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Fermentation
Volume 11
Issue 2
10.3390/fermentation11020055
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This is an early access version, the complete PDF, HTML, and XML versions will be available soon.
Article

Microbiota Composition in Raw Drinking Milk from Vending Machines: A Case Study in Croatia

by
Nataša Mikulec
1,
Jasminka Špoljarić
1,
Dijana Plavljanić
1,
Monica Darrer
1,
Fabijan Oštarić
1,
Jasenka Gajdoš Kljusurić
2,
Khan Mohd. Sarim
3,
Nevijo Zdolec
4 and
Snježana Kazazić
3,*
1
Faculty of Agriculture, University of Zagreb, 10000 Zagreb, Croatia
2
Faculty of Food Technology and Biotechnology, University of Zagreb, 10000 Zagreb, Croatia
3
Ruđer Bošković Institute, Bijenička cesta 54, 10000 Zagreb, Croatia
4
Faculty of Veterinary Medicine, University of Zagreb, 10000 Zagreb, Croatia
*
Author to whom correspondence should be addressed.
Fermentation 2025, 11(2), 55; https://doi.org/10.3390/fermentation11020055
Submission received: 27 December 2024 / Revised: 16 January 2025 / Accepted: 23 January 2025 / Published: 24 January 2025
(This article belongs to the Special Issue Fermented Dairy Products: Processing Technology, Microbiology and Health Benefits: 2nd Edition)
Versions Notes

Abstract

According to the Regulation on the Quality of Fresh Raw Milk, up to 100,000 microorganisms/mL are allowed in milk obtained by the hygienic milking of healthy cows, which represents the natural microbiota of milk and has no negative impact on the overall quality of milk. However, with unprofessional handling during and after milking, milk is easily contaminated and becomes a potential medium for the growth and reproduction of microorganisms, some of which can be harmful to human health. Since the number of aerobic mesophilic bacteria in milk is one of the indicators of the hygienic quality of milk, their number and identification are fundamental in the control of raw milk from milk vending machines. From five different milk vending machines, 35 samples were collected, from which the total number of aerobic mesophilic bacteria was determined using the flow cytometry method and the classic method of counting colonies on a nutrient medium. Randomly selected colonies based on morphological differences (n = 700) were identified by comparing MALDI-TOF mass spectra with reference spectra stored in the microorganism library and processing using the MALDI Biotyper computer program. Thirty-eight genera and eighty-one bacterial species and five genera and seven fungal species were successfully identified. The species that predominate are Lactococcus lactis, Hafnia alvei, Escherichia coli, Leuconostoc mesenteroides, and Kluyveromyces lactis. By integrating advanced methods like flow cytometry and MALDI-TOF MS for precise microbial identification, this study highlights the need for enhanced monitoring and adherence to hygienic standards in raw milk vending machines. This approach not only safeguards public health but also supports consumer confidence in milk quality from vending machines.
Keywords: raw milk; vending machine; hygiene quality; microbiological hazards raw milk; vending machine; hygiene quality; microbiological hazards

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MDPI and ACS Style

Mikulec, N.; Špoljarić, J.; Plavljanić, D.; Darrer, M.; Oštarić, F.; Gajdoš Kljusurić, J.; Sarim, K.M.; Zdolec, N.; Kazazić, S. Microbiota Composition in Raw Drinking Milk from Vending Machines: A Case Study in Croatia. Fermentation 2025, 11, 55. https://doi.org/10.3390/fermentation11020055

AMA Style

Mikulec N, Špoljarić J, Plavljanić D, Darrer M, Oštarić F, Gajdoš Kljusurić J, Sarim KM, Zdolec N, Kazazić S. Microbiota Composition in Raw Drinking Milk from Vending Machines: A Case Study in Croatia. Fermentation. 2025; 11(2):55. https://doi.org/10.3390/fermentation11020055

Chicago/Turabian Style

Mikulec, Nataša, Jasminka Špoljarić, Dijana Plavljanić, Monica Darrer, Fabijan Oštarić, Jasenka Gajdoš Kljusurić, Khan Mohd. Sarim, Nevijo Zdolec, and Snježana Kazazić. 2025. "Microbiota Composition in Raw Drinking Milk from Vending Machines: A Case Study in Croatia" Fermentation 11, no. 2: 55. https://doi.org/10.3390/fermentation11020055

APA Style

Mikulec, N., Špoljarić, J., Plavljanić, D., Darrer, M., Oštarić, F., Gajdoš Kljusurić, J., Sarim, K. M., Zdolec, N., & Kazazić, S. (2025). Microbiota Composition in Raw Drinking Milk from Vending Machines: A Case Study in Croatia. Fermentation, 11(2), 55. https://doi.org/10.3390/fermentation11020055

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