Assessment of Seminal Plasma Trace Elements among Infertile Sudanese Males in Khartoum State, 2019

Abstract

Background: Male infertility is a multifaceted state and overlaps a lot of factors and affects infertility in about 8-5 % of the people in the world and the man is responsible for 40% of these cases. Dietary insufficiency of trace element, zinc and copper may play a role in male infertility as trace element plays an important role not only in normal testicular improvement, but also in spermatogenesis and sperm motility. Objective: The current study was intended to analyze the level of seminal plasma trace elements mainly zinc and copper amongst dissimilar groups of infertile men. Method: The concentrations of zinc and copper were measured in 160 semen samples from normozoospermic, oligoathenospermic, and azoospermic men using the atomic absorption spectroscopy, data was analyzed using the statistical software package SPSS version 17. Result: Outcomes showed that the mean values of seminal plasma zinc concentrations were significantly decreased in the two groups of infertile male subjects, azoospermic (P.value=0.000), and oligozoospermic (P.value=0.013) compared with fertile males, while there was significant decrease in seminal plasma copper concentration of azoospermic patients compared to control (P.value=0.000), and significant decrease in oligozoospermic patients compared to control (P.value=0.000).

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© 2020 |Published by Scholars Middle East Publishers, Dubai, United Arab Emirates 9
Saudi Journal of Biomedical Research
Abbreviated Key Title: Saudi J Biomed Res
ISSN 2518-3214 (Print) |ISSN 2518-3222 (Online)
Scholars Middle East Publishers, Dubai, United Arab Emirates
Journal homepage: https://saudijournals.com/sjbr
Original Research Article
Assessment of Seminal Plasma Trace Elements among Infertile
Sudanese Males in Khartoum State, 2019
Mutaz Ibrahim Hassan1*, Abdelrhman Mohamed Sid Ahmed2, Mosab Nouraldein Mohammed Hamad3, Gamal Mahmoud
Elimairy4
1Faculty of Medical Laboratory Science, Shendi University, Shendi, Sudan
2Clinical Chemistry Department, Faculty of Medicine, Kassala University, Kassala, Sudan
3Medical Laboratory Science Department, Faculty of Health Science, Elsheikh Abdallah Elbadri University, Sudan
4Faculty of Medicine, Elrazi University, Azhari Road, Khartoum North, Sudan
DOI: 10.36348/sjbr.2020.v05i01.003 | Received: 01.01.2020 | Accepted: 16.01.2020 | Published: 30.01.2020
*Corresponding author: Mutaz Ibrahim Hassan
Abstract
Background: Male infertility is a multifaceted state and overlaps a lot of factors and affects infertility in about 8–5 % of
the people in the world and the man is responsible for 40% of these cases. Dietary insufficiency of trace element, zinc
and copper may play a role in male infertility as trace element plays an important role not only in normal testicular
improvement, but also in spermatogenesis and sperm motility. Objective: The current study was intended to analyze the
level of seminal plasma trace elements mainly zinc and copper amongst dissimilar groups of infertile men. Method: The
concentrations of zinc and copper were measured in 160 semen samples from normozoospermic, oligoathenospermic,
and azoospermic men using the atomic absorption spectroscopy, data was analyzed using the statistical software package
SPSS version 17. Result: Outcomes showed that the mean values of seminal plasma zinc concentrations were
significantly decreased in the two groups of infertile male subjects, azoospermic (P.value=0.000), and oligozoospermic
(P.value=0.013) compared with fertile males, while there was significant decrease in seminal plasma copper
concentration of azoospermic patients compared to control (P.value=0.000), and significant decrease in oligozoospermic
patients compared to control (P.value=0.000).
Keywords: Seminal Plasma, Trace Elements Infertile Male.
Copyright @ 2020: This is an open-access article distributed under the terms of the Creative Commons Attribution license which permits unrestricted
use, distribution, and reproduction in any medium for non-commercial use (NonCommercial, or CC-BY-NC) provided the original author and source
are credited.
INTRODUCTION
Infertility is complicated and has manifold
reasons and outcomes depending on the sex, sexual
history, life style and cultural environment [1].
Infertility have an effect on about 8% to 12% of the
world’s population and in about half of cases, men are
either the single reason or add to couple’s infertility [2].
Seminal plasma is very vital for sperm metabolism,
function, survival, and transport in the female genital
tract. Cations such as Na, K, Ca, and Mg set up osmotic
balance, as necessary trace elements are components of
several essential enzymes in the seminal plasma [3].
The probable effect of the trace elements particularly
Cu and Zn on male infertility is a topic of great interest
[1], rising proof of a direct association of zinc was
found with seminal parameters [4]. Zinc is the second
major element, after iron, in seminal plasma. It
maintains the cell membrane and nuclear chromatin of
spermatozoa [5]. It may also have an antibacterial role
[6], defends testis from the degenerative alterations [7].
It regulates mechanism of capacitation and acrosome
reaction [8]. Zinc has a significant function in normal
testicular growth, spermatogenesis and sperm motility
[9]. It is a cofactor for a number of metalloenzymes in
human semen, concerned in DNA transcription and
protein production. Shortage of zinc in the reproductive
system leads to hypogonadism and gonadal hypo
function [6, 10, 11] reported that zinc in seminal plasma
is implicated in nuclear chromatin decondensation and
acrosin activity. Zinc insufficiency in the nucleus may
destabilize the quaternary structure of chromatin; a
characteristic essential for the fertilizing capability of
the spermatozoa [12, 13] conducted an experiment in
adult males and reported that production of testosterone
b [14, 8]. Preceding study discussed the role of zinc in
exchange of testosterone into its biologically active
form 5α-dihydrotestosterone (DHT) and mentioned that
reduction of dietetic zinc May reduce semen volume
and serum testosterone levels [15]. Zinc content in
seminal plasma is mostly secreted by the prostate gland
and may reflect prostatic function. Copper is an

Mutaz Ibrahim Hassan et al., Saudi J Biomed Res, Jan., 2020; 5(1): 9-13
© 2020 |Published by Scholars Middle East Publishers, Dubai, United Arab Emirates 10
imperative element for many metalloenzymes and
metalloproteins that are concerned in energy
metabolism. It works in diverse ways in order to
preserve normal environment for spermatozoa for
normal fertilization to occur [16]. Though, a higher
level is toxic to a variety of cells, including human
spermatozoa [16]. In vitro studies, established that
utilize of Cu in intrauterine devices stop conception
[16]. The recent study was intended to assess seminal
plasma levels of zinc and copper and to correlate their
concentrations with a variety of semen parameters
between fertile and infertile Sudanese male.
Rationale
The determination of spermatozoa
concentration, morphology and motility remains the
primary clinical means for the evaluation of male
infertility. Nutritional deficiency of trace element, zinc
and copper may engage in recreation a role in male
infertility.
This study aimed to enrich data about the level
of these vital trace elements between infertile Sudanese
men.
OBJECTIVES
General objective
To assess the seminal plasma trace elements
among infertile Sudanese males in Khartoum state.
Specific Objectives
 To estimate seminal plasma zinc & copper
among infertile males.
 To compare between Azoospermia and
Oligoasthenospermia in the level of seminal
plasma trace elements.
MATERIALS AND METHODS
Study design
Case control hospital based study.
Study area
Reproductive care center in Almuk Nemer
Street in Khartoum state.
Study Duration
The study was carried out during the period
from December 2016 to September 2019.
Study Population
Infertile Sudanese male referred to the study
setting, by various fertility centers and hospitals in
Khartoum state during study period.
Inclusion Criteria
Males with oligoasthenospermia and
azoospermia as test group and normal males (age group
24 – 78 years) belong to the same socioeconomic status
were selected as control group.
Exclusion Criteria
Infertile men under hormonal treatment and
diabetic patients.
Sample Size and Study population
One hundred and sixty blood samples were
collected in this study.
Case group: 80infertile male patients
(oligoasthenospermia and azoospermia).
Control group: 80 apparently healthy
individuals.
Blood Sample Collection and Analysis
Before collection, a local antiseptic (70%
alcohol) was used to clean the skin, venous blood
(about 4 ml) was be taken from each participant in
lithium heparin container, then the centrifugation will
be done at 3000 rpm for 3-5 minute to obtain plasma,
used for measurement of zinc and copper.
Collection of Semen Samples and analysis
Semen was collected by masturbation into a
sterile plastic specimen container at the hospital.
Subjects were instructed to abstain from ejaculation for
at least 72 hours prior to producing the semen sample.
The sample was liquefied for at least 20 minutes, but no
longer than 1 hour prior to performing a routine semen
analysis, which included measurements of volume, pH,
sperm concentration, sperm motility and morphology
and direct microscopic examination.
Estimation of sperm counting will be done
using the Neubauer chamber. Sperm analysis was
carried out according to the World Health Organization
guidelines, based on the sperm concentration the
infertile subjects were classified as follows:
 Normozoospermia (> 20 million sperm /ml
and normal semen profile)
 Oligoasthenospermia (<20 million sperm/ml
and motility grade C or D)
 Azoospermia (no spermatozoa)
In proven fertile controls, the sperm count
ranged from 20 – 120 million sperm /ml
Seminal Plasma Collection and Analysis
Sample Preparation
Seminal plasma specimens were diluted 1:10
with 0.5% v/v HNO3 to determine the concentrations of
zinc and copper.

