Biol Trace Elem Res (2011) 144:454462 DOI 10.

1007/s12011-011-9138-2

Effect of Zinc and Selenium Supplementation on Serum Testosterone and Plasma Lactate in Cyclist After an Exhaustive Exercise Bout
Leila Shafiei Neek & Abas Ali Gaeini & Siroos Choobineh

Received: 19 June 2011 / Accepted: 29 June 2011 / Published online: 9 July 2011 # Springer Science+Business Media, LLC 2011

Abstract Zinc and selenium are essential minerals and have roles for more than 300 metabolic reactions in the body. The purpose of this study was to investigate how exhaustive exercise affects testosterone levels and plasma lactate in cyclists who were supplemented with oral zinc and selenium for 4 weeks. For this reason, 32 male road cyclists were selected equally to four groups: PL group, placebo; Zn group, zinc supplement (30 mg/day); Se group, selenium supplement (200 g/day); and ZnSe group, zincselenium supplement. After treatment, free, total testosterone, and lactate levels of subjects were determined before and after exhaustive exercise. Resting total, free testosterone, and lactate levels did not differ significantly between groups, and were increased by exercise (P>0.05). Serum total testosterone levels in Zn group were higher than in Se group after exercise (P<0.05). Serum-free testosterone levels in the Zn group were higher than the other groups (P<0.05).There was an insignificant difference between levels of lactate in the four groups after exercise (P>0.05). The results showed that 4-week simultaneous and separately zinc and selenium supplementation had no significant effect on resting testosterone and lactate levels of subjects who consume a zinc and selenium sufficient diet. It might be possible that the effect of zinc supplementation on free testosterone depends on exercise. Keywords Exhaustion exercise . Lactate . Road cyclists . Testosterone . Selenium supplementation . Zinc supplementation

Introduction Many researchers concentrate on the relation between exercise and antioxidant supplementation; therefore, it can be stated that there is an increasing interest about evaluation of the ergogenic effects of minerals such as zinc and selenium on exercise.
L. Shafiei Neek (*) : A. A. Gaeini : S. Choobineh Faculty of Physical Education and Sport Sciences, University of Tehran, Tehran, Iran e-mail: shafieineek@gmail.com

Effect of Zinc and Selenium Supplementation on Serum Testosterone and Plasma Lactate

455

Zinc is the fourth most abundant intercellular metal, and a biologically essential trace metal is found in over 200 enzymes and proteins [1]. Since zinc is necessary for the activity of a number of enzymes in the energy metabolism, low muscle zinc levels may lead to a reduction in endurance capacity [2]. Zinc plays a key role in reproductive physiology [3, 4]. There was a positive relation between zinc and testosterone [5]. Lactic dehydrogenase is a Zn-containing enzyme. Adequate muscle zinc concentration may facilitate the reduction of lactic acid to pyruvate through the action of lactic dehydrogenase in exercising skeletal muscle and, therefore, may decrease muscle fatigue [6]. Kaya et al. [7] reported that zinc supplementation leads to a significant increase in testosterone levels and a significant decrease in lactate levels in response to exhaustive exercise. Also, selenium, which is commonly found in nature, is an essential trace element required for the normal development of human and animal organisms [8, 9]. The increase in oxidative stress caused by exercise and the recognition of the stimulation of antioxidant activity by selenium inevitably entails a relation between selenium and exercise [10]. Selenium is also needed for normal testosterone metabolism and testicular morphology, which may explain the presence of several other selenoproteins in the male gonads [11]. Akil et al. [10] reported that the increase in free radical production and lactate levels due to acute swimming exercise in rats might be offset by selenium supplementation. A relationship between exercise and testosterone, which has important effects on energy metabolism, seems inevitable. From a large number of studies exploring the relationship between exercise and testosterone, no definite conclusion can be drawn [1214]. Bosco et al. [15] reported that short-term strenuous exercise increased total and free testosterone levels by 12% and 13%, respectively. But Kilic et al. [16] reported that exhaustive exercise decreased thyroid hormone and testosterone concentrations in elite athletes. Studies on the relation between selenium and exercise and zinc and exercise mainly focus on the antioxidant role of selenium and the distribution of zinc in the body on response to exercise. There is limited information about the effect of zinc and selenium supplementation especially around their relationship with testosterone hormone and exercise. Accordingly, this topic needs to be studied. The purpose of this study was to examine how exhaustion exercise affects testosterone and lactate levels in athletes who consume a zinc and selenium sufficient diet, and without any lack of selenium and zinc, who are supplemented oral zinc and selenium for 4 weeks.

