The incidence of infertility is approximately 15%. Out of this percentage males and females contribute equally in 50% of cases respectively. In our country there are some data, for the western part of North Macedonia, which suggest that male factor infertility is solely responsible in approximately 44.2% of cases. We aimed to evaluate the efficacy of antioxidant therapy in amelioration of semen parametres the reduction of oxidative stress. An antioxidant formula was used in a male, age of 30, previously diagnosed with oligoasthenozoospermia, in a period of six months, from May to October 2019. Semen analysis with microscope phase-contrast was done in the department of Physiology, and measurement of oxidative stress markers such as malonedialdehyde (MDA) and Protein Carbonyl (PC) in the department of Biochemistry, Medical Faculty, University of Tetova, with 3 days of abstinence. Difference can be seen in the increase of Total sperm Motility from 20% in the first analyse to 50% in the last one. Concentration improved from 10x106 spermatozoa per millilitres to 25 x 106 spermatozoa per ml. Significant decrease was seen in the levels of MDA, 5.057 in the beginning becoming to 3.431 by the end, while there was seen a small decrease of PC from 0.671 to 0.659 by the end of antioxidant intake. This case study is an encouragement for further research involving a larger sample size, which can help in determining the appropriate antioxidant compounds as well as a good study design that will confirm the positive effect of antioxidant intake.

1. RELATIONSHIP OF OXIDO-
REDUCTION POTENTIAL AND SPERM
MORPHOLOGY DISORDERS OF
INFERTILE MEN
1. Department of Physiology, Faculty of Medical Sciences, University of Tetova, North Macedonia
2. Department of Biochemistry, Faculty of Medical Sciences, University of Tetova, North
Macedonia
VEGIM ZHAKU1, SHEQIBE BEADINI2
4th International Istanbul
Scientific Research Congress

2. Infertility
≠
January
February
March
April
May
June
July
August
September
October
November
December
12

3. Male Infertility
INFERTILITY JUSTA PROBLEM

4. Prevalence

5. Male infertility in North Macedonia
1. Zhaku V, Beadini Sh, Beadini N, Xhaferi V, Golaboska J (2019). The role of semen analysis in the expression of male infertility in southwesternpart of North Macedonia (Experiences from 7
municipalities). UNIVERSI–Int J of Ed Sci Tech Innov Health and Env. 5(2):96-104.
2. Beadini Sh, Beadini N, Durmishi N, Asani L, Zhaku V, Beadini A, Bexheti E, Denjalli I (2014). The impact of testosterone, spermogram and sperm culture in the appearance of male infertility
in Western Macedonia. UNIVERSI–Int J of Ed Sci Tech Innov Health and Env. 1(1):123-129.
http://www.stat.gov.mk/publikacii/2019/RegioniteVoRM.2019.pdf

6. Risk factors
Emotional
Stress
Smoking
Alcohol &
Drugs
Age
Inappropriate
food
Pollution
Radiation
Cancer Inactivity &
Obesity

7. Diagnostic categories in male
infertility

8. Oxidative Stress & Male Infertility
Catalase
Glutathione
peroxidase
Superoxide
dismutase
Free radicals are like robbers, deficient in Energy
They attack and snatch energy from other cells
To satisfy themselves
Oxidation
Antioxidant
effect

9. ROS Paradox (delicate balance)
ROS act as double edged sword

10. Aim of the study
• To evaluate differences on sperm morphology,
MDA Levels, TAC levels and Sperm Deformity
Index between fertile and infertile group.
• Correlation between sperm morphology in
infertile patients with levels of MDA, and TAC.
• Correlation between SDI in infertile patients
and levels of MDA and TAC.

