Composition and Significance of Bacterial Microbiota and Volatile Organic Compounds of Swiss-Dutch-Type Cheese as Determined by PCR-DGGE and HS-GC
More details
Hide details
Department of Industrial and Food Microbiology, Faculty of Food Sciences, University of Warmia and Mazury in Olsztyn, Pl. Cieszyński 1, 10-726 Olsztyn, Poland
Department of Immunology and Food Microbiology, Institute of Animal Reproduction and Food Research, Polish Academy of Sciences, Tuwima 10, 10-748 Olsztyn, Poland
Department of Dairy Science and Quality Management, Faculty of Food Sciences, University of Warmia and Mazury in Olsztyn, Oczapowskiego 7, 10-719 Olsztyn, Poland
Beata Nalepa   

Department of Industrial and Food Microbiology, Faculty of Food Sciences, University of Warmia and Mazury in Olsztyn, Pl. Cieszyński 1, 10-726, Olsztyn, Poland
Online publish date: 2019-06-28
Submission date: 2019-02-13
Final revision date: 2019-05-15
Acceptance date: 2019-06-06
This study aimed to determine seasonal differences in the composition of bacterial microbiota and volatile organic compounds (VOCs) in Swiss-Dutch-type cheese (manufactured between 2012 and 2014). Bacterial diversity and VOCs (acetaldehyde; ketones: acetone, diacetyl, acetoin; alcohols: methanol, ethanol; esters: ethyl acetate, ethyl propionate, ethyl butyrate; fatty acids: acetic acid, propionic acid, isobutyric acid, butyric acid, isovaleric acid, valeric acid, isocaproic acid, caproic acid, heptanoic acid) were determined by polymerase chain reaction - denaturing gradient gel electrophoresis (PCR-DGGE), and headspace gas chromatography (HS-GC), respectively. Season influenced the composition of both bacterial microbiota and VOCs in cheese. Counts of starter bacteria (Lactococcus, Leuconostoc and Propionibacterium – 6.51-7.14, 3.6-3.96 and 2.88-4.72 log CFU/g, respectively) were higher in the first year of the study, likewise these of the non-starter Lactobacillus (4.12-5.69 log CFU/g). The total VOC content was substantially lower in the summer-autumn 2012 (0.73228-3.34111 mg/g) than in the other seasons (63.28810-131.27690 mg/g). Differences in bacterial microbiota and the VOC profiles were observed between cheeses manufactured in winter-spring and summer-autumn seasons. Winter- and spring-manufactured cheeses were also characterized by a lower number of bacterial species (average 8.7-10.5 species/sample) than the cheeses produced in the summer and in the autumn (average 10-13 species/sample). The results of the study indicate that the cheese-making process has to be continuously monitored to minimize differences across manufacturing seasons.
We are grateful to Dr. Marta Mikš for calibration of the gas chromatograph for quantitative identification volatile compounds of microbiological origin in samples of milk and ripened chees.
This study was supported by the National Science Center, Poland (grant no. N N312 484140) and by University of Warmia and Mazury in Olsztyn (17.610.015-300).
Abriouel, H., Martin-Platero, A., Maqueda, M., Valdivia, E., Martinez-Bueno, M. (2008). Biodiversity of the microbial community in Spanish farmhouse cheese as revealed by culture-dependent and culture-independent methods. International Journal of Food Microbiology, 127(3), 200−208.
Alegría, A., Alvarez-Martin, P., Sacristan, N., Fernandez, E., Delgado, S., Mayo, B. (2009). Diversity and evolution of the microbial populations during manufacture and ripening of Casin, a traditional Spanish, starter-free cheese made from cow’s milk. International Journal of Food Microbiology, 136(1), 44−51.
AOAC International 2005. Method 926.08. Official Methods of Analysis. 18th ed. AOAC International, Gaithersburg, MD, USA: AOAC International; 2005.
Ayad, E.H.E., Verheul, A., De Jong, C., Wouters, J.T.M., Smit, G. (1999). Flavour forming abilities and amino acid requirements of Lactococcus lactis strains isolated from artisanal and non-dairy origin. International Dairy Journal, 9, 725–735.
Caridi, A., Micari, P., Caparra, P., Cufari, A., Sarullo, V. (2003). Ripening and seasonal changes in microbial groups and in physico-chemical properties of the ewes’ cheese Pecorino del Poro. International Dairy Journal, 13, 191−200.
De Freitas, I., Pinon, N., Thierry, A., Lopez, C., Maubois, J.-L., Lortal, S. (2007). In depth dynamic characterization of French POD Cantal cheese made from raw milk. Le Lait, 87, 97−117.
Dolci, P., Alessandria, V., Rantsiou, K., Rolle, L., Zeppa, G., Cocolin, L. (2008). Microbial dynamics of Castelmagno PDO, a traditional Italian cheese, with a focus on lactic acid bacteria ecology. International Journal of Food Microbiology, 122(3), 302−311.
Drinan, F.D., Cogan, T.M. (1992). Detection of propionibacteria in cheese. Journal of Dairy Research, 59(1), 65-69.
Duru, I.C., Laine, P., Andreevskaya, M., Paulin, L., Kananen, S., Tynkkynen, S., Auvinen, P., Smolander O.-P. (2018). Metagenomic and metatranscriptomic analysis of the microbial community in Swiss-type Maasdam cheese during ripening. International Journal of Food Microbiology, 281, 10–22.
Duthoit, F., Callon, C., Tessier, L., Montel, M.-C. (2005). Relationships between sensorial characteristics and microbial dynamics in “Registered Designation of Origin” Salers cheese. International Journal of Food Microbiology, 103(3), 259−270.
Ercolini, D., Moschetti, G., Blagiotta, G., Coppola, S. (2001). The potential of a polyphasic PCR-DGGE approach in evaluating microbial diversity of natural whey cultures for water-buffalo Mozzarella cheese production: bias of culture-dependent and culture-independent analyses. Systematic and Applied Microbiology, 24(4), 610–617.
Felicio, T.L., Esmerino, E.A., Vidal, V.A.S., Cappato, L.P., Garcia, R.K.A., Cavalcanti, R.N., Freitas, M.Q., Conte Junior, C.A., Padilha, M.C., Silva, M.C., Raices, R.S.L., Arellano, D.B., Bollini, H.M.A., Pollonio, M.A.R., Cruz, A.G. (2016). Physico-chemical changes during storage and sensory acceptance of low sodium probiotic Minas cheese added with arginine. Food Chemistry, 196, 628–637.
Flórez, A.B., Mayo, B. (2006). PCR–DGGE as a tool for characterizing dominant microbial populations in the Spanish blue-veined Cabrales cheese. International Dairy Journal, 16, 1205–1210.
Fox, P.F., McSweeney, P.L.H., Cogan, T.M., Guinee, T.P. (2004). Cheese: Chemistry, Physics and Microbiology. 3nd edition, Elsevier Ltd., London, UK.
Franciosi, E., Settanni, L., Cavazza, A., Poznanski, E. (2009). Biodiversity and technological potential of wild lactic acid bacteria from raw cows’ milk. International Dairy Journal, 19, 3−11.
Frece, J., Vrdoljak, M., Filipčić, M., Jelić, M., Čanak, I., Jakopović, Ž., Pleadin, J., Gobin, I., Landeka Dragičević, T., Markov, K. (2016). Microbiological quality and variability of natural microbiota in Croatian cheese maturing in lambskin sacks. Food Technology and Biotechnology, 54(2), 129–134.
Gala, E., Landi, S., Solieri, L., Nocetti, M., Pulvirenti, A., Giudici, P. (2008). Diversity of lactic acid bacteria population in ripened Parmigiano Reggiano cheese. International Journal of Food Microbiology, 125(3), 347−351.
Gobbetti, M., De Angelis, M., Di Cagno, R., Mancini, L., Fox, P.F. (2015). Pros and cons for using non-starter lactic acid bacteria (NSLAB) as secondary/adjunct starters for cheese ripening. Trends in Food Science and Technology, 45(2), 167−178.
Granato, D., Putnik, P., Bursać Kovačević, D., Sousa Santos, J., Calado, V., Silva Rocha, R., Gomes Da Cruz, A., Jarvis, B., Ye Rodionova, O., Pomerantsev, A. (2018). Trends in chemometrics: Food authentication, microbiology, and effects of processing. Comprehensive Reviews in Food Science and Food Safety, 17(3), 663-677.
IDF 222: 2008. ISO 3433:2008. Cheese - determination of fat content - Van Gulik method. International Standard Organization; 2008.
IDF 88:2006. ISO 5943:2006. Cheese and processed cheese products - Determination of chloride content -- Potentiometric titration method. International Standard Organization; 2006.
IDF 50:2008. ISO 707:2008. Milk and milk products — Guidance on sampling; International Standard Organization; 2008.
Januszkiewicz, J., Sabik, H., Azarnia, S., Lee, B. (2008). Optimization of headspace solid-phase microextraction for the analysis of specific flavors in enzyme modified and natural cheddar cheese using factorial design and response surface methodology. Journal of Chromatography A, 1195(1−2), 16–24.
Johnson, M.E. (2017). A 100-Year Review: Cheese production and quality. Journal of Dairy Science, 100(12), 9952–9965.
Jordan, K.N., Cogan, T.M. (1993). Identification and growth of non-starter lactic acid bacteria in Irish cheddar cheese. Irish Journal of Agricultural and Food Research, 32, 47−55.
Joux, F., Lebaron, P. (2000). Use of fluorescent probes to assess physiological functions of bacteria at single-cell level. Microbes and Infection, 2(12), 1523−1535.
Klijn, N., Nieuwenhof, F.F.J., Hoolwerf, J.D., van der Waals, C.B., Weerkamp, A.H. (1995). Identification of Clostridium tyrobutyricum as the causative agent of late blowing in cheese by species-specific PCR amplification. Applied and Environmental Microbiology, 61, 2919–2924.
Lindberg, A.-M., Christiansson, A., Rukke, E.-O., Eklund, T., Molin, G. (1996). Bacterial flora of Norwegian and Swedish semi-hard cheese after ripening, with special reference to Lactobacillus. Netherlands Milk and Dairy Journal, 50, 563−572.
Mangia, N.P., Fancello, F., Deiana, P. (2016). Microbiological characterization using combined culture dependent and independent approaches of Casizolu pasta filata cheese. Journal of Applied Microbiology, 120, 329–345.
Marino, M., Maifreni, M., Rondinini, G. (2003). Microbiological characterization of artisanal Montasio cheese: analysis of its indigenous lactic acid bacteria. FEMS Microbiology Letters, 229, 133−140.
Mauriello, G., Moio, L., Genovese, A., Ercolini, D. (2003). Relationships between flavoring capabilities, bacterial composition, and geographical origin of natural whey cultures used for traditional water-buffalo Mozzarella cheese manufacture. Journal of Dairy Science, 86, 486–497.
McSweeney, P.L.H., Sousa, M.J. (2000). Biochemical pathways for the production for flavour compounds in cheese in during ripening: a review. Le Lait, 80(3), 293−324.
Milosavljević, N.P., Blagojević, P.D., Savić, D.S., Radulović, N.S. (2012). Application of HS-SPME–GC-MS-derived variables for monitoring ripening-induced changes in cheese volatile compounds. Dairy Science & Technology, 92(4), 321–333.
Mondello, L., Costa, R., Tranchid, P.Q., Chiofalo, B., Zumbo, A., Dugo, P., Dugo, G. (2005) Determination of flavour components in Sicilian goat cheese by automated HS-SPME-GC. Flavour and Fragrance Journal, 20(6), 659–665.
Nalepa, B., Markiewicz, L.H. (2017). PCR-DGGE markers for qualitative profiling of microbiota in raw milk and ripened cheeses. LWT - Food Science and Technology, 84, 168−174.
Ogier, J.-C., Lafarge, V., Girard, V., Rault, A., Maladen, V., Gruss, A., Leveau, J.-Y., Delacroix-Buchet, A. (2004). Molecular fingerprinting of dairy microbial ecosystems by use of temporal temperature and denaturing gradient gel electrophoresis. Applied and Environmental Microbiology, 70(9), 5628−5643.
Pastink, M.I., Sieuwerts, S., de Bok, F.A.M., Janssen, P.W.M., Teusink, B., van Hylckman Vlieg, J.E.T., Hugenholtz, J. (2008). Genomics and high-throughput screening approaches for optimal flavor production in dairy fermentation. International Dairy Journal, 18(8), 781−789.
Pérès, C., Viallon, C., Berdague, J.-L. (2001). Solid-phase microextraction-mass spectrometry: a new approach to the rapid characterization of cheeses. Analytical Chemistry, 73, 1030–1036.
Porcellato, D., Skeie, S.B. (2016). Bacterial dynamics and functional analysis of microbial metagenomes during ripening of Dutch-type cheese. International Dairy Journal, 61, 182−188.
Randazzo, C.L., Pitino, I., Ribbera, A., Caggia, C. (2010). Pecorino Crotonese cheese: Study of bacterial population and flavour compounds. Food Microbiology, 27(3), 363−374.
Randazzo, C.L., Vaughan, E.E., Caggia, C. (2006). Artisanal and experimental Pecorino Siciliano cheese: Microbial dynamics during manufacture assessed by culturing and PCR-DGGE analyses. International Journal of Food Microbiology, 109(1-2), 1−8.
Rehfeld, I.S., Fraiha, A.L.S., Matos, A.C.D., Guedes, M.I.M.C., Costa, E.A., de Souza, M.R., Cavalcante, L.F.L., Lobato, Z.I.P. (2017). Survival of Vaccinia virus in inoculated cheeses during 60-day ripening. Journal of Dairy Science, 100(9), 7051–7054.
Ricciardi, A., Guidone, A., Ianniello, R.G., Cioffi, S., Aponte, M., Pavlidis, D., Tsakalidou, E., Zotta, T., Parente, E. (2015). A survey of non-starter lactic acid bacteria in traditional cheeses: Culture dependent identification and survival to simulated gastrointestinal transit. International Dairy Journal, 43, 42−50.
Santiago-López, L., Aguilar-Toalá, J.E., Hernández-Mendoza, A., Vallejo-Cordoba, B., Liceaga, A.M., González-Córdova, A.F. (2018). Bioactive compounds produced during cheese ripening and health effects associated with aged cheese consumption. Journal of Dairy Science, 101(5), 3742–3757.
Sienkiewicz, J. (2010). Concepts of biodiversity - their dimensions and measurements in the light of the subject literature. Ochrona Środowiska i Zasobów Naturalnych, 45, 7−29 (in Polish).
Skelin, A., Mrkonjić Fuka, M., Čanžek Majhenič, A., Redžepović, S., Samