Project title (Full title and Acronym) - WBC-INCO Net

Conservation and standardisation of traditional technologies of fermented milk
products based on autochthonous lactic acid bacteria
Potentials for the commercialisation of traditional dairy
technologies and autochthonous starter cultures in
WBC
Natasa Golic, Ph.D.
University of Belgrade
Institute of Molecular Genetics and Genetic Engineering
Laboratory for Molecular Genetics of Industrial Microorganisms,
Serbia

Conservation and standardisation of traditional technologies of fermented milk
products based on autochthonous lactic acid bacteria
INTRODUCTION
- Western Balkan Countries (WBC) harbours a variety
of traditional, spontaneously fermented foods
manufactured in small scale at specific ecologica l
localities
- Artisanal products harbour a rich diversity of yeasts,
LAB and acetic acid bacteria
- The market share of the traditional fermented foods in
Europe is growing as consumers have an increasing
awareness for artisan handling, gastronomic quality, and
healthy food status.
- The tightened (European) legislation on food safety
(HACCP rules) resulted in the disappearance of number
of regional artisanal fermented products.

Conservation and standardisation of traditional technologies of fermented milk
products based on autochthonous lactic acid bacteria
Laboratory for Molecular Genetics of Industrial
Microorganisms (LMGIM)
Institute of Molecular Genetics and Genetic Engineering,
(IMGGE)
University of Belgrade
LMGIM collection include over 4000 LAB isolates from various
spontaneously fermented foods
Cheese
Yogurt
Kajmak
Meat products
LAB of human origin
The expansion of the existing collection of natural LAB strains
Selection of new, potential probiotic strains

Conservation and standardisation of traditional technologies of fermented milk
products based on autochthonous lactic acid bacteria
SCIENTIFIC OBJECTIVE
Isolation and characterisation of lactic acid bacteria
and yeasts from artisanal dairy products manufactured at
different geographical regions in Western Balkan.
Designation of functional starter cultures based on
selected autochthonous LAB and yeast strains for production
of cheeses with geographical origin.

Conservation and standardisation of traditional technologies of fermented milk
products based on autochthonous lactic acid bacteria
Partners:
WG1:. University of Belgrade, Institute of Molecular Genetics and Genetic
Engineering (IMGGE), Laboratory for Molecular Genetics of Industrial
Microorganisms (LMGIM), Belgrade, Serbia
Prof. Ljubiša Topisirović, PhD, Nataša Golić, PhD – Secretary of PSALAB
Coordinator of PSALAB Project Project Milan Kojić, PhD -
Amarela Terzić-Vidojević, PhD Ivana Strahinić, PhD
Jelena Begović, PhD Jelena Lozo, PhD
Branko Jovčić, PhD Sanja Mihajlović, PhD
Katarina Veljović, PhD
Maja Tolinački, MSci
Milica Nikolić, Msci Jovanka Lukić, PhD student
Brankica Filipić, PhD student
Goran Vukotić, PhD student
Gordana Uzelac, PhD student
Head of LMGIM

Conservation and standardisation of traditional technologies of fermented milk
products based on autochthonous lactic acid bacteria
WG2: University of Zagreb, Faculty of Food Technology and Biotechnology
(FFTB), Laboratory for Antibiotic, Enzyme, Probiotic and Starter Culture
Technology, Zagreb, Croatia
Prof. Jagoda Šušković, PhD
Blaženka Kos, PhD
Jasna Beganović, PhD
Andreja Leboš Pavunc, BSc
Ksenija Uroić,
BSc
WG3: University of Ljubljana, Biotechnical Faculty (UL-BF), Department of Food
Science and Technology, Ljubljana, Slovenia
Prof. Peter Raspor, PhD
Neža Čadež, PhD
Golob, BSc
Hana Šuranska, BSc
Petra Doušova, BSc
WG4: Probiotik d.o.o., Zagreb, Croatia (SME)
Prof. Ljerka Gregurek, PhD
Željka Krušev Šurić,BSc Katarina
Gregor