Mutaz Ibrahim Hassan et al., Saudi J Biomed Res, Jan., 2020; 5(1): 9-13
© 2020 |Published by Scholars Middle East Publishers, Dubai, United Arab Emirates 11
Instrument
Atomic absorption spectrophotometer (Buck
Scientific, model 210 VGP).
Principle of Atomic Absorption Spectrophotometer
Atomic absorption spectrophotometer utilizes
the phenomenon that atoms absorb, radiation of
particular wavelength. When the light from a hollow
cathode lamp shines into the flame, ground state atoms
of the element wanted to be measured absorb some of
the light resulting in a decrease of the light (atomic
absorption).
Data Collection
Direct questionnaire was done to obtain
clinical data for each participant and sample.
Data Analysis
Data was analyzed using Statistical Package
for Social Science Software (SPSS).
Ethical Considerations
This study was approved by the research
committee – College of Medical Laboratory Sciences -
Shendi University. Informed consent was obtained from
each participant before taking the samples.
RESULTS
This is a case control hospital based study
conducted in Khartoum state in the Reproductive Care
Center during the period from December 2016 to
September 2019. This study included 160 samples, 80
from these samples were collected from infertile males
as case group (40 of them collected from
Azoospermia25% and the rest from
Oligoathenospermia25%) and the rest of the samples
collected from normal male (Normozoospermia 50%)
as control group. The results of the study were
presented in texts and tables.
Table-1: Independent sample T.test showed mean and std. deviation of seminal plasma Copper among case group
(Azoospermia and Oligoasthenospermia) and Control group
Azoospermia
Oligoasthenospermia
Case
Control
Case
Control
No
40
80
40
80
Mean (mg/dl)
0.05
0.13
0.057
0.123
Std. Dev
0.038
0.037
0.035
0.037
P. Value
0.000
0.000
P. value ≤ 0.05 is considered significant.
Table-2: Independent sample T. test showed mean and std. deviation of seminal plasma Zinc among case
(Azoospermia and Oligoasthenospermia) group and Control group
Azoospermia
Oligoasthenospermia
Case
Control
Case
Control
No
40
80
40
80
Mean (mg/dl)
16.0
22.1
20.7
22.1
Std. Dev
6.55
4.44
5.76
4.44
P. Value
0.000
0.13
P. value ≤ 0.05 is considered significant.
Table-3: Paired sample T. test showed Mean of seminal Trace elements among (Oligoasthenospermia) and
Azoospermia
Paired Group
Mean (mg/dl)
N
Std. Deviation
P. Value
Copper level AZO
0.05
40
0.038
0.474
Copper level OAS
0.056
40
0.035
Zinc level AZO
16.0
40
6.55
0.006
Zinc level OAS
20.6
40
5.76
P. value ≤ 0.05 is considered significant.
DISCUSSION
Statistical analysis of the gathered data shows
significant decrease in mean of seminal plasma Copper
among case (Azoospermia) group compared to Control
group with P. Value (0.000). The results revealed a
marked decrease in seminal plasma copper in case
(oligoasthenospermia) group, mean (0.057 mg/l) while
it is (0.123mg/l) in control group. Agree with [17, 18]
The laboratory analysis for seminal plasma
samples demonstrate very low level of zinc in patients
with azoospermia compared to control group with mean
of (16.0 mg/l) and (22.1) respectively. also the results
show insignificant decrease in seminal plasma zinc
level among patients with oligoasthenospermia
compared to control group, with mean of (20.7 mg/l)
and (22.1mg/l) respectively. This result agrees with
Yosra M et al 2015and Mohammad Shoaib Khan 2011.

Mutaz Ibrahim Hassan et al., Saudi J Biomed Res, Jan., 2020; 5(1): 9-13
© 2020 |Published by Scholars Middle East Publishers, Dubai, United Arab Emirates 12
The trace elements Zn and magnesium (Mg)
found in seminal plasma originate primarily from the
prostate gland and may reflect prostatic secretary
function. Studies have suggested that Mg may play a
role in spermatogenesis, particularly in sperm motility
[19]. The same is true for Zn, although the results of
several studies are still contradictory [20]. Zn, B
complex vitamins (B6, B12 and folic acid), vitamin C,
and antioxidants are critical nutrients in the male
reproductive system for proper hormone metabolism,
sperm formation, and motility [21]. Moreover, a
positive correlation has been observed between the
sperm count and seminal plasma Zn concentration in
oligozoospermic and azoospermic patients [22].
CONCLUSION
According to our result, we conclude that there
was significant decrease in seminal zinc and copper in
patients with azoospermia and oligoasthenospermia as
compared with control group.
RECOMMENDATIONS
According to our outcomes we recommend:
 Molecular based study to confirm effect of
zinc deficiency on the spermatozoa integrity.
 Experimental supplementation of zinc &
copper for patients with Azoospermia and
Oligoasthenospermia.
 Health education plan should be achieved to
aware population for the significance of
micronutrient.
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