G Model

PHYMED-51608; No. of Pages 5 ARTICLE IN PRESS

Phytomedicine xxx (2014) xxx–xxx

Contents lists available at ScienceDirect

Phytomedicine
journal homepage: www.elsevier.de/phymed

Effects of Nigella sativa L. seed oil on abnormal semen quality in

infertile men: A randomized, double-blind, placebo-controlled clinical
trial
M. Kolahdooz a , S. Nasri a , S. Zadeh Modarres b , S. Kianbakht c , H. Fallah Huseini c,∗
a
Department of Biology, Payamenoor University, Tehran, Iran
b
Department of Obstetrics and Gynecology, Mahdieh Hospital, Shahid Beheshti University of Medical Sciences, Tehran, Iran
c
Pharmacology and Applied Medicine Department of Medicinal Plants Research Center, Institute of Medicinal Plants, ACECR, Alborz, Karaj, Iran

a r t i c l e i n f o a b s t r a c t

Article history: In recent years, wide utilization of herbal drugs has encouraged scientists to determine their impressive
Received 27 August 2013 effects on health. Since Nigella sativa L. seed (N. sativa) has many uses including infertility in traditional
Received in revised form 1 December 2013 medicine, the effects of Nigella sativa L. seed oil on abnormal semen quality in infertile men with abnormal
Accepted 22 February 2014
semen quality are of interest. This study was conducted on Iranian infertile men with inclusion criteria of
abnormal sperm morphology less than 30% or sperm counts below 20 × 106 /ml or type A and B motility
Keywords:
less than 25% and 50% respectively. The patients in N. sativa oil group (n = 34) received 2.5 ml N. sativa oil
Nigella sativa seed oil
and placebo group (n = 34) received 2.5 ml liquid paraffin two times a day orally for 2 months. At baseline
Male infertility
Abnormal semen quality
and after 2 months, the sperm count, motility and morphology and semen volume, pH and round cells
Traditional medicine as primary outcomes were determined in both groups. Results showed that sperm count, motility and
morphology and semen volume, pH and round cells were improved significantly in N. sativa oil treated
group compared with placebo group after 2 months. It is concluded that daily intake of 5 ml N. sativa oil
for two months improves abnormal semen quality in infertile men without any adverse effects.
© 2014 Published by Elsevier GmbH.

Introduction Ranunculaceae and commonly is known as black seed (Malhotra

2004). N. sativa seed is often used as a spice, food preservative and
Infertility resulting in involuntary childlessness is a social and medicine by people in Asia, Middle East and Africa (Goreja 2003).
clinical problem affecting couples worldwide. In a nation-wide N. sativa is one of the components of Iranian traditional remedies
study in France infertility was reported in 11% and 24% of couples for treatment of infertility (Salehi Surmaghi 2008). The efficacy
after 12 and 24 months of unprotected intercourse (Slama et al. of N. sativa oil has been reported on some reproductive traits of
2012). Among infertile couples, male factors, contribute to more male chickens (Samir Bashandy 2007) and male rat’s spermato-
than half of all cases of childlessness (Araoye 2003). Infertility does genesis and fertility (Al-Sa’aidi et al. 2009; Mukhallad et al. 2009;
not affect the couples’ life only, but it also affects the healthcare Abdulkarim and Al-Sardary 2009), but no clinical study has been
services and social environment (Zafar and Mohsin 2001). Several done to investigate the effects of N. sativa oil on infertile men so
factors such as congenital diseases, hormonal imbalance, genetic far. The N. sativa oil has a wide margin of safety (Zaoui et al. 2002)
and nutritional defects are among disorders in the normal function and has been administered to abnormal metabolic syndrome as
of testicle which causes men’s infertility (Whitman-Elia and well as diabetic patients at the dosage of 5 ml daily for two months
Baxley 2001). A number of antioxidants, nutritional therapies, without any side effects (Najmi et al. 2008; Mohtashami et al.
and botanical medicines have proven beneficial in treating male 2011). Thus in the light of the above findings, the present study
infertility (Sinclair 2000; Agarwal et al. 2008; Chen et al. 1999). N. was conducted to determine the effects of consumption of 5 ml
sativa is a medicinal plant which belongs to the botanical family daily N. sativa oil on infertile men having abnormal semen quality.

