Full Paper

CHARACTERIZATION OF LACTIC ACID BACTERIA

ISOLATED FROM HORSE MILK OF BIMA

Nyoman Semadi Antara*

I Nyoman Dibia
Wayan Redi Aryanta
*Corespondence Author:

nsantara@ftp.unud.ac.id
Laboratory of Bioindustri
Agroindustrial Technology
Faculty of Agricultural Technology
Udayana University
Kampus Bukit Jimbaran
Bali, INDONESIA

INTERNATIONAL JOINT SEMINAR AND WORKSHOP

LACTIC ACID BACTERIA AND OTHER IMPORTANT MICROBES
Their Role in Food, Health, and Industry
January 16-17, 2009
Yogyakarta
Published on

Agritech. 29(1): 1-9. 2008 --- ISSN: 0216-0455

ABSTRACT
The aim of the research was to characterize of the six species of Lactic Acid
Bacteria (LAB) indigenous (Lactobacillus acidophilus, Lb. salivarius, Lb. brevis, Lb.
delbrueckii, Lb. plantarum and Lactococcus lactis) from the fermented horse milk of
Bima as probiotic. The tests done to cover safety aspect, ability to survive on low pH
condition, ability of bile salt tolerance, ability to adherence and colonization, and
effect toward cholesterol level reduction. The six LAB species were not pathogen, not
toxic and not invasive and were able to grow well on media which have bile salt at
0.750%. Lactobacillus acidophilus and Lb. brevis had ability to survive on media pH
2.5 for 3 hours and were able to adherence and colonization on gut mucosa epithel.
Lactobacillus acidophilus and Lb. brevis had ability to reduce cholesterol level of
blood serum of rabbits on hypercholesterolemia condition significantly at 53.74% and
51.70%, respectively. Based on the characterization of the six species LAB, only
Lactobacillus acidophilus and Lb. brevis can be used as probiotic.
Key words : Lactic acid bacteria, characterization, probiotic, horse milk.

INTRODUCTION
Traditionally, fermented milk products have already been known and
consumed by Indonesian for long time, such as dadih in West Sumatera (Sugitha,
1996; Sugitha, 2004), dangke in South Sulawesi (Suryono, 2003), fermented horse
milk in West Nusa Tenggara (Hermawati et al., 2004). The indigenous
microorganisms that play important role in fermentation process are lactic acid
bacteria (Oberman, 1985; Aryanta, 1995).
Lactic acid bacteria (LAB) play important role in food fermentation, primarily
by causing the characteristic flavor changes and contributing a preservative effect on
the fermented product. These bacteria are used widely in fermented food as starter
cultures, such as in dairy products (Parente et al. 1997; Fitzsimons et al. 1999),
fermented vegetables (Sanchez et al. 2000; Kalac et al. 2000), alcoholic beverages
(Patarata et al. 1994; Pattison et al. 1998), and fermented meat (Hammes and Hertel
1998; Antara et al. 2002). Some species of LAB have been used in industry scale as
culture starter and also as probiotic (Goldin, 1998; Salminen et al., 1998).
Exploration of LAB from fermented food Indonesia origin and using them as
probiotik could contribute in the development of science and technology. Widiada, et
al. (2006) have isolated and identified 6 species of LAB from fermented horse milk
of Bima namely Lactobacillus acidophilus, Lb. brevis, Lb. plantarum, Lb. salivarius,
Lb. delbrueckii dan Lactococcus lactis. The research in order to characterize snd
selection of the strain as probiotic has not been conducted.
The objective of this research was to characterize the indigenous LAB isolated
from fermented horse milk of Bima. The characteristics of LAB investigated in this
research were pathogenity of the strain, survival of the strain in low pH, growth
ability in bile salt, adherence and colonization ability on intestine mucosa epithelia,
and effect on decreasing of blood serum cholesterol concentration.

