A O RTICLE

2778
Advances in Environmental Biology, 6(10): 2778-2783, 2012
ISSN 1995-0756
This is a refereed journal and all articles are professionally screened and reviewed
ORIGINAL ARTICLE
The effects of hydro alcoholic extract of Ceratonia siliqua L. seeds on pituitary – testis
hormones and spermatogenesis in rat
Mokhtari M Ph.D, Sharifi E MSc, Azadian Sh , MSc.
Islamic Azad University, Kazerun Branch, Department of Biology, kazerun, Iran.
Mokhtari M Ph.D, Sharifi E MSc, Azadian Sh, MSc.: The effects of hydro alcoholic extract of
Ceratonia siliqua L. seeds on pituitary – testis hormones and spermatogenesis in rat
ABSTRACT
In traditional medicine, Carob tree has been used as antidiarrheal medicine and to treat digestion diseases.
In this research, the possible effects of Carob seeds extract on levels of LH, FSH, testosterone and
dihydrotestosterone hormones and testicular tissue as well as improving fertility in male rats were investigated.
Fifty adult male rats were divided into five groups of tens as follow: The control group was left on treated, the
sham group received only distilled water and experimental groups which received 150,300 and 600 (mg/kg)
extract respectively for 31 days. Blood samples were taken to measure LH, FSH, testosterone and
dihydrotestosterone concentrations and testicular histological changes was investigated. The consumption of
Carob seeds extract caused a significant increase in testosterone and DHT concentrations were as the level of
LH declined in experimental groups receiving 150 and 300 (mg/kg) extract and the FSH concentrations did not
show any significant changes. Furthermore, sperm density in seminiferous tubules was increased in response to
medium and high dosages of the extract. The seed extract of Carob may have some effects on the synthesis of
cAMP and the activities of enzymes involved in steroidogenesis in leydig cells thereby leading to an increase in
serum concentration of testosterone.
Key words: Carob seed, LH, FSH, Testosterone, Dihydrotestosterone, Rat.
Introduction
About half century ago, plants were considered
as the most important medicinal resources for
treating diseases. Fertility disorder and its related
ramifications have been identified as one of the most
important problems in married life [10]. According
to present statistics, about 35% of married infertility
is related to men, and the most prevalent reason for
men infertility, is their inability to produce sufficient
number of intact and active sperms [2,1,22]. Studies
indicate that various factors can affect sperm
production and contribute to infertility incidence.
Among these factors are taking special anti cancer
medicines, antibiotics, toxic substances, pesticides,
radiation, stress, air pollution and vitamin deficiency.
These factors can decrease sperm counts by inducing
free radicals and germinal cells oxidation in testicular
tissues [1,31]. Previous studies have indicated that
infertility incidence in men is on the increase.
According to Eskenazib et al., (2005), for reforming
semen quality and infertility rate decrease, American,
men need to receive vitamin and nutritional
supplements [28]. Evidence indicates that usage of
medicinal plants could have positive effects on
fertility increase, as well as reducing such disorders
as hormonal imbalance, sexual weakness,
oligospermia, varicocele and prostate inflammation.
Carob or Ceratonia siliqua L. is a beautiful tree
belonging to Legominasae family and is about 7 to
12 meters tall. It has compound leaves and its red,
yellow or purple flowers has no petal. Its arch pod
fruit is bright brown with, 10 to 30 cm long and
contain 12 to 16 hard seeds. Carob is native to
Mediterranean regions and is found in south of Syria,
India and most of Mediterranean areas as well as in
California. It grows wildly in shapoor, Fars, Iran
[19,44,16].
