Microbiological Characteristic and Antimicrobial Activity of
Koumiss Against
Salmonella
typhimurium
and
Mycobacterium tuberculosis
R. Yahya*, R. R. A. Maheswari, & R. H. Mulyono
Faculty of Animal Science, Bogor Agricultural University Bogor 16680 Indonesia
*e-mail: rithoh yahya@yahoo.com
Abstract
Koumiss is a traditional fermented milk product originated from the Central Asian steppes and mostly produced from mare milk by spontaneous fermentation of lactose to lactic acid and alcohol. Koumiss’s starter cultures consist of lactic acid bacterias (Lc. lactis and Lb. acidophilus) and yeast (Sc. cereviceae). Koumiss is believed to pose health promoting properties, which is mainly related to the ability of the starter to produce vitamins of the B-group and antimicrobials. Koumiss could also served as functional food that was showed by its capability to produce the antimicrobial substrate that inhibit pathogenic bacterias. The objectives of this research were to identify microbiological characteristics of koumiss and to study its antimicrobial activity towards pathogenic bacterias such as S. Typhimurium ATCC 14028 and M. tuberculosis H37RV. The experimental design used on this research were non parametric of cohran test for M. tuberculosis H37RV and descriptive analysis for S. Typhimurium ATCC 14028. Variables observed were diameter of inhibition zone of the antagonistic assay used well diffusion method agar for S. Typhimurium ATCC 14028. In addition, the Lowenstein Jensen (LJ) modification agar was used for study the inhibition of M. tuberculosis H37RV. The result showed, koumiss could decrease the total of coliform. The average of koumiss’s inhibition zone in different storage time toward S. Typhimurium ATCC 14028 was ± 7.801 mm. It was bigger than filtrate which was ± 6.002 mm. The average of diameter showed the antimicrobial activity of koumiss against S. Typhimurium ATCC 14028. The result of Cohran test showed the growth of M. tuberculosis H37RV could obstructed with modification of LJ extra koumiss stored for 4, 6, and 8 days. The conclusion of this research that koumiss was effective to against pathogenic bacterias such as S. Typhimurium ATCC 14028 and M. tuberculosis H37RV.
Introducton
Mlk s anmal products contan a varety of potental nutrents that the body needs. Physcal and chemcal composton of mare’s mlk s dfferent from cow’s mlk, goat, buffalo and camels. Mare’s mlk has a low fat content s 1.6% and hgh
lactose of 6.1% Chandanet al.(2008). n Indonesa mare mlk called wld horse mlk
s wdely produce n West Nusa Tenggara (NTB). Ths mlk s a naturally fermented mlk product that has a lqud consstency wthout pasteurzaton treatment.
Koumss s made by fermentaton wth a mxed mcroflora, whch contans dfferent lactc acd bactera and yeasts that use for the treatment of tuberculoss n Russa. Bactera are commonly used as starter cultures are producng antmcrobal
substrates that have antagonstc propertes aganst pathogenc bactera.The number
of hgh antmcrobal substrates wll play a more powerful n nhbtng pathogenc
bactera, especally Salmonella typhimurium and Mycobacterium tuberculosis bac-tera.
Materals and Methods
Preparation of Koumiss starter culture
The frst step for makng a starter koumss s pasteurzed mare mlk at 65 °C for 30 mnutes, then cooled to a temperature of 28 oC. Koumss starter culture made
by dvdng the three equal parts, one part mlk wth Lc. lactis D-01, one part mlk
noculated wth Lb. acidophilus Y-01 and then ncubated at 37 °C for seven hours
and one part mlk noculated wth Sc. cereviceae at 25 °C for fve hours. Lc. lactis
D-01, Lb. acidophilus Y-01 and Sc. cereviceae as much as 3%-5% (v/v) mved nto the mare pasteurze mlk. The results of a mxture of mlk and starter cultures were ncubated at 28 °C for 24 hours to form the desred starter (modfed Rahman et al., 1992).
Koumiss manufacture
Koumss made by pasteurzed at 65 °C for 30 mnutes and after the tempera-ture reached 28 °C were noculated wth a starter (30%). Incubaton was performed agan at 28 °C for 42 hours (Rahman et al, 1992).
Characteristics of microbiological koumiss
The antimicrobial activity of Koumiss against Salmonella Typhi-murium
The nhbton of antmcrobal actvty aganst the Salmonella typh-murum
made by the well agar dffuson method (Wryawan et al., 2009). Ths method
per-formed by spreadng the bactera standard 0.5 Mc. Farland wthout equally dluted, cut well wth a hole punch or cork borer (5 mm), coated wth Bacterologcal meda koumss used to avod seep at the bottom of the well. A total of 50 µl koumss ppet-ted nto the well, then the cup s placed n the refrgerator and then ncubappet-ted at 37 °C for 24 hours.
The antimicrobial activity of Koumiss against Mycobacterium tuberculosis
Inoculaton of bacteral suspenson begns wth the preparaton of
Mycobac-terium tuberculosis H37RV wth a standard concentraton of 0.5 Mc. Farland and
dluted to 103 cfu/ml. Dluton made by addng 1% of bactera (v/v) nto fve ml of
NaCl sterle. A total of 100 ml bacteral suspenson was noculated nto the meda of resstance that has been prepared. Incubaton meda tubes wth horzontal poston wth the angle of 30o to the ncubator at 37 °C for one nght wth the ld loose. After
ncubaton, the tube caps are tghtened and enforced tube nto a vertcal poston. Colony growth readngs performed at day 28 and 42 (Sjahrurachman, 2008).Sjahrurachman, 2008)..
