The Effect of Combination Sweet Orange Peel Essential Oil with Trish

Yolk and Streptomycin Extender on the Quality Boer Goat Liquid Semen

Sukma Aditya Sitepu

, Zaituni Udin

, Jaswandi

and Hendri

University of Pembangunan Panca Budi Medan

Andalas University

Keywords: Boer Goat, Essential oil, Liquid semen, Streptomycin, Sweet orange peel.

Abstract: The purpose of this research is to know the quality of Boer Goat liquid semen after being given combination

of sweet orange peel essential oil with streptomycin on tris yolk extender. The research used completely

randomized design with 5 treatments and 5 replications. Treatment research is the addition sweet orange peel

essential oil to tris yolk and streptomycin extender are 0%, 0,25%; 0.5%; 0.75% and 1%. Parameters were

Motility, viability, Membrane Integrity, and Acrosome Integrity. Observations are made before and after

equilibration. The results showed that the addition of sweet orange peel essential oil was significantly

influenced (P <0.01) to percentage of motility, viability, Membrane Integrity, and Acrosome Integrity liquid

cement of Boer Goat. The best research treatment is the addition 1% sweet orange peel essential oil.

1 INTRODUCTION

The need for meat in Indonesia is very high because

of the rising population, the awareness of the

importance protein needs and the increase of people's

income. To meet these needs may be done by

livestock producers of meat.

Goat cattle is one effort that can be done to meet

the demand of meat. Goats are small ruminants cattle

that are kept by livestock farmers in the village. Goats

have the advantage of being able to adapt in extreme

conditions, resistant to disease, rapidly proliferate and

proliferate so that goats are very potential to be

developed (Mulyono and Sarwono, 2008).

To develop a goat farm can be done by improving

genetic quality and population. Genetic quality of

goats can be improved through crossbreeding with

superior goats, such as the Boer Goats. The Boer

Goats have the superiority of having a high body

weight, fast growth and high litter size (Ginting and

Mahmilia, 2008). Boer goat good quality still hard to

find in Indonesia. It is necessary for the optimal use

of the Boer Goat males.

Artificial Insemination is a reproductive

technology that can be done to increase the

population and genetic quality of livestock. Artificial

Insemination has the advantage that the spread of goat

livestock can be done cheaply, easily and quickly

because the breeders do not need to buy and maintain

the Boer Goat male.

Artificial Insemination program's success

depends on the quality of liquid semen is used. Good

quality semen is obtained from superior goats, as well

as semen processing from shelter to dilution. The

materials used in the manufacture of liquid semen

must be considered. Selection of the type of semen

extender greatly affects the quality of the resulting

liquid semen. In goat semen extender used is still

being developed. Tris yolk extender is very

commonly used in the preservation and

cryopreservation process of semen of various types of

animals and livestock. Tris yolk extender are

composed of: tris, citric acid, fructose and akuabidest

(Rizal and Herdis, 2008).

The decreasing quality of Boer Goats liquid

semen is caused by the development of bacteria

contained therein. Bacteria enter the liquid semen

from the shelter process and the manufacture of less

hygienic semen. To inhibit bacterial growth can be

accomplished by the addition of antibiotics, one of

which is streptomycin (Rizal and Herdis, 2008).

However, the addition of streptomycin is still lacking

inhibit bacterial growth so we need a combination

with other materials that contain antibacterial.

One of the ingredients that contain antibacterial is

essential oil. Essential oils widely used fragrance

chemical industry, adds flavor to beverages and

Sitepu, S., Udin, Z., Jaswandi, . and Hendri, .

The Effect of Combination Sweet Orange Peel Essential Oil with Trish Yolk and Streptomycin Extender on the Quality Boer Goat Liquid Semen.

DOI: 10.5220/0010044404370441

In Proceedings of the 3rd International Conference of Computer, Environment, Agriculture, Social Science, Health Science, Engineering and Technology (ICEST 2018), pages 437-441

ISBN: 978-989-758-496-1

437

foods. In the pharmaceutical and health industries are

used as, antiinfectives, antibacterials, viruses and

fungi (Lutony, 1994). Sweet orange essential oil

contains the main components such as limonene,

linalool which is toxic in bacteria (Fisher and Phillips,

2006).

Combining streptomycin and essential oils of

sweet oranges are expected to minimize bacterial

growth, improve the quality and survival of

spermatozoa in frozen semen and increase the fertility

rate of Boer Goats. Boer Goat liquid semen produced

can be used in the development and improvement of

the goat population through artificial insemination.

2 RESEARCH METHODS

2.1 Materials and Equipment

The research materials used are fresh cement of Boer

Goat, streptomycin, Sweet orange essential oil and

tris yolk extender, citric acid, fructose, 2% eosin,

formolsaline, aquabidestillata, warm water (45-55 º

C), HOST (Hipo Osmotic Swelling Test ) and tissues.

The research tool used is 400 ml beaker glass,

deck glass, tissue, stirrer, an artificial vaginal set, test

tube, water bath, electric microscope, thermometer,

dropper drop, object glass, cover glass, pH meter,

bunsen burner, denominator , container cage and mat.

2.2 Methods

The research method used is a complete randomized

design trial method using 5 treatments and 5

replications.

The treatment provided is:

P0 = Streptomycin + Essential Oils 0%

P1 = Streptomycin + Essential Oils 0.25%

P2 = Streptomycin + Essential Oils 0.5%

P3 = Streptomycin + Essential Oils 0.75%

P4 = Streptomycin + Essential Oils 1%.

2.3 Evaluation Parameters

Motility: Assessment of motility percentage is done

by observing progressive moving spermatozoa

forward. The evaluation was performed using a

microscope that observed spermatozoa in eight

different fields of view with 400 times magnification

light (Toelihere, 1993).

Viability: Observations were made by using eosin

staining on the sample (Toelihere, 1993). Live

spermatozoa characterized by the head of

spermatozoa which do not absorb the color, while the

die is characterized by a head to absorb the color. The

evaluation was performed using a 400 times

magnification light microscope. Spermatozoa were

observed at least 200.

Membrane integrity: The observations were

performed on the integrity of the plasma membrane

of spermatozoa using hypoosmotic swelling test

method by mixing 0.1 ml of semen and 9.9 ml of

hypoosmotic medium, then incubating at waterbath at

37 ° C for 30 minutes (Rodriquezgil et al., 1994). The

evaluation was performed using a microscope at 400

times magnification.

Acrosome integrity: The observations were

performed on the spermatozoa acrosomal whiteness

of the head against the thick black spermatozoa by

mixing 0.1 ml of semen and 9.9 ml a physiologic

NaCl solution containing 1% formalin (Saacke dan

White, 1972). The evaluation was performed using a

microscope at 400 times magnification. Spermatozoa

were observed at least 200.

3 RESULTS AND DISCUSSION

3.1 Research Result

The results of Boer goat semen research before and

after equlibrasi by using a combination of

streptomycin and sweet orange essential oil in the

diluent can be seen in Table 1.

Table 1: Recapitulation of research results of semen quality

of Boer Goats before and after equilibration.

Paramete

s Treatments

Observation

Before

Equilibration

After

Equilibration

Motilit

0% 67 69

0,25% 70 72

0,5% 71 75

0,75% 73 75

1% 75 77

Viabilit

0% 80 77

0,25% 85 80

0,5% 87 83

0,75% 88 84

1% 89 86

ICEST 2018 - 3rd International Conference of Computer, Environment, Agriculture, Social Science, Health Science, Engineering and

Technology

438

Table 1: Recapitulation of research results of semen quality

of Boer Goats before and after equilibration (cont.).

Paramete

s Treatments

Observation

Before

uilibration

After

uilibration

Membrane

inte

rit

0% 72 70

0,25% 77 75

0,5% 79 76

0,75% 80 77

1% 83 81

Acrosome

inte

rit

0% 68 65

0,25% 72 70

0,5% 74 72

0,75% 75 74

1% 77 76

Description: Different superscript on the column showed a

highly significant difference (P<0.01)

3.2 Discussion

Boers liquid semen quality is a determining factor for

the success of fertilization after artificial

insemination. determinants of the quality of

microscopically viable spermatozoa for artificial

insemination ie motility, viability, membrane

integrity percentage and the percentage of acrosome

integrity.

3.2.1 Percentage Motility of Boer Goat

Spermatozoa

The results of observational studies of fresh semen

motility after preparation using tris diluent yolk and

essential oil and observations after equilibrasi show

different results each treatment. According to Hafez

(1987), the motility of spermatozoa is one of the

determinants of successful spermatozoa to reach the

ovum of the fallopian tube and the simplest way of

sperm assessment for artificial insemination.

The results showed the average observation of

each motility treatment on all treatments meet the

standards for use as liquid semen for Artificial

Insemination because the above motility of 67% and

the number has been eligible for Artificial

Insemination. Sufficient semen requirements

Artificial Insemination has a motility that is less than

40% (Evans and Maxwell, 1987).

The best spermatozoa motility that can be used

and meet the standard in this research is on the

addition of 1% essential oil as diluent and the higher

the essential oil level given to the diluent will further

decrease sperm motility either after dilution and after

equilibration. The quality of semen will decrease if

storage is not coupled with the right diluent (Hafez,

2000).

