Relationship Between Testosterone Plasma Concentration and
Semen Parameters in the Guys in the Aral Sea Ecological Disaster
Region
Sergey Lytaev
1a
, Valerii Erkudov
1b
, Andrey Pugovkin
1c
, Kenjabek Rozumbetov
2d
,
Azat Matchanov
2e
and Adilbay Esimbetov
3f
1
Department of Normal Physiology, St. Petersburg State Paediatric Medical University,
194100, Litovskaya str., 2, St. Petersburg, Russian Federation
2
Department of General Biology and Physiology, Berdakh Karakalpak State University, Faculty of Biology,
230100, Ch. Abdirov str, 1, Nukus, Republic of Karakalpakstan, Uzbekistan
3
Administrative Department, Nukus branch of the Samarkand Institute of Veterinary Medicine,
230102, A. Utepov St. 31, Nukus, Republic of Karakalpakstan, Uzbekistan
Keywords: Sperm Quality, Testosterone, Aral Sea Ecological Disaster, Endocrine-Disrupting Chemicals.
Abstract: This paper was aimed to study of the sperm quality and assess the possible relationship between the
spermogram parameters and the plasma total testosterone concentration in the local guys in the Nukus city
(Uzbekistan). Methods. 302 medical profiles, which included data from the analysis of ejaculate and total
testosterone plasma level in apparently local healthy subjects in the Aral Sea ecological disaster were
analyzed. According to the World Health Organization guidelines, study participants into «group A» and
«group B» (spermogram parameters less and more than the 25
th
centile respectively) were divided. Total
testosterone plasma concentration in subjects from these groups was compared. The proportion of included
patients of both groups also was calculated. Outcomes. The proportion and total testosterone level of patients
included in «group A» was lower than the proportion of study participants in «group B». The results of the
study exposed the problem of fertility disorders in men living in this South Aral Sea region. Disruption of
testosterone function is probably a consequence of the antiandrogenic effect of environmental pollutants. The
data obtained can be useful in the complex monitoring of the state of men's health among residents of
environmentally problematic regions.
1 INTRODUCTION
Southern Kazakhstan, western Uzbekistan and
northern Turkmenistan, named the Aral Sea region, is
infamous because of the ecological disaster that has
existed there since the middle of the 20
th
century
(Krivonogov, 2014; Opp, 2017; Nazhmetdinova,
2017). The Aral Sea basin became the major cotton
producer for the Soviet Union (Opp, 2017). This was
the reason for the almost complete disappearance of
the Aral Sea. Between 1918 and 1960, water
a
https://orcid.org/0000-0002-1291-0381
b
https://orcid.org/0000-0001-7351-0405
c
https://orcid.org/0000-0001-5936-6232
d
https://orcid.org/0000-0001-5967-4219
e
https://orcid.org/0000-0001-6066-1327
f
https://orcid.org/0000-0002-6958-1873
withdrawals for irrigating cotton fields increased by
about 40%, from 1960 to 2008 by more than 200%
(Opp, 2017). Thus, by 2008 almost 90% of the lake's
water volume and 74.3% of its former surface area
were lost (Opp, 2017). Salting out of heavy metals to
the bottom of the former Aral Sea, environmental
pollution by products of the metallurgical industry led
to the eolian process of the spread of micro-particles
with precipitated compounds of lead, chromium,
cadmium, mercury and etc. throughout the Aral Sea
region (Rzymski, 2019).The use of pesticides in
326
Lytaev, S., Erkudov, V., Pugovkin, A., Rozumbetov, K., Matchanov, A. and Esimbetov, A.
Relationship Between Testosterone Plasma Concentration and Semen Parameters in the Guys in the Aral Sea Ecological Disaster Region.
DOI: 10.5220/0012045100003536
In Proceedings of the 3rd International Symposium on Water, Ecology and Environment (ISWEE 2022), pages 326-330
ISBN: 978-989-758-639-2; ISSN: 2975-9439
Copyright
c
2023 by SCITEPRESS – Science and Technology Publications, Lda. Under CC license (CC BY-NC-ND 4.0)
agriculture is the cause of the accumulation of
organochlorine compounds in the environment due to
the disturbance of their elimination under these
environmental conditions (Bapayeva,
2018).Organochlorine pesticides and heavy metals
enter the human organism with food and water, where
they have an endocrine-disrupting chemicals (EDC)
effect (Gore, 2015). They interfering with all
physiological processes of action and regulation of
sex steroids and they are the cause unsatisfactory
sperm quality in in men living, for example, in
different regions of China (Ren, 2020; Zeng, 2022)
and Russia (Abou, 2020; Williams, 2022).
