Relationship Between Testosterone Plasma Concentration and

Semen Parameters in the Guys in the Aral Sea Ecological Disaster

Region

Sergey Lytaev

, Valerii Erkudov

, Andrey Pugovkin

, Kenjabek Rozumbetov

Azat Matchanov

and Adilbay Esimbetov

Department of Normal Physiology, St. Petersburg State Paediatric Medical University,

194100, Litovskaya str., 2, St. Petersburg, Russian Federation

Department of General Biology and Physiology, Berdakh Karakalpak State University, Faculty of Biology,

230100, Ch. Abdirov str, 1, Nukus, Republic of Karakalpakstan, Uzbekistan

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

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

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

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https://orcid.org/0000-0001-6066-1327

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

 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

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

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

-75

centile

for semen volume: 2.3-4.2 ml, for sperm

concentration: 36-11010

, 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

centile), and «B» – with average high (more than 25

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

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

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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