effect of queracetin (onion juice) and zinc sulphate … · 2017-10-27 · levels of onion or zinc...

Egypt. Poult. Sci. Vol (33) (II): (331-347) (1428)

Egyptian Poultry Science Journal

http://www.epsaegypt.com

ISSN: 1110-5623 (Print) – 2090-0570 (On line)

EFFECT OF QUERACETIN (ONION JUICE) AND ZINC SULPHATE

ON REPRODUCTIVE PERFORMANCE OF MALE RABBITS UNDER

HOT SUMMER CONDITIONS

M. E. El-Speiy and A. M. El-Hanoun Anim. Prod. Rese. Institute, Agric. Rese. Center, Ministry of Agric., Egypt

Received: 05/03/2013 Accepted: 29/04/2013

ABSTRACT: The objective of this work was to study the effect of queracetin (onion juice)

or zinc sulphate or their combination supplementations on some reproductive performance

of rabbit males exposed to high temperature during summer season conditions. Forty- two

male V-line rabbits 7 months old were randomly divided into six groups for 7males/ each.

Group 1was served as control without any supplementations. Groups 2 and 3 were

supplemented orally with 3 and 6 ml of onion juice /rabbit/day, group 4 supplemented

orally with 50 mg zinc sulphate /rabbit/day, groups 5 and 6 supplemented orally with 50 mg

zinc sulphate +3 and 6 ml onion juice /rabbit /day, respectively. The supplementations

persisted 10 weeks before semen collection. Highest significant (P<0.01) values for each

of ejaculate volume, sperm motility, total sperm output and total motile sperm were

observed for group of males supplemented with zinc sulphate +higher level of onion

(6ml/rabbit/day) as compared to those for other groups. Reverse response was detected for

reaction time (libido) by seconds as the highest (P<0.01) value was observed for control

group and lowest ones were detected for groups of zinc separately and zinc sulphate+ onion

supplementation groups. Supplementation the males with combination of zinc sulphate+6

ml of onion/rabbit/day recorded the highest (P<0.05) values of alkaline phosphates (AIP),

acid phosphates (AcP) and lowest ones for seminal plasma aspartate aminotransferase

(AST) and alanine aminotransferase (ALT) compared to other supplemented groups and

control. Seminal plasma lipid peroxidation as indicated by thiobarbituric acid-reactive

substances was decreased (P<0.05), while seminal plasma antioxidant enzymes were

increased (P<0.05) due to onion juice and zinc sulphate supplementation. Values of FSH

and LH hormones were increased (P<0.05) for males supplemented separately with higher

levels of onion or zinc sulphate or their combination compared to those for control or lower

level of onion. Besides, rabbits supplemented with combination between zinc sulphate +

both levels of onion represented the highest (P<0.05) value of testosterone hormone

Key words: Rabbits, queracetin, zinc sulphate, seminal plasma and fertility.

Corresponding author: [email protected] & [email protected]

mailto:[email protected]

mailto:[email protected]

M. E. El-Speiy and A.M.El-Hanoun

332

compared those for control and other supplemented groups. All parameters of reproductive

performance such as fertility rate ,litter size at birth and at weaning, bunny weight at birth

and at 28 day had been significantly (P<0.01)influenced and decreased for rabbits exposed

to hot summer condition without any dietary experimented supplementations (control)

compared to other groups.

INTRODUCTION

Allium cepa (onion) has a beneficial

effect on disease treatment worldwide and

has been used since ancient times as a

medicinal and food source. Recently

several reports have shown that onion has

high antioxidant activity. Onion is one of

the important Allium species commonly

used in our daily diet, and has recently been

the source of much interest because of its

antithrombotic, hypolipidaemic,

hypotensive, diaphoretic, antibiotic, anti-

diabetic, anti atherogenic and anticancer

medicinal properties (Lee et al., 2008). The

amount of queracetin in fresh onion was 12

mg/100 g onion bulb (Arash et al., 2012).

The biological action of Allium products is

ascribed to organosulfur and phenolic

compounds (Kumari et al., 1990). Several

studies have shown that onion contains

exogenous and endogenous antioxidants

such as selenium, glutathione, vitamins A,

B and C and flavonoids such as queracetin

and isorhamnetin (khaki et al., 2009).

These antioxidants protect DNA and other

important molecules from oxidation and

damage, which improve sperm health

parameters and increasing the rate of

fertility (Yang et al., 2006).

Studies on the effect of queracetin

on oxidative damage in cultured chicken

spermatogonial cells showed that

queracetin have no deleterious effect on

spermatogonial cells. Queracetin (1 mg/ml)

increased the number of spermatogonial

cells, enhanced motility ability and

decreased the mortality of aroclor-induced

oxidative damage. (Aral et al., 2011).

Trace elements are essential for the

function of various enzymes and other

proteins. The effects of trace element

biochemistry and physiology on parameters

of fertility are presented for zinc, selenium,

iodine, copper and manganese (Leonhard–

Marek, 2000). On the cellular level, the

function of zinc can be divided into three

categories: Catalytical, Structural and

Regulatory. Zinc also plays a role in cell

signaling and has been found to influence

hormone release and nerves impulse

transmission (Debjit et al., 2010).

Intracellular zinc can function as a

temporary inhibitor for sperm lipid

peroxidation, sperm oxygen uptake, sperm

nuclear chromatin decondensation, sperm

capacitation, acrosome reaction and for the

in vitro fertilizing ability of spermatozoa

(Stephenson and Brackett, 1999). A

mechanism by which zinc may function as

cellular antioxidant, is through its

involvement in synthesis of metallothionein

(zn-mt ) a metal binding protein that may

scavenge hydroxide radicals and provides

effective protection against lipid

peroxidation (Prasad et al., 2004).

Therefore, the aim of the present

study was to evaluate the androgenic

effects of different doses of queracetin

(onion juice) or zinc sulphate or their

combination supplementation on sperm

characteristic, hormone measurements and

reproductive performance of male V.Line

rabbits under hot summer conditions of

Alexandria governorate, Egypt.

MATERIALS AND METHODS

The present study was carried out at

El-Sabahia Poultry Research Station

(Alexandria), Animal Production Research

Institute, Agricultural Research Center,

Ministry of Agriculture, Egypt during

summer season conditions from June to

August.

Rabbits, queracetin, zinc sulphate, seminal plasma and fertility.

333

Forty- two males V-line rabbits 7

months old with average initial body

weight of 3.45 Kg were used in this

experiment. Rabbits housed in a naturally

ventilated building and kept in individual

Italian wire galvanized cages (60 × 55 × 40

cm). Rabbits were fed ad libitum with a

commercial pelleted diet containing 18.12

% CP, 13.26% CF and 2670 kcal DE/kg

diet. Fresh tab water was automatically

available all the time in each cage. All the

experimented animals were healthy and

clinically free from internal and external

parasites and kept under the same

managerial and hygienic conditions. Males

were randomly divided into six groups and

supplemented orally with the following

doses of fresh onion juice and zinc sulphate

diluted in distilled water.

Rabbit groups and treatments as follows:

Group 1: control (without any

supplementation).

Group 2: supplemented with 3 ml fresh

onion juice /rabbit/day.


onion juice /rabbit/day.

Group 4: supplemented with 50 mg zinc

sulphate /rabbit/day.


onion juice + 50 mg zinc



onion juice + 50 mg zinc


Extraction of Onion Juice:

Fifty grams of finely chopped red

onions (Alium cepa L.) were thoroughly

agitated with 500 ml of preheated water

(90ºC) for 60 min in order to extract

maximum content of phenolic compounds.

The mixture was cooled at room

temperature and homogenized in a blender.

The homogenate was centrifuged at 10,000

rpm for 20 min and the resulting

supernatant was used as onion juice (Khaki

et al., 2009).

Temperature-Humidity Index (THI):

Air temperature (oC) and relative

humidity (%) inside the rabbitry building

were daily recorded using electronic digital

thermo-hygrometer under hot summer

conditions of Alexandria, Egypt. Averages

of ambient temperature, relative humidity

and temperature humidity index inside

building were 30.8+0.22oC, 81.4+1.05%

and 29.9 respectively, which indicate heat

stress during the experimental period. The

temperature humidity index (THI) was

calculated using the equation modified by

Marai et al. (2001).

THI= db oC- [(0.31-0.31x RH) x (db

oC -

14.4)],

Where THI= Temperature humidity

index, db oC =Dry bulb temperature in

Celsius, RH= Relative humidity

percentage/100. The THI values classified

as follow : >27.8= absence of heat stress,

27.8 ->28.9 moderate heat stress, 28.9 -

>30.0= server heat stress and 30.0 and

more = very sever heat stress.

Semen was collected from rabbit

bucks through two weeks after 10 weeks

from oral supplementations of onion juice

and zinc sulphate. One hundred and sixty

eight ejaculates were obtained (7 rabbit

bucks x 6 groups x 4 ejaculates). Ejaculates

were collected using an artificial vagina

maintained at a teaser doe (45- 46°C).

Reaction time (RT) was the time interval

from introducing of the teaser doe into the

male’s cage to ejaculation and measured in

seconds with a stop watch and considered as

an indication of libido. Immediately after

collection, semen was kept at 35 °C in water

bath in order to be evaluated. Semen volume

of each ejaculate was recorded after removal

of the gel mass. Immediately following

semen collection, for measuring mass

motility rate two drops of fresh semen were

placed on a warmed slide and covered with

a cover slip (20×20 mm). Mass motility

from at least three fields was

examined at 37ºC under a phase microscope

at 40 x and assessed from 0 to 100%. A


334

weak eosin solution was used at a rate of 1:

99 before counting the cells, for evaluation

of sperm concentration (×106/ml) by the

haemocytometer slide according to the

method of Smith and Mayer (1955). Total

sperm output was calculated through

multiplying semen ejaculate volume by

semen concentration. Assessment of live

and abnormal spermatozoa was performed

using an eosin-nigrosine blue staining

mixture (Blom, 1950). The percentage of

live spermatozoa was determined by using

stains that penetrate cells with damaged

membranes. Total number of motile

sperm was calculated as multiplying

percentage of motile sperm by total sperm

outputs. The total functional sperm fraction

(TFSF) was calculated as the product of

total sperm output multiplied by percent of

motile sperms times percent normal sperms

(Correa and Zavos, 1996).

Seminal Plasma:

Seminal plasma was obtained by

centrifugation of semen samples at 4000

rpm for 20 minutes at 4 ºC and stored at -20

ºC until analysis. Acrosomal damage was

determined by using a Giemsa stain

procedure as described by Watson (1975).

Samples were analyzed for the activities of

aspartate aminotransferase (AST) and

alanine aminotransferase (ALT), alkaline

phosphatase (AlP) activity and acid

phosphatase (AcP) activity. Thiobarbituric

acid-reactive substances (TBARS) were

measured in the seminal and blood serum

using the method of Tappel and Zalkin

(1959). Seminal and blood serum glutathione

contents (GSH) were determined using

commercial glutathione reduced kits

according to the method of Beutler et al.

(1963). Glutathione peroxidase (GPx)

activity was assayed using the method of

Chiu et al. (1976). Superoxide dismutase

(SOD) activity was assayed according to

Misra and Fridovich (1972). Glutathione S-

transferase (GST) activity was determined

according to Habig et al. (1974) using P-

nitrobenzylchlorid as a substrate. Catalase

(CAT) activity was measured according to

Aebi (1984).

Blood Analysis:

Blood samples were collected from

the ear vein of each buck through two

weeks after supplementation period (7

rabbits × 5 collections/group). The blood

samples were centerifugated at 4000 rpm

for 20 minutes to separate the serum and

stored in a deep freezer at -20 C until

biochemical analysis.

Serum FSH, LH and Testosterone

Hormones Measurements:

Serum concentrations of FSH and

LH were determined in duplicated samples

using radioimmunoassay (RIA). FSH and

LH kits obtained from Biocode Company-

Belgium, according to the protocol

provided with each kit of serum. The

sensitivities of hormones detected per assay

tube were 0.2ng/ml and 0.14ng/ml for FSH

and LH respectively (Khaki et al., 2009).

Serum testosterone concentration was

measured using immunoassay (Biosource-

Europe S.A. 8, rue de L’Lndustrie.B-1400

Nivelles. Belgium).

Reproductive Performance:

For evaluation of males fertility

criteria, sixty receptive nulliparous V.Line

female rabbits were divided into six groups

and housed in individual cages. Females

from each group were artificially

inseminated from the pooled semen of each

males group. Parameters of kindling rate

and litter size at birth (total born and total

born alive) and at weaning were monitored.

Also, litter weight at birth and 28 days were

recorded.

Statistical Analysis:

Data were analyzed as a one-way

analysis of variance with semen collection

time as replicates using the General Linear

Model procedure of SAS (2002).

Significant differences among treatment

means were tested using Duncan multiple


335

range test (Duncan, 1955).

RESULTS AND DISCUSSION

Semen Characteristics:

Semen characteristics of V-line

male rabbits supplemented orally with

freshly onion juice and zinc sulphate or

their combination are shown in Table 1.

Highest significant (P<0.01) values for

each of ejaculate volume, sperm motility,

total sperm output, total motile sperm and

TFSF were observed for group of males

supplemented with zinc sulphate +higher

level of onion (6ml/rabbit/day) compared to

those for other groups. Males supplemented

with zinc sulphate +lower level of onion

(3ml/rabbit/day) recorded the same highest

significant value for sperm concentration

and TFSF. Also, control males without any

supplementations recorded the worst

(P<0.05) values of ejaculate volume, sperm

motility, sperm concentration, total sperm

output, total motile sperm, live sperm,

normal sperm and TFSF as compared with

those for other groups. Reverse response

was detected for reaction time (libido) by

seconds as the highest (P<0.01) value was

observed for control group and lowest ones

were detected for groups of zinc sulphate

separately and zinc sulphate + onion

supplementation groups and the delay

between the highest and lowest values was

17.35 sec.

The delay of reaction time as

indication of sexual desire for males could

be due to high temperature exposure,

therefore orally supplementation with

onion or and zinc sulphate decreased the

reaction time and increased the sexual

desire. This conclusion is in harmony with

those reported by different research

workers. Tharwat et al. (1994) found that

libido was delayed in NZW rabbit bucks

exposed to 40 ºC with relative humidity 60-

65%. This delay may be due to the decrease

in testosterone concentration, minimal

spermatogenesis (Zeidan et al., 1997) and/

or the low quality semen (El-Kelawy et al.,

1997), occurring in a hot climate. Seleem

(2003) found that addition of zinc

methionine to balanced feed mixtures and

fed to rabbits improved libido and

positively influenced both quantitative and

qualitative of seminal traits.

Supporting to our results regarding

the increase in sperm concentration and

viability of semen, Khaki et al. (2009)

reported that Allium cepa (onion) has a

good effect on spermatogenesis in rats and

the results showed that administration of

onion juice (1g/rabbit/day) for 20

consecutive days caused a marked increase

in sperm count and viability. Moreover, our

findings about the harmful effect of high

temperature on morphological abnormality,

dead sperm and acrosomal damages are in

accordance with the findings of Finzi et al.

(1995). Also, Setchell et al. (2001)

indicated that brief exposure of the scrotum

to the heat results in damage to the most

heat sensitive germ cells, the pachytene

spermatocytes and early spermatids. The

results of improvement in rabbit sperm due

to administration of zinc sulphate in the

current study are in accordance with those

reported by El-Tohamy et al. (2012) who

mentioned that adequate reproductive

performance in NZW rabbit bucks can be

achieved in summer months due to zinc

sulphate supplementation and it is not

obligatory to stop breeding in this period of

year.

Biochemical Constituents of Seminal

Plasma:

Biochemical evaluation of seminal

plasma constituents of male rabbits

supplemented orally with onion and zinc

sulphate under summer season condition is

presented in Table 2. Supplementations the

bucks with onion or zinc sulphate or

combinations between them increased

(P<0.05) the values of AcP, ALP for male

rabbits compared with those for control

one, besides the highest value of AcP was

recorded for group supplemented with zinc

sulphate + high level of onion. Moreover,


336

highest values of AIP were recorded for

groups supplementation with combinations

of zinc sulphate +higher or lower levels of

onion. Reverse response was detected for

AST and ALT values as they decreased

(P<0.05) for all supplemented buck groups

compared to those for control. Furthermore,

males for zinc sulphate+ higher level of

onion recorded the lowest value of AST

(25.69 IU) and ALT (18.72 IU) compared

with the rest of values for all groups. As

can be seen from data of this table that

supplementation the males with

combination of zinc sulphate + 6ml of

onion /rabbit/day were recorded the highest

values of AcP and AIP besides lowest ones

for AST and ALT compared to those for

others.

The improvement in biochemical

constituent concentrations of seminal

plasma resulting from onion or zinc

sulphate supplementations in this study

could be due to the antioxidant activity in

these materials. These results and

conclusion are keeping with those reported

by Chimienti et al. (2003 ) who stated that

onion juice or zinc have strong antioxidant

and antiepileptic. Also, there are multiples

pieces of evidence that injury caused to

spermatozoa by reactive oxygen species

(ROS) plays a key role in the etiology of

male infertility (Aitken and Sawyer, 2003).

Spermatozoa have large quantities of

polyunsaturated fatty acids in their plasma

membrane and low levels of scavenging

enzymes in their cytoplasm. Oxidative

stress has been identified as an imbalance

between the production of ROS and

antioxidant-scavenging activities, in which

the generation of ROS overcomes (Sikka,

2001). Human spermatozoa generate ROS

in physiologic amounts from activities such

as capacitation, the acrosome reaction and

oocyte fusion (Saleh and Agarwal, 2002).

However, too much ROS production when

it exceeds the limited antioxidant defenses

in semen, results in spermatozoa

dysfunction (Agarwal et al., 2003). Seminal

oxidative stress results in decreased semen

quality by several mechanisms, including

injury of the sperm membrane and/or DNA

(Lewis et al., 1995). Sub fertile males with

high levels of ROS have 1/7 the likelihood

of initiating a pregnancy compared with

those with low ROS levels (Sharma el al.,

1999).

The decrease in values of AST and

ALT as shown in Table 2 due to onion and

zinc sulphate supplementations is a good

indicator of semen quality improvement as

Corteel (1980) mentioned that transaminase

activities (AST and ALT) in semen are

good indicators of semen quality because

they measure sperm membrane stability.

Thus, increasing the percentage of

abnormal spermatozoa in ejaculate causes

high concentration of transaminase enzyme

in the extra cellular fluid due to sperm

membrane damage and ease of leakage of

enzymes from spermatozoa (Gundogan,

2006). Al-Daraji et al. (2002) reported that

both of alkaline and acid phosphatase

enzymes were involved in the metabolism

of spermatozoa via the hydrolysis of

carbohydrates and positive correlation

between ALP activity and spermatozoa

concentration and liveability and the

number of spermatozoa per ejaculate.

Seminal Plasma Antioxidants

Constituent:

Seminal plasma antioxidants for

males orally supplementations of onion and

zinc sulphate under summer season

condition are presented in Table 3. Each of

oral supplementation level of onion juice or

zinc sulphate or their combinations

increased (P<0.05) the values of GSH, GPx

and SOD activities for male rabbits

compared with those for control. Whereas,

the GST and CAT were significantly

increased for males supplemented with zinc

sulphate + lower or higher doses of onion

compared with other supplemented groups

and control. Besides the highest values of

GSH, GPx, SOD, GST and CAT were

recorded for males supplemented with zinc

sulphate +higher level of onion compared


337

the other groups. Reverse response of the

experimented supplementations was

detected for TBARS value as they

decreased (P<0.05) for all male groups of

onion levels, zinc sulphate and zinc+ lower

and higher levels of onion compared with

those for control.

Thermal stress is often accompanied

by oxidative stress in which ROS

compounds are produced in greater

amounts (Heise et al., 2003). One of the

most important factors contributing to poor

quality semen has been reported to be

oxidative stress (Bucak et al., 2010). The

increase in production of ROS determines

semen characteristics and sperm-oocyte

fusion (Akiyama, 1999). Also, antioxidant

mechanism is necessary to prevent free-

radical damage to the sperm cell as a result

of secrete hydrogen peroxide and the

superoxide ion by rabbit spermatozoa

(Holland et al., 1982).

Zinc sulphate plays an important

role in the physiology of spermatozoa, in

sperm production and/or viability in the

prevention of spermatozoa degradation, and

in sperm membrane stabilization (Lewis-

Jones et al., 1996 ) and antioxidative

properties may also act to reduce the ROS

leading to an increase in male fertility

(Powell, 2000).

Therefore, Zinc ions act in terms of

membrane stabilization, as well as the

protective role of zinc methionine as a

cellular antioxidant. Zinc intake has been

reported to produce an antiatherogenic

effect as it may have a protective effect on

lipid metabolism consisting in its ability to

prevent hyperlipidemia including especially

hypercholesterolemia and to protect from

lipid peroxidation (Joanna Rogalska et al.,

2009).

Blood Serum Testosterone, FSH and LH

Concentrations:

Data in Table 4 revealed that values

of FSH and LH hormones were increased

(P<0.05) for males supplemented

separately with higher level of onion or

zinc sulphate or their combinations

compared to those for control or lower

level of onion. Besides, rabbits

supplemented with combination between

zinc sulphate + both levels of onion

represented the highest (P<0.05) value of

testosterone hormone compared with those

for control and other supplemented groups.

Supporting to the results herein as FSH, LH

and testosterone hormones decreased with

elevated heat exposure in control group,

Graves (1978) showed that exposure to

hyperthermia is harmful for

spermatogenesis and also decreases

testosterone and gonadotrophins (FSH and

LH) levels. Normal testicular function

requires hormonal stimulation by pituitary

gonadotrophins (LH and FSH) which are in

turn controlled by pulsatile secretion of

gonadotrophin-releasing hormone (GnRH)

from the hypothalamus (Garner and Hafez,

2000).

El-Tohamy et al. (1997) reported

that zinc plays an important role in the

synthesis and secretion of luteinizing (LH)

and follicle stimulating (FSH) hormones

and it is essential in the production of many

sex hormones including testosterone and

gonadotrophin-releasing hormone. In zinc

deficiency, cells are unable to form sex

steroids, leading to arrest of

spermatogenesis and impairment of fertility

(Om and Chung, 1996). Also, Khaki et

al.(2009) revealed that serum total

testosterone was significantly increased in

the male rats supplemented with onion.

Moreover, Jamashid et al.(2012) indicated

that onion had a good effect on LH and

FSH hormones in rats.

Reproductive Performance:

Data of Table 5 presented that

fertility rate of females mated with rabbit

males supplemented orally with either level

of onion juice or zinc sulphate or their

combination were improved (P<0.01)

compared with the control group. In

addition, all experimented

supplementations in this study increased


338

(P<0.01) the values of litter size, live kits,

litter size at weaning and bunny weight at

28 days as compared to those from control.

Whereas, bunny weight at birth realized the

same significant increase due to

experimented supplementations except that

of lower level of onion. Moreover, highest

values of litter size, live kits, litter size at

weaning and bunny weight at 28 days is

observed for group of males supplemented

with zinc sulphate + higher level of onion

compared those for all supplemental groups

and control. Also, it can be observed from

data of control group in this Table for

bucks exposed to hot summer conditions

without any supplementations had

detrimental and bad influence on all

parameters of reproductive performance.

The reduction in fertility rate in

control group is related to the low sperm

count, motility and high percentage of

abnormal spermatozoa level as presented in

Table 1. This conclusion is in accordance

with those reported by Raji et al. (2003).

Also, Brun et al. (2002) found that the mass

motility was significantly influenced the

kindling rate. Furthermore, the same

authors found that litter size (total born)

was significantly influenced by

concentration and number of total motile

sperms.

The improvement of reproductive

performance in the current results as

kindling rates, litter size up to weaning for

groups supplemented with onion and zinc

sulphate compared to control were

accompanied with seminal plasma

constituent improvement for bucks of these

supplemented groups. Ulkowski et al.

(2005) confirmed this statement as they

mentioned that the positive effect of onion

and zinc as an enhancer of reproductive

capacity of rabbit bucks could be attributed

to its ability to protect mammal cells from

oxidation.

Correlation between Sperm Quality

Parameters and Seminal Plasma

Antioxidants Status:

Table 6 showed the correlation

coefficient between GSH, SOD, GPx and

CAT with each of TBARS, abnormal

sperm, dead sperm, sperm motility,

acrosomal damage and sperm concentration

for V.Line male rabbits. Highly significant

negative correlations were detected

between each of GSH, SOD, GPx and CAT

as seminal plasma antioxidant with each of

TBARS and contents of seminal plasma as

abnormal and dead sperms besides

acrosomal damage of sperms. Whereas,

there were strong significant negative

correlations between each one of

antioxidants with sperm motility and

concentration. This observation suggest

that GSH, SOD, GPx and CAT activity of

seminal plasma could play a role in the

protection against lipid peroxidation in

seminal plasma. Moreover, results herein

are in harmony with those reported by

Khoserwbeyvgi and Zarghami (2007) who

mentioned that GSH, SOD, GPx and CAT

activities in seminal plasma from fertile

and infertile men can act coordinately to

protect spermatozoa from lipid

peroxidation. Since lipid peroxidation leads

to loss of motility in human spermatozoa,

the possibility exists that

asthenozoospermic sperm suffer from the

lack of protection against lipid peroxidation

due to lack of adequate or non-coordination

between GSH, SOD, GPx and CAT activity

in seminal plasma (Zini et al., 2000). Also

sperm membrane has been reported to be

adversely affected by peroxidation of

polyunsaturated fatty acid and

accumulation of organic hydroperoides

(Zunjarrao et al., 2011).

Results in Tables 1 and 2 are in

agreement with those reported by Al-Daraji

(2001) who found significant positive

correlation between percentages of dead

spermatozoa, abnormal spermatozoa and


339

acrosomal abnormalieies with seminal

plasma AST and ALT enzymes.

CONCLUSION

Oral supplementation of male

rabbits with 50 mg zinc sulphate + 6 ml

fresh onion juice /rabbit/day through period

of ten weeks under hot summer conditions

improved semen characteristic,

reproductive hormones and performance

compared to control.

M. E

. El-S

peiy

an

d A

.M.E

l-Han

ou

n

Table (1): Overall means of semen characteristics of V.Line male rabbits supplemented orally with onion, zinc sulphate and their

combination

Items Control Onion Zinc

50 mg/rabbit/day

Zinc + Significant

L H L. Onion H. Onion

Reaction time (sce) 34.19a±0.44 25.05

b±0.35 25.13

b±0.51 17.15

d±0.56 16.98

d±0.51 16.84

d±0.47

**

Ejaculate volume (ml) 0.53d±0.021 0.63

c±0.015 0.67

c±0.016 0.75

b±0.030 0.76

b±0.028 0.85

a±0.31

**

Sperm motility (%) 62.14e±0.65 67.86

d±0.61 72.67

c±0.63 75.23

b±0.69 77.42

b±0.43 81.23

a±0.59

**

Sperm concentration

(x106/ml)

214.33e±4.2 325.41

d±4.0 382.46

c±4.3 435.61

b±5.9 468.55

a±6.0 479.85

a±4.6

*

Total sperm output

(x106)

114.23f±8.3 211.47

e±8.8 262.11

d±7.2 329.45

c±9.1 365.16

b±7.3 412.74

a±8.7

**

Total motile sperm

(x106)

72.36f±7.3 143.62

e±8.6 191.43

d±6.9 252.63

c±8.8 285.14

b±7.8 339.45

a±6.3

**

Live sperm (%) 75.92c±0.64 79.61

b±0.69 82.45

ab±0.55 83.72

a±0.61 83.91

a±0.59 84.71

a±0.62

*

Normal sperm (%) 80.68c±0.53 83.36

b±0.48 84.14

b±0.22 86.56

a±0.53 87.12

a±0.66 87.44

a±0.39

*

Dead sperm (%) 21.58a±0.27 19.36

b±0.30 16.57

c±0.25 15.21

d±0.33 13.04

e±0.40 13.11

e±0.27

*

Abnormal (%) 16.51a±0.26 14.38

b±0.29 13.27

c±0.24 12.37

d±0.40 11.53

e±0.41 11.38

e±0.43

*

Acrosomal damage (%) 13.19a±0.33 12.54

ab±0.39 11.24

b±0.32 11.46

b±0.37 10.65

c±0.41 10.24

c±0.34

*

TFSF (x 106) 94.9

e±3.7 214.7

d±4.2 229.6

c±5.4 253.5

b±5.5 356.9

a±9.4 360.5

a±8.2

**

a-f Means within the same row bearing different superscripts, differ significantly

* P<0.05;

** P<0.01;

TFSF= Total functional sperm fraction; L= Lower dose (3ml/rabbit/day); H= Higher dose (6ml/rabbit/day).

