ZANCO Journal of Pure and Applied Sciences

The official scientific journal of Salahaddin University-Erbil

ZJPAS (2017), 29 (s4); s150-s156
http://dx.doi.org/10.21271/ZJPAS.29.s4.18

Impact of Trace Elements in the Seminal Plasma on Sperm Quality

in Infertile Men

Ayad F. Palani1, Azad H.A. Alshatteri2

1-Department of Chemistry, College of Science, University of Garmian, Kalar, Iraq
2-Department of Chemistry, College of Education, University of Garmian, Kalar, Iraq

ARTICLE INFO ABSTRACT

Article History: In the modern world about 15% couples are unable conceive due to infertility.
Received: 01/06/2017 Male factor infertility contributes half of all infertility cases. Poor sperm quality is
Accepted: 05/08/2017 the main cause in male infertility. Recent researches focused on the chemical
Published: 20/12/2017 composition of seminal plasma to explain the causes of low sperm quality. In the
Keywords: present study, semen samples from 106 infertile men [oligoasthenozoospermia
OSZ ((n=38), asthenozoospermia ASZ (n=27) and normozoospermia NOZ (n=41)]
Sperm Quality and 25 fertile men were analyzed using computer-aided sperm analysis (CASA)
Trace elements system. concentrations of selected trace elements viz. Ca, Cu, Fe, Zn, Mg, Mo, Ni
ICP and Se in all the samples were determined using inductively coupled plasma
Nickel optical emission spectroscopy , and the associations between the concentrations of
Molybdenum. trace elements with that of sperm concentration, motility and morphology were
investigated. Results were analyzed statistically via ANOVA using Minitab
statistic program version 15. Results revealed that while Mo was significantly
*Corresponding Author:
lower among the infertile groups when compared with that of control (P < 0.05).
Ayad f. Palani Significantly low concentration of Ni was also observed in both the OSZ and NOZ
Ayad.palani@garmian.edu.krd
groups than that of controls (P < 0.05).