Materials and methods

∗ Corresponding author at: Institute of Medicinal Plants, ACECR, Karaj-Qazwain
Freeway, Supa Boulevard, Jahade-Daneshgahi Research Society, Alborz, Karaj, Iran.
N. sativa oil and mineral oil were purchased from local market in
Tel.: +98 26 34764010 20; fax: +98 26 34764010 20. the Tehran city. N. sativa oil was the product of Barij Essence Com-
E-mail address: huseini [email protected] (H.F. Huseini). pany, Kashan, Iran. N. sativa oil had been prepared by cold press

http://dx.doi.org/10.1016/j.phymed.2014.02.006
0944-7113/© 2014 Published by Elsevier GmbH.

Please cite this article in press as: Kolahdooz, M., et al., Effects of Nigella sativa L. seed oil on abnormal semen quality in infertile men:
A randomized, double-blind, placebo-controlled clinical trial. Phytomedicine (2014), http://dx.doi.org/10.1016/j.phymed.2014.02.006
G Model
PHYMED-51608; No. of Pages 5 ARTICLE IN PRESS
2 M. Kolahdooz et al. / Phytomedicine xxx (2014) xxx–xxx

Enrollment Assessed for eligibility (n=104)

Excluded (n=24)
♦ Not meeting inclusion criteria (n=10)
♦ Declined to participate (n=12)
♦ Other reasons (n=0)

Randomized (n=80)

Allocation
Allocated to intervention (n=40) Allocated to intervention (n=40)
♦ Received allocated intervention (n=40) ♦ Received allocated intervention (n=40)
♦ Did not receive allocated intervention (give ♦ Did not receive allocated intervention (give
reasons) (n=0) reasons) (n=0)

Follow-Up
Lost to follow-up (they did not come for Lost to follow-up (they did not come for
monthly visit due to personal reasons) (n=6) monthly visit due to personal reasons) (n=6)

Discontinued intervention (give reasons) (n=0) Discontinued intervention (give reasons) (n=0)

Analysis
Analysed (n=34) Analysed (n=34)
♦ Excluded from analysis (give reasons) (n=0) ♦ Excluded from analysis (give reasons) (n=0)

Fig. 1. The CONSORT flow diagram describing the progress of the patients through the trial.

procedure as indicated in its brochure. 0.1 ml of a mixture consist- Exclusion criteria

ing of chlorophyll and red chili pepper extract in equal proportions
dissolved in oil was added to 100 ml of mineral oil and N. sativa oil Men with azoospermia, gonad abnormalities, varicocelle and
in order to give color and taste to placebo. The N. sativa and mineral urinary retention and infections; those who have taken hormone
oils were filled in 150 ml colored bottles and marked as A and B. treatment; men with genital accessory gland infection; men with
recent medical or surgical illnesses; those who have time limitation
Determination of total fatty acid content of N. sativa oil (e.g. those whose wives are above 35) and cigarette smokers.

The concentration of fatty acids in N. sativa oil was measured by Protocol

the gas–liquid chromatography method (Duchateau et al. 1996).
Eighty Iranian infertile men 20–45 years old, referred to the
Infertility Center of Mahdieh Hospital in Tehran, Iran were selected.
Determination of chemical composition of the volatile oil of N.
All patients were initially visited by urologist, interviewed and
sativa oil
questioned about their sexual behavior, history of prior surgical
interventions or childhood diseases such as cryptorchidism that
25 ml of the N. sativa oil were hydro-distilled for 4 h. The dis-
affect fertility and family history of infertility. A written informed
tillate or volatile oil was diluted using n-hegxane and analyzed by
consent was obtained from each participant. The ethics committee
the GC–MS using Agilent 6890 system (Adams 2007; McLafferty
of the Infertility Center of Mahdieh Hospital approved the protocol.
and Stauffer 1989).
The study was double-blind and block randomization was used for
allocation of the patients to the N. sativa oil and placebo groups.
Participants Forty patients participated in each group of the study. However,
34 patients in each group finished the study (Fig. 1). The sample
Inclusion criteria size calculated to detect 21 × 106 sperm/ml difference of sperm
count between N. sativa oil and placebo groups, considering type 1
Healthy men 20–45 years of age with infertility lasting more error = 0.05 and 80% power was 34 patients per group. The patients
than one year; men not receiving any drugs for infertility during in N. sativa oil and placebo groups received N. sativa oil and mineral
the last 3 months; men without any history of renal, hepatic or oil respectively each at the dose of 2.5 ml, two times a day orally,
other chronic illnesses; men having semen samples abnormality for 2 months. All participants were requested to report any adverse
(sperm abnormal morphology less than 30% or sperm count blow effects.
20 × 106 ml or type (A) motility less than 25% and type (B) motility At baseline and after 2 months, the semen samples were
less than 50%) according to WHO standard guidelines (WHO 1999). obtained from the patients in both groups, 48–72 h after the