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Table (2): Overall means of seminal plasma biochemical (AcP, AIP, AST and ALT) for V.Line male rabbits supplemented

orally with onion, zinc sulphate and their combination

Items

Control Onion Zinc

50 mg/rabbit/day

Zinc + Significant

L H L. Onion H. Onion

AcP (U/L) 33.11e±0.21 33.73

d±0.32 34.19

d±0.18 34.83

c±0.25 35.76

b±0.22 36.29

a±0.36

*

AIP (U/L) 51.78e±0.42 53.24

d±0.39 56.63

c±0.51 60.67

b±0.36 66.57

a±0.44 67.34

a±0.55

*

AST (IU) 30.77a±0.39 29.17

b±0.41 27.85

c±0.42 27.72

c±0.34 26.89

cd±0.46 25.69

d±0.37

*

ALT (IU) 24.58a±0.28 23.83

b±0.32 21.63

c±0.27 20.47

d±0.60 19.43

e±0.19 18.72

f±0.26

*

a-f Means within the same row bearing different superscripts, differ significantly

* P<0.05;

AcP=Acid phosphatase; AIP=Alkaline phosphatase; ALT= Alanine aminotransferase; AST=Aspartate aminotransferase;

L= Lower dose (3ml/rabbit/day); H= Higher dose (6ml/rabbit/day).

Table (3): Overall means of seminal plasma antioxidants (TBARS, GSH, GPx, SOD, GST and CAT) for V.Line male rabbits

supplemented orally with onion, zinc sulphate and their combination

Items Control Onion Zinc

50 mg/rabbit/day

Zinc + Significant

L H L. Onion H.Onion

TBARS

(nmol/ml) 1.74

a±0.051 1.25

b±0.047 1.05

c±0.036 1.023

c±0.051 1.031

c±0.049 1.013

c±0.052

*

GSH(g/dl) 13.13d±0.19 14.56

c±0.24 16.87

b±0.31 18.73

a±0.18 18.71

a±0.22 18.81

a±0.26

*

GPx (mg/l) 4.37d±0.037 4.80

c±0.063 4.77

c±0.051 5.00

b±0.044 5.08

b±0.036 5.29

a±0.047

*

SOD(IU) 7.24b±0.025 7.49

a±0.036 7.50

a±0.047 7.52

a±0.025 7.51

a±0.019 7.57

a±0.027

*

GST(mol/hr) 1.26c±0.062 1.30

c±0.044 1.30

c±0.052 1.33

c±0.063 1.42

b±0.57 1.82

a±0.046

*

CAT (/ml) 14.30d±0.12 14.96

d±0.14 16.40

c±0.12 16.90

c±0.13 17.93

b±0.11 19.11

a±0.12

*

a-d Means within the same row bearing different superscripts, differ significantly

* P<0.05;

TBARS = Thiobarbituric acid–relative substances; GSH= Glutathione content; GPx= Glutathione peroxidase; SOD= Superoxide

dismutase; GST= Glutathione S-transferase; CAT= Catalase; L= Lower dose (3ml/rabbit/day); H= Higher dose 6ml/rabbit/day).

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Table (4): Overall means of reproductive hormones for V.Line male rabbits supplementated orally with onion, zinc sulphate and

their combination

Items

Control Onion Zinc

50 mg/rabbit/day

Zinc + Significant


FSH 20.37b±0.095 20.39

b±0.086 21.61

a±0.078 21.65

a±0.091 21.72

a±0.088 21.70

a±0.076

*

LH 18.78b±0.076 18.79

b±0.056 19.97

a±0.073 19.96

a±0.059 20.02

a±0.078 20.06

a±0.048

*

Test. 4.24e±0.046 4.45

d±0.055 4.77

c±0.062 5.05

b±0.049 5.30

a±0.062 5.28

a±0.051

*


* P<0.05;

FSH= Follicle-stimulating hormone; LH= Luteinizing hormone; Tes= Testosterone hormone;

L= Lower dose (3ml/rabbit/day); H= Higher dose (6ml/rabbit/day).

Table (5): Overall means of reproductive performance for V.Line male rabbits supplemented orally with onion, zinc sulphate and their

combination

Items Onion Zinc

50 mg/rabbit/day

Zinc + Significant


Fertility (%) 56.68 e±1.33 71.89

d±1.18 79.70

c±1.24 82.94

b±1.31 84.34

b±1.43 89.73

a±1.29

**

Litter size at

birth (n)

5.57c±1.02 7.30

b±1.14 8.90

a±0.89 8.85

a±1.19 9.28

a±1.01 9.85

a±1.13

**

Live litter at

birth (n)

4.35c±0.76 6.71

b±0.92 8.28

a±1.03 8.42

a±0.79 8.86

a±0.83 9.29

a±0.74

**

Litter size (n)at

weaning

3.58d±0.85 6.28

c±0.96 7.28b

c±0.74 7.85

b±0.81 8.29

ab±0.69 8.86

a±0.91

**

Bunny weight

(g) at birth

44.57c±1.39 46.14

c±1.51 56.44

b±1.25 56.57

b±1.46 57.53

b±1.44 59.73

a±1.62

**

Bunny weight

(g) at 28 d

422.00d±21.1 559.00

c±27.15 606.60

b±28.18 639.43

ab±29.1 647.29

a±26.71 652.86

a±28.4

**


** P<0.01;

L= Lower dose (3ml/rabbit/day); H= Higher dose (6ml/rabbit/day)


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Table (6): Correlation coefficient between GSH, SOD, GPx and CAT with TBARS,

abnormal sperm, dead sperm, sperm motility, acrosomal damage and sperm

concentration for V.Line male rabbits.