Ceratonia siliqua fruits have been used as food
sweetener and thickener [38]. According to some
researches, its consumption can decrease serum
cholesterol and glucose level [12,11,37,45]. In
traditional medicine, carob fruits have been used as
antidiarrhetic medicine and used for treating
digestive diseases [21]. The presence of fiber, tannan
and polyphenols in its fruits could lower potential
risks of colon cancer [27]. In addition, presence of
pectin and lignin in carob could function as
regulators of digestive system and could act as a
toxin excretor in digested foods [27]. Ceratonia
siliqua L pods contain antioxidants and vitamin E,
and have anti asthma properties. Its tannans, function
Corresponding Author
Mokhtar Mokhtari, Islamic Azad University, Kazerun Branch, Department of Biology, kazerun,
Iran.
E-mail: [email protected]
2779
Adv. Environ. Biol., 6(10): 2778-2783, 2012
as antioxidant, tranquilizer, antibacterial and anti
allergy and can help to improve bone fractures
because the seeds are rich in phosphorus and
calcium. Some studies indicate that carob, can help
to treat mellitus diabetes and can decrease blood
glucose level [37,42]. Since fibers of this plant are
complex, containing many substances with various
properties, they can improve glucose regulation and
lower the risks of diabetes 2 and cardiovascular
diseases [18]. Studies on chemical contents of
Ceratonia siliqua seed suggests that it contains plenty
of fibers, polyphenolic compounds, arachidonic acid,
lignin, fats, proteins, carbohydrates, calcium,
potassium, and phosphorus[34,30,25,4]. They also
contain aspartic acid, glutamic acid, linolenic acid,
linoleic acid, vitamin E, betasitosterol, [9] silica, iron
and magnesium [44]. These compounds can have
different effects on animal bodies. The Carob seeds
also considered as a potential source of natural
antioxidants [17] and Carob antioxidant activity is
related to phenolic compounds (Owen, 2003;Ayaz,
2007)and tannans substances existence [29].
In recent decades,human resort to medicinal
plants, because chemical medicines have side effects
and complications. Twenty-First century is called the
century of returning back to nature. Many attempts
have been made to exploit medicinal plants.
Considering the limited number of studies on the
effects of carob seed extract on male reproductive
activities, this study has made an attempt to
investigate the probable effect of hydro - alcoholic
extract of carob seed on the rate of FSH,LH,
testosterone and dihydrotestosterone hormones and
testis histological changes. The possible results could
be useful in fertility and family planning.
groups were kept in separate cages at 22  2c .
Water and dry food from Fars birds nutrition stock
company were available to rats. The polycarbonate
cages were sterilized twice a week and debris of
wood was changed every two days. The experiment
was started one week after the accommodation of
animals in the lab environment.
Extract Preparations:
Ripen fruits of carob tree were collected around
Kazeroun in the middle of spring 2010. Pods were
crushed , seeds were separated, washed, dried and
grinded until a fine powder was obtained. The
powder was solved a 50/50 ratio of water and %96
alcohol for 72 hours during which was shaken
several times, then it was filtered and hydro –
alcoholic extract was obtained. It was dried in oven
at 40c and a concentrated brown extract was
obtained [23,15].
Hormones measurement and tissue study:
At the end of experimental period, rats were
weighed, and blood samples were taken from heart
under light anesthetic. Blood samples were kept in
the lab conditions for 20 minutes and were
centrifuged for 15 minutes at 2000 RPM and
Materials And Methods
collected and frozen at  20 C to be used for
hormone measurement. The concentration of FSH,
LH, testosterone and dihydrotestosterone were
measured using RIA. Testis were also removed,
washed, dried, weighed, fixed in 10% formalin,
sectioned and got ready for microscopic examination.
Data were analyzed by ANOVA and complementary
Tukey test. The P values was sat at P<0/05.
Experimental Animals:
Results:
In this experimental study, 50 adult male wistar
rats, each weighing about 210  10 g and 2.5 -3
months old were used. Animals were divided into 5
groups of 10 as follow: The control group was left on
treated, the sham group received only distilled water
and experimental groups which received 150,300 and
600 mg/kgbw extract respectively for 31 days. These
Statistical studies and comparison of LH, FSH,
testosterone and dihydrotestosterone hormones were
made among experimental , control and sham groups,
following consumption of hydro – alcoholic extract
of Ceratonia siliqua seeds and the results are shown
in table 1.