Please cite this article in press as: Kolahdooz, M., et al., Effects of Nigella sativa L. seed oil on abnormal semen quality in infertile men:
A randomized, double-blind, placebo-controlled clinical trial. Phytomedicine (2014), http://dx.doi.org/10.1016/j.phymed.2014.02.006
G Model
PHYMED-51608; No. of Pages 5 ARTICLE IN PRESS
M. Kolahdooz et al. / Phytomedicine xxx (2014) xxx–xxx 3

patients last sexual contact for determination of semen volume, Table 1

Fatty acid contents of fixed oil component of N. sativa oil.
round cells and pH and sperm count, motility and morphology
according to WHO guidelines (WHO 1999). Fatty acids Percentage

Myristic acid 0.2090

Semen quality assessment Palmitic acid 12.84
Palmitoleic acid 0.2812
All semen parameters were primary outcome. Heptadecanoic acid 0.1047
Stearic acid 3.3884
Oleic acid 22.5836
Sample collection Linoleic acid 58.2370
Arachidic acid 0.2250
The patients’ semen samples were collected (after 72 h absti- Eicosenoic acid 0.3839
Behenic acid 0.0356
nence from ejaculation) in the laboratory.
Lignoceric acid 0.0414

Sperm count
Table 2
Semen smear was prepared from mixture of 10 ␮l semen and Chemical compositions of volatile oil component of N. sativa oil.

10 ␮l of Giemsa stain on slide, dried in air and washed under run- Compound Percentage
ning tap water and then dried in air. Sperm count (sperm density) ␣-Thujene 13.95
was determined by routine laboratory procedure and expressed as ␣-Pinene 3.74
million/ml of semen. 2,4,10,Thujaden 0.08
Camphene 0.05
2-Heptenal(z) 0.18
Sperm motility
Sabinene 1.52
␤-Pinene 3.27
Quantitative motility (expressed as percentage) was deter- ␳-Cymene 51.62
mined by counting both motile and immotile spermatozoa per unit Limonene 1.95
area. Sperm motility was expressed as class A (fast moving for- 1,8-Cineole 0.07
␥-Terpinene 0.17
ward), class B (slow moving forward), class C (moving at same
cis-Thujon 0.06
place), Immotile (with no moving) and class A + B (moving fast (−)-cis-Sabinol 0.11
forward and moving slow forward) motilities. 4-ol-Terpineol 0.27
Dodecane(n) 0.07
Thymoquinone 14.48
Sperm morphology
Isobornyl acetate 0.09
Carvacrol 0.96
The semen smear was prepared from mixture of 10 ␮l semen ␣-Longipinene 0.28
and 10 ␮l of Giemsa stain on slide, dried in air and washed under ␤-Longipinene 0.82
running tap water and then dried in air. Sperm morphology (nor-
mal/abnormal) expressed as percentage was assessed under oil Table 3
(Immersion oil) at 100× magnification of microscope using ocu- The demographic and baseline data in N. sativa oil and placebo groups (mean ± SE).
lar micrometer (ocular micrometer should be calibrated with stage
N. sativa seeds Placebo p-Value
micrometer to measure the exact size).
oil (N = 34) (N = 34)