Materials and Methods Subjects The study was performed at the National Olympic & Paralympics Academy of Islamic Republic of Iran (NOPA.I.R.Iran). Thirty-two male road cyclists volunteered to participate in this experiment. The characteristics of the participants are given as meanSD in Table 1. First of all, the subjects provided written informed consent. The cyclists had 34 years of exercise experiences, and they were members of the Tehran Traffic team. The study groups were exercised for 120180 min, 5 days a week. One week to study, the subjects reported to the NOPA.I.R.Iran after an overnight fast. In this session, the subjects' age, height, mass, body composition (via in body 220), and maximal aerobic power (Moark ergometer 894Ea) were determined. Also, blood samples were taken for assessment of zinc and selenium status.

456 Table 1 Anthropometric data and baseline zinc and selenium intake of the cyclist Group Height (cm) Weight (kg) BMI (kg/m2) Aerobic power (Watt) 291.544.56 299.8830.17 304.0631.99 296.1241.54

Neek et al.

Baseline dietary Baseline dietary zinc (mg d1) selenium (mg d1) 15.670.89 16.401.02 14.981.08 16.230.45 64.0515.02 59.9813.29 56.8714.67 62.5612.34

PL Zn Se

176.878.02 176.878.2 177.754.23

66.156.4 68.118.66 61.664.75 63.178.45

21.120.98 21.752.5 20.51.26 21.572.1

ZnSe 174.814.3 Values are meansSD

Supplementation Protocol None of the subjects had ingested zinc and selenium or any other dietary supplements, for a minimum of 2 weeks before the initiation and during the study. The subjects were assigned base on body composition, and they were equally divided into four groups by using a double-blind design. (a) Placebo (PL, 30 mg of dextrose; n=8), (b) zinc (Zn, 30 mg zinc sulfate), (c) selenium (Se, 200 g sodium selenite), (d) zincselenium (ZnSe, 30 mg zinc sulfate200 g selenium selenite). According to their group assignment, all subjects ingested one capsule of zinc, selenium, zincselenium, and placebo each day for 4 weeks. Diets' Zinc and Selenium Assay In the course of trial, the daily intake of zinc and selenium was determined on the base of diet history interviews (24-h recall and food frequency questionnaire), which were conducted with Dorosty Food Processor (version 2.1). The interviews were performed by an experimental nutritionist, and were designed to reflect the habitual dietary intake of the subjects in the month before and during the trial period. The daily zinc and selenium intake of all subjects (16/341/23 mg d1 and 64/4519/03 mg d1, respectively) were higher than the recommended daily allowance of 11 mg d1 and 55 g d1 [17], respectively for zinc and selenium intake, so all subjects were considered to be not zinc and selenium deficient; therefore, the usage of supplements was more than their daily needs. Exercise Tests One week before the actual experiment started, subjects had to perform a graded exercise test in order to determine their maximal power output (Wmax) [18]. For 3 days before the experiments, the subjects were obliged to abstain from strenuous exercise. No caffeine, only tea was permitted during the 48 h before the experiment. On the experimental days, subjects reported to the NOPA.I.R.Iran after an overnight fast. Exercise test started with a 10-min warm-up period at a workload of 50% Wmax. Thereafter, the subjects were instructed to cycle 2-min block periods at alternating workloads of 90% and 50% of Wmax, respectively. This was continued until the subjects were no longer able to complete the 2 min at 90% Wmax. That moment was defined as the time at which the subject was unable to maintain cycling speed at 60 revolutions per minute. At that moment, the high-intensity block was reduced to 80% Wmax. Again, the subjects had to cycle until they were unable to complete a 2-min block at 80% Wmax, after which the high-intensity block was reduced to 70%

Effect of Zinc and Selenium Supplementation on Serum Testosterone and Plasma Lactate

457

Wmax. The subjects were allowed to stop when pedaling speed could not be maintained at 70% Wmax. Water was provided ad libitum during the exercise protocol [19]. To remain blood samples were taken from all subjects pre and post exercise. Laboratory Procedures Blood samples collected from the subjects were centrifuged for 10 min at 3,000 rpm and were kept frozen at 20C until analysis. Free and total testosterone analyses were conducted on the serum samples. Lactate, zinc, and selenium were assessed on the plasma samples. Zinc and Selenium Analysis Plasma zinc and selenium concentrations were assessed on a Younglin AAS 8020 atomic absorption spectrometer (AAS). Values were expressed as micrograms per deciliter. Plasma Lactate Lactate analyses were carried out in the separated plasma samples according to colorimetric method (using Roche Diagnostic lactate kits) in a Cobas Integra 400 autoanalyzer. Plasma lactate levels (read at 552 nm wavelength) are expressed as milligrams per deciliter. Free Testosterone Measurements Serum-free testosterone analyses were done in Elisa test kit (LDN Company) by enzymelinked immunosorbent assay method; the results are expressed as picogram per milliliter. Total Testosterone Measurements Serum total testosterone was measured with chemiluminescence immunoassay on a LIAISON (Diasorin) analyzer. The results are expressed as nanograms per milliliter.

Statistical Analysis Statistical analysis was performed with SPSS version 16 program (SPSS Inc., Chicago, IL, USA). Statistical evaluation was done by KolmogrovSmirnov test at first to examine the normal distribution and Leven's test for homogeneity. One-way ANOVA was performed in order to compare between groups in anthropometric data and aerobic power. Two-way ANOVA was also performed to assess differences between intra and intergroups followed by Bonferroni's test for multiple comparisons. All results were shown as meansSD in all statistical comparisons P<0/05 was used as the criterion for statistical significance.

Results The results of statistical analysis showed that there was an insignificant difference about physical characteristic between groups (p>0.05). Intra and intergroup comparison, respectively indicated that resting levels of plasma lactate and free and total testosterone

458

Neek et al.

did not differ significantly differ between groups, and were increased significantly by exercise. There was a significant difference between effects of exhaustion exercise on serum total testosterone in Zn group higher than Se group (p<0.05) (Fig. 1). But there was no significant difference between Zn group with PL and ZnSe groups (p<0.05). It was resulted that in this case, there was no significant difference between PL, Se, and ZnSe groups (p<0.05). There was also a significant difference between effects of exhaustion exercise on serum-free testosterone in Zn group in contrast with other groups (p<0.05) (Fig. 2). In this case, there was no significant difference between PL, Se, and ZnSe groups (p<0.05). There was an insignificant difference between effects of exhaustion exercise on plasma lactate in the four groups (p>0.05) (Fig. 3).

Discussion Considering that the results of this study about two different sides of the nutritionexercise effects (Zn, Se, and ZnSe groups) and exercise effects (PL group) are comparable with the related obtained results, therefore, the results of this research will be discussed in these two fields separately. In results of our study, resting free and total testosterone levels measured were significantly lower than those measured after an exhaustive exercise bout. This result reveals that there is a positive correlation between testosterone and exhaustive exercise. In fact, results of studies examining the relation between exercise and testosterone illustrates that there is no agreement on this topic. In other words, there are contradictions about the cause and effect of physical exercise and the amount of testosterone in different researches [7, 16, 2022]. Besides the studies reporting that free and total testosterone levels did not change with exercise [2325], there are also those noting that free and total testosterone levels significantly decreased with exercise [26 28]. The increase we obtained in free and total testosterone levels immediately after exhaustion is in contrast with the findings of these researchers. Ransen et al. [29] showed that there was an important increase in levels of testosterone, epinephrine, norepinephine, ACTH, cortisol, and growth hormone of the subjects following hightintensity endurance exercise on a cycle ergometer. The results of this study are consistent with our findings.

Total Testosterone (ng/dl)

Fig. 1 Variations of total testosterone in research groups. (MeanSD). Asterisks indicate p<0.05 pre-test vs. post-test; Euro sign indicates p<0.05 Zn group vs. Se group

900 800 700 600 500 400 300 200 100 0 Placebo Zn

* * *

Se
Post-Test

Zn-Se

Pre-Test

Effect of Zinc and Selenium Supplementation on Serum Testosterone and Plasma Lactate Fig. 2 Variations of free testosterone in research groups. (MeanSD). Asterisks indicate p<0.05 pre-test vs. post-test; Euro sign indicates p<0.05 Zn group vs. Se group; cross indicates p<0.05 Zn group vs. PL group

459

30

Free Testosterone (pg/dl)

25 20 15 10 5 0
Placebo Zn

* *

Se
Post-Test

Zn-Se

Pre-Test

The mechanisms responsible for the increase in serum testosterone levels during exercise are controversial, and may include decreased testosterone clearance, hemoconcentration, increased SHBG, catecholamine, precursor molecule DHEA sulfate, and testosterone production due to stimulation by factors other than LH and the slight decrease in progesterone [3035]. Probably, the reason of contradiction around this issue is about the difference between physical fitness of the subjects, exercise intensity, type and duration, and the work load. Jezova et al. [36] reported that both plasma testosterone and catecholamine responses to physical effort depend more on work intensity than on work duration or total work output. Zinc enhances human chorionic gonadotropin-induced production of cAMP and consequently testosterone in rat testes [37]. Additionally, zinc may increase the conversion of androstenedione to testosterone in the periphery tissue [22]. Zinc also interferes with the metabolism of testosterone by decreasing its hepatic clearance and reducing hepatic 5 alpha-reductase activities [38]. The findings of our study demonstrated that resting free and total testosterone levels after a 4-week zinc supplementation had no variation relative to PL group. Exercise increased total testosterone levels in the Zn group than Se group, and also free testosterone levels in Zn group than other groups. Kilic et al. [16] showed that 4-week zinc supplementation increased resting levels of both thyroid hormones and testosterone concentrations, and exhaustion exercise led to a significant inhibition of those hormones,

Plasma Lactate (mg/dl)

Fig. 3 Variations of plasma lactate in research groups. (MeanSD). Asterisks indicate p<0.05 pre-test vs. post-test

60 50 40 30 20 10 0 Placebo Zn
Pre-Test

* * *

Se
Post-Test

Zn-Se

460

Neek et al.

but that 4-week zinc supplementation prevented this inhibition in wrestlers. Similarly, Kilic [39] reported mentioned results about sedentary males in response to fatiguing bicycle exercise. Kilic concluded that administration of a physiologic dose of zinc can be beneficial to performance. Contradictory, Koehler et al. [40] noted that zinc supplementation may reverse lowered testosterone levels and restore disturbed testosterone metabolism in cases of mild or severe zinc deficiency; it is not capable of further increasing serum testosterone when sufficient zinc is provided by the regular diet. The disparity in results in the aforementioned studies with our findings might be due to variations in the consumed dose of zinc supplement, status of zinc in subjects, amount of zinc intake on diet, as well as the improper time duration for the effect of supplementation does in our study. Previous studies demonstrated that significant decrease in polymorphonuclear leukocyte and lymphocyte activity, and high-density lipoprotein cholesterol after 6 weeks of 150 mg Zn per day [41, 42]. Similarly, that consumption of zinc supplements in excess of 50 mg/day have been linked to impair copper absorption [43]. For these reasons, use of Zn supplements should be limited to those containing no more than 30 mg/d [44]. Therefore, in the present study, the considered supplementation of consumed dose 30 mg/d was performed by its practical order. In the Leydig cells, glutathione peroxidise (se-dependent) has been localized immunocytochemically in the cytoplasm in close relationship to the smooth endoplasmic reticulum, and it is possible that the metabolic pathway of testosterone biosynthesis requires protection against peroxidation and is thus affected by a decrease in the activity of this enzyme [45]. In our study, 4-week selenium supplementation had no effect on resting levels of total and free testosterone and plasma lactate in cyclist. Probably, the normal status of selenium in cyclists and the enough amount of intake selenium from diet can be the other reasons of the results about the selenium group. It is known that plasma lactate concentration increases together with increased exercise [46]. This study showed that 4-week zinc and selenium supplementations had no effect on plasma lactate in pre and post-exhaustive exercise. There are only a limited number of studies about how selenium, which is known to reduce oxidative damage in exercise, affects glucose metabolism, lactate levels, and tiredness in physical activity [47]. Akil et al. [10] investigated the effects of selenium on lipid peroxidation and lactate levels in rats subjected to acute swimming exercise. He reported that the increase in free radical production and lactate levels due to acute swimming exercise in rats might be offset by selenium supplementation. Selenium supplementation may be important in that it supports the antioxidant system in physical activity. Baltaci et al. [48] demonstrated that 4-week zinc deficiency increased plasma lactate in rats after acute swimming and zinc supplementation (3 mg/kg weight) has the opposite effect. In summary, even though supplementation may reverse negative effects of nutritional deficiencies (and consequently improve athletic performance), this cannot be transferred directly to non-deficient athletes. In most cases, if energy intakes are sufficient, the mineral needs of athletes are analogous to healthy individuals. Some athletes, however, may have greater requirements as a consequence of disproportionate losses of I nutrients in sweat and urine. For these athletes, supplementation may need to be considered on an individual basis to maintain normal body stores, not for ergogenic purposes. A systematic approach to the study of minerals and exercise performance is needed. This approach needs to use the same protocol to evaluate whether minerals can be effective ergogenic aids. It would require longer supplementation periods, control of exercise settings, multi-center trials, men and women participants, elite and recreational athletes, and precise measures of mineral status.

Effect of Zinc and Selenium Supplementation on Serum Testosterone and Plasma Lactate

461

References
1. Flinn JM, Hunter D, Linkous DH et al (2005) Enhance zinc consumption causes memory deficits and increased brain levels of zinc. Physiol Behav 83:793803 2. Cordova A, Alvarez-Mon M (1995) Behaviour of zinc in physical exercise: a special reference to immunity and fatigue. Neurosci Biobehav Rev 19:439445 3. Stallard L, Reeves PG (1997) Zinc deficiency in adult rats reduces the relative abundance of testisspecific angiotensin-converting enzyme mRNA. J Nutr 127:2529 4. Favier AE (1992) The role of zinc in reproduction. Biol Trace Elem Res 32:363 5. Fuse H, Kazama T, Ohta S, Fujiuchi Y (1999) Relationship between zinc concentrations in seminal plasma and various sperm parameters. Int Urol Nephrol 31:401408 6. VanLoan MD, Sutherland B, Lowe NM, Turnland JR, King JC (1999) The effects of zinc depletion on peak force and total work of knee and shoulder extensor and flexor muscles. Int J Sport Nutr 9:125135 7. Kaya O, Gokdemir K, Kilic M, Baltaci AK (2006) Zinc supplementation in rats subjected to acute swimming exercise: its effect on testosterone levels and relation with lactate. Neuroendocrinol Lett 27(1 2):267270 8. Shamberger RJ (1986) Selenium metabolism and function. Clin Physiol Biochem 4:4249 9. Shu H (1989) Human selenium deficiency during total parenteral nutrition support (a case report). Zhongguo Yi Xue Ke Xue Yuan Xue Bao 11:7476 10. Akil M, Gurbuz U, Bicer M, Sivrikaya A, Mogulkoc R, Baltaci AK (2011) Effect of selenium supplementation on lipid peroxidation, antioxidant enzymes, and lactate levels in rats immediately after acute swimming exercise. Biol Trace Elem Res. (in press) 11. Behne D, Weiler H, Kyriakopoulos A (1996) Effects of selenium deficiency on testicular morphology and function in rats. J Reprod Fertil 106:291297 12. Duma E, Orbai P, Derevenco P (1998) Blood levels of some electrolytes and hormones during exercise in athletes. Rom J Physiol 35:5560 13. Huang WS, Yu MD, Lee MS, Cheng CY, Yang SP, Chin HM, Wu SY (2004) Effect of treadmill exercise on circulating thyroid hormone measurements. Med Princ Pract 13:1519 14. Rosolowska-Huszcz D (1998) The effect of exercise training intensity on thyroid activity at rest. J Physiol Pharmacol 49:457466 15. Bosco C, Tihanyl J, Rivalta L, Parlato G, Tranquilli C, Pulvirenti G, Foti C, Viru M, Viru A (1996) Hormonal responses in strenuous jumping effort. Jpn J Physiol 46:9398 16. Kilic M, Baltaci AK, Gunay M et al (2006) The effect of exhaustion exercise on thyroid hormones and testosterone levels of elite athletes receiving oral zinc. Neuroendocrinol Lett 27(12):247252 17. Institute of Medicine. Food and Nutrition Board (2001) (2000). Dietary reference intakes for vitamin A, vitamin K, arsenic, boron, chromium, copper, iodine, iron, manganese, molybdenum, nickel, silicon, vanadium, and zinc. Dietary reference intakes for vitamin C, vitamin E, selenium, and carotenoids. National Academy Press: Washington, DC 18. Kuipers H, Verstappen FT, Keizer HA, Geurten P, Van kranenburg G (1985) Varability of aerobic performance in the laboratory and its physiological correlates. Int J Sports Med 6(4):197201 19. Kuipers H, Keizer HA, Brouns F, Saris WHM (1987) Carbohydrate feeding and glycogen synthesis during exercise in man. Pflugers Archives 410(6):662665 20. Cinar V, Polat Y, Baltaci AK, Mogulkoc R (2011) Effects of magnesium supplementation on testosterone levels of athletes and sedentary subjects at rest and after exhaustion. Biol Trace Elem Res 140(1):1823 21. Cinar V, Baltaci AK, Mogulkoc R, Kilic M (2009) Testosterone levels in athletes at rest and exhaustion: effects of calcium supplementation. Biol Trace Elem Res 129(13):6569, Summer 22. Bermudez JA, Perex-Pasten E, Villalpando S et al (1986) Low plasma zinc and androgen in insulin dependent diabetes mellitus. Arch Androl 16(2):151 23. Berchtold P, Berger M, Cuppers HJ, Herrman J, Nieschlag E, Rudroff K et al (1978) Nonglucoregulatory hormones (T4, T3, rT3, TSH, testosterone) during physical exercise in juvenile type diabetic. Horm Metab Res 10:269273 24. Ravaglia G, Forti P, Mailoi F, Pratell L, Vettori C, Bastagli L et al (2001) Regular moderate intensity physical activity and blood concentrations of endogenous anabolic hormones and thyroid hormones in aging men. Mech Aging Dev 122:191203 25. Woody CJ, Weber SL, Laubach HE, Ingram-Willey V, Amini-Alashti P, Sturbaum BA (1998) The effects of chronic exercise on metabolic and reproductive functions in male rats. Life Sci 62(4):327332 26. Hackney AC, Sinning WE, Bruot BC (1988) Reproductive hormonal profiles of endurance-trained and untrained males. Med Sci Sports Exerc 28:180189 27. Hackney AC, Fahrner CL, Stupnicki R (1997) Reproductive hormonal response to maximal exercise in endurance-trained men with low resting testosterone levels. Exp Clin Endocrinol Diabetes 105:291295

462

Neek et al.

28. Hekonen M, Naveri H, Kuoppasalmi K, Huhtaniemi I (2001) Pituitary and gonadal function during physical exercise in the male rat. J Steroid Biochem 35(1):127132 29. Ronsen O, Huge E, Klarlund PB, Bahr R (2001) Increased neuroendocrine response to a repeated bout of endurance exercise. Med Sci Sports Exerc 33(4):568575 30. Cadoux-Hudson TA, Few JD, Imms FJ (1985) The effect of exercise on the production and clearance of testosterone in well trained young men. Eur J Appl Physiol 154(3):321325 31. Caballero MJ, Mena P, Maynarm M (1992) Changes in sex hormone binding globulin, high density lipoprotein cholesterol and plasma lipids in male cyclists during training and competition. Eur J Appl Physiol 64(1):913 32. Cumming DC, Brunsting LA, Strich G, Ries AL, Rebar RW (1986) Reproductive hormone increase response to acute exercise in men. Med Sci Sports Exerc 18(4):369373 33. Lin H, Wang SW, Wang RY, Wang PS (2001) Stimulatory effect of lactate on testosterone production by rat Leydig cells. J Cell Biochem 83(1):147154 34. Raastad T, Bjoro T, Hallen J (2000) Hormonal responses to high and moderate intensity strength exercise. Eur J Appl Physiol 82(12):121128 35. Vaamonde D, Da Silva ME, Poblador MS, Lancho JL (2005) Reproductive profile of physical active men after exhaustive endurance exercise. Int J Sports Med 26:110 36. Jezova D, Vigas M, Tatar P, Kvetnansky R, Nazar K, Kaciuba-Uscilko H, Kozlowski S (1985) Plasma testosterone and catecholamine responses to physical exercise of different intensities in men. Eur J Appl Physiol 54(1):6266 37. Nishi Y, Hatano S, Aihara K et al (1984) Effect of zinc ion on human chorionic gonadotropin stimulated in vitro production of cAMP and testosterone by rat testis. Pediatr Res 18:232 38. Grumbach MM, Coute FA (1992) Disorders of sex in differentiation. In: Wilson JD, Foster WD (eds) Williams textbook of endocrinology, 8th edn. WB Saunders Co, Philadelphia, p 853 39. Kilic M (2007) Effect of fatiguing bicycle exercise on thyroid hormone and testosterone levels in sedentary males supplemented with oral zinc. Neuroendocrinol Lett 28(5):181185 40. Koehler K, Parr MK, Geyer H et al (2007) Serum testosterone and urinary excretion of steroid hormone metabolites after administration of a high-dose zinc supplement. Eur J Clin Nutr 63:6570 41. Chandra RK (1984) Excessive intake of zinc impairs immune responses. JAMA 252:14431446 42. Hooper PL, Visconti L, Gary PJ, Jahnson GE (1980) Zinc lowers high-density lipoprotein cholesterol levels. JAMA 244:19601961 43. Haymes EM (1991) Vitamin and mineral supplementation to athletes. Int J Sport Nutr 1:146169 44. Benardot RD (2006) Advanced sports nutrition. Human Kinetics PP, United State, p 92 45. El-Sisy GA, Abdel-Rezek AMA (2008) Effect of dietary zinc or selenium supplementation on some reproductive hormone levels in male Baladi goats. Global Vet 2(2):4650 46. Grant S, McMillan K, Newell J, Wood L, Keatley S, Simpson D, Leslie K, Fairlie-Clark S (2002) Reproducibility of the blood lactate threshold, 4 mmol.l(1) marker, heart rate and ratings of perceived exertion during incremental treadmill exercise in humans. Eur J Appl Physiol 87:159166 47. Thomson CD (2004) Assessment of requirements for selenium and adequacy of selenium status: a review. Eur J Clin Nutr 58:391402 48. Baltaci AK, Ozyurek K, Mogulkoc R, Kurtoglu E, Ozkan Y, Celik I (2003) Effects of zinc deficiency and supplementation on the glycogen contents of liver and plasma lactate and leptin levels of rats performing acute exercise. Biol Trace Elem Res 96(13):227236