Skip to main content
Log in

Identification and evaluation of thermotolerance of yeasts from milk in natura exposed to high temperature and slow and fast pasteurization

  • Food Microbiology - Research Paper
  • Published:
Brazilian Journal of Microbiology Aims and scope Submit manuscript

Abstract

Milk is considered one of the basic raw materials of animal origin; it must present hygienic quality and physical–chemical properties suitable for processing and human consumption. Thus, the ingestion of milk in natura when not properly treated can be characterized as an opportunistic route of transmission of possible microbial pathogens, which can offer risks to public health. The present study aimed the yeast identification, to analyze the thermo-resistance of yeasts isolated from fresh milk, and to trace the susceptibility profile of the isolates to antifungal agents. For this, 23 samples of fresh milk type B, collected by manual or mechanical milking, were stored in collective refrigeration tanks of farms located in the Metropolitan Region of Natal and nearby, State of Rio Grande do Norte (RN), Brazil. Twenty samples of fresh milk commercially traded in the city of Ceará-Mirim RN were also analyzed. The yeasts were quantified by count of colony-forming units (CFU). All isolated species were treated by slow pasteurization (62–64 °C for 30 min) and fast (72–75 °C for 20 s), as well as by boiling (100 °C). Fifty yeast strains were obtained, and the species were identified as Candida tropicalis (28%), Candida parapsilosis (14%), Candida albicans (12%), Candida glabrata (10%), Candida krusei (10%), Kluyveromyces marxianus (10%), Candida guilliermondii (8%), Candida rugosa (2%), Candida orthopsilosis (2%), Pichia manshurica (2%), and Kodamaea ohmeri (2%). Five isolates showed resistance to the antifungal agents tested. Among all the isolates submitted to heat treatment, 80% were resistant to fast pasteurization and 60% to boiling, but none of them resisted the slow pasteurization. The milk collected through mechanical milking and stored in collective cooling tanks, presented higher rates of yeast contamination, compared to milk samples collected by manual milking and kept under the same storage conditions.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  1. Spanamberg A, Sanches EMC, Santurio JM, Ferreiro L (2009) Mastite micótica em ruminantes causada por leveduras. Ciência Rural 39(1):282–290

    Article  Google Scholar 

  2. Brasil (2011) Ministério da Agricultura, Pecuária e Abastecimento. Gabinete do ministro. Instrução normativa. 62, Brasília, p 24

  3. Beloti V, Barros MAF, Souza JA, Nero LA, Santana EHW, Balaria O, Yassuo C (1999) Avaliação da qualidade do leite cru comercializado em Cornélio Procópio, Paraná: controle do consumo e da comercialização. Semina Ciências Agrárias 20(1):12–15

    Google Scholar 

  4. Oliveira CAF, Fonseca LFL, Germano PML (1999) Aspectos relacionados à produção, que influenciam a qualidade do leite. higiene alimentar. São Paulo 62(13):10–21

  5. Agarwal A, Awasthi V, Dua A, Ganguly S, Garg V, Marwaha SS (2012) Microbiological profile of milk: impact of household practices. Indian J Public health 56(1):88–94

    Article  PubMed  Google Scholar 

  6. Zambonim MC (2014) Caracterização de leveduras promotoras de estufamento em iogurte com polpa de fruta. 2014. Dissertação (Mestrado em Engenharia de Alimentos) – Setor de Tecnologia, Universidade Federal do Paraná, Curitiba, PR, Brasil, p 74

  7. Suriyarachchi VR, Fleet GH (1981) Occurrence and growth of yeasts in yogurts. Appl Environ Microbiol 42(3):574–579

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Venturini KS, Sarcinelli MF, Silva LC (2007) Características da Carne de Frango. Universidade Federal do Espírito Santo. UFES. Pró-Reitoria de Extensão Programa Institucional de extensão

  9. Tondo EC, Bartz S (2019) Microbiologia e sistemas de gestão da segurança de alimentos. 2a ed. Porto Alegre: Sulina, RS. Brasil, p 407

  10. Schukken Y, Wilson D, Welcome F, Garrison-Tikofsky L, Gonzalez R (2003) Monitoring udder health and milk quality using somatic cell counts. Vet Res 34:579–596

    Article  PubMed  Google Scholar 

  11. Alhussien MN, Dang AK (2018) Milk somatic cells, factors influencing their release, future prospects, and practical utility in dairy animals: an overview. Vet World 11(5):562–577

    Article  PubMed  PubMed Central  Google Scholar 

  12. Richard JL, McDonald JS, Fichtner RE, Anderson AJ (1980) Identification of yeasts from infected bovine mammary glands and their experimental infectivity in cattle. Am J Vet Res 41:1991–1994

    CAS  PubMed  Google Scholar 

  13. Riddell RW (1950) Permanent stained mycological preparations obtained by slide culture. Mycologia 42:265–270

    Article  Google Scholar 

  14. Pfaller MA, Huston A, Coffman S (1996) Application of chromagar candida medium for rapid screening of clinical specimens for Candida albicans, Candida tropicalis, Candida krusei, and Candida (torulopsis) glabrata. J Clin Microbiol 34:56–61

    Article  Google Scholar 

  15. Looke M, Kristhuhan K, Kristhuhan A (2011) Extration of genomic DNA from yeasts for pcr-based applications. Biotecniques 50(5):325–328

    Article  CAS  Google Scholar 

  16. Godinho VM, Furbino LE, Santiago IF, Pellizzari FM, Yokoya N, Pupo D, Alves TMA, Junior PAS, Romanha AJ, Zani CL, Cantrell CL, Rosa CA, Rosa LH (2013) Diversity and bioprospecting of fungal communities associated with endemic and cold-adapted macroalgae in Antarctica. ISME 7:1434–1451