Duration of Infertility (year) 4.3 ± 0.6 3.9 ± 0.6 1.631

Other semen parameters
Age (year) 31.5 ± 1.1 32.1 ± 0.8 0.711
BMI (kg m−2 ) 26.3 ± 0.6 26.6 ± 0.8 0.806
Semen volume was measured by Falcon tube and expressed as
ml. Semen pH was determined by using pH meter. Semen round
cells was determined by counting round cells such as white blood Results indicate that 12 patients in N. sativa and 15 patients
cells or germinal cells and expressed as percentage. in placebo group had sperm counts below 20 × 106 ml and 15
The trial was registered in Iranian Registry of Clinical Trial with patients in N. sativa and 15 patients in placebo group had abnormal
the number IRCT201011205204N1. sperm morphology less than 30%. In N. sativa group 17 patients and
placebo group 20 patients had type A motility less than 25% and in
Statistical analysis each groups 21 patients had B motility less than 50%.
In N. sativa oil group sperm count, morphology and motility
Chi squared and t-tests were used for data analyses and p < 0.05 improved significantly (p = 0.001, p = 0.011 and p = 0.001 respec-
was considered as significant. tively) compared with placebo group at the end of study (Table 4).
Furthermore, in N. sativa oil group semen round cells, volume and
Results pH also improved significantly (p = 0.025, p = 0.001 and p = 0.001
respectively) compared with placebo group at the end of study
The values for fatty acid contents of the N. sativa oil are presented (Table 4). Also, different classes of sperm motility also improved
in Table 1. significantly in N. sativa oil group compared with placebo group at
The N. sativa oil contained 1.1% volatile constituents. The chem- the end of study (Table 5).
ical compositions of the volatile oil component of N. sativa oil are
presented in Table 2. Discussion
No adverse effects were reported. The groups were matched
in regard to demographic data including age, duration of infer- The results indicate that daily intake of 5 ml N. sativa oil
tility and body mass index (Table 3). The baseline blood levels of improves semen quality including sperm count, morphology and
all parameters were not significantly different between the two motility and semen volume, pH and round cells in infertile men
groups (Table 4). having abnormal semen parameters. The effects of N. sativa oil

Please cite this article in press as: Kolahdooz, M., et al., Effects of Nigella sativa L. seed oil on abnormal semen quality in infertile men:
A randomized, double-blind, placebo-controlled clinical trial. Phytomedicine (2014), http://dx.doi.org/10.1016/j.phymed.2014.02.006
G Model
PHYMED-51608; No. of Pages 5 ARTICLE IN PRESS
4 M. Kolahdooz et al. / Phytomedicine xxx (2014) xxx–xxx

Table 4
Effects of 2 months N. Sativa and mineral oils consumption on semen quality in infertile men. The semen quality values of fertile men are given for reference. Values are
expressed as mean ± SE.

Semen quality parameters Fertile men Groups at baseline Groups after treatment

Placebo N. sativa oil p-Value Placebo N. sativa oil p-Value

Sperm count (million/ml) 98.23 ± 8.31 46.68 ± 5.11 43.53 ± 4.94 0.660 42.88 ± 4.46 60.18 ± 5.16 0.014
Sperm motility (%) 89.46 ± 3.62 62.83 ± 1.56 63.45 ± 2.27 0.812 61.66 ± 2.01 69.75 ± 2.25 0.015
Sperm morphology (normal/abnormal) 73.23 ± 2.12 40.56 ± 3.27 39.53 ± 2.54 0.805 37.21 ± 3.04 49.09 ± 2.27 0.003
Semen volume (ml) 4.31 ± 0.23 3.73 ± 0.44 3.77 ± 0.33 0.947 3.06 ± 0.25 4.21 ± 0.34 0.025
Semen pH 8.00 ± 0.3 8.44 ± 0.07 8.38 ± 0.07 0.261 8.26 ± 0.02 8.01 ± 0.09 0.001
Semen round cell (%) 0.83 ± 0.01 3.83 ± 0.69 3.32 ± 0.44 0.525 4.23 ± 0.83 0.97 ± 0.25 0.001

p-Values are the results of comparison with the placebo group at baseline and after treatment.
p < 0.05 was considered as significant.

Table 5
Effects of 2 months N. Sativa and mineral oils consumption on sperm motility in infertile men. The semen quality values of fertile men are given for reference. Values are
expressed as mean ± SE.

