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Fluorescence microscopy methods for the determination of somatic cell count in raw cow's milk

Author

Listed:
  • P. Zajac

    (Veterinary and Food Institute Bratislava, Bratislava, Slovak Republic)

  • S. Zubricka

    (Veterinary and Food Institute Bratislava, Bratislava, Slovak Republic)

  • J. Capla

    (Slovak Agriculture University in Nitra, Nitra, Slovak Republic)

  • L. Zelenakova

    (Slovak Agriculture University in Nitra, Nitra, Slovak Republic)

Abstract

The direct fluorescence microscopy method with ethidium bromide staining can be used for somatic cell counting in raw cow's milk. However, this method has some limitations that may influence the results of the analysis. We therefore aimed at improving the procedure of somatic cell nuclei staining. We tested the hypothesis that ethidium bromide can better penetrate into the DNA of cells with degraded somatic cell walls or into dead cells. Therefore, we increased the temperature of the sample to 100 °C in order to disrupt the somatic cell wall membrane and to improve ethidium bromide penetration to somatic cell nuclei. In all, 90 samples of raw cow's milk were analysed in this experiment. Three parallel measurements were performed using each of the microscopic methods and the routine flow cytometry method. In all, 810 microscopic smears were analysed. The somatic cells were counted using fluorescence microscopic methods and flow cytometry. The increased temperature during the sample preparation improved (P < 0.005) the penetration of ethidium bromide into the somatic cell nuclei. It is concluded that the direct fluorescence microscopy method is suitable for precise laboratory analysis of somatic cell in raw cow's milk.

Suggested Citation

  • P. Zajac & S. Zubricka & J. Capla & L. Zelenakova, 2016. "Fluorescence microscopy methods for the determination of somatic cell count in raw cow's milk," Veterinární medicína, Czech Academy of Agricultural Sciences, vol. 61(11), pages 612-622.
    • Handle: RePEc:caa:jnlvet:v:61:y:2016:i:11:id:222-2015-vetmed
    DOI: 10.17221/222/2015-VETMED
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    References listed on IDEAS

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    1. D. Cervinkova & H. Vlkova & I. Borodacova & J. Makovcova & V. Babak & A. Lorencova & I. Vrtkova & D. Marosevic & Z. Jaglic, 2013. "Prevalence of mastitis pathogens in milk from clinically healthy cows," Veterinární medicína, Czech Academy of Agricultural Sciences, vol. 58(11), pages 567-575.
    2. D. Rysanek & V. Babak & M. Zouharova, 2007. "Bulk tank milk somatic cell count and sources of raw milk contamination with mastitis pathogens," Veterinární medicína, Czech Academy of Agricultural Sciences, vol. 52(6), pages 223-230.
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    1. D. Cervinkova & H. Vlkova & I. Borodacova & J. Makovcova & V. Babak & A. Lorencova & I. Vrtkova & D. Marosevic & Z. Jaglic, 2013. "Prevalence of mastitis pathogens in milk from clinically healthy cows," Veterinární medicína, Czech Academy of Agricultural Sciences, vol. 58(11), pages 567-575.

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