    Article  CAS  Google Scholar 

  17. White TJ, Bruns TD, Lee SB (1990) Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In: Innis NA, Gelfand J, Sninsky J, White T (eds) PCR Protocols: a guide to methods and applications. Academic Press, San Diego, pp 315–322

  18. Melville PA, Watanabe ET, Benites NR, Ribeiro AR, Silva JAB, Garino JF, Costa EO (1999) Evaluation of the susceptibility of Prototheca zopfii to milk pasteurization. Mycopathologia 146:79–82

    Article  CAS  PubMed  Google Scholar 

  19. Roostita LB, Fleet GH, Wendry SP, Apon ZM, Gemilang LU (2011) Determination of yeasts antimicrobial activity in milk and meat products. J Food Sci Technol 3(6):445–445

    Google Scholar 

  20. Sanguinetti M, Posteraro B, Lass-flõrl C (2015) Antifungal drug resistance among Candida species: mechanisms and clinical impact. Mycoses 58(2):2–13

    Article  PubMed  Google Scholar 

  21. Santos RC, Marin JM (2005) Isolation of Candida spp. from mastitic bovine milk in Brazil. Mycopathologia 159:251–253

  22. Mbuk EU, Kwaga JKP, Bale JOO, Umoh JU (2016) Molecular identification of yeasts associated with raw cow milk from peri-urban farms in Kaduna State, Nigeria. J Yeast Fungal Res 7(5):39–46

    Article  CAS  Google Scholar 

  23. Alangaden GJ (2011) Nosocomial fungal infections: epidemiology, infection control, and prevention. Infect Dis Clin Am 25(1):201–225

    Article  Google Scholar 

  24. Du J, Wang X, Luo H, Wang Y, Liu X, Zhou X (2018) Epidemiological investigation of non-albicans Candida species recovered from mycotic mastitis of cows in Yinchuan, Ningxia of China. BMC Vet Res 14:251–260

    Article  PubMed  PubMed Central  Google Scholar 

  25. Eldesouky I, Mohamed N, Khalaf DD, Salama AA, Elsify AM, Ombarak RA, El-ballal SS, Effat M, Al-Shabrawy M (2016) Candida mastitis in dairy cattle with molecular detection of Candida albicans. Kafkas Universitesi Veteriner Fakultesi Dergisi 22(3):461–464

    Google Scholar 

  26. Menezes MFC, Simeoni CP, Etchepare MA, Huerta K, Bortoluzzi DP, Menezes CR (2014) Microbiota e conservação do leite. Revista Eletrônica em Gestão, Educação e Tecnologia Ambiental 18:76–89

    Article  Google Scholar 

  27. Santana EHW, Beloti V, Muller EE, Ferreira MA, Moraes, LB, Pereira MS, Gusmão VV (2004) Milk contamination in different points of the dairy process. mesophilic, psychrotrophic and proteolytic. Semina: Ciências Agrárias: Londrina 25(4):349–358

  28. Filho WFL, Souza AR, Prado RS (2015) Análise microbiológica de leite cru refrigerado e industrializado proveniente de propriedades dos municípios de Carmo do Rio Verde, Itapaci, Ipiranga de Goiás, Nova América, Rubiataba e São Patrício-Goiás. 2 Congresso Internacional de Pesquisa, Ensino e Extensão v. 1, p 2

  29. Hahne J, Isele D, Berning J, Lipski A (2019) The contribution of fast growing, psychrotrophic microorganisms on biodiversity of refrigerated raw cow’s milk with high bacterial counts and their food spoilage potential. Food Microbiol 79:11–19

    Article  CAS  PubMed  Google Scholar 

  30. Qvirist LA, De Filippo C, Strati F, Stefanini I, Sordo M, Andlid T, Felis GE, Mattarelli P, Cavalieri D (2016) Isolation, identification and characterization of yeasts from fermented goat milk of the Yaghnob Valley in Tajikistan. Front Microbiol 7:1690

    Article  PubMed  PubMed Central  Google Scholar 

  31. Ruz-Peres M, Nilson RB, Eugenio Y, Priscilla AM (2010) Resistência de fungos filamentosos e leveduras isolados de leite in natura bovino à pasteurização e fervura. Periódicos Brasileiros em Medicina e Zootecnia 17(1):62–70

    Google Scholar 

  32. Montanari C, Tabanelli G, Zamagna I, Barbieri F, Gardini A, Ponzetto M, Redaelli E, Gardini F (2019) Modeling of yeast thermal resistance and optimization of the pasteurization treatment applied to soft drinks. Int J Food Microbiol 301(10):1–8

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

The authors are grateful to the CNPq and CAPES by structural and financial support to this work.

Author information

Authors and Affiliations

Authors

Contributions

JSC and ACVJ executed, analyzed, and interpreted the data for the manuscript. DB, DLS, and UGPL executed and analyzed the data for the antifungal tests and fungi identification. JVF, MARS, and VSA coordinated all the research and corrections and suggestions in the preparation of the manuscript.

Corresponding author

Correspondence to Douglas Boniek.

Ethics declarations

Conflict of interest

The authors declare no conflict of interest.

Additional information

Responsible Editor: Mariza Landgraf

Publisher's note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

da Silva Campos, J., Júnior, A.C.V., Boniek, D. et al. Identification and evaluation of thermotolerance of yeasts from milk in natura exposed to high temperature and slow and fast pasteurization. Braz J Microbiol 54, 1075–1082 (2023). https://doi.org/10.1007/s42770-023-00975-7

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s42770-023-00975-7

Keywords

Navigation