METHODS
Bacterial Strains and Trial Animals
Strains used in this research were Lb. plantarum, Lb. acidophilus, Lb.
salivarius, Lb. delbrueckii dan Lc. Lactis, which were indigenous LAB isolated from

horse milk of Bima. Bacteria of Pasteurella multocida tipe B2 322 dan Streptococcus
zooepidemicus EPI 560 were used as positive control of pathogenic test.
The animal used for in vivo trial of the ability of LAB to adherence and
colonize on the surface of intestinal mucosa epithel and reducing the blood serum
cholesterol were mice (Mus musculus) and 5 months age of local rabbit (Oryctolagus
cuniculus) (Kusumawati, 2004), respectively.
Pathogenic in vivo Test of LAB
One ose of LAB were inoculated in BHI broth (Oxoid, CM225) and then incubated at
37oC for overnight. Overnight cultured of LAB were injected in to the trial mice with
dose of 0.1 ml (109 cfu/ml) once intraperitoneally per mice and continued to
administrate 0.1 ml of LAB (109 cfu/ml) orally per mice per day for 14 days. As
positive control two pathogenic bacteria were used namely Pasteurella multocida
type B2 332 (Priadi dan Natalia, 2000) and Streptococcus zooepidemicus EPI 560
(Dibia

et al., 1995a), and uncultured BHI broth was used as negative control.

Clinical observation of trial mice was conducted every day for 14 days.
All death mice within observation were done necroption and life mice on 15th
day observation were killed and necroption were done. Samples of lever, lung, limp
and hearth were collected aseptically. Isolation and identification of injected bacteria
were done to proof that whether there was invasion or not. Part of the samples were
submerged in fixation solution (10% formalin buffer solution) and observed
pathological change of the organ cells.
Resistance of LAB in Low pH
One ose of LAB isolate were inoculated into GYP broth with vary in pH (1.5; 2.0;
and 2.5) and incubated at 37oC for 2, 3, and 4 h. After incubation the growth of LAB
isolates were observed on MRS agar with anaerobic condition (10%CO2) at 37oC for
48 h incubation.
Ability of LAB Growth in Bile Salt Medium
The isolates of LAB were grown on MRS agar which supplemented by bile salt vary
in concentration (0,750%; 1,875%; 3,750%; 5,625% dan 7,500%). One ose of the

isolate were stricked on MRS agar supplemented by bile salt and incubated
aerobically (10% CO2) at 37oC for 48 h.
Adherence and Colonization Ability of LAB on Mice Intestine
In this research, 6 weeks old of mice (Mus musculus) was used as trial animal. During
the experiment the mice were fed standard concentrate 551 (Charoen Pokphand)
mixed with banana and then sterilization at 121oC for 15 min. The drink of mice was
physiological solution (0.85% NaCl). The mice were administrated by 5 g
antibacterial (Ampicol, Medion) per liter of drinking water (ad libitum) for 3 days in
order to kill the LAB in mice intestine before using them as animal trial. To make
sure that the mice were LAB free, the feces of mice was collected and checked it for
the existence of LAB. The mice that negative growth of LAB from 4th day feces were
used as animal trial.
The LAB isolates used in this experiment were LAB which had probiotic
characteristics selected from the previous experiment. The characteristics were (1) not
pathogenic bacteria, (2) be able to grow in low pH (lower or equal to 2.5) for at least
2 h, and (3) be able to grow in medium with bile in concentration of more than or
equal to 0.75%.
The selected LAB were grown on MRS agar plate. Overnight colonies were
then suspended in physiological solution (about 1011 cfu/ml). Each experimented
isolate were administrated into 5 mice (0.5 ml for each mouse) orally every day for 2
weeks. On 14th day the administration of LAB was stopped and the feces of mice
were started to collect everyday for a weeks. From the feces were isolated and
identified the administrated LAB. On 21st day the mice were killed and encropted,
and the LAB were isolated from the mucosa of mice intestine and identified them.
Effect of LAB on the Serum Blood Cholesterol
The trial animals used in this experiment were rabbits (Orytolagus cuniculus) with
about 1.5 kg body weight (Kusumawati, 2004). The rabbit were grouped into three
groups and each group had three rabbits. Group I was the rabbit with
hypercholesterolemia and administrated orraly by the selected LAB (2ml of 1011
cfu/ml). Group II was positive control that the hypercholesterolemia rabbits without

administrated by LAB. Group III was negative control that the normal rabbit without
administrated by LAB. To make the rabbits to be hypercholesterolemia, the rabbits
was treated by feeding them with 6 ml/rabbit/day yolk of chicken for 7 days, while
the negative control of rabbits were not fed by yolk of chicken (Suarsana et al.,
2004).
The blood all of rabbit groups were sampled every week after administrated
by LAB for 4 week. The collected blood were kept for 4 h at room temperature and
then centrifugated at 3.000 rpm for 15 min to separate the blood serum and the
coagulated blood. To mesure the blood serum cholesterol was used kit of cholesterol
liquicolor (Human GmbH, Cat. 10017) and the procedure followed the protocol.

