Journal of Veterinary and Animal Science pISSN 2550-1283 February 2019 Vol.2 No.1

JOURNAL OF VETERINARY AND ANIMALS SCIENCE Institute for Research and Community Service, Udayana University E-mail: jvas@unud.ac.id

EDITOR IN CHIEF

I Wayan Suardana (Udayana University, Indonesia)

CO EDITOR IN CHIEF I Nyoman Suarsana (Udayana University, Indonesia)

Ni Ketut Suwiti (Udayana University, Indonesia)

I.G.M. Krisna Erawan (Udayana University, Indonesia)

EDITORIAL BOARDS

A.A. Ayu Mirah Adi (Udayana University, Indonesia) Aris Haryanto (Gadjah Mada University, Indonesia) Christian Bauer (Justus Liebig, German) Alan P. Dargantes (Central Mindanao University, Philipines) Zdenek Knotek (Brno University, Cekoslovakia) Dyah Ayu Widiasih (Gadjah Mada University, Indonesia)

Komang G. Wiryawan (Bogor Agriculture University, Indonesia) Michael Haryadi Wibowo (Gadjah Mada University, Indonesia) Omar Akineden (Justus Liebig University, German) Saleha Abd. Aziz (University of Putra Malaysia, Malaysia) Duangporn Pichpol (Chiang Mai University, Thailand) Yasunobo Matsumoto (The University of Tokyo, Japan)

TECHNICAL EDITORS

Kadek Karang Agustina (Udayana University, Indonesia)

I Wayan Nico Fajar Gunawan (Udayana University, Indonesia)

Putu Henrywaesa Sudipa (Udayana University, Indonesia)

I Made Merdana (Udayana University, Indonesia)

PUBLISHER Institute for Research and Community Service Udayana University

Campus of Bukit Jimbaran, Badung, Bali.

Email: jvas@unud.ac.id

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SCOPE OF JOURNAL

Journal of Veterinary and Animal Sciences (JVAS) with pISSN 2550-1283 is a peer-reviewed journal which devoted to the advancement and dissemination of scientific knowledge concerning veterinary and animal science which includes research findings, case report, experimental design, and their application for the treatment of diseases in birds, wild and domestic animals. This journal published in English twice a year on Pebruary and August by Institute for Research and Community Service, Udayana University. It covers all the scientific and technological aspects of veterinary medicine in general, anatomy, physiology, biochemistry, pharmacology, microbiology, pathology, public health, parasitology, infectious diseases, clinical sciences, biotechnology, alternative veterinary medicine and other biomedical fields. In the field of animal science, the journal receives original manuscripts covering breeding and genetics, reproduction and physiology, nutrition, feed sciences, animal products, biotechnology, behavior, livestock farming system, sosio-economic, and policy

ABOUT JOURNAL

Journal of Veterinary and Animal Sciences (JVAS) is a scientific journal published since 2017. The journal consistently two times a year in Pebruary, and August. The journal is registered in DOAJ. JVAS already used Cross Check to prevent any suspected plagiarism in the manuscripts.

SECRETARIAT OF JVAS

Institute for Research and Community Service, Udayana University Bukit Jimbaran, Badung, Bali.IndonesiaPhone / Fax: +62-361-704622 / 703367

e-mail: jvas@unud.ac.id

Journal of Veterinary and Animal Sciences pISSN 2550-1283 August 2017 Vol. 1 No.2

Journal of Veterinary and Animal Science pISSN 2550-1283 February 2019 Vol.2 No.1

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JOURNAL OF VETERINARY AND ANIMALS SCIENCE Institute for Research and Community Service, Udayana University E-mail: jvas@unud.ac.id

Volume 2-Issue 1- February 2019

Table of Contents

Original Article

The Growth Hormone Level of Bali Cattle’s Post Treatment

with Ethinil Esthradiol and Progesteron Hormones in Combination 1-9

with Mineral. Aletha Yuliana Mandala, Ni Ketut Suwiti, I Wayan Suardana

Morphometry of Liver Fluke (Fasciola Gigantica) Infecting 10-17

Balinese Cattle. Putu Anna Oktaviana, Nyoman Adi Suratma, I Nengah Wandia

Studies on Species of Lactic Acid Bacteria Isolate Sr 12 From Bali Cattle 18-23

Gastric with Conventional Method and Api Kit 50ch. Made Radwika,

I Wayan Suardana, I Dewa Made Sukrama

Probiotic Potential Test for Lactic Acid Bacterial Isolate from Labi Cattle 24-30