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3
3 1 2
Conservation and standardisation of traditional technologies of fermented milk
products based on autochthonous lactic acid bacteria
Serbia
1
Croatia
Collection of cheese samples
18 cheese samples (1–10 days old) collected from:
- 3 Serbian localities (South Morava region, Mountain Golija
and Eastern Serbia mountain region)
- 3 Croatian localities (Prigorje, Bilogorsko-Podravska region
and Zagorje)

Conservation and standardisation of traditional technologies of fermented milk
products based on autochthonous lactic acid bacteria
Enumeration and isolation of LAB and yeast strains
from cheese samples
- A total of 828 LAB and 433 yeast strains were isolated from 18
cheese samples
- 88 LAB isolates were determined by 16S rDNA sequencing
- 69 yeast isolates were determined by ITS DNA region sequencing

Conservation and standardisation of traditional technologies of fermented milk
products based on autochthonous lactic acid bacteria
LAB and yeast diversity

Conservation and standardisation of traditional technologies of fermented milk
products based on autochthonous lactic acid bacteria
Characterization of LAB and yeast isolates

Conservation and standardisation of traditional technologies of fermented milk
products based on autochthonous lactic acid bacteria
Probiotic properties of LAB and yeast isolates

Conservation and standardisation of traditional technologies of fermented milk
products based on autochthonous lactic acid bacteria
•
•
88
69
Culture collection management
strains of LAB were deposited in ZIM
strains of yeasts were deposited in ZIM
Preservation methods for microorganisms.

Conservation and standardisation of traditional technologies of fermented milk
products based on autochthonous lactic acid bacteria
Starter culture preparation
- Nine LAB isolates selected for the sensor evaluation;
- Eight combinations of starter cultures were designed

Conservation and standardisation of traditional technologies of fermented milk
products based on autochthonous lactic acid bacteria
Production of white brined cheese with selected LAB
isolates

Conservation and standardisation of traditional technologies of fermented milk
products based on autochthonous lactic acid bacteria
Production of fresh soft cheese with selected LAB isolates

Conservation and standardisation of traditional technologies of fermented milk
products based on autochthonous lactic acid bacteria
Future prospective
The results of the project provided:
An inventory of Serbian and Croatian traditionall y
manufactured white-pickled and fresh soft cheeses and their
microbial diversity;
A well characterized bacterial culture collection that can
serve as a source of bacteria for the construction of starter
cultures with well-defined functional properties;
Production of white-pickled and fresh soft cheeses with
autochthonous LAB as functional starter cultures could
attract SMEs interested in production of innovative dairy food
products.

Conservation and standardisation of traditional technologies of fermented milk
products based on autochthonous lactic acid bacteria
Commercialization of autochthonous starter cultures
Experience
- Previous attempts to implement autochthonous starter
cultures into industrial sector in Serbia - experimental stage.
- Five innovation projects financed by Ministry of Education,
Science and Technological Development, Republic of Serbia
- Pilot cheese production in dairy plants in Sremska Mitrovica,
Zrenjanin, Subotica, Bac, Zlatar, Kopaonik
Problems
- Aggressive market strategies of larger companies for
production of commercial starter cultures
- Lack of governmental support for the implementation of
autochthonous starter cultures

Conservation and standardisation of traditional technologies of fermented milk
products based on autochthonous lactic acid bacteria
Commercialization of autochthonous starter cultures
Possibilities
- Dissemination of knowledge by internet site and flyers
distributed at conferences and other relevant events.
Opportunities
- Increasing number of small dairy companies interested in
using autochthonous starter cultures; “Ivancic and sons”clinical
trials
- Breaking down barriers to get in touch with producers
(SMEs) of artisan food products, will allow stimulation of
their innovation drive.
-Full knowledge of the microbial diversity of traditional
fermented food products would make SMEs eventually
stronger on the EU market.

Conservation and standardisation of traditional technologies of fermented milk
products based on autochthonous lactic acid bacteria
CONCLUSIONS
The results obtained in this study indicates that the
cheese samples traditionally manufactured in different
regions in Serbia and Croatia seem to be diverse in the
LAB and yeast composition.
These results have important impact on designation of
starter cultures for dairy products with geographical
origin.
Knowledge gained during this project would be
essential for the food industry in order to start marketing
the food with specially defined functional properties .