Table 1: The comparison of mean serum concentrations of testosterone, LH, FSH, DHT hormones among groups receiving different
dosages of hydro – alcoholic extract of carob seeds (P ≤ 0/05). Values are based on X ± SEM and asterisk shows significant
difference among groups.
(X±SEM)
LH
FSH
Testosterone
DHT
Groups
(IU/L)
(mIU/L)
(ng/ml)
(mIU/L)
Control
0.25 ± 0.03
17.97 ± 1.65
1.024 ± 0.21
0.211 ± 0.007
Sham
0.24 ± 0.02
17.97 ± 1.77
1.141 ± 0.19
0.212 ± 0.005
150
2780
Adv. Environ. Biol., 6(10): 2778-2783, 2012
Experimental
groups
(mg/Kg)
0.19 ± 0.002
18.72 ± 1.40
2.12 ± 0.91
0.218 ± 0.005
300
(mg/Kg)
0.15 ± 0.003 ‫٭‬
19.1 ± 1.75
3.97 ± 0.47 ‫٭‬
0.221 ± 0.005
600
(mg/Kg)
0.14 ± 0.002 ‫٭‬
16.5 ± 1.45
4.1 ± 0.27 ‫٭‬
0.222 ± 0.003 ‫٭‬
Hormonal measurements indicate a significant
increase in serum testosterone level in groups
receiving medium and maximum dosages of the seed
extract in comparison to control and sham groups (P
< 0/05) while it shows no significant difference in
group receiving minimum dosage (Table 1). In
contrast, the serum LH concentration declined
significantly in groups taking medium and maximum
dosages of extract and showing no meaningful
difference in group receiving minimum doses (Table
1). At the same time, serum FSH levels did not
confirm any significant differences among various
groups (Table 1). Also dihydrotestosterone levels
Fig. 1: photomicrograph of seminiferous tubules
in control group ×200
Fig. 3: photomicrograph of seminiferous tubules in
medium group ×200
increased significantly only in group receiving
maximum dosage (Table 1).
According to photomicrograph and comparison
of seminiferous tubule cross sections among
experimental, control and sham groups, there was no
tissue changes in spermatogonium , primary
spermatocyte and other cells in terms of size,
numbers, cytoplasm characteristics , nucleus and
their staining. However, the results of histology
indicate that consumption of medium and maximum
dosages of seed extract can result in sperm density
raise [Figures 1,2,3,4] .
Fig. 2: photomicrograph of seminiferous tubules in
sham group ×200
Fig. 4: photomicrograph of seminiferous tubules in
maximum group ×200
2781
Adv. Environ. Biol., 6(10): 2778-2783, 2012
Discussion:
Many studies are performed on the use of
medicinal plants for treating sexual impotent and
according to traditional medicine, it is possible to use
herbs for improving fertility. Hydro – alcoholic seed
extract of Ceratonia siliqua contains such compounds
as phytosterols, and can cause a decrease in total
cholesterol , LDL and triglycerides, through an
increase in lipoproteins and hepato lipases as well as
reducing HMG – COA reductase activity [20].
Results of this study are indicative of Carob
ability to make changes in pituitary- testis hormonal
axis and these changes can affect on male
reproductive activities.Reports indicate that carob
seeds are rich in phytosterol. These compounds are
structural analogues of cholesterol and can prevent
from many cancers (such as breast and prostate
cancers) [5]. According to our results, daily
consumption of 300,600 (mg/kg) hydro – alcoholic
seed extract of Ceratonia siliqua could elevate serum
testosterone concentration. It seems that the increase
in testosterone levels by higher dosages of extract is
due to its direct effect on leydig cells and on
testosterone biosynthesis. These effects are probably
performed through stimulating PGE2 synthesis.
