboratory (n=203). The sperm concentration was counted in an improved Neubauer haemacytometer (Marienfeld, Lauda-Königshofen); sperm morphology was classified according to 1999 World Health Organization criteria.

Sperm counts and sex hormone concentrations were positively skewed, and we transformed them to their cubic root to normalise their distributions. Morphology measurements were logit transformed. All potential confounders were included in the multiple linear regression.

The crude median sperm count was 19% higher among monozygotic twins and 9% lower among dizygotic twins than among singletons (table). The groups did not differ significantly with respect to any of the measures of semen quantity and quality. Inhibin B concentrations were significantly higher for singletons than for monozygotic twins.

We did not expect selection bias due to differential fertility to explain our findings: we found no difference between men who agreed to participate and those who did not in terms of the number of children they had (1.8 v 1.9).

	Comment

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Higher prenatal concentrations of oestrogen are not related to reduced sperm counts in adulthood. In particular, we did not find lower sperm counts in twin brothers: both the concentration and potency of oestrogens during pregnancy with twins are greater than for most environmental oestrogens.⁵

The low inhibin B concentration in monozygotic twins is surprising and could reflect slower multiplication of Sertoli's cells in fetal life. If so, high sperm counts indicate a compensatory mechanism which should be present in both monozygotic and dizygotic twins. Inhibin B is a new marker of testicular function, and deeper insight about its importance for male reproduction is needed.

	Acknowledgments

We thank Gorm Dancher, research technician Kirsten Lunding, and technicians at the laboratory at the Institute of Anatomy, Aarhus University.

Contributors: JO and JPB had the idea for the study. EE was the laboratory adviser. CYA analysed inhibin B. KOK withdrew twins from the Danish twin registry. LS designed the study. JO, JPB, and LS collected and analysed the data. All of the authors wrote the paper.

	Footnotes

Funding: Danish Research Council (Jnr 9802562), Danish Ministry of Health: Research Centre for Environmental Health Found (Jnr 383-4-1999), Danish Health Insurance Fund (Jnr 11/207-98 and 2000B514), and Danish Epidemiology Science Centre, University of Aarhus, Denmark.

Competing interests: None declared.

Figure A shows how the study groups were established and table A gives characteristics of the participants.

	References

Top Participants, methods, and... Comment References

1.	Sharpe RM, Skakkebaek NE. Are oestrogens involved in falling sperm counts and disorders of the male reproductive tract? Lancet 1993; 341: 1392-1395[CrossRef][ISI][Medline].
2.	Gill WB, Schumacher GF, Bibbo M, Straus FH, Schoenberg HW. Association of diethylstilbestrol exposure in utero with cryptorchidism, testicular hypoplasia and semen abnormalities. J Urol 1979; 122: 36-39[ISI][Medline].
3.	Johnson MR, Bolton VN, Riddle AF, Sharma V, Nicolaides K, Grudzinskas JG, et al. Interactions between the embryo and corpus luteum. Hum Reprod 1993; 8: 1496-1501[Abstract/Free Full Text].
4.	Kappel B, Hansen K, Moller J, Faaborg-Andersen J. Human placental lactogen and dU-estrogen levels in normal twin pregnancies. Acta Genet Med Gemellol (Roma) 1985; 34: 59-65[Medline].
5.	Daston GP, Gooch JW, Breslin WJ, Shuey DL, Nikiforov AI, Fico TA, et al. Environmental estrogens and reproductive health: a discussion of the human and environmental data. Reprod Toxicol 1997; 11: 465-481[CrossRef][ISI][Medline].

(Accepted 22 January 2002)