The scientific literature accumulates information
on the impact of environmental pollutants on the
reproductive health of the population of different
countries. Recent studies involving volunteers from
Russia and China report a direct effect of
organochlorine pesticides found in plasma on
spermatogenesis (Abou, 2020; Williams, 2022),
genetic polymorphism of enzymes contained in
spermatozoa (Miao, 2022), and hormonal profile
(Lin, 2021). These studies were carried out with the
involvement of a contingent living in ecologically
unfavorable areas where the state of the environment
is not recognized as catastrophic (Abou, 2020;
Williams, 2022). Due to the catastrophic
environmental conditions of the Aral Sea region and
the design of this work, the results obtained can be
considered unique. Present outcomes were obtained
during examination of men living under conditions of
long-term exposure to pollutants potentiated by
climatic factors. Previous studies related to the
reproductive health of the inhabitants of the Aral Sea
region are extremely limited and were carried out on
the inhabitants of Kazakhstan, located more than
1000 km from the epicenter of the Aral Sea
catastrophe (Kislitskaya, 2015; Kultanov, 2016).
On this basis the paper was aimed to study of the
sperm quality and assessment the possible
relationship between the spermogram parameters and
the plasma total testosterone concentration in guys
living in the Nukus city (Uzbekistan) – the epicenter
of the Aral Sea environmental disaster.
2 METHODS
Following the study design, we determined
sperogram parameters and plasma testosterone
concentrations in volunteers living in the Aral
ecological disaster region. Using methods of
mathematical statistics, the level of testosterone was
compared in volunteers from group «A» consisted of
patients with low and «B» – with average high values
of spermogram parameters. A correlation between the
values of the hormone and the spermogram
parameters was also calculated.
2.1 Patient Selection Procedure
The study was conducted according to the guidelines
of the Declaration of Helsinki and was approved by
the local ethics committee of the St. Petersburg State
Pediatric Medical University (protocol no. 17/3, 10
May 2018). The research was included patients
(guys) aged 23-30 years who lived in Nukus city or in
the suburbs from birth, students or office workers
who do not practice heavy physical work or are
employed in hazardous manufacturing establishment
with normal body mass index. Anamnesis for all
persons did not contain information about
inflammatory diseases, including sexually
transmitted diseases and genital trauma, varicocele,
vas deferens strictures, infectious diseases, including
mumps and COVID-19. Smokers, alcohol and drug
addicts were excluded from the study. If necessary,
all persons by a urologist to clarify the diagnosis were
examined.
2.2 Research Limitations
Thus, we received 439 anonymous patient profiles,
which included data on health status, lifestyle,
spermogram parameters and total testosterone plasma
level. 341 profiles of study participants who had no
pathology and adhered to a healthy lifestyle were
selected. Another 33 patients were excluded due to
aspermia, which may indicate a deletion of the Y
chromosome, and 6 volunteers due to the
inflammatory ejaculate – yellow color and high white
blood cell count on microcopy. Thus, the final
homogeneous sample consisted of 302 profiles of
apparently healthy subjects.
2.3 Semen Collection and Spermogram
Parameters Analysis
Collection of semen samples, analysis of spermogram
parameters, and assessment of sperm quality were
carried out according to the World Health
Organization (WHO) guidelines (Campbell, 2021).
At the request of the attending physician, patients
have excluded alcohol, overheating, including fever,
medication, physiotherapy, X-ray diagnostics, and
prostate massage, physical and emotional stress for
ten days before ejaculate examination. Semen
samples were collected by masturbation in a separate
equipped laboratory room in a special non-toxic
plastic container after 2-7 days of abstinence. The
Relationship Between Testosterone Plasma Concentration and Semen Parameters in the Guys in the Aral Sea Ecological Disaster Region
327
resulting material was placed in a thermostat and kept
at a temperature of 37
0
until liquefaction. The analysis
of spermogram parameters was carried out in all
subjects within one hour after collection. As
additional criteria for inclusion of patients in the
study, the milky-white color of the sperm and its pH,
which in healthy people should not be lower than 7.2,
were evaluated. Semen volume was measured with a
pipette, then a smear was made, which was
microscopically counting sperm concentration (10
6
per ejaculate), total motility (%), vitality (%). Sperm
quality was assessed according to the centile
distribution of spermogram parameters (Campbell,
2021). In accordance with the methodology proposed
by WHO expert’s reference intervals 25
th
-75
th
centile
for semen volume: 2.3-4.2 ml, for sperm
concentration: 36-11010
6
, for total motility: 55-
73%, for vitality: 69-88% (Campbell, 2021). Thus,
the volunteers were divided into two groups: «A»
consisted from patients with low (less than 25
th
centile), and «B» – with average high (more than 25
th
centile) values of spermogram parameters.