RESULT AND DISCUSSION

Pathogenity of Lactic Acid Bacteria Isolates

Clinical observatioan of the mice administrated by LAB showed that the mice were
not sick or death after 14 days LAB administration. On the other hand, the mice
treated by pathogenic bacteria (positive control) were death after 3th day (Table 1).
Result of observation also showed that the LAB administrated did not invade the
lever, kidney, lung and hearth of mice, and also there was not pathological change of
the organ. On the group of mice that orally administrated by pathogenic bacteria
(Past. multocida and Strep. zooepidemicus) found that the demage and infiltration of
netrophilium cell, villi erotion, necrosis of epithelium cel, congestion, and there were
colonies of the bacteria found in lever tissue.
Clinical observation on trial mice showed that there was not found death or
sick mouse and demage of organ, and there was no reisolated LAB from mice organ.
This evident proofed that the strain LAB isolated from Bima horse milk were safe
and could be used as safe food.

Survival of LAB on Low pH Medium

The acidity (pH) and survival time is the indicator used to select microorganism
(bacteria) as probiotic. Result of this experiment showed that all of the LAB strain

tested could not survive on extremely acid medium (pH 1.5). Only Lb. brevis could
survive on pH 2.0 medium for 2 and 3 h. And on the pH 2.5 medium two strains of
Lb. brevis and Lb. acidophilus could grow for 2 and 3 h, and the others were death.

Table 1. Safety of LAB isolates on mice (Mus musculus)

Group
Positive control
Past. Multocida

Mice
status
Death

Hystopathological
changes
Lever

: congestion, netrophile cell

infiltration, detection of
colony

Reisolated
bacteria

Isolated

Kidney : congestion on glomerulus

Intestine

Strep. zooepidemicus

Death

Lever

: villi erotionerosi,
netrophilium cell infiltration,
and necrosis of epithelium cell
: congestion, netrophile cell
infiltration, detection of
colony

Isolated

Kidney : congestion on glomerulus

Intestine

Negative control
Lb. acidophilus
Lb. plantarum
Lb. salivarius
Lb. brevis
Lb. delbrueckii
Lc. Lactis

Life
Life
Life
Life
Life
Life
Life

: villi erotionerosi, netrophile

cell infiltration, and necrosis of
epitel cell

Normal, there was no demage

Normal, there was no demage
Normal, there was no demage
Normal, there was no demage
Normal, there was no demage
Normal, there was no demage
Normal, there was no demage

Not isolated
Not isolated
Not isolated
Not isolated
Not isolated
Not isolated
Not isolated

Growth of LAB in Medium With Bile Salt

Determination of growth ability of LAB in medium contain bile salt was based on the
condition of human the intestinal track. In general, all six LAB strains could grow
well in the medium which contain 0.750% and 1.875% bile salt. In medium with
3.75% bile salt only Lb. acidophilus and Lb. brevis can grow well. One strain LAB
tested, Lb. acidophilus, could grow in extremely bile salt concentration (5.625%)
(Table 3).