11. Material &
Methods

12. STUDY
PROTOCOL
• Sample size
(28)
• Inclusive and
Exclusive
criteria
• Written
informed
consent of the
nature of the
study
• GCP &
Helsinki
Declaration
DIAGNOSTIC
PROCEDURE
• Spermogram
evaluation
• Sperm
morphology
• MDA and TAC
evaluation
• Sperm Deformity
Index (SDI)
STATISTICAL
ANALYSIS
• Mean and SD
values
• Pearson
correlation
• Newmann Kuels
Test
• P<0.001
statistically
significant
𝑺𝑫𝑰 =
𝑻𝒐𝒕𝒂𝒍 𝒏𝒐. 𝒐𝒇 𝒎𝒐𝒓𝒑𝒉𝒐𝒍𝒐𝒈𝒊𝒄𝒂𝒍 𝒂𝒏𝒐𝒎𝒂𝒍𝒊𝒆𝒔
𝑻𝒐𝒕𝒂𝒍 𝒏𝒐. 𝒐𝒇 𝒔𝒑𝒆𝒓𝒎 𝒊𝒏𝒗𝒆𝒔𝒕𝒊𝒈𝒂𝒕𝒆𝒅

13. Results
0%
10%
20%
30%
40%
50%
60%
70%
Sperm
morphology
(%)
Normozoospermic Teratozoospermic
Mean and SD values of sperm morphology in
control group and infertile group
P<0.001
0.00
0.20
0.40
0.60
0.80
1.00
1.20
1.40
Sperm
Deformity
Index
Score
Normozoospermic Teratozoospermic
Mean and SD values of SDI in control group
and infertile group
P<0.001
0.00
10.00
20.00
30.00
40.00
50.00
60.00
70.00
80.00
Level
of
TAC
(U/ml)
Normozoospermic Teratozoospermic
Mean and SD values of TAC level in control
group and infertile group
P<0.001
0.00
0.20
0.40
0.60
0.80
1.00
1.20
1.40
Sperm
Deformity
Index
Score
Normozoospermic Teratozoospermic
Mean and SD values of SDI in control group
and infertile group
P<0.001

14. y = 0.1649x + 0.1624
R² = 0.9335
0.6
0.7
0.8
0.9
1
1.1
1.2
1.3
1.4
2.500 3.500 4.500 5.500 6.500
Sperm
Deformity
Index
Score
MDA Level (nmol/mL)
Relation between SDI and MDA level
(r= 0.96)
y = -0.0439x + 1.5119
R² = 0.7545
0.6
0.7
0.8
0.9
1
1.1
1.2
1.3
1.4
7 12 17
Sperm
Deformity
Index
Score
TAC Level (U/ml)
Relation between SDI and TAC level
(r= - 0.86)
y = -0.0802x + 0.5874
R² = 0.8637
0%
5%
10%
15%
20%
25%
30%
35%
40%
3.000 4.000 5.000 6.000 7.000
Sperm
Morphology
(%)
MDA Level (nmol/mL)
Relation between sperm morphology
and MDA level (r= - 0.92)
y = 0,0233x - 0,0909
R² = 0,831
0%
5%
10%
15%
20%
25%
30%
35%
40%
7 12 17
Sperm
Morphology
(%)
TAC Level (U/ml)
Relation between sperm morphology
and TAC level (r= 0.91)

15. Discussion and Conclusion
• Differences in sperm morphology, levels of MDA, levels of
TAC and SDI, between the control group and infertile patients
were significant (p<0.001).
• Sperm Morphology in infertile patients had negative correlation
with MDA levels (r= -0.92) and positive correlation with TAC
Levels (r= 0.91) .
• Positive correlation was observed between SDI and MDA level
(r= 0.96), and inversely related to TAC (r= - 0.86) which indicates
that high level of ROS may be responsible for the
morphological deformities of the sperm cells.
• The rationale for antioxidant therapy in infertile patients should
be based on raised OS status and decreased semen
parameters.
• Further randomized controlled trial.

16. Thank you for your attention!

17. References
1. WHO Laboratory Manual for the Examination and Processing of Human Semen. 5th ed. Geneva: World Health Organization; 2010.
2. Aziz N., Buchan I., Taylor C., Kingsland C.R. and Lewis-Jones I: The sperm deformity index: a reliable predictor of the outcome of oocyte
fertilization in vitro. Fertil. Steril. 1996; 66: 1000-1008.
3. Villaverde, A.I.S.B.; Netherton, J.; Baker, M.A. From Past to Present: The Link Between Reactive Oxygen Species in Sperm and Male
Infertility. Antioxidants 2019, 8, 616.
4. Ritchie C, Ko EY. Oxidative stress in the pathophysiology of male infertility. Andrologia.2020;00:e13581. https://doi.org/10.1111/and.13581
5. Ayala A, Muñoz MF, Argüelles S. Lipid peroxidation (2014): Production, metabolism, and signaling mechanisms of malondialdehyde and 4-
hydroxy-2-nonenal. Oxidative Medicine and Cellular Longevity, vol.2014; 1-31 https://doi.org/10.1155/2014/360438
6. Sulagna Dutta, Ahmad Majzoub & Ashok Agarwal (2019) Oxidative stress and sperm function: A systematic review on evaluation and
management, Arab Journal of Urology, 17:2, 87-97, https://doi.org/10.1080/2090598X.2019.1599624
7. Du Plessis SS, Agarwal A, Halabi J, et al. Contemporary evidence on the physiological role of reactive oxygen species in human sperm
function. J Assist Reprod Genet. 2015;32:509–520. https://doi.org/10.1007/s10815-014-0425-7
8. Agarwal A., Sengupta P. (2020) Oxidative Stress and Its Association with Male Infertility. In: Parekattil S., Esteves S., Agarwal A. (eds) Male
Infertility. Springer, Cham. https://doi.org/10.1007/978-3-030-32300-4_6
9. Beadini Sh, Beadini N, Durmishi N, Asani L, Zhaku V, Beadini A, Bexheti E, Denjalli I (2014). The impact of testosterone, spermogram and
sperm culture in the appearance of male infertility in Western Macedonia. UNIVERSI–Int J of Ed Sci Tech Innov Health and Env. 1(1):123-
129.
10. Zhaku V, Beadini Sh, Beadini N, Xhaferi V, Golaboska J (2019). The role of semen analysis in the expression of male infertility in
southwesternpart of North Macedonia (Experiences from 7 municipalities). UNIVERSI–Int J of Ed Sci Tech Innov Health and Env. 5(2):96-
104.
11. Strategjia Nacionale për Shëndet Seksual dhe Riprodhues në Republikën e Maqedonisë 2010-2020 (draft verzioni). Ministira e Shëndetësisë
së Republikës së Maqedonisë, Shkup, 2009.
12. Zhaku V, Beadini Sh, Beadini N, Murtezani B. Combination of Maca, Korean ginseng extract and antioxidant therapy for male with
oligoasthenozoospermia: case study. Journal of Hygienic Engineering and Design (2019). Vol.26, pp.28-35
13. Durmishi N. Veçoritë morfologjike dhe funksionale të spermatozoidëve si faktorë të infertilitetit Human, Disertacion i doktoraturës, Universiteti
i Mjekësisë, Fakulteti i Shkencave Mjekësore Teknike – Tiranë, 2014
14. C. Martinez, C. Mar, M. Azcarate, P. Pascual,J.M. Aritzeta , A. Lopez- Urrutia : Sperm Motility Index: A quick screening parameter from sperm
quality analyzer IIB to rule out ologo and asthenozoospermia in male fertility study: Human Reproduction 2000;vol.15 no. 8 :1727-1733
15. D. Pannadis, J. Matalliotakis, S. Skiadopoulos, D. Rousso, E. Koumantakis, M. Mamopoulos: The Sperm Deformity and Multiple Anomalies
Indices: Are They Reliable In The Identification Of Fertile and Infertile Semen? Int. J. Fertility 1998; 43(3): 159-164.
16. Gomez E, Buckingham DW, Brindle J, Lanzafame F, Irvine DS, Aitken RJ. Development of an image analysis system to monitor the retention
of residual cytoplasma by human spermatozoa: correlation with biochemical markers of the cytoplasmic space, oxidative stress, and sperm
function. J Androl 1996;17:276–87.