Furthermore, carob seeds contain gammalinolenic
acids and alpha linolenic acid that can be
transformed into dihomogamma linolenic acid and
then into arachidonic acid which is the precursor of
all prostaglandins, like PGE2.
In addition, these seeds contain own arachidonic
acid [7]. It appears that arachidonic acid plays a
major role in testis steroidogenesis. Studies indicate
that this organic acid and its metabolite (PGE2) can
increase the production of cyclic adenosine
monophosphate
(cAMP),
thereby
elevating
cholesterol side chain break-down, and stimulating
testosterone production [38]. Some studies indicate
that arachidonic acid and it’s metabolite (PGE2)
stimulate prostate growth which is more influenced
by testosterone. Eicosanoids are so involved in
esteroidogenesis
thus,
insufficient
use
of
alphalinolenic acid and abnormal production of
eicosanoids can lead to infertility [14].
Another compound found in carob seeds is
aspartic acid [9] which can function as a second
messenger in leydig cells through increasing
synthesis of cAMP and testosterone release. In
addition, this fatty acid along with a second
messenger production in pituitary gland give rise to
synthesis and release of LH; thereby elevating the
concentration of testosterone [13].
Similarly, treatment with hydro – alcoholic
extract of Ceratonia siliqua seeds in high dosages
induces LH decrease. It seems that this decline in
hormone level is the result of negative feed back
control of testosterone secretion which has a direct
effect on hypothalamus and reduction of GnRH. At
last, LH secretion is balanced by anterior part of
pituitary. Therefore, testosterone, has a direct
negative and weak feedback effect on anterior of
pituitary and this feedback lowers LH secretion [36].
Considering the results of FSH concentration,
indicated that there was no significant changes in
experimental groups receiving extract compared to
control and sham groups. Feedback mechanisms on
FSH are not exerted just by testis steroids, but also
by inhibin, activin and folistatin. These factors have
central effects on GnRH to regulate FSH
concentration, and perhaps lack of any significant
change in FSH level is the result of adjusting effects
of these factors.
Presence of vitamin E in carob seeds has been
demonstrated [9]. One of the most important
chemical properties of vitamin E is its high
sensitivity to oxidants and this vitamin has been used
as an antioxidant in pharmaceutical industry. In
addition, the lack of vitamin E in some mammals; for
example rats, brings about animal sterilization. Some
researches have indicated that vitamin E is helpful in
decreasing cadmium effects on testicular tissues and
on spermatogenesis [41]. Moreover some studies
have been performed on the usefulness of vitamin E
and the other antioxidant such as glutathione and coenzyme Q10 in treating male infertility brought about
stresses, environment pollution and malnutrition. The
results of these studies are indicative of its
helpfulness and can preserve cell's nuclear DNA
[33,24,32,3]. Similarly, therapeutics effects of
vitamin E on testis has shown that as an antioxidant,
it could regenerate seminiferous tubules after being
damaged by ozone gas; it also reduces the destructive
effects of this gas on the tissues [24]. Based on the
above studies, one can infer a safe role for vitamin E
in comparison to vitamin C (an effective antioxidant
element) in testicular tissues against exterior toxic
elements. Sperms are very sensitive to ROS injected
into semen by leukocytes and ROS can cause
infertility in men [22]. Furthermore, according to
some Studies, usage of antioxidants and vitamin B,E
and C can lower infertility through reducing damages
caused by free radicals, sperms DNA regeneration
and preservation, and through strengthening bloodtestis barrier [2,8,26]. In present study, the
experimental groups receiving high doses of extract
show increase in sperm density within seminiferous
tubules, which probably the result of a boost in
testosterone synthesis and the presence of
antioxidants and vitamin E.