Table 2. Survival of LAB on low pH mediuma

Strain of LAB
Lb.
Lb.
brevis
plantarum

Lb.
acidophilus

Lb.
salivarius

Lb.
delbrueckii

Lc.
lactis

2h

3h

4h

2h

3h

4h

2h

3h

4h

Survival on pH
1.5 for:

Survival on pH
2.0 for:

Survival on pH
2.5 for:

+ = survive

= not survive

Tabel 3. Growth ability of LAB in medium contain bile salta

Bile salt concentration (%)

Isolat BAL indigenous

Lb
Lb
salivarius
brevis
+
+

0,750

Lb
plantarum
+

Lb
acidophilus
+

Lb
delbrueckii
+

Lc
lactis
+

1,875

3,750

5,625

7,500

+ = grow; = not grow

Ability of LAB Adhere and Colonize on Mucose Intestinal Epithelium

The ability of LAB adhere and colonize on mucosa intestinal epithelium was tested in
vivo using mice as trial animal. Reisolated administrated LAB was used as indicator

on this experiment. The experiment result showed that Lb. acidophilus and Lb.brevis
could be reisolated form mice feces and wall of intestinal mucosa after 7 days
administration (Table 4).

Table 4. Ability of LAB Adhere and Colonize on Mucose Intestinal Epithelium

Reisolated LAB
Lb. acidophilus
Feces
Intestinal wall
Feces
+
Nd
+

Day of
observation
15

Lb. brevis
Intestinal wall
Nd

16

Nd

Nd

17

Nd

Nd

18

Nd

Nd

19

Nd

Nd

20

Nd

Nd

21

+ = reisolated; Nd= not done

Reduce of Blood Serum Cholesterol of Rabbit (Oryctolagus cuniculus)

Effect of LAB administration for 4 weeks on blood serum cholesterol is shown in
Table 5.. Lb. acidophilus and Lb. brevis could reduce rabbit blood cholesterol
significantly (P<0,05) up to 53,74% and 51,70%, respectively.

Table 5. Effects of LAB administration on blood serum cholesterol of rabbit

Treatment

Cholesterol concentrationy,z (mg/dl)

SP

HK
a

35,3

M1
a

34,3

M2
a

33,3

M3
a

34,0

M4
a

36,0a

KN

35,7

KP

37,3b

303,3a

294,7a

289,0a

284,0a

280,7a

LA

36,0e

278,0a

255,3ab

218,3bc

171,3cd

128,7d

LB

29,7d

296,7a

260,7a

195,7b

158,0b

143,3c

KN: no treatment control; KP: hypercholesterol control; LA: hypercholesterol and Lb.acidophilus
administration; LB: hypercholesterol and Lb.brevis administration.
y
SP: before treatment; HK: hypercholesterol treatment; M (week).
z
Value followed by different character on the same row states significantly different (P<0,05).

CONCLUSION
Six of LAB strains isolated from Bima horse milk (Lactobacillus acidophilus, Lb.
brevis, Lb. plantarum, Lb. salivarius, Lb. delbrueckii and Lactococcus lactis) were
safe microorganisms used in food. They could grow in medium with bile salt in
concentration of 0.75%. Two species out of six, namely Lb.brevis dan Lb.acidophilus,
could survive in low pH medium (pH2.5) for 3 h. These two species could adhere and
colonize on mice intestinal mucosa epithelium, and could reduce blood serum
cholesterol of rabbit in hypercholestrolmia condition up to 53,74% dan 51,70% for
Lb.brevis and Lb.acidophilus, respectively. Lactobacillus acidophilus and Lb. brevis
fulfil requirement used as probiotic.

REFERENCES
Antara, N.S., Sujaya, N., Yokota, A., Asano, K., Aryanta, W.R., and Tomita, F. 2002.
Identification and Succession of Lactic Acid Bacteria during Fermentation of
Urutan, a Balinese Indigenous Fermented Sausage. World J. Microbiol and
Biotechnol. 18: 255-262.
Antara, N.S. 2004. Isolation and Identification of Indigenous Lactic Acid Bacteria,
Their Role and Aplication in Production of Urutan, A Balinese Fermented
Sausage. Disertasi. Laboratory of Applied Microbiology, Department of
Molecular Bioscience, Graduate School of Agriculture, Hokkaido University,
Tokyo, Japan.
Aryanta, W.R. 1995. Peranan Bakteri Asam Laktat dalam Industri Pengolahan Bahan
Pangan. Pidato Pengenalan Jabatan Guru Besar Tetap dalam Bidang
Mikrobiologi Pangan pada Program Studi Teknologi Pertanian Universitas
Udayana. Disampaikan dalam Rapat Senat Terbuka UNUD Denpasar, 27
Oktober 1995.
Brashears, M.M., Gilliland, S.E., and Buck, L.M. 1998. Bile Salt Deconjugation and
Cholesterol Removal from Media by Lactobacillus casei. J. Dairy Sci.
81:2103-2110.
Buxton, A. and Fraser, G. 1977. Animal Microbiology. Vol. 1. Black Well Scientific
Publications.