Classes of sperm motility (%) Fertile men At baseline After treatment

Placebo N. sativa oil p-Value Placebo N. sativa oil p-Value

Class A 64.73 ± 4.12 20.18 ± 1.88 21.79 ± 2.31 0.590 19.29 ± 2.25 30.47 ± 3.503 0.010
Class B 51.43 ± 3.91 43.26 ± 1.41 39.54 ± 2.27 0.311 38.33 ± 2.19 37.35 ± 2.38 0.600
Class C 8.67 ± 1.28 9.93 ± 0.76 9.27 ± 0.70 0.182 11.26 ± 1.02 10.91 ± 1.02 0.213
Class A + B 81.32 ± 4.35 62.23 ± 1.62 61.73 ± 2.41 0.314 58.71 ± 2.21 69.93 ± 2.37 0.002
Immotile 12.74 ± 1.21 37.85 ± 1.56 42.54 ± 2.27 0.310 42.91 ± 0.53 36.26 ± 2.25 0.001

p-Values are the results of comparison with the placebo group at baseline and after treatment.
p < 0.05 was considered as significant.

on abnormal semen parameters in the present study agree with content of semen fluid we could not find out the possible mecha-
the previous study indicating the favorable effects of N. sativa nisms of action of N. sativa oil. Furthermore, due to short duration
oil on male fertility in normal and hyperlipidemic rats (Samir of study we could not find out the possible effects of N. sativa oil on
Bashandy 2007). Furthermore favorable effects of N. sativa oil also reproductive ability of infertile men.
have been reported on spermatogenesis, infertility and some repro- It conclusion, N. sativa oil improves semen quality in infertile
ductive parameters in male rats and chickens (Al-Sa’aidi et al. men having abnormal semen parameters; further research seems
2009; Mukhallad et al. 2009; Abdulkarim and Al-Sardary 2009). The necessary to elucidate possible mechanisms of action of N. sativa
mechanisms underlying these effects are not clear, but antioxidant oil semen parameters and its effects on the ability of infertile men
properties of N. sativa oil may be involved. In normal conditions, having abnormal semen parameters to reproduce.
the reactive oxygen species (ROS) are neutralized by antioxidants
of ejaculatory fluid (Lamirande and Gagnon 1999), but in case of Conflict of interest
reduction of the antioxidant property or increased production of
ROS in semen for any reason, increased level of oxidative stress All authors have no conflict of interest to disclose.
would be harmful to sperm parameters (Schulte et al. 2010; Aitken
and Krausz 2001). There is also report indicating that ROS level Acknowledgements
is increased in semen of infertile patients (Zini et al. 1993). Thus,
the favorable effects N. sativa oil on sperm parameters in infertile This study was supported by the ACECR (Iranian Academic
men observed in the present study may be due to excellent ability Center for Education, Culture and Research), and Payame Noor
of N. sativa oil to inactivate free radicals (Burits and Bucar 2000). University and Infertility Center of Mahdieh Hospital Tehran, Iran.
In support to this hypothesis it should be noted that the thymo- Thanks are due to Dr. Kaveh Sultanzadeh urologist from Royan
quinone (Al-Wafai 2013; Ghosheh et al. 1999; Houghton et al. 1995) Research Institute and Clinical Center for Infertility and Reproduc-
with antioxidative properties, present in of N. sativa oil may neu- tive Health in Tehran, for his valuable assistance.
tralize the ROS in semen and positively influence abnormal sperm
parameters. In agreement with this hypothesis the positive effects References
of antioxidants like vitamin E, selenium and nutritional supple-
ments on sperm motility of infertile men have been documented Abdulkarim, S.M., Al-Sardary, S.Y., 2009. Effect of black seed (Nigella sativa L.) on
some reproductive traits in Ross broiler breeder male chickens. Journal Bombay
(Suleiman et al. 1996; Vézina et al. 1996; Sinclair 2000). Further-
Veterinary College 17, 19–28.
more, the favorable effects of N. sativa oil on abnormal sperm Adams, R.P., 2007. Identification of Essential Oil Components by Gas Chromatogra-
parameters in the present study may be due to unsaturated fatty phy/Mass Spectrometry, 4th ed. Allured Publishing Corporation Carol Stream,
acids content of N. sativa oil (Nickavar et al. 2003). This hypothesis IL, USA, pp. 22–25.
Agarwal, A., Makker, K., Sharma, R., 2008. Clinical relevance of oxidative stress in
is supported by studies indicating significant positive correlation male factor infertility: an update. American Journal of Reproductive Immunol-
between the unsaturated fatty acid supplementation and enhanced ogy 59, 2–11.
sperm count, motility, and normal morphology in infertile men Aitken, R.J., Krausz, C., 2001. Oxidative stress, DNA damage and the Y chromosome.
Reproduction 122, 497–506.
(Nissen and Kreysel 1983; Safarinejad 2011). Although in the Al-Wafai, R.J., 2013. Nigella sativa and thymoquinone suppress cyclooxygenase-2
present study we quantified the unsaturated fatty acid and volatile and oxidative stress in pancreatic tissue of streptozotocin-induced diabetic rats.
constituents of N. sativa oil that agree with previous studies, but Pancreas 42, 841–849.
Al-Sa’aidi, J.A.A., Al-Khuzai, A.L.D., Al-Zobaydi, N.F.H., 2009. Effect of alcoholic extract
due to study limitations such as lack of determination of male of Nigella sativa on fertility in male rats. Iraqi Journal of Veterinary Sciences 23,
sex hormone levels, antioxidant activity and unsaturated fatty acid 123–128.

