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Effect of Avena sativa (Oats) on Spermatogenesis and Reproductive Health

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Infertility is a major problem of the day amongst men and women and marked by no pregnancy even after one year of unprotected intercourse. Almost 30% of infertility has been related with male factors-low sperm concentration, motility, viability and morphology. Factors like pollution, drugs, stress, life style changes, toxicants and insufficient nutrition have harmful effects on spermatogenesis and reproductive health. Fluorine is one such potent toxicant to which humans are exposed and the problem of fluorosis was known for long in India, especially in Andhra Pradesh. It was reported in several studies that fluorine interferes with the structural and functional integrity of the male reproductive system resulting in male factor infertility. The present research has been undertaken to evaluate the efficacy of Avena sativa as a nutrient and food supplement to lower the fluoride induced infertility in male rats as, A. sativa has a wide range of chemical and mineral constituents. In this study, fluorine at a dose of 0.01 g/kg body weight, administered through oral route induced infertility by causing damage to histoarchitecture of the testis and decreased the levels of serum testosterone, FSH, and LH. Treatment with hydroalcholic extract of A. sativa resulted in decreased damage to the reproductive organs and lesser impact on sperm parameters-sperm count, viability, morphology, motility etc. The results were analyzed through one way ANNOVA using graph pad prism 5 software. Form the present study it is concluded that oats has the ability to decrease the toxic effect of fluorine.
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Journal of Endocrinology and Reproduction, Vol 20(2), 118-125, 2016, DOI: 10.18311/jer/2016/15471
ISSN (Print) : 0971-913X
ISSN (Online) : 2455–1244
Eect of Avena sativa (Oats) on Spermatogenesis
and Reproductive Health
Vara Prasad Saka*1, Siva Reddy Challa2 and Akondi Butchi Raju3
1*Department of Pharmacology, Vignan Pharmacy College, Vadlamudi, Guntur District,
Andhra Pradesh, India.
2Department of pharmacology, K.V.S.R.Siddhartha College of Pharmaceutical sciences, Vijayawada,
Andhra Pradesh, India.
3Department of Clinical Pharmacy and Pharmacology, Ibn Sina National College for Medical Studies;
drraju2020@gmail.com
Keywords: Avena sativa(oats), Male Infertility, Spermatogeresis, Fluoride
Abstract
Infertility is a major problem of the day amongst men and women and marked by no pregnancy even after one year of
unprotected intercourse. Almost 30% of infertility has been related with male factors, concerning sperm-low concentration,
poor motility, decreased viability, and deformities. Factors like pollution, drugs, stress, life style changes, toxicants and
            
                  
        
of the male reproductive system resulting in male factor infertility. Oat, Avena sativa, has a wide range of chemical and
           

administered through oral route, induced infertility by causing damage to histoarchitecture of the testis and decrease in
the levels of plasma testosterone, FSH, and LH. Treatment with hydroalcoholic extract of oats resulted in decreased damage
to the reproductive organs and lesser impact on sperm parameters- sperm count, viability, morphology, motility, etc. Form

1. Introduction
Infertility can be dened as a lack of pregnancy aer a year
of unprotected regular intercourse1. e number of infer-
tility cases around the world is increasing and, currently,
between 15% and 20% of couples of reproductive age are
infertile. Recent reports suggest that approximately 30%
of infertility is due to a male factor2 wherein low sperm
count is the main cause. e total sperm count, motility
and normal morphological characteristics are generally
measured in men and, in most cases, these indices are
clearly dened4. A couple of conditions can interfere with
spermatogenesis and cause decline in sperm counts and
quality. Factors such as therapeutics, in particular anti-
cancer and antimicrobial drugs, toxins, polluted water or
air, and malnutrition, can all produce deleterious eects
on sperm production and fertility1. Many environmental
toxins and irradiation are well studied worldwide in rela-
tion to deterioration of semen quality5,6.
Fluoride is one of the vulnerable toxicants to which
human beings are exposed7. e problem of uorosis has
been quite well known in India. When the uoride con-
centration exceeds 1.5 mg per liter of water, it aects the
development of teeth during the stage of calcication in
children. Fluoride toxicity can damage the lungs and fetus.
Latest reports arm the relationship between uoride lev-
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119
Eect of Avena sativa (Oats) on Spermatogenesis and Reproductive Health
els in drinking water and increasing infertility8. Fluoride
is believed to produce abnormalities in the structural and
functional integrity of the testis, epididymis and vas def-
erens. Animal studies have shown the eect of uoride on
morphology and metabolism of spermatozoa. High uoride
intake can lead to Zn deciency in the testes and male re-
production9,10 leading to reduction in testosterone levels that
is critically necessary for testicular development and main-
tenance9,11. High uoride content is also reported to increase
oxidative stress in the testes aecting spermatogenesis9,11,12.
e grass/Poaceae family of oats (Avena sativa L.), is
an annual green grass3,14,15. A. sativa originated in England,
France, Poland, Germany and Russia and is now grown
all around the world. e word Avena is derived from the
Sanskrit word “avi, which means “sheep” or “avasa” meaning
“food”. In traditional medicine, oat is used to treat nervous
debility, insomnia and nerve weakness. It is considered as
a good antispasmodic, antitumoral, cyanogenic, demulcan,
diuretic, neurotonic, stimulant, and tonic. Several stud-
ies have reported that oat and its constituents have various
pharmacological eects such as reduction of blood cho-
lesterol and sugar levels, immunomodulation, anti-cancer,
antioxidant, antiatherogenic, and anti-inammatory besides
control of childhood asthma, body weight, etc. Oatmeal is
rich in protein, has many benecial minerals like iron (Fe),
calcium (Ca), potassium (K), magnesium (Mg), copper
(Cu), zinc (Zn), and silicon (Si), fatty acids, selenium (Se)
and a number of vitamins such as Vitamin B1, B2, B6, B12,
Niacin, Vitamin C, Vitamin A, and Vitamin E.
Selenium works with Vitamin E in various essen-
tial cancer prevention agent frameworks all through the
body16. Likewise, it also works with glutathione peroxide
(GSH-PX), a protein that ensures the internal structures
of cells against free radicals. It is a cancer prevention agent
by acting at the level of lipids of the cell membrane17.
Zn plays a key role in spermatogenesis18–23 and reverses
uoride toxicity9,10. Also, vitamin E has been proved to
reverse uoride toxicity in mice7. As oats are rich in sele-
nium, vitamin E and zinc, the present study focused on
nding if uoride induced infertility in male albino rats
could be reversed by oats.
2. Materials and Methods
2.1 Plant Material and Preparation
Avena sativa (oat) whole grains were purchased from local
grains shop and dried under shade.Oat grains were milled
and passed through #10 mesh screen. e oat powder was
mixed with the solvent in 1:4 ratio and then added to 50%
v/v aqueous ethanol solution at 400C. e resultant blend
was then mixed by swirling for 60 minutes, then cooled to
room temperature. is blend was centrifuged for 7 min
to separate the supernatant. e pellet was resuspended
in solvent and centrifuged. e supernatant was then
gathered, dried and pulverized24.
2.2 Chemicals
Sodium uoride from Loba Chemie (Mumbai, India) was
used as infertility inducing agent. Testosterone propionate
from Sigma Aldrich was used as standard medication.
Physiological saline was utilized as vehicle for oat extract
and sodium uoride, and arachis oil was used as vehicle
for testosterone propionate.
2.3 Animals, Maintenance and
Experimentation
e protocol for the experimentation was approved by
IAEC (approval no: 007/IAEC/NCPA/M. Pharm/2012-
13). Male Wistar albino rats were obtained from Gosh
and Gosh Enterprises, Kolkata, and maintained in animal
house at 25±2 °C temperature with 12 hr dark and light
cycle with standard pellet diet and water ad libitum.
Male albino rats were divided into 6 groups of 6 each.
Rats in group I served as control and received normal
saline. Rats in group II were administered with sodium
uoride, dissolved in normal saline, at a daily dose of 10
mg/kg body weight through oral route for 30 days. e
dose was mainly based on the LD50 for rat, 250 mg/kg
body weight, and the present dose is 1/25th of the LD50
25.
Rats in group III were administered with oat extract, dis-
solved in normal saline, at a daily dose of 400 mg/kg body
weight dose, through oral route. Rats in groups IV andV
were administered with oats extract at 200 and 400 mg/
kg body weight along with NaF at 10 mg/kg body weight
through oral route. Rats in group VI were administered
with testosterone propionate at a dose of 0.5 mg/kg body
weight through subcutaneous route26. All the groups were
treated for 28 days1,27 and sacriced 24 hr aer the last
dose.
2.4 Analyses
At the time of sacrice the rats were rst weighed and
then subjected to cervical dislocation28,29. e abdominal
cavity was dissected to expose the reproductive organs.
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Vara Prasad Saka, Siva Reddy Challa and Akondi Butchi Raju
e testes were removed and cleared free of mucus and
surrounding tissue28.
e testes were weighed in an electronic balance,
while the testis volumes were measured using immersion
method28.
e content of cauda epididymal duct was released
in 0.5 ml of normal saline taken in a petri dish and the
temperature was maintained at 37 °C1,30. e sperm count
was carried out using a haemocytometer29,31. e sperm
in four corner squares were counted29,31. A drop of sperm
suspension was placed on a clear glass slide and then cov-
ered with a cover slip. e slide was then examined in a
microscope at 400x, and the motility was scored in the
dierent elds of view. Spermatozoa showing any degree
of movement were considered to be motile. e spermato-
zoa (motile as well as immotile, separately) were counted.
Sperm viability was assessed using a supravital staining
technique based on the principle that cells with damaged
plasma membrane take up the stain, whereas viable ones
do not. All glassware as well as the eosin–Nigrosine stain
were maintained at 37 °C. Spermatozoa appearing pinkish
(stained) were considered as dead, whereas those appear-
ing colorless (unstained) were counted as viable. Using the
same preparation, sperm morphology was determined.
Abnormalities were classied as headless sperm, banana
head, bent neck and bent tail5. e sperm were classied
into normal and abnormal. e total sperm abnormality
was expressed as percentage incidence33.
Blood was collected from rats by cardiac puncture,
centrifuged plasma was separated and stored for hor-
monal assays. Plasma testosterone, luteinizing hormone,
and follicle stimulating hormone levels were determined
by chemilumin escence-immunoassay using immuno-
assay kits and ADVIA Centaur immuno-analyzer.
e testis was xed in Bouin’s uid for 24 hr for his-
tological analysis. e tissues were dehydrated in an
ascending series of alcohol, treated with xylene, and
embedded in paran wax. Sections of 6-µm thickness
were cut in a rotary microtome and stained with hema-
toxylin and eosin. e permanently mounted sections
were viewed in a microscope, and images were captured
ina digital camera1.
2.5 Statistical Analysis
Data were analyzed by one-way ANOVA with Tukey’s post
hoc test using graph pad prism soware, and expressed as
mean ± SEM. P≤0.01 was considered as statistically sig-
nicant.
3. Results
3.1 Body and Organ Weights
As seen in Table 1, in the uoride treated rats body as
well as organ weights decreased signicantly. In the rats
treated uoride accompanied by oats extract at 200 mg/
kg body weight there was no signicant dierence when
compared with disease control group. But when uoride
treated rats were also treatedwith oat extract at 400 mg/
kg body weight there was signicant increase in the body
weight as well as organ weight. is indicates that oat
Table 1: Eect of oat extract on uoride induced changes in weight parameters on male rats.
Group/parameter Body weight (gm.) Testis weight (mg)
Initial Final
Normal control 200.2±4.672 211.3±3.293 0.933±0.0123
NaF (10 mg/kg) treated 200.7±2.753 181.0±6.506c0.826±0.0203b
Oat extract (400 mg/kg) only 203.3±4.410 217.3±2.871 0.986±0.00232
NaF - oat extract (200 mg/kg) 202.2±3.609 194.8±2.482 0.833±0.0195
NaF + oat extract (400 mg/kg) 201.3±2.186 202.3±0.9545#0.970±0.00404$
Testosterone (0.5 mg/kg) 202.2±3.167 221.0±2.840c1.07±0.0319b
Values are expressed as mean ±SEM of each group (n=6) and are signicant when done ONE WAY ANOVA with
Tukey’s post hoc test. ‘b’ P<0.01; ‘c’ p<0.001 when compared with normal control; ‘#’p<0.01; $p<0.001 when compared
with disease control.
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121
Eect of Avena sativa (Oats) on Spermatogenesis and Reproductive Health
extract at 400 mg/kg treatment obviated the weight loss
caused by uoride treatment.
3.2 Semen Parameters
In the uoride treated animals sperm count, viability and
motility were signicantly reduced at p<0.001 when com-
pared with the respective controls (Table 2). However,
co-administration of oat extract with uoride attenu-
ated the decrease in sperm count, viability and motility.
ere was a signicant increase (P < 0.001) in sperm with
abnormal morphologies in uoride treated rats; however,
co-administration of oat extract reduced (p<0.001) the
percentage abnormal morphologies, although the num-
ber did not reach the saline control value.
3.3 Hormone Levels
Levels of FSH, LH and testosterone decreased in the
uoride treated rats (Table 3). Co-administration of oat
extract to uoride-treated rats increased the levels of FSH
and testosterone above the control, whereas the LH level
also was signicantly increased but slightly lesser than
control value.
3.4 Histopathological Changes
Sections of testis of healthy control rats revealed normal
histoarchitecture. ere were uniform, well-organized
seminiferous tubules with all stages of spermatogenesis
and normal interstitial tissue (Figure 1A). Testicular
tissue of rats that received only uoride showed degen-
erative changes in the majority of the seminiferous
tubules (Figure 1B). ese changes were characterized
by shrunken, disorganized seminiferous tubules with
irregular, buckled basement membrane and incomplete
spermatogenesis. Moreover, the seminiferous tubules
were almost devoid of spermatids and spermatozoa.
Vacuolar degeneration of spermatogonia and Sertoli cells
was evident. Degenerated germinal epithelial cells were
sloughed into the lumina of most seminiferous tubules.
e histological ndings of the testicular sections of rats
that received only oat extract (Figure 1C) showed normal
and healthy histoarchitecture in which well-developed
seminiferous tubules with complete spermaotgenesis
and intact interstitial tissue were seen along with the
uniformly distributed Leydig cells which were absent in
uoride treated animals. Testicular sections of the rats
treated oat extract (200 mg/kg body weight) along with
uoride (Figure 1D) showed well-organized seminifer-
ous tubules with incomplete spermatogenesis, and the
seminiferous epithelium was sloughed into the lumen of
the seminiferous tubules. Testicular sections of rats that
received oat extract (400 mg/kg) (Figure 1E) along with
uoride showed the well-organized seminiferous tubules
with complete spermatogenesis. Well-developed sper-
matids and uniform connective tissue were seen in the
interstitial space and resembled the histoarchitecture of
the normal and testosterone-treated rats. Well organized
and completely developed seminiferous tubules with sper-
matids, spermatozoa and complete spermaotgenesis and
also with uniformly distributed Leydig cells in the con-
Table: 2 Eect of oat extract on uoride induced changes in semen parameters of male rats.
Group/parameter Sperm count
(millions/ml)
Sperm motility (%) Sperm viability
(%)
Sperm morphology
(%)
Normal control 29.17±0.4773 30.50±0.6708 61.83±1.67 29.50±1.088
NaF (10 mg/kg)
treated
22.67±1.145a11.83±1.721c32.67±1.453c62.33±1.054c
Oat extract (400 mg/
kg) only
40.83±1.302c32.83±1.302c64.33±2.261c29.33±0.8028c
NaF + oat extract (200
mg/kg)
31.83±1.108$21.17±1.302$49.83±2.638$51.83±1.167$
NaF + oat extract (400
mg/kg)
38.50±1.478$32.67±1.145$61.33±1.358$43.00±1.528$
Testosterone (0.5 mg/
kg)
50.83±1.641c70.83±1.641c71.00±1.414c20.50±1.310c
Values are expressed as mean ±SEM of each group (n=6) and are signicant when done ONE WAY ANOVA with Tukey’s
post hoc test. ‘a’ P<0.05; ‘c’<0.001 when compared with normal control; $p<0.001 when compared with disease control.
Journal of Endocrinology and Reproduction 122
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Vara Prasad Saka, Siva Reddy Challa and Akondi Butchi Raju
nective tissue lling the interstitial space were observed
in the testicular sections of the rats treated with testos-
terone (0.5 mg/kg) as reference standard (Figure 1F). e
histological ndings showed that oat extract at a dose
of 400 mg/kg signicantly reversed the uoride toxicity
and maintained the histoarchitecture of the reproductive
organ of the rats by marked improvement in spermato-
genesis evidenced by the presence of well-organized
seminiferous tubules and well developed Leydig cells.
3.4.1 Histological Examination
Figure 1. Histopathological observation of the eect of
hydro-alcoholic extract of Avena sativa on uoride induced
testicular toxicity; L indicates eumen of seminiferous
tubule; ST indicates seminiferous tubule. A: Normal; B:
Fluoride treated; C: Oats alone; D: Fluoride+Oats 200 mg/
kg; E: Fluoride + Oats 400 mg/kg; F: Fluoride+Testosterone
treated.
4. Discussion
In the present study, the role of Avena sativa in preventing
the infertility induced by sodium uoride was studied. e
model employed in this work has been used previously by
several investigators to assess the toxicity of uoride on
fertility and reproduction in laboratory animals7–9,25,34,35,37.
e concentration of NaF used in this study was chosen
according to previous studies25. e drinking water route
of exposure was chosen to mimic human exposure and to
reect the impact on fertility of the sustained blood lev-
els of uoride that would occur from water consumption
throughout the day35.
e results of present study show that sodium uoride
treatment would cause a signicant decrease in the body
weight. Similar results were reported by other workers in
rats and mice fed dierent concentrations of uoride78
and it is due to oxidative damage38 to the body tissues.
However, the extract of Avena sativa when administered
along with sodium uoride maintainedthe body and
organ weights at normal signicantly and it might be the
result of antioxidant activity of Avena sativa13,15,16.
In the present study the count, motility and viability
of epididymal sperm in NaF treated group of rats declined
signicantly and increased in abnormal morphologies.
e decrease could be correlated with the testicular sper-
matogenic arrest and decline in fertility following uoride
ingestion in mice, rats and rabbits7,8,9,25,39. e hormones,
like testosterone, FSH and LH were decreased signi-
cantly due to uoride intoxication.
Fluoride toxicity led to the decline in sperm count
via hormonal imbalance9, structural and functional
defects in spermatozoa40,41 and alterations in epididymis
and accessory reproductive glands40. Fluoride toxicity
resulted in sloughing o the spermatogenic cells in semi-
niferous tubules of testes leading to disorganization
of their epithelium42, and hampered steriodo genesis
and spermatogenesis. Fluoride intoxication causes oxi-
dative damage38, and zinc deciency, disturbs signal
transduction9 and suppresses testosterone level43–46.
Fluoride directly interferes with spermatogenesis by
depressing EGF and EGFR47, modifying G-protein signal-
ing48, diminishing the levels of Androgen Receptor (AR),
disturbing the levels of estradiol and increasing the
levels of FSH and LH9,44,45. ough decrease in estrogen
levels is known to increase FSH acting in the negative
feedback loop, an F-induced decrease in estradiol level
may reduce FSH, leading to decreased spermatogenesis9.
In previous studies, it has been reported that high u-
oride intake can lead to zinc deciency in testes and the
accessory male reproductive organs9,49,50,. Zinc deciency
may suppress the testosterone levels critically neces-
sary for testis development9,46 and, more importantly, it
increases oxidative stress in testes leading to poorer qual-
ity spermatozoa9,12,46,51.
e mechanism by which uoride aects sperm
motility has not been clearly elucidated. However, it has
been postulated that uoride could decrease the level of
Journal of Endocrinology and ReproductionVol 20 (2) | December 2016 | http://www.informaticsjournals.com/index.php/jer/index
123
Eect of Avena sativa (Oats) on Spermatogenesis and Reproductive Health
fructose, the sugar, which provides energy to sperm for
motility7, by inhibiting many enzymes like enolases, and
acid and alkaline phosphatases by binding with cofactors
like Mg, Ca, Zn, and Se52 and, thus, inhibit glycolysis, res-
piration and motility of sperm9. e testicular oxidative
stress caused by uoride has been shown to result in dam-
age of sperm cell membrane which might accountable for
inhibition of spermatogenesis and impairment of sperm
activity53.