100

Casla, D., Requena, T., and Gomez, R. 1996. Antimicrobial Activity of Lactic Acid
Bacteria Isolated from Goats Milk and Artisanal Cheeses: Characteristics of a
Bacteriocin Produced by Lactobacillus curvatus IFPL105. J. Appl. Bacteriol.
81: 35-41.
Chan, H.Y., Chiu, H.H., Lin, F.M., Liu, Y.J, Hung, H.Y., OuYang, H., Yu, C.K.,
Wang, Y.J., and Liao, C.C. 2005. Characterization of Lactobacillus gasseri
B38T38 with Cholesterol Assimilation and Antitumor Activities. 9th National
Congress of Indonesian Society for Microbiology & 3rd Asian Conference for
Lactic Acid Bacteria. August 25-26, 2005, Bali.
Chou, L and Weimer, B. 1999. Isolation and Characterization of Acid and Bile
Tolerant Isolates from Strains of Lactobacillus acidophilus. J.Diary Sci 82:
23-31.
Cowan, S.T. and Steel. 1979. Manual for Identification of Medical Bacteria. Second
edition . Cambrige University Press, London.
Davidson, P.M. and Hoover, D.G. 1998. Antimicrobial Components from Lactic Acid
Bacteria. In: S. Salminen and A. von Wright (Eds.). Lactic Acid Bacteria.
Marcel Dekker Inc., New York.
Dibia, N., Soeharsono, dan Dartini, N.L. 1995a. Keganasan Isolat Streptococcus
zooepidemicus pada Kera, Mencit dan Babi. Seminar Nasional Peternakan dan
Veteriner, 7 8 Nopember 1995, Bogor.
Dibia, N., Amintorogo, S., Putra, A.A.S., Dartini, N.L., dan Supartika, K.E. 1995b.
Epidemiologi dan Gejala Klinis Streptococcosis pada Babi di Propinsi Bali.
Bulletin Veteriner VIII (43) : 1-17.
Djide, M.N. 2005. Pengaruh Formulasi Granul Kedelai terfermentasi terhadap Bakteri
Asam Laktat dan Pengikatan Kolesterol secara Invitro. 9th National Congress
of Indonesian Society for Microbiology & 3rd Asian Conference for Lactic
Acid Bacteria. August 25-26, 2005, Bali.
Duniaji, A.S. 2000. Peranan Bakteri Asam Laktat pada Silase Limbah Ikan
Terfermentasi. Gitayana 6 (1): 57-63.
Fardiaz, S. 1992. Petunjuk Laboratorium Mikrobiologi Pengolahan Pangan. PAUIPB, Bogor.
Gilmour, A. and Rowe, M.T. 1981. Microorganisme Associated With Milk. In: The
Microbiology of Milk. (Editor: Robinson R.K.). Vol. 1. Applied Science
Publishers.
Goldin, B.R. 1998. Health Benefit of Probiotic. Department of Family Medicine and
Community Health, Boston.
Guyton, A.C. 1995. Fisiologi Manusia dan Mekanisme Penyakit. Edisi III.
(Penerjemah : Petrus Andrianto). Penerbit Buku Kedokteran EGC, Jakarta.
Harimurti, S., Rahayu, E.S., and Kurniasih. 2005. Application of Lactic Acid Bacteria
Probiotic to Broiler Chicken and Its Adherence Mechanism to the Gut
Epithelia Cells. 9th National Congress of Indonesian Society for Microbiology
& 3rd Asian Conference for Lactic Acid Bacteria. August 25 -26, 2005, Bali.
Harmayani, E. 2004. Peranan Probiotik untuk Menurunkan Kolesterol. Seminar
Nasional Probiotik dan Prebiotik sebagai Makanan Fungsional, 30 Agustus
2004, Denpasar.