1. INTRODUCTION specific etiology could explain the

abnormalities of sperm production (De Kretser,
Human ability to fertilize obviously
Baker, 1999). Sperm ability to fertilize
declined from the last few decades (Zafar et al.,
acquired in the seminal plasma (De Silva et al.,
2015) and (Wang et al., 2005). Semen quality
2016), at the level of molecules many studies
is being reduced by 3% each year; this
have been concerned with the chemistry and
indicates that infertile couples increase in time
the biochemistry of seminal plasma (Abou-
(Sultan, 2016). In the modern world about 15%
Shakra,et al.,1989). Many molecules have vital
of all couples are unable to conceive due to
role in quality of seminal fluid including trace
infertility, approximately half of the infertility
elements (Sultan, 2016). In the chemistry of
cases are due to male factor (Agarwal et al.,
life trace elements considered important to cell
2004) and (Zalata et al., 2016). Inadequate
metabolism and function (Sağlam et al., 2015).
sperm numbers or quality is a major factor
Trace elements form part of enzymes, involved
contributes male factor infertility (Lewis et al.,
in the synthesis of 4hormones and in the
1997) and (Marzec-Wróblewska et al.,
transport of oxygen, and act as antioxidants
2011).Unfortunately until recently there is no
151 Palani, A. and ALshatteri, A. /ZJPAS: 2017, 29 (s4): s150-s156
(Forrer et al., 2001) and (Aguiar et al., 2015).
The decrease in trace elements results in
various cellular abnormalities because of their
specific biochemical changes, while increase
makes them toxic to cells (Sundaram et al.,
2013).
The role of trace elements in the male
reproductive has been investigated in many
studies. Trace elements have been found to be
essential for testicular growth and development
(Sağlam et al., 2015). It has been reported that
intracellular calcium (Ca) is essential for sperm
motility (Colagar et al., 2009) , magnesium
(Mg) have a role in semen transport and
ejaculation (Omu et al.,2001), Zinc (Zn) is
critical to spermatogenesis and influence sperm
motility (Sørensen et al., 1999) and (Saaranen
et al.,1987), Copper (Cu) plays an indirect role
in male infertility via increasing or decreasing
of other elements, which consequently, may
have a positive or negative influence on male
fertility (Akinloye et al., 2011). High level of
iron (Fe) may cause decrease of spermatozoa
motility (Slivkova et al., 2009). Selenium (Se)
is essential for testosterone biosynthesis and
the formation and normal development of
spermatozoa (Mistry et al., 2012) . Nickel (Ni)
enhances reproductive performance, and
involved in the physiological characteristics of
spermatozoa(Yokoi et al.,2003), and high
molybdenum (Mo) concentration reduces
sperm concentration and morphology (Meeker
et al., 2003). In the present study, we assessed
the relationships between concentrations of
different trace elements in the seminal plasma
and sperm quality in infertile and fertile men.
2. MATERIALS AND METHODS
2.1. Experimental design
This study included 106 infertile men aged
(32.3 ± 6.9) admitted to fertility clinics in
sulaymaniyah city. In addition 25 normal
fertile men aged (36.3 ± 6.9) who have one
child or more were included in the study as
control group. Infertile group was then
subdivided into subgroups according to semen
quality [oligoasthenozoospermia OSZ (n=38),
asthenozoospermia ASZ (n=27) and
normozoospermia NOZ (n=41)].
2.2. Sample collection and routine analysis
Samples were collected by
masturbation after 3-5 days of sexual
abstinence and incubated at (37 ºC) until
routine semen analysis. Semen analyses were
performed within 1 hour after sample delivery
according to (WHO, 2010) guidelines (WHO,
2010). The sperm concentration and motility
were assessed using computer-aided sperm
analysis (CASA), and sperm morphology was
evaluated by light microscopy using
papanicolaou's stain (WHO, 2010).
2.3. ICP analysis
After routine semen analysis samples
were centrifuged for 10 minutes and seminal
plasma has been separated and preserved in
deep freeze (-80 ºC) for further element
analyses. Elements analyzed using ICP, 1 mL
were taken from each sample and digested by
addition of 1 mL concentrated HNO 3 and 0.5
ml H 2 O 2 (30%) and left for 1 hour to complete
the digestion, (the inclusion of H 2 O 2 in the
digestive process is essential for completing
decomposition of the organic matrix). After 1
hour samples were sonicated to achieve a
homogeneous solution. An aliquot of clear
solutions was diluted to 10 mL by means of
deionized water as a diluent and analyzed using
ICP (Spectro Arcos, Germany) was used for
analysis of 8 metals. The instrument conditions
used were: Spray chamber is Scott spray;
Nebulizer: crossflow; RF power/W: 1400;
pump speed: 30 RPM; Coolant flow (L/min):
14; Auxiliary flow (L/min): 0.9; nebulizer gas
152 Palani, A. and ALshatteri, A. /ZJPAS: 2017, 29 (s4): s150-s156

flow (L/min): 0.8; Preflush (s): 40; Measure
time (s): 28; replicate measurement: 3; multi-
elements stock solutions containing 1000 mg/L
were obtained from Bernd Kraft (Bernd Kraft
GmbH, Duisburg, Germany); standard
solutions were diluted by several dilution into
0.1, 0.5, 2 ppm in 0.5% nitric acid ad diluent
(Hsiung et al., 1997).
2.4. Statistical analysis
Results were analyzed statistically via
ANOVA using Minitab statistic program
version 15. Differences value of p ≤ 0.05 used
to indicate a statistical significance. Also the
correlation between two parameters was taken,
correlation value (0.25-0.50) was considered
weak, (0.5-0.75) was moderate and (0.75-1.0)
was strong. Correlation between two
parameters was taken, correlation value (0.25-
0.50) was considered weak, (0.5-0.75) was
moderate and (0.75-1.0) was strong.
3. RESULTS AND DISCUSSION
Results of semen analysis parameters of
infertile and control groups showed in (table 1).
Results showed a significant decrease motile
sperm (grade A and grade C) and an increase in
grade D in OSZ group and a decrease in (grade
C) and an increase in (grade D) for ASZ group,
Also an increase was seen in (grade A, B and
C) and all parameters were normal in NOZ
groups as compared to normal fertile men.
Normal sperm morphology was decreased
significantly in OSZ and ASZ and non-
significantly in NOZ as compared to normal
fertile men.