is study reveals the therapeutic eect of Avena
sativa extract in signicantly preventing these changes
from occurring. is might be due to the presence of zinc,
vitamin E, selenium, and antioxidants like aventhramides
in Avena sativa15. Zinc23 is involved in initiation of sper-
matogenesis by ribonuclease activity54, by participating
in spermatozoa maturation54 and preserving seminifer-
ous tubules and the germinal epithelium55, and also by
enhancing sperm motility54. Vitamin E is also important
in maintaining the male reproductive health. e role of
Vitamin E in the reproductive system is elaborately stud-
ied and it has been demonstrated that Vitamin E also
facilitates reproductive health by supporting testicular
and epididymal structure and function15,23,56–58.
is study leads to the conclusion that Avena sativa,
which is in vitamin E, aventhramides rich like antioxi-
dants and several minerals like zinc, selenium etc., has the
potential to be protective and preventive in the context of
uoride-induced impairment of male reproductive struc-
ture and function.
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Table: 3 Eect of oat extract on uoride induced changes in hormonal parameters of male rats.
Group/parameter Serum testosterone (ng/dl) FSH (mIU/ml) LH (mIU/ml)
Normal control 57.51±1.570 0.1900±0.005774 0.2350±0.01176
NaF (10 mg/kg) treated 51.51±0.8232b0.1450±0.009916a0.1650±0.01668c
Oat extract (400 mg/kg) only 64.47±1.133c0.2400±0.01065b0.2370±0.01269c
NaF + oat extract (200 mg/kg) 58.86±1.378$0.1950±0.009916#0.2050±0.009916
NaF + oat extract (400 mg/kg) 63.32±0.9058$0.2250±0.007638$0.2150±0.009916
Testosterone (0.5 mg/kg) 70.39±0.534c0.3100±0.007303c0.3983±0.01662c
Values are expressed as mean ±SEM of each group (n=6) and are signicant when done ONE WAY ANOVA with Tukey’s
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The Present investigation covers the scientific comprehension of the potential effects of fluoride toxicity on body and testes weight and the spermatozoal activities in male albino rat and the beneficial effect of novel medicinal plant Tamarindus indica therapeutic treatment (10gm/kg body weight) after fluoride water intoxication. Three groups of male albino rats five of each were intoxicated with ground water fluoride (14.29 mg/L fluoride ion concentration) collected from fluoride zone of Agra. The other three groups were given the subsequent treatment of Tamarindus indica after fluoride water intoxication. The fluoride water exposure led to the significant reduction in the body and organ weight, whereas the sperm count, motility and sperm sluggishness were the decline of sperm count and its activities. However, the therapeutic treatment of Tamarindus indica pulp extract manifested the improvement in the recovery and mitigation of devastating toxic effects of fluoride on the concerned parameters in male albino rat.
Article
The effects of zinc, magnesium and calcium in seminal plasma on time-to-pregnancy (TTP) in healthy couples, on conventional semen parameters and computer-assisted semen analysis (CASA) parameters were evaluated. The localization of chelatable zinc ions in seminal plasma and spermatozoa were assessed by autometallography (AMG). Differences in chelatable zinc localization in samples with high and low total zinc were evaluated. Semen samples from 25 couples with short TTP and 25 couples with long TTP were subjected to conventional semen analysis, CASA, zinc and magnesium measurements by inductively coupled plasma mass spectrometry, and calcium by flame atomic absorption spectrometry. The cations were strongly inter-correlated, but no correlation with TTP or conventional semen parameters was found. Semen samples with high zinc concentrations exhibited statistically significant poorer motility assessed by the CASA parameters straight line velocity and linearity than samples with low zinc content. Calcium concentration also showed statistically significant differences for the same parameters, but the effect was removed by entering zinc concentration into a multiple regression model. Semen samples with high total zinc exhibited stronger staining of the seminal plasma at AMG. It is suggested that high seminal zinc concentrations have a suppressing effect on progressive motility of the spermatozoa (`quality of movement'), but not on percentage of motile spermatozoa (`quantity of movement').