Hermawati, D., Sudarwanto M., Soekarto S.T., Zakaria F.R., Sudrajat S., dan Tjatur
R.F.S. 2004. Aktivitas Antimikroba pada Susu Kuda Sumbawa. Jurnal
Teknologi dan Industri Pangan XV (1) : 47-53.
Jay, J.M. 1996. Modern Food Microbiology. Fifth Edition. Chapman and Hall, New
York.
Ketchum, P.A. 1988. Microbiology: Concepts and Applications. John Wiley and
Sons, New York.
Kim, H.Y. and Lee, H.J. 2005. Anticancer and Immunopotentiating Activity of
Cellular Components of Latic Acid Bacteria. 9th National Congress of
Indonesian Society for Microbiology & 3rd Asian Conference for Lactic Acid
Bacteria. August 25 -26, 2005, Bali.
Kimoto, H., Ohmomo, S., and Okamoto, T. 2002. Cholesterol Removal from Media
by Lactococci. J. Dairy Sci. 85: 3182-3188.
Kusumaningtyas, R.W., Susanti, I., and Illaningtyas, F. 2005. Selection of Lactic
Acid Bacteria Isolates for Probiotics. 9th National Congress of Indonesian
Society for Microbiology & 3rd Asian Conference for Lactic Acid Bacteria.
August 25-26, 2005, Bali.
Kusumawati, D. 2004. Bersahabat dengan Hewan Coba. Gajah Mada University
Press, Yogyakarta.
Lay, B.W. 1994. Analisis Mikroba di Laboratorium. Cetakan I. Raja Grafindo
Persada, Jakarta.
Mitsuoka, T. 1990. A Profile of Intestinal Bacteria. Yakult Honsha Co. Ltd., Japan.
Mukartini, S. 2002. Penyakit yang Ditularkan Melalui Produk Pangan Asal Hewan.
Direktorat Kesmavet, Direktorat Jenderal Peternakan, Departemen Pertanian,
Jakarta.
Mukherjee, K.L. 1988. Medical Laboratory Technology. Vol. II. Tata McGraw Hill,
New Delhi.
Muwakhid, B., Ardyanti, T., Soebandrio, dan Chizaemi, S., 2005. Isolasi dan Seleksi
Bakteri Asam Laktat dari Sampah Organik untuk Pembuatan Silase. 9th
National Congress of Indonesian Society for Microbiology & 3rd Asian
Conference for Lactic Acid Bacteria. August 25-26, 2005, Bali.
Nishida, S,. Inoue, S., Fujii, T., Nkajima, K., Fujiwara, D., and Iino, H. 2005. The
Selection of Lactobacilus paracasei strain KW3110 Having Antiallergy
Effect, Confirmation of Antiallergy Effect in the Animal Model, and
Evaluation of the Character as a Probiotic. 9th National Congress of
Indonesian Society for Microbiology & 3rd Asian Conference for Lactic Acid
Bacteria. August 25-26, 2005, Bali.
Nousiainen, J. and Setala, J. 1998. Lactic Acid Bacteria as Animal Probiotics. In: S.
Salminen and A. von Wright (Eds.). Lactic Acid Bacteria. Marcel Dekker
Inc., New York.
Oberman, H. 1985. Fermented Milks. In: Microbiology of Fermented Foods. Vol. 1.
(Editor: Brian J.B. Wood). Elsevier Applied Science Publishers, London.
Ohashi,Y., Nakai, S., Tsukamoto. T., Masumori, N., Akaza, H., Miyanaga, N.,
Kitamura, T., Kawabe, K., Kotake, T., Kuroda, M., Naito, S., Koga, H., Saito,
Y., Nomata, K., and Kitagawa, M. 2002. Habitual Intake of Lactic Acid
Bacteria and Risk Reduction of Bladder Cancer. J.Urol.Int. 68:273-280.