Conclusion:
Based on the results of this study, the use of high
dosages hydro – alcoholic extract of Ceratonia
siliqua seeds probably have some influences on
cAMP production and activity of enzymes involved
in rat's testicular steroidogenesis and can increase
testosterone synthesis. Some LH effects on leydig
cells are effected by the extract content, modifying
2782
Adv. Environ. Biol., 6(10): 2778-2783, 2012
the release rate of this hormone from anterior of
pituitary. Similarly, consumption of extract in high
doses increases sperm density in seminiferous
tubules. Finally, it should be mentioned that more
studies are necessary in this field.
12.
References
13.
1.
Aitken, R.J., 1999. The Amoroso Lecture The
human spermatozoon-a cell in crisis?J Reprod
Fertil., 115: 1-7.
2. Amin, A., AAA. Hamz, 2006. Effects of Roselle
and Ginger on Cisplatin-induced reproductive
toxicity in rats. Asian J Androl., 8(5): 607-12.
3. Amr, A., E. Alaa, A. Hamza, 2006. Effects of
Roselle and Ginger on ciplatin-induced
reproductive toxicity in rats.Asian J Androl.,
8(5): 607-612.
4. Avallone, R., M. Olessi, M. Baraldi and A.
Monzani, 1997. Determination of chemical
composition of carob (Ceratonia siliqua):
Protein, Fat, Carbohydrates , and Tannins,
Journal of food composition and Analysis,
10(2): 166-172.
5. Awad, A.B., C.s. Fink, 2000. Phytosterols as
anticancer dietary component.J Nutr., 130(9):
2127-30.
6. Ayaz, F.A., H. Torun, S. Ayaz, P.J. Correia, M.
Alaiz, C. Sanz, J. Gruz, M. Strand, 2007.
Determination of chemical composition of
Anatolian carob pod (Ceratonia siliqua L.) :
sugar,Amino and organic Acids,Minerals and
phenolic compounds.J Food Qual., 30(6): 10401055.
7. Bengoechea, C., M.C. Puppo, A. Romero, F.
Cordobes and A. Guerrero, 2008. Linear and
non-linear
Viscoelasticity
of
emulsions
containing carob protein as emulsifier. journal of
Food Engineering, 87(1): 124-135.
8. Brooks, D.E., 1976. Activity of androgenic
control of glycolytic enzymes in the epididymal
spermatozoa of the rats.J Biochem., 156(3): 52737.
9. Duke, J., 2009.
Phytochemical and
Ethnobotanical Databases Ceratonia siliqua,
Green Farmacy Garden 8210 Murpht Road
Fulton,MD 20759. Agricultural Research
Service, Beltsville Area: http://www.aragrin.gov/duke/Ceratonia siliqua information
from NPGS/GRIN.
10. Ebisch, I.M.W., C.M.G. Thomas, W.H. Peters,
D.D.M. Braat, R.P.M. Steeger-Theunissen,
2007. The importance of folate, Zinc and
antioxidants in the pathogenesis and prevention
of subfertility.J Hum Repord Update., 13(2):
163-74.
11. Edwards, C.A., N.A. Blackburn, L. Craigen, P.
Davison, J. Tomlin, K. Sugden and et al. 1987,
Viscosity of food gums determined in vitro
14.
15.
16.
17.
18.
19.
20.
21.
22.
23.
related to their hypoglycemic actions . Am J
Clin Nutr., 46(1): 72-77.
Elsenhans, B., U. Sufke, R. Blume and W.F.
Caspary, 1980. The influence of carbohydrate
gelling agents on rat intestinal transport of
monosaccharides and neutral amino acid in vitro
. Clin Sci(Lond). 59(5): 373-80.
Enza Topo, Andrea Soricell,Antimo D Aniello,
Salvatore Ronsini and Gemma D Aniello, 2009.
The role and molecular mechanism of D-aspartic
acid in the release and synthesis of LH and
testosterone in humans and rats ;Journal
Reproductive Biology and Endocrinology, 7:
120.