Gastric Again Low Acidity and Sodium Deoxycholate. I Made Rahma Sanjaya,

I Wayan Suardana, I Nyoman Suarsana

Studies on Lactic Acid Bacteria Isolate Sr 13 From Bali Cattle Gastric 31-38

Bagus Nanda Govinda Muria Siddhi, I Wayan Suardana, Nyoman Semadi Antara

The Antibiotics Group Aminoglycosides and Macrolides on Beef in Bali 39-45

Siswanto Siswanto, I Nyoman Sulabda

9Journal of Veterinary and Animal Sciences pISSN 2550-1283

DOI: https://doi.org/10.24843/JVAS.2019.v02.i01.p03 February 2019 Vol. 2 No.1: 18 - 23

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Studies On Species Of Lactic Acid Bacteria Isolate Sr 12 From Bali Cattle

Gastric With Conventional Method And Api Kit 50ch

Made Radwika1*, I Wayan Suardana2, I Dewa Made Sukrama3

1Student at Faculty of Veterinary Medicine;2Laboratory of Veterinary Public Health, Faculty

of Veterinary Medicine; 3Laboratory of Clinical Microbiology, Faculty of Medicine,

University of Udayana, Bali

*Corresponding Author: radwikam@yahoo.com

Abstract. Lactic acid bacteria (LAB) are Gram positive bacteria that characterized by non-

motil, non-spore, negative catalase, coccus or bacillus cell shape, and live in microaerophilic

condition with CO2 aeration 5-7%. The purpose of study was to identifyy LAB isolate SR 12

isolated from gastrointestinal tract of bali cattle that previously known have potency as a

source of probiotic. The study initiated by cultivation of isolate on de Man, Rogosa, Sharpe

(MRS) medium, confirmation of isolate with Gram staining and catalase test as a main

procedure for Gram positive indentification. The next identification in order to know the

species of isolate was conducted both conventional test and kit API 50CH test. The result of

study showed the LAB SR12 isolate as a genus of Pediococcusor Enterococcus according to

its growth in temperature 10oC, pH9,6 and it can’t grow in halophilic condition. On the other

hands, the use of ofAPI kit 50CH test showed the LAB SR12 was detected as Pediococcus

pentosaceus.

Keyword: Bali cattle, Identification, Lactic Acid Bacteria, SR 12.

I. INTRODUCTION

Food processing through

fermentation has been done by humans

since prehistoric times. Pure culture of

Lactic Acid Bacteria was found by J.

Lister in 1873 and use in cheese making as

fermenter bacteria in 1890. Lactic Acid

Bacteria (LAB) is a bacterium that can

fermentate carbohydrate to lactic acid.

Lactic Acid Bacteria can be divided into

two groups, such as homofermentative

LAB produce 95% lactic acid as a result of

its fermentation and heterofermentative

LAB produce not only lactic acid but

ethanol and carbon dioxide as

byproduct[1].It is known that cows

gastrointestinal track are the habitat of

several types of lactic acid bacteria which

have good potential as probiotics[2-

5].Based on the results of previous

research conducted by Suardanaet al in

2012 (unpublished data), it was found that

from some strains of rumen origin of the

Bali cattle, especially SR 12 is known to

have the potential to be developed as a

source of probiotics. This is based on the

ability of isolate SR 12 to inhibit

pathogenic bacteria with large inhibitory

zones.

II. RESEARCH METHOD

Cultivation of LAB SR 12 isolate

The cultivation of BAL isolate SR

12 was done by isolate reculture that

characterized by turbidity on MRS broth

media.

Radwika et al., JVAS

19

Gram Staining

Gram stain on SR 12 isolate was

done by coloring preparations that have

been fixed with crystal violet 2%, lugol

solution, acetone alcohol, and safranin.

The staining results can be seen under a

microscope with 1000x magnification.

Catalase Test

The catalase test was conducted to

determine the activity of catalase enzyme.

Positive results are characterized by

bubbles.

Conventional Test

Growth Test on 10oC Temperature

A 10oC temperature test is

conducted to determine the ability of

bacteria to grow in low temperatures.

Isolates that grow at this temperature are

classified as Pediococcus, Lactobacillus,

Enterococcus or Tetragonococcus. On the

contrary, isolate that do not grow at this

temperature are classified as Streptococcus.

Growth Test on NaCl 15%

A 15% NaCl test is conducted to

determine the ability of bacteria to grow in

hypersaline conditions. Isolates that grow

at this condition are classified as

Tetragonococcus. On the contrary, isolate

that do not grow at this condition are

classified as Pediococcus.

Growth Test on pH 9,6

A pH 9.6 test was conducted to

determine the ability of bacteria to grow in

alkaline conditions. Isolates that grow at

this pH are classified asEnterococcus. On

the contrary, isolate that do not grow at

this pH are classified as Lactococcus.

API Kit 50CH Test

The API test kit aims to identify

species of lactic acid bacteria by utilizing

the ability of LAB to ferment

carbohydrates. The test was carried out

with the Analytical Profiler Index kit.