Ouwehand, A.C. 1998. Antimicrobial Components from Lactic Acid Bacteria. In: S.
Salminen and A. von Wright (Eds.). Lactic Acid Bacteria. Marcel Dekker
Inc., New York.
Ouwehand, A.C., Salminen, S. Tolkko, S., Roberts, P., Ovaska, J., and Salminen, E.
2002. Resected Human Colonic Tissue: New Models for Characterizing
Adhesion of Lactic Acid Bacteria. J. Clinical and Diagnostic Laboratory
Immunol. 9 (1) :184-186.
Parente, E., Grieco, S., and Crudele, M.A. 2001. Phenotypic Diversity of Lactic Acid
Bacteria Isolated from Fermented Sausages Produced in Basilicata (Southern
Italy). J. Appl. Microbiol. 90: 943-952.
Pato, U. 2003. Potensi Bakteri Asam Laktat yang diisolasi dari Dadih untuk
Menurunkan Resiko Penyakit Kanker. Jurnal Natur Indonesia 5(2): 162-166.
Priadi, A. dan Natalia, L. 2000. Patogenesis Septicaemia Epizootica (SE) pada Sapi/
Kerbau : Gejala Klinis, Perubahan Patologis, Reisolasi, Deteksi Pasteurella
multocida dengan Media Kultur dan Polymerase Chain Reaction (PCR).
Jurnal Ilmu Ternak dan Veteriner 5 (1) : 65-71.
Price, S.A dan Wilson, L.M. 1995. Patofisiologi. Edisi 4. Buku I. (Penerjemah :
Anugerah, P.). Penerbit Buku Kedokteran EGC, Jakarta.
Rahayu, E.S. 2004. Makanan Probiotik untuk Kesehatan. Seminar Nasional Probiotik
dan Prebiotik Sebagai Makanan Fungsional, 30 Agustus 2004, Denpasar.
Rahman, A., Fardiaz, S., Rahayu W.P., Suliantari, dan Nurwitri C.C. 1992.
Teknologi Fermentasi Susu. PAU-IPB, Bogor.
Ray, B. 1996. Lactic Acid Bacteria : Current Advances in Metabolisme, Genetic and
Application. Springer-Verlag, Germany.
Ray, B. 2004. Fundamental Food Microbiology. 3rd Ed. CRC Press, Boca Raton,
Florida.
Reksohadiwinoto, B.S., Sugama, Y., dan Zakiyah, U.Y. 2005. Penelitian,
Pengembangan dan Aplikasi Probiotik proChick LAB pada Peternakan Ayam.
9th National Congress of Indonesian Society for Microbiology & 3rd Asian
Conference for Lactic Acid Bacteria. August 25-26, 2005, Bali.
Ressang, A.A. 1983. Patologi Khusus Veteriner. Penerbit Percetakan Bali, Denpasar.
Rohim, A. dan Soebijanto. 2002. Probiotik dan Flora Normal Usus. Dalam: Ilmu
Penyakit Anak. (Editor: Soegianto, S.). Penerbit Salemba Medika, Jakarta.
Sadikin, M. 2002. Biokimia Darah. Cetakan I. Penerbit Widya Medika, Jakarta.
Salminen, S., Deighton, M.A., Benno, Y., and Gorbach, S.L. 1998. Lactic Acid
Bacteria in Health and Disease. In: S. Salminen and A. von Wright (Eds.).
Lactic Acid Bacteria. Marcel Dekker Inc., New York.
Suarsana, N., Utama, I.H., dan Suartini, N.G.A.A. 2001. Aktivitas Invitro Senyawa
Antimikroba dari Streptococcus lactis. Jurnal Veteriner 2 (1): 25-31.
Suarsana, N., Suartini, G.A.A., dan Utama, I.H. 2004. Pengaruh Yoghurt terhadap
Kadar Kolesterol Total dan Profil Lipoprotein Serum Kelinci. Jurnal Veteriner
5 (1):14-19.
Sudarmo, S.M. and Ranuh, R. 2004. Protective and Therapeutic Effect of Probiotics
in Gastrointestinal Disease. International Symposium Probiotic for Human
and Immunity, September 7-8, 2004, Bali.

Sugitha, I.M. 1996. Dadih: Olahan Susu Kerbau, Manfaat, Kendala dan Prospeknya
dalam Era Industrialisasi. Jurnal Peternakan dan Lingkungan 2(2):31-38.
Sugitha, I.M., Mukhtar H., Kasrad, dan Yuherman. 1999. Rekayasa Dadih dengan
Starter Streptococcus lactis dan Lactobacillus acidophilus untuk Mencegah
Kanker dan Mengurangi Kolesterol Darah. Laporan Penelitian HB VI.
Faterna Universitas Andalas, Padang.