Please cite this article in press as: Kolahdooz, M., et al., Effects of Nigella sativa L. seed oil on abnormal semen quality in infertile men:
A randomized, double-blind, placebo-controlled clinical trial. Phytomedicine (2014), http://dx.doi.org/10.1016/j.phymed.2014.02.006
G Model
PHYMED-51608; No. of Pages 5 ARTICLE IN PRESS
M. Kolahdooz et al. / Phytomedicine xxx (2014) xxx–xxx 5

Araoye, M.O., 2003. Epidemiology of infertility: social problems of the infertile cou- Nissen, H.p., Kreysel, H.W., 1983. Polyunsaturated fatty acids in relation to sperm
ples. West African Journal of Medicine 22, 106–109. motility. Andrologia 115, 264–269.
Burits, M., Bucar, F., 2000. Antioxidant activity of Nigella sativa essential oil. Phy- Safarinejad, M.R., 2011. Effect of omega-3 polyunsaturated fatty acid supplementa-
totherapy Research 14, 323–328. tion on semen profile and enzymatic anti-oxidant capacity of seminal plasma
Chen, J.C., Xu, M.X., Chen, L.D., 1999. Effect of Panax notoginseng extracts on inferior in infertile men with idiopathic oligoasthenoteratospermia: a double-blind,
sperm motility in vitro. American Journal of Chinese Medicine 27, 123–128. placebo-controlled, randomized study. Andrologia 43, 38–47.
Duchateau, G., Van Oosten, H.J., Vasconcellos, M.A., 1996. Analysis of cis-fatty acid Salehi Surmaghi, M.H., 2008. Nigella sativa. In: Herbal Medicine and Herbal Therapy,
and trans-fatty acid isomers in hydrogenated and refined vegetable oils by cap- 2nd ed. Donyay Taghziah Press, Tehran, Iran, pp. 216–219.
illary gas-liquid chromatography. Journal of the American Oil Chemists’ Society Samir Bashandy, A.E., 2007. Effect of fixed oil of Nigella sativa on male fertility
73, 275–282. in normal and hyperlipidemic rats. International Journal of Pharmacology 3,
Ghosheh, O.A., Houdi, A.A., Crooks, P.A., 1999. High performance liquid chro- 27–33.
matographic analysis of the pharmacologically active quinones and related Schulte, R.T., Ohl, D.A., Sigman, M., Smith, G.D., 2010. Sperm DNA damage in male
compounds in the oil of the black seed (Nigella sativa L.). Journal of Pharma- infertility: etiologies, assays, and outcomes. Journal of Assisted Reproduction
ceutical and Biomedical Analysis 19, 757–762. and Genetics 27, 3–12.
Goreja, W.G., 2003. Black seed. In: Nature’s Miracle, Remedy. Amazing Herbs Press, Sinclair, S., 2000. Male infertility: nutritional and environmental considerations.
New York, pp. 1–64. Alternative Medicine Review 5, 28–38.
Houghton, P.J., Zarka, R., de las Heras, B., Hoult, J.R., 1995. Fixed oil of Nigella sativa and Slama, R., Hansen, O.K., Ducot, B., Bohet, A., Sorensen, D., Giorgis, Allemand, L., Eijke-
derived thymoquinone inhibit eicosanoid generation in leukocytes and mem- mans, M.J., Rosetta, L., Thalabard, J.C., Keiding,.N., Bouyer, J., 2012. Estimation of
brane lipid peroxidation. Planta Medica 61, 33–36. the frequency of involuntary infertility on a nation-wide basis. Human Repro-