III. RESULT AND ANALYSIS

Confirmation of Isolate BAL SR 12

Gram Staining

Gram staining results (Figure 1)

show that LAB isolates SR 12 are Gram

positive bacteria, and bacterial cells are

coccus shaped. Based on its form, lactic

acid bacteria SR 12 isolates belong to the

genus Lactococcus, Aerococcus,

Enterococcus, Streptococcus, Pediococcus,

Leuconostoc [6].

Figure 1. The results of Gram stain LAB isolate SR 12. The results of Gram staining bacteria

showed that bacteria were Gram positive bacteria with a coccus shape

9Journal of Veterinary and Animal Sciences pISSN 2550-1283

DOI: https://doi.org/10.24843/JVAS.2019.v02.i01.p03 February 2019 Vol. 2 No.1: 18 - 23

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Catalase Test

The principle of the catalase test is to

detect active enzymes in bacteria that are

useful in breaking down hydrogen

peroxide (H2O2) into 2 molecules of water

(H2O) and 1 molecule of oxygen (O2).

Catalase test results on LAB SR 12

isolates are presented in Figure 2.

Figure 2. Result of catalase test on LAB SR 12 isolates. SR 12 isolates showed negative

results.

Isolate Identification of LAB SR 12

Conventional test

Conventional tests are carried out

with the aim of detecting BAL through its

physiological characteristics. Conventional

test results on LAB SR 12 isolates are

presented in table 1.

TABLE 1. CONVENTIONAL TEST RESULT OF LAB ISOLATE SR 12

No Type of Test Result

1. Gram Staining Positive

2. Shape Coccus

3. Catalase Test Negative

4. 10oC Temperature Test Positive

5. NaCl 15% Test Negative

6. pH 9,6 Test Positive

Conclusion Pediococcus/Enterococcus

Based on the results of conventional tests,

the LAB SR 12 isolates can be identified

as the genus Pediococcus or Enterococcus

based on the LAB classification through

morphological and physiological

characteristics [7].

API Kit 50CH Test

The API kit test was carried out to

detect LAB isolates SR 12 by utilizing

biochemical characteristics of isolate by

testing 50 type of sugar. The API 50CH kit

test results are presented in Figure 3.

Radwika et al., JVAS

21

Figure 3. Results of API 50CH kit test on LAB SR 12isolates. (A). Positive results. (B).

Negative results. (C). Positive control.

(A). Positive results (microtube no. 11) are marked by changes in color from blue to

yellow; (B). Negative results (micotube no. 20) are marked with no discoloration; (C). On

microtube no. 25 positive results were marked by reddish-black discoloration, this was due to

hydrolysis of esculin ferric citrate to esculitin and glucose. Color changes caused by exculitin.

The test results in Figure 3 are briefly as shown in Figure 4.

Figure 4. Test results of API Kit 50CH on apiweb

B

A

C

9Journal of Veterinary and Animal Sciences pISSN 2550-1283

DOI: https://doi.org/10.24843/JVAS.2019.v02.i01.p03 February 2019 Vol. 2 No.1: 18 - 23

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Based on the results of the API

50CH kit test, LAB isolate SR 12 was

identified as Pediococcus pentosaceus 2.

The results of this study were the same as

conventional tests. Pediococcus

pentosaceus bacteria have various benefits,

especially as probiotics. Borges et al in his

study said Pediococcus pentosaceus in the

form of frozen dry powder has the ability

to inhibit pathogenic bacteria such as

Listeria monocytogenes [8]. These bacteria

can also be used as anti-obesity drugs by

reducing the Body mass Index (BMI),

waist circumference, and fat in the body

[9]. Pediococcus pentosaceus bacteria is

very useful in the process of fermentation

of mackerel fish sausages by suppressing

the growth of Enterobacteriaceae and

Staphylococcus bacteria [10]. Sadeghi et al

in his study found that Pediococcus

pentosaceus has antifungal abilities [11].

Pediococcus pentosaceus bacteria also

have benefits as a growth promoter in

tilapia (Oreochromisniloticus) [12].

Pediococcus pentosaceus can also increase

growth, quality of meat, and reduce the

content of ammonia in the digestive tract

of wild chickens [13].

IV. CONCLUSION

The results showed that LAB SR

12isolates in conventional tests as

Pediococcus / Enterococcus whereas in the

API 50CH test kit showed Pediococcus

pentosaceus 2.

SUGGESTION

Identification of LAB isolates SR

12 by utilizing physiological and

biochemical characteristics still has some

disadvantages, namely identification is

limited to conventional tests and API

50CH kits. Therefore, it is necessary to

carry out advanced conventional tests with

more parameters.

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