Statistical Analysis
Testng treatment for M. tuberculosis H37RV s day 0, 2, 4, 6 and 8. Analyss of data for M. tuberculosis H37RV usng non-parametrc desgn, Cohran test. Statstcal models are used as follows:
Koumss mcrobologcal charac-terstcs observed a total colform, total m-croorgansms (TPC), total lactc acd bactera and yeast total. Mcrobologcal cha-racterstcs of koumss n ths study had 9.67 log10 cfu TPC/ml, colform> 1 log10
cfu/ml. LAB and yeasts grew together form a symboss n the koumss lke kefr grans. Yeasts n the kefr grans serves to mantan the ntegrty and vablty of mcroflora populatons. Essental amno acds and growth factors for lactc acd bactera produced by yeast, whereas the metaboltes of LAB s used as an energy source. Symboss between the LAB and the yeast s makng kefr nto a stable product (Farnworth and Manvlle, 2003).
The inhibitory activity of antimicrobial Koumiss againts S. typhimurium ATCC 14028
The dameter nhbton of koumss aganst S. typhimurium ATCC 14028
ncreased on the day of the storage (H8). Koumss has a pH of 3.87 ± 1.77 0.004 ±
0.032 and TAT. The optmum pH value for growth of S. typhimurium s 6.5 to 7.5
(Cox, 2000), the growth of S. typhimurium ATCC 14028 koumss been restraned by
a low pH. Dameter nhbton of koumss agan S. typhimurium ATCC 14028 wth
a spread plate method s smaller than the pour plate method. The populaton of S.
typhimurium ATCC 14028 on a spread plate method s 108 cfu/ml, whereas the pour
plate method 106 cfu/ml.
Antmcrobal actvty can be observed n the dffuson test wells nfluenced by several factors, such as: (1) the type and sze of the tube, (2) the type of agar, pH and salt content, (3) the ablty of substances to dffuse nto the agar, (4) characterstcs of the meda and (5) type of test bactera used (Branen, 1993).
Inhbton of the fltrate and koumss by category Morales et al. (2002) ncluded the ntermedate category. Intermedate category s the category wth nhbtory response aganst bacteral pathogens that need to be treated wth hgh doses of antmcro-bals. One way s by dong regular therapy wth these antmcrobals.
The inhibitory activity of antimicrobial Koumiss againts M. tuberculosis H37RV on variety storage times
Control ndcates the growth of M. tuberculosis wth means proporton of
1.00 M. tuberculosis H37RV grown on Lowensten Jensen meda controls that are
genune growth medum M. tuberculosis. Growth of M. tuberculosis H37RV n the
8th week of observaton s presented n Fgure 2.
Growth of M. tuberculosis H37RV n the 8th week of observaton for the
con-trol treatment hghly sgnfcant (P<0.01) wth the addton of koumss treatment on day zero storage tme (H0), four days (H4), sx days (H6) and eght days (H8). Adtama (1999) suggest that the nhbtory propertes of fermented compounds are
bacterostatc, because M. tuberculosis stll can grow when the acdty s removed
control (n=6) koumiss H8 (n=6)
Fgure 2. Growth of M. tuberculosis H37RV n the Storage Koumss Days (H2), (H4), (H6) and (H8)
Concluson
Koumss wth storage treatment can nhbt bacteral growth of S. typhimurium
ATCC 14028 n the range of nhbton zones varyng, whle the antmcrobal
actv-ty of koumss effectve n nhbtng the growth of bactera M. tuberculosis H37RV
after the product has a mnmum of four days of storage.
Acknowledgment
The authors are grateful to Andranjsah, Prawoto, Asyah, Dev Murtn and Sukma Wjaya for techncal assstances n preparng the expermental products and the analyses.
References
Chandan R. C., Arun K & Nagendra P. Shah. 2008. Dary Processng and Qualty Assurance. Wley-Blackwell, USA.
Cox, J. 2000. Salmonella thypimurium. In: Robnson, R.K, Carl A. Batt, & Pradp D. Patel. (eds.) Encyclopeda of Food Mcrobology. Academc Press. New York, USA.
Danel, W. W. 1990. Appled Nonparametc Statstcs. 2nd edton. PWS-KENT
Publshng Company, Boston.
Fardaz, S. 1992. Food Mcrobology vol. I. Grameda Press n collaboraton wth the Food and Nutrton PAU-IPB Jakarta.
Farnworth, E. R., & I. Manvlle. 2003. Kefr: A fermented mlk product. In: E. R. Farnworth (ed). Handbook of Fermented Functonal Foods. CRC Press, London.
antmcrobal actvty and botoxcty aganst. Artema salna. J. Chle. Chem. Soc. 48 (2).
Rahman, A. S. Fardaz, W.P. Rahaju, Sulantar & C.C. Nurwtr. 1992. Mlk Fer-mentaton Technology. Center of nter unversty for food and nutrton., BogorCenter of nter unversty for food and nutrton., Bogor Agrcultural Unversty, Bogor.
Sjahrurachman, A. 2008. Culture and senstvty testng of M. tuberculosis Drugs
Aganst Tuberculoss frst lne. Mnstry of Health of the Republc of Indonesa ,Jakarta.