Lamirande, E., Gagnon, C., 1999. The dark and bright sides of reactive oxygen species duction 27, 1489–1498.
on sperm function. In: Gagnon, C. (Ed.), The Male Gamete: From Basic Science Suleiman, S.A., Ali, M.E., Zaki, Z.M., el-Malik, E.M., Nasr, M.A., 1996. Lipid peroxidation
to Clinical Applications. Cache River Press, IL, pp. 455–467. and human sperm motility: protective role of vitamin E. Journal Andrology 17,
Malhotra, S.K., 2004. Nigella. In: Peter, K.V. (Ed.), Handbook of Herbs and Spices, vol. 530–537.
2. CRC Press/Woodhead Publishing Limited, Abington Hall, Abington/Cambridge Vézina, D., Mauffette, F., Roberts, K.D., Bleau, G., 1996. Selenium-vitamin E sup-
CB1 6AH, England, pp. 218–226. plementation in infertile men. Effects on semen parameters and micronutrient
McLafferty, F.W., Stauffer, D.B., 1989. The Wiley/Nbs Registry of Mass Spectral Data. levels and distribution. Biological Trace Element Research 53, 65–83.
Wiley, New York. Whitman-Elia, G.F., Baxley, E.G., 2001. A primary care approach to the infertile
Mohtashami, R., Amini, M., Fallah, Huseini, H., Ghamarchehre, M., Sadeqhi, Z., Haji- couple: clinical review. Journal of the American Board of Family Medicine 14,
agaee, R., Fallah, H.A., 2011. Blood glucose lowering effects of Nigella Sativa L. 33–45.
seeds oil in healthy volunteers: a randomized, double-blind, placebo-controlled World Health Organization, 1999. WHO Laboratory Manual for the Examination
clinical trial. Journal of Medicinal Plants 10, 90–94. of Human Semen and Sperm–Cervical Mucus Interaction, 4th ed. Cambridge
Mukhallad, A.M., Mohamad, M.J.M., Dradka, H., 2009. Effects of black seeds (Nigella University Press, UK, pp. 1–138.
Sativa) on spermatogenesis and fertility of male albino rats. Research Journal of Zafar, M.A.F., Mohsin, A., 2001. Advancement in treatment of male infertility. Annals
Medicine and Medical Sciences 4, 386–390. King Edward Medical College 7, 224–226.
Najmi, A., Haque, S.F., Naseeruddin, M., Khan, R.A., 2008. Effect of Nigella Sativa oil Zaoui, A.Y., Cherrah, A., Mahassini, K., Aloui, H., Amarouch, M.H., 2002. Acute and
on various clinical and biochemical parameters of metabolic syndrome. Inter- chronic toxicity of Nigella sativa fixed oil. Phytomedicine 9, 69–74.
national Journal of Diabetes in Developing Countries 28, 11–14. Zini, A., de Lamirande, E., Gagnon, C., 1993. Reactive oxygen species in semen of
Nickavar, B., Mojab, F., Javidnia, K., Amoli, M.A., 2003. Chemical composition of the infertile patients: levels of superoxide dismutase- and catalase-like activities
fixed and volatile oils of Nigella sativa L. from Iran. Zeitschrift fur Naturforschung in seminal plasma and spermatozoa. International Journal of Andrology 16,
58, 629–631. 183–188.

Please cite this article in press as: Kolahdooz, M., et al., Effects of Nigella sativa L. seed oil on abnormal semen quality in infertile men:
A randomized, double-blind, placebo-controlled clinical trial. Phytomedicine (2014), http://dx.doi.org/10.1016/j.phymed.2014.02.006