Table 1: Seminal fluid analysis parameters [Mean±SD] of fertile and different infertile groups

Fertile group Infertile groups

Semen
n=25 Oligoasthenoz- Asthenoz- Normoz- P
parameter
oospermia n=38 oospermia n=27 oospermia n=41 value
Volume 3.6±1.6 4.2±1.8 3.8±1.5 3.7±1.1 n.s

Concentration 62.0±40.0 5.1±4.4* 56.7±20.1 102.3±62.5 0.001

Activity
Grade A 18.21±6.39* 7.43±9.72* 15.26±10.97 34.37±12.95 0.001

Grade B 12.85±6.31* 9.81±7.78 11.66±5.79 20.35±6.12 0.001

Grade C 31.73±13.08* 8.31±10.05* 10.26±6.51* 19.57±9.21 0.001

Immotile 37.21±13.55* 73.96±17.52* 63.29±14.728* 25.70±11.26 0.001

Morphology
Normal 53.66±15.01* 17.00±13.14* 28.66±12.43* 38.79±19.98 0.001

Abnormal Head 39.36±11.13 66.24±12.11 56.30±13.07 53.65±20.27 n.s

Abnormal midpiece 5.19±2.59 11.31±6.31 9.26±6.41 4.89±3.52 n.s

Abnormal Tail 1.78±1.03 5.43±3.51 5.774±2.29 2.658±1.52 n.s

ANOVA analyisis for ferile and infertile groups: n.s: non-significant, *; significant at p ≤0.001
153 Palani, A. and ALshatteri, A. /ZJPAS: 2017, 29 (s4): s150-s156

rate of the testes, spermatozoa density in the

Trace elements are essential for testicular
growth and development (Sağlam et al.,
2015).The concentration of trace elements in
the seminal plasma is much more than their
concentrations in the blood (Omu et al., 2001),
this is due to that sperm have proteins and
enzymes, thus, trace elements are important for
normal physiology (Abou-Shakra,et al., 1989).
The role of trace elements in sperm and whole
semen quality may have significant influence
upon men infertility (Marzec-Wróblewska et
al., 2011). Nickel (Ni) concentration was
significantly low in OSZ and NOZ infertile
men and molybdenum (Mo) was significantly
low in OSZ, OLZ and NOZ infertile men as
compared to fertile control (table 2). Conducted
experimental studies were proved the
beneficial of Ni for reproductive performance;
deficiency of Ni diminished physiological
characteristics of spermatozoa, which most
likely could result in impaired reproductive
ability (Yokoi et al., 2003). Ni deficiency
significantly decreases spermatozoa production
Table 2: Concentrations of trace elements [Mean±SD] in the seminal plasma of fertile and different infertile groups
epididymis, epididymal transit time, and
spermatozoa motility (Slivkova et al., 2009)
and (Yokoi et al., 2003). It is likely that Ni
affects sperm motility by changing CNG cation
channel function (Yunsan, Wanxi, 2011), CNG
cation channel involved in spermatozoa
capacitation through providing the influx of
Ca2+ to cytoplasm during capacitation.
Deficiency in Ni concentration leads to
impaired CNG channel and insufficiency in
capacitation (Cisneros‐Mejorado et al.,
2014). A weak negative correlation was found
between Ni and immotile sperm (r=-0.189,
p≤0.03), suggesting its positive role in sperm
motility.
Conducted experimental studies were proved
the beneficial of Ni for reproductive
performance; deficiency of Ni diminished
physiological characteristics of spermatozoa,
which most likely could result in impaired
reproductive ability (Yokoi et al., 2003).