Sugitha, I.M., Mulyani, Abdi, D., dan Sumaryati, S. 2002. Aktivitas Bakteriosin yang
Dihasilkan Lactococcus lactis mutan ssp. lactis pada Dadih sebagai
Penghambat Bakteri Kontaminan. Jurnal Peternakan dan Lingkungan 8
(2):57-65.
Sugitha, I.M. 2004. Ability of Dadih as Probiotic Supplement for Supporting Health
Care at Rural Communities of West Sumatra. International Symposium
Probiotic for Human and Immunity, September 7-8, 2004, Bali.
Supar, Setiadi, Y., Djaenuri, Kurniasih, N., dan Poerwadikarta, B. 2000. Patogenesis
Pasteurella multocida Isolat Lokal pada Mencit dan Ayam. Jurnal Ilmu
Ternak dan Veteriner 5(1): 53-58.
Suriawiria, U. 1983. Pengawetan Ikan secara Biologis dan Peranan Bakteri Laktat di
Dalamnya. Dalam: Mikrobiologi di Indonesia (Editor: Suharno J., Robert, U.,
dan Usman C.W.). Perhimpunan Mikrobiologi Indonesia, Jakarta.
Surono, I.S., Koesnandar, Zakaria, F.R., Koestomo, F., Novitasari, N., Dharmawan,
J., Lee, Y.K., Nurani, D. 2005. Immunomodulatory Properties of Dadih
Probiotic Bacteria in Indonesian Undernourished Children. 9th National
Congress of Indonesian Society for Microbiology & 3rd Asian Conference for
Lactic Acid Bacteria. August 25-26, 2005, Bali.
Suryohudoyo, P. 2000. Kapita Selekta Ilmu Kedokteran Molekuler. Penerbit Sagung
Seto, Jakarta.
Suryono. 2003. Dadih: Produk Olahan Susu Fermentasi Tradisional Yang Berpotensi
Sebagai Pangan Probiotik. Diambil dari URL:
http://www.rudyct.topcities.com/pps702_71034/suryono.htm
Tannock, G.W. 1999. Probiotics A Critical Review. Horizon Scientific Press, New
Zealand.
Tizard, I.R. 1988. Pengantar Immunologi Veteriner. (Penerjemah: Masduki
Partodiredjo). Universitas Airlangga Press, Surabaya.
Utama, I.H. 1998. Ekspresi Fenotip dan Aktivitas Biologis Streptococcus Grup C
Isolat Asal Babi dan Kera. Disertasi. Pascasarjana IPB, Bogor.
Utama, I.H. 2001. Pengantar Kimia Biofisika. UPT Penerbit Univesitas Udayana,
Denpasar.
Widiada, G.N. 2006. Isolasi dan Identifikasi Bakteri Asam Laktat Indigenous dari
Susu Kuda Liar Bima Selama Penyimpanan dan Aktivitas Antibakterinya.
Tesis. Program Pascasarjana Universitas Udayana, Denpasar.
Widodo. 2003. Bioteknologi Industri Susu. Lacticia Press, Yogyakarta.
Widyastuti, P. 2000. Penyakit Bawaan Makanan. Penerbit Buku Kedokteran EGC,
Jakarta.
Wilson, G.S. and Miles, A.A. 1966. Principles of Bacteriologi and Immunity. Vol. I
5th Edition. Butler & Tanner Ltd., London.
Winarno, F.G. 1997. Probiotik dan Keamanan Pangan. Seminar Nasional
Biopreservasi dan Probiotik Dalam Industri Pangan, 12 Juni 1997,
Yogyakarta.
Yan, T.R. and Lou, D.W. 2005. Effect of Survival of Lactic Acid Bacteria on
Adhesion onto Gastrointestinal Cells. 9th National Congress of Indonesian
Society for Microbiology & 3rd Asian Conference for Lactic Acid Bacteria.
August 25-26, 2005, Bali.