Hydroxynonenal is one of lipid peroxidation that

can inhibit glycolysis and sperm motility. In addition

to the damage caused by lipid peroxidation, decreased

motility can also occur due to several factors White

(1993). According Tounere (1993) factors that can

decrease motility that changes pH Medium, osmotic

pressure and the effects of electrolyte and non

electrolyte. Decreased sperm motility may be due to

high levels of essential oils used in diluents. Lipid

peroxidation which occurs as a result of increasing oil

content in the diluent is too large.

3.2.2 Percentage Viability of Boer Goat

Spermatozoa

The immobile spermatozoa may not necessarily die

so as not to absorb the color, whereas the

interpretation of the moving and immobile base is

considered immotile. At the spermatozoa are alive

and moving, but there are defects in the cell wall, can

absorb the color is presumed dead, while the other

interpretation is considered not die Partodihardjo

(1982)

The result of variance analysis showed that the

effect of the addition of combination of streptomycin

with essential oil of sweet orange as a diluent had a

very significant effect (P <0.01) on the viability of

spermatozoa both before and after equlibration.

Further test results showed that the best viability was

found in 1% addition of 89% before equlibration and

86% after equlibration.

The National Standardization Agency determines

the quality of semen after the freezing process should

show live spermatozoa (viability) of at least 40%

(Anonimous, 2005). Decreased spermatozoa quality

after equlibration caused by spermatozoa experience

cold shock (cold shock) (Toelihere, 1993).

3.2.3 Percentage Membrane Integrity of

Boer Goat Spermatozoa

An intact plasma membrane integrity of spermatozoa

that was instrumental in the process of fertilization to

the success of artificial insemination. Spermatozoa

with intact membrane will retain hypoosmotic fluid

inside the cell so that it does not suffer damage

(Hafez, 2000).

The Effect of Combination Sweet Orange Peel Essential Oil with Trish Yolk and Streptomycin Extender on the Quality Boer Goat Liquid

Semen

439

The result of analysis of intact plasma membrane

variety of Boer goat showed a very real effect (P

<0.01) on the addition of essential oil as diluent.

Viewed from the average value of the highest intact

plasma membrane of Boer Goats at P4 treatment ie

83% before equilibrium and 81% after equilibration.

Sweet orange essential oil serves as an antioxidant

very useful for intact intact plasma membrane Boer

Goat. This is in accordance with the opinion of

Maxwell and Watson (1996) which states that the

spermatozoa plasma membrane is rich in unsaturated

fats and therefore susceptible to lipid peroxidation.

The result of lipid peroxidation is the formation of

lipid peroxides, which will react as free radicals and

induce an autocatalytic reaction, resulting in the

destruction of the plasma membrane (Sinha et al.,

1996).

The results showed that the percentage of intact

plasma membranes was also associated with viability

of spermatozoa. Spermatozoa with a high percentage

of live shows a high percentage of intact plasma

membranes as well. Spermatozoa that have a high

percentage of life indicate that the plasma membrane

is still intact physically, so the organelles of

spermatozoa cells will be protected, the need for food

substances and ions for metabolic processes is

available (Tambing et al., 1999).

Higher levels of sweet orange essential oil

administration it will increase the percentage of intact

plasma membrane. This may be due to a higher

lactose concentration causing a change in osmotic

pressure in the diluent toward the hypertonic.

Hypertonic diluents indicate that the molecules or

particles outside the cell are more than in the cell. As

a result there is discharge of water from within the

cell to thin out the molecules outside the cell, so that

the cell will contract (Supriatna and Pasaribu, 1992).

3.2.4 Percentage Acrosome Integrity of Boer

Goat Spermatozoa

Acrosome integrity role in the fertilization process to

the success of artificial insemination. The changes

that occur in acrosom are more often associated with

fertility than sperm motility.

The results showed the highest percentage of

intact acrosomal cap on P4 treatment after dilution of

77% and 76% after equilibration. The results showed

that increasing the concentration of essential oils in

the diluent will cause the average percentage of intact

sperm acrosome covering the lower Boers.

The low percentage of intact acrosome hood and

intact plasma membrane is associated with low

percentage of motility, viability and intact plasma

membrane. According to Salisbury and Van DeMark

(1985) there is a physiological relationship between

motility and the integrity of the plasma membrane

and the survival of the spermatozoa. Plasma

membrane damage will cause the loss of the

necessary enzymes

4 CONCLUSIONS

The best combination of streptomycin and sweet

orange essential oil with percentage of motility,

viability, membrane integrity and acrosome integrity

of Boer Goat's cement is at 1% level.

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The Effect of Combination Sweet Orange Peel Essential Oil with Trish Yolk and Streptomycin Extender on the Quality Boer Goat Liquid

Semen

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