Eritsland, J., 2000. Safety considerations of
polyunsaturated fatty acids . Am J Clin Nutr.,
71(1): 197S-201S.
Germanò, M.P., V. D'Angelo, R. Sanogo, A.
Morabito, S. Pergolizzi, R. De Pasquale, 2001.
Hepatoprotective activity of Trichilia roka on
carbon tetrachloride-induced liver damage in
rats. J Pharm Pharmacol., 53(11): 1569-74.
Ghahreman, A., 1998. Plant Systematics
Cormophytes of Iran. .2 sted,Tehran: Unive Med
Sci., pp: 508-510. Persian
Gohar, A., S.R. Gedara and H.N. Baraka,
2009,New acylated flavonol glycoside from
Ceratonia siliqua L. seed, Journal of medicinal
plants research, 3(5): 424-428.
Gruendel, S., B. otto, A.L. Garcia, K. Wagner,
C. Mueller and et al. 2007. Carob pulp
preparation rich inisoluble dietary fibre and
polyphenols increases plasma glucose and serum
insulin responses in combination with a glucose
load in humans . The British Journal of
Nutrition, 98(1): 101-105.
Haji sharifi, A., 2008. The Encylopedia of
medicinal plant.1th ed , Tehran:Hafeze novin, pp:
384-385.
Hendriks, H.F., J.A. Westrate, T. Rliet, G.
Metter 1999. Spreads enriched with three
different levels of vegetable oil sterols and the
degree
of
cholesterol
lowering
in
normocholesterolameic
and
mildy
hypercholesterolameic
subjects,European
Journal of Clinical Nutrition, 53(4): 319-327.
Hostettler, M., R. Steffen, A. Tschopp, 1995.
Efficacy and tolerance of insoluble Carob
fraction in the treatment of travelers ,diarrhea . J
Diarrhoel Dis Res., 13(3): 155-8.
Imai, H., K. Suzuki, K. Ishizaka, S. Ichinose, H.
Oshima, I. Okayasu, et al., 2001. Failure of the
expression of phospholipid hydroperoxid
glutathione peroxidase in the spermatozoa of
human infertile males.J Biol Repord., 64: 67483.
Jain, H., S.D. Parial, E. Jarald, A.S. Daud, S.
Ahmad, 2010. Extraction of Ashwagandha by
conventional extraction method and evaluation
2783
Adv. Environ. Biol., 6(10): 2778-2783, 2012
24.
25.
26.
27.
28.
29.
30.
31.
32.
33.
of its anti-stress activity, International Journal of
Green pharmacy, 4(3): 183-185.
Jedlinska-Krakowska, M., G. Bomba, K.
Jakubowski, T. Rotkiewicz, B. Jana,
A.
Penkowski, 2006. Impact of oxidative stress and
supplementation with Vitamins E and C on testis
morphology in rats.J Reprod Dev., 52(2): 203-9.
Karabulut, A., O. Canbolat and A. Kamalak,
2006. Evaluation of carob,Ceratonia siliqua Pods
as a feed for sheep, Departement of animal,
Nutrition,
Kahramanmaras,
TURKEY,
Livestock
Research
for
Rural
Development18(7).
Khaki, A., F. Fathiazad, M. Nouri, et al,
2009,The effects of Ginger on spermatogenesis
and sperm parameters of rat,Iranian Journal of
Reproductive Medicine; 7(1): 7-12.
Klenow, S., M. Glei, R. Owen and B. PoolZobel, 2008. Carob Fibre compounds modulate
parameters of cell growth differently in human
HT29 colon adenocarcinoma cell than in LT97
colon adenoma cell,Food and Chemical
Toxicology, 46(4):1389-1397.
Kumar, R., G. Gautam, N.P. Gupta, 2006. Drug
Therapy for Idiopathic male infertility :rationale
Versus evidence.J Urol. 176(4pt1):1307-12.