2.4 Blood Sample Total Testosterone
Assessment
During the cross-sectional study, blood samples were
collected from 302 subjects selected by simple
random sampling. Venous blood samples were taken
on an empty stomach until 10 am before ejaculate
collection on the same day. Blood samples were
centrifuged at 1000 rpm for 10 min to obtain the
serum. Total testosterone concentrations in serum
were measured by the direct solid-phase
chemiluminescent enzyme immunoassay
(«sandwich» method) with commercial test kits (MR-
96A Mindray microplate reader, Shenzhen Mindray
Bio-Medical Electronics Co., Ltd, China). In
accordance with the technical specifications of the
device, the reference value of total testosterone
plasma concentrations for the assay was 9.03 nmol/l
to 38.19 nmol/l, which corresponds to the values
accepted in the previous studies (Qin, 2012;
Mezzullo, 2020). Values of total testosterone plasma
concentrations below 9.03 nmol/l were not found in
patients who participated in the present study.
2.5 Statistical Analysis
The comparison of total testosterone plasma
concentrations, spermogram parameters between
patients from «group A» and «group B» was
statistically measured using Mann–Whitney U test.
Data were presented as mean valued of total
testosterone, spermogram parameters and lower or
upper limits of 95% confidence intervals (95% CI).
The Spearman's rank correlation coefficient
(Spearman's ρ) and its 95% CI (Kelley, 2019;
Pugovkin, 2021; Lytaev, 2021) between total
testosterone values and semen volume, sperm
concentration, total motility and vitality were
calculated in all patients, regardless of the
comparison group. At a value of equal to 0, the
statistical relationship was considered absent; from
0.01 to 0.29 (from -0.01 to -0.29) – weak direct
(reverse); from 0.3 to 0.69 (from -0.3 to -0.69) – the
average direct (reverse); from 0.7 to 0.99 (from -0.7
to -0.99) – strong direct (reverse); 1 (-1) – full forward
(reverse). In addition, the proportion and their 95%
confidence intervals of patients included in «group
A» and «group B» were calculated. The results were
considered to be significant at p<0,001. Estimations
were carried out using statistical programmer
(version 2.17, Norway, Oslo, 2012) and algorithm of
statistical data processing StatXact-8 with Cytel
Studio software shell version 8.0.0.
3 RESULTS AND DISCUSSION
It was found that the proportion of patients from
group A was higher than the proportion of study
participants from group B (table 1).
Table 1: The proportion and their research patients.
Group Proportion
A 0.54 (0.47; 0.60)
B 0.46 (0.40; 0.52)
Data analysis revealed statistically significantly
lower total testosterone values, sperm concentration,
total motility and vitality in patients from «group A»
compared to «group B»
The study found an average positive correlation
of total testosterone plasma level and sperm
concentration, total motility and vitality (table 2).
The
values of total testosterone and semen volume did not
correlate (table 3).
ISWEE 2022 - International Symposium on Water, Ecology and Environment
328
Table 2: The mean and of total testosterone plasma level
and spermogramm parameters in patients from «group A»
compared to «group B».
Parameters/Groups A B p
Total testosterone,
nmol/l
15.10
(14.30;
15.91)
19.47
(18.86;
20.10)
9.95
×10
-13 ***
Semen volume, ml
2.56
(2.46; 2.67)
2.75
(2.65; 2.84)
0.02
154
Sperm
concentration, 10
6
p
e
r
ejaculate
26.36
(25.16;
27.56)
72.09
(70.06;
74.13)
3.36
1×10
-6 ***
Total motility, %
29.50
(27.23;
31.38)
63.09
(61.80;
64.37)
6.22
×10
-49 ***
Vitality, %
49.28
(46.66;
51.89)
81.66
(80.59;
81.62)
3.16
2×10
-50
***
Note. P < 0.001
Table 3. The Spearman's ρ and of total testosterone plasma
level and spermogram parameters in all patients, regardless
of the comparison group.