Fertile group Infertile groups

Trace elements
n=25 Oligoasthenoz- Asthenoz- Normoz- P
[ppm]
oospermia n=38 oospermia n=27 oospermia n=41 value

Ca 100.12±1.88 100.98±7.67 103.45±5.25 100.12±1.88 n.s

Mg 56.06±33.54 72.89±51.66 65.00±31.91 69.21±34.60 n.s

Zn 125.18±5.92 122.18±92.61 120.49±59.12 125.74±60.33 n.s

Cu 0.095±0.01 40.126±0.084 0.136±0.169 0.160±0.160 n.s

Fe 2.31±0.04 2.47±1.45 2.43±0.92 2.76±4.27 n.s

Se 0.706±0.101 0.347±0.907 0.255±0.313 0.203±0.307 n.s

Ni 0.056±0.023* 0.035±0.014* 0.042±0.018 0.033±0.012* 0.001

Mo 0.148±0.009* 0.097±0.038* 0.108±0.028* 0.093±0.041* 0.001

ANOVA analyisis for ferile and infertile groups: n.s: non-significant, *; significant at p ≤0.001
154 Palani, A. and ALshatteri, A. /ZJPAS: 2017, 29 (s4): s150-s156
Ni deficiency significantly decreases
spermatozoa production rate of the testes,
spermatozoa density in the epididymis,
epididymal transit time, and spermatozoa
motility (Slivkova et al., 2009) and (Yokoi et
al., 2003). It is likely that Ni affects sperm
motility by changing CNG cation channel
function (Yunsang, Wanxi, 2011), CNG cation
channel involved in spermatozoa capacitation
through providing the influx of Ca2+ to
cytoplasm during capacitation. Deficiency in
Ni concentration leads to impaired CNG
channel and insufficiency in capacitation
(Cisneros‐Mejorado et al., 2014). A weak
negative correlation was found between Ni and
immotile spermatozoa (r = -0.189, p≤0.03),
suggesting its positive role in sperm motility.
Spermatozoal acrosome reaction induced by
oligosaccharide ligands on the zona surface and
sperm surface sugar receptors [mannose-ligand
receptor]; surface receptors are involved in the
prevention of acrosome reaction (Benoff et al.,
1997). Ni is involved in the activation of
mannose-ligand receptor binding (Benoff et al.,
1997), Decreased Ni concentration may
inactivate mannose-ligand receptor and prevent
acrosome reaction, this may explain why NOZ
group is infertile even they have normal sperm
parameters (table 2).
Molybdenum (Mo) is a trace element essential
for human health. It has been identified as part
of the active sites of over 50 enzymes, and may
promote normal cell function possibly by
catalyzing a variety of hydroxylations and
oxidation-reduction reactions (Zhai et al.,
2013) and ( Wirth, Mijal 2010.) . Mo has direct
effects on biological processes controlling
growth and reproductive performance (Pandey,
Singh, 2002.), Mo deficiency decreases male
libido, reduced spermatogenesis and causes
sterility (Talukdar et al., 2016) and (Upadhyay
et al., 2006).
Mo is interdependent with Cu, lower level of
one occurring in presence of high level of
another (Kumar et al., 2011). The deficiency in
Mo is a result of elevation of Cu level and vice
versa (Strachan, 2010) and (Mendel and
Bittner, 2006). Copper is a significant risk for
sperm morphology low Mo concentration is
correlated to high Cu concentration; this may
be the cause of deterioration of sperm
morphology in OSZ and ASZ groups (Sakhaee
et al., 2012). A moderate negative correlations
between (Mo and Ni) and sperm tail
abnormality were found [r=-0.325, p ≤0.001]
for Ni, and (r = -0.325, p≤0.001) for Mo,
suggestion their role in normal
spermatogenesis. Results showed a moderate
positive correlation between Ni and Mo
[r= 0.554, p ≤0.001], this results demonstrate
their positive role both in sperm quality and
male fertility.
4. CONCLUSIONS
We concluded that the trace elements
have a vital role in sperm quality because of
their role in the sperm activity and sperm
morphology. Low Ni concentration reduces
sperm motility and low Mo concentrations
have a role in the deterioration of sperm
morphology, and both (low sperm motility and
normal morphology) increase male infertility.
Conflict of Interest
We have no conflict of interest to declare.
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