Makris, D.P., p. Kefalas, 2004. carob
pods(Ceratonia siliqua L.)as a source of
polyphenolic
antioxidants.Food
Technol.
Biotechnol., 42(2): 105-108.
Moh'D Khaiv, J., EL-shatnawi and Khalil I.
Ereifej, 2001. Chemical Composition and
Livestock Ingestion of carob (Ceratonia siliqua
L.)seeds. Journal of Range Management, 54(6):
669-673.
Mosher, W.D., W.F. Pratt, 1991. Fecundity and
infertility in the United States:incidence and
trends. J Fertil steril., 56(2):192-3.
Mozdarani, H., M. Salimi, 2006. Numerical
chromosome abnormalities in 8-cell embryos
generated from gamma-irradiated male mice in
the absence and presence of Vitamine E.Int J
Radiat Biol., 82(11): 817-22.
Murugesan,
P.,
T.
Muthusamy,
K.
Balasubramanian, J. Arunakaran, 2005. studies
on the protective role of Vitamin C and E
against polychlorinated biphenyl (Aroclor
1254)-induced Oxidative damage in leydig cell.J
Free radic Res., 39(11): 1259-72.
34. Ortiz, L.T., M.L. Rodriguez, C. Alzueta, A.
Rebole, C. Centeno, & J. Trevino, 2004. Effect
of carob seed (Ceratonia siliqua L.)in broiler
chick diet on nutrient digestibility and intestinal
Viscosity. EAAP publication NO.110, Toledo,
spain, .Wageningen Academic publishers, pp:
239-242.
35. Owen, R.W., R. Haubner, W.E. Hull, G. Erben,
B. Spiegelhalder, H. Bartsch, B. Haber, 2003.
Isolation and structure elucidation of the major
individual polyphenols in carob fibre. Food
Chem Toxicol, 41(12): 1727-38.
36. Shadan Farrokh, M.D., 1996. Textbook of
Medical Physiology,vol2, Tehran,Ninth Edition,
Tchehr CO. pp: 1506.
37. Tsai, A.C. and B. Peng, 1981. Effects of locust
bean gum on glucose tolerance suger, digestion ,
and gastric motility in rats. J Nutr., 111(12):
2152-2156.
38. Wadw, M.G., G. Van der kraak, 1993.
Arachidonic acid and PGE2 stimulate testostron
production by goldfish testis in vitro.Gen Comp
Endocrinal., 90(1): 109-18.
39. Wang, Y., P.S. Belton, H. Bridon, E. Garanger,
N. Wellner, M.L. Parker et al. 2001. Physicoch emical studies of caroubin:A gluten like
protein.J Agric Food chem, 49(7): 3414-9.
40. World Health organization, 1999. WHO
Laboratory Manual for the Examination of
Human semen and sperm- cervical mucus
Interactions.4th
ed.
Cambridge,United
Kingdom:Cambridge; University press.
41. Yang Hs, Han D.K., J.R. Kim, J.C. Sim, 2006.
Effects of alpha-tocopherol on cadmium-induced
toxicity in rat testis and spermatogenesis .J
Korean Med Sci., 21(3): 445-51.
42. Yaniv, Z., A. Dafni, J. Friedman and D.
Palevitch, 1987. plants used for the treatment of
diabetes J Ethnopharmacol., 19(2): 145-151.
43. Yazilim, Y., Do You Know What Carob is?Have
You ever seen a carob or tasted one? Anglo
Jaoanese
AmericN
reg
No,059-A.
http://www.carobmolasses.com/.
44. Zargari, A., 1996. medicinal Plants.2 thed,
University publications,Tehran,Iran, p: 114-116.
Persian
45. Zunft, H.J., W. Luder, A. Harde, B. Haber, H.J.
Graubaum and J. Gruenwald, 2001. carob pulp
preparation
for
treatment
of
hypercholesterolemia, J Adv Ther, 18(5): 230-6.