Semen
volume
Sperm
concentrati
on
Tota
l motility
Vital
ity
Total
testosterone
0.11 (-
0.0008;
0.15)
0.45 (0.35;
0.47)
0.41
(0.31;
0.43)
0.39
(0.28;
0.41)
p-values 0.06761
2.00×10
-16
***
1.58×10
-
13 ***
2.93×10
-
12 ***
Recent studies involving volunteers from Russia
and China report a direct effect of organochlorine
pesticides found in plasma on spermatogenesis
(Abou, 2020; Williams, 2022), genetic
polymorphism of enzymes contained in spermatozoa
(Miao, 2022), and hormonal profile (Lin, 2021).
These studies were carried out with the involvement
of a contingent living in ecologically unfavorable
areas where the state of the environment is not
recognized as catastrophic (Abou, 2020; Williams,
2022). Due to the harsh catastrophic environmental
conditions of the Aral Sea region and the design of
this work, the results obtained can be considered
unique. They were obtained during examination of
men living under conditions of long-term exposure to
pollutants potentiated by climatic factors. Studies of
reproductive health among the inhabitants of the Aral
Sea region are extremely limited and were performed
on the inhabitants of Kazakhstan, located more than
1000 km from the epicenter of the Aral Sea
catastrophe. The research outcomes are not only
consistent with the data presented earlier in the
analysis of sperm from local men of the Aral Sea
region of Kazakhstan, but also complement them
(Kislitskaya, 2015; Kultanov, 2016). Scientific
articles are focused on the possible involvement of
environmental pollutants in disrupting the integrity
of the hereditary material of spermatozoa
(Kislitskaya, 2015; Kultanov, 2016), which was also
shown early in other studies (Pilsner, 2018; Williams,
2022). Thus, these works do not take into account the
EDC effect of heavy metals and organochlorine
pesticides, which disrupts the action of sex steroids
(Zeng, 2022; Ren, 2020; Williams, 2022) and their
mechanisms of regulation of their activity (Williams,
2018; Abou, 2020). At the same time, in our work
and in other papers (Keskin, 2015; Yerkudov, 2020;
Suslov, 2022), an association of quantitative
characteristics of spermogram parameters and
testosterone plasma level were reported. In the
present investigation, the proportion of healthy men
with reduced values of spermogram parameters was
54% (table 1) and the concentration of testosterone in
this group was significantly reduced compared to
study participants with satisfactory sperm quality
(table 2). Thus, the results of the present work
suggest an anti-androgenic effect of environmental
chemical pollutants on the role of anti-androgenic
EDC substances (Gore, 2015; Kislitskaya, 2015;
Williams, 2022).
4 CONCLUSION AND FUTURE
RESEARCH
The present outcomes exposed the problem of fertility
disorders from local men of the Aral Sea ecological
disaster region. More than half of the practically
healthy participants in the study had a reduced sperm
concentration, total motility and vitality. Their
unsatisfactory sperm quality was combined with a
reduced testosterone plasma level. Disruption of
testosterone function is probably a consequence of the
anti-androgenic effect of EDC in the environment of
the South Aral See region, found in previous studies.
The data obtained can be useful in the complex
monitoring of the men's health state of among
residents of environmentally problematic regions.
The analysis of the data obtained opens up the
possibility for expanding the tasks of future research
in the development of the designated project. In
particular, it is planned to compare the parameters of
the spermogram in men living in the Aral Sea region
and in the city of Tashkent, which is located at the
maximum distance from the epicenter of the Aral
ecological catastrophe. Sperm quality assessment will
Relationship Between Testosterone Plasma Concentration and Semen Parameters in the Guys in the Aral Sea Ecological Disaster Region
329
be complemented by a comparison of the hormonal
profile in subjects living in both geographic areas.
ACKNOWLEDGEMENTS
We thank for technical support of the research to Mr.
Polat Muhammatdinov and Dr. Gul'bahar
Tleumuratova, who provided invaluable assistance in
organizing the examination of patients at all stages of
work from the collection of blood and sperm samples
to the presentation of the result
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