Items TBARS Abnormal

sperm

Dead

sperm

Sperm

motility

Acrosomal

damage

Sperm

concentration

GSH -0.855 -0.917 -0.929 0.907 -0.644 0.941

SOD -0.790 -0.599 -0.652 0.665 -0.574 0.747

GPx -0.795 -0.782 -0.816 0.851 -0.585 0.878

CAT -0.702 -0.805 -0.775 0.800 -0.629 0.803

TBARS = Thiobarbituric acid–relative substances; GSH= Glutathione content; GPx=

Glutathione peroxidase; SOD= Superoxide dismutase; CAT= Catalase.

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الملخص العزبى

لذكور األرانب األداء التناسلىتأثيز استخذام الكيوراستين )عصيز البصل( وكبزيتات الزنك على

الحارهظزوف الصيف تحت

و على محمد الحنون محمد السيد السبيعى لبحوث الزراعيو ، وزاره الزراعو ، مصرمعهد بحوث االنتاج الحيوانى ، مركز ا

تذف تهك انذساس ان دساس تأحيش استخذاو انكيساستي )عصيش انبصم( أ كبشيتات انضك أ يخهغ ي

زكس األساب انعشظ نذسجات حشاس عاني خالل فصم انصيف. حيج تى استخذاو عذد ن األداء انتاسهكهيا عه

ركس. انجع 7بكم يا يجاييع يتساي 6تى تقسيا عشائيا ان انف اليأساب شس ي 7ركش عش 24

بانتجشيع بانفى . انجاييع انخاي انخانخ تى يعايهتا )كتشل( بذ أ يعايالت أعتبشت يجع يقاسكات األن

بانتجشيع بانفى انجع انشابع تى يعايهتايهه/ األسب/ انيو عه انتان. 6 3بعصيش انبصم بجشعات

05بـ أيعا بانتجشيع بانفى تى يعايهتا انخايس انسادسبيا انجاييع .األسب/انيويهجى/ 05بكبشيتات انضك بعذل

نذ . انعايالت استشتانتانيهه ي عصيش انبصم/األسب/ انيو عه 6 3كبشيتات صك باالظاف ان يهجى

كات أى انتائج يايه:يتتاني قبم انبذء ف تجيع انسائم ان أسابيع 05

، عذد انحياات انحشك انتقذيي نهحياات اني حجى انقزف ،ف كم ي (P<0.01) حذحت صياد يعي

، عذد انحياات اني انحي بانقزفعذد االسبشيات ف انقزف ، عذد انحياات اني انتحشك اني بانقزف ،

يهه/ األسب/ 6انطبيعي نجع انزكس انت تى يعايهتا بخهغ ي انضك انجشع انعاني ي عصيش انبصم )

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يشاذ نجع انكتشل أقم قي نهجاييع انعايه بانضك فقػ أ بانضك يعافا اني (P<0.01)يعي قي

عصيش انبصم.

يهه ي عصيش انبصم/األسب/انيو أد ان حذث صياد 6يعافا اني يعايه انزكس بخهغ ي انضك

بجاب حذث اخفاض يع نكال AIPانقاعذ AcPنكال ي تشكيض اضيى انفسفيت انحايع (P<0.05)يعي

seminal plasma aspartate aminotransferase (AST), alanine aminotransferase (ALT)ي

تعتبش يؤشش الخفاض أكسذ انهيبيذات ف انبالصيا اني يجع انكتشل.بانجاييع انعايه األخش بانقاس

صياد تحذحيه بعصيش انبصم كبشيتات انضك. انشاسد انحش بيا صادت االضيات انعاد نألكسذ تيج نهعا

نهزكس ف كال ي يست شي انتستيستيش شي انتبيط شي عج انحيصالت (P<0.05)يعي

عصيش انبصم أ ي انضك أ يخهغ ي كالا بانقاس بجع انكتشل انت تى يعايهتا بانجشع انعاني ي

ألساب انت تى يعايهتا بخهغ ي انضك اانجع انعايه بست يخفط ي عصيش انبصم. ان جاب رنك

يستيستيش ف قي شي انت (P<0.05)كال انستيي ي عصيش انبصم أد ان صياد يعي يعافا اني

بانقاس بجع انكتشل انجاييع انعايه األخش. جيع انصفات انخاص باألداء انتاسه يخم يعذل انخصب ،

بشكم كبيش حذث (P<0.01)يو قذ تأحشت 42حجى انخهف عذ انيالد عذ انفطاو ، ص انخهف عذ انيالد عذ

ف انصيف انحاس نى يتى يعايهتا بأ ي يعايالت انتجشب )انكتشل( بانقاس با اخفاض نألساب انعشظ نظش

بانجاييع األخش.

effect of queracetin (onion juice) and zinc sulphate … · 2017-10-27 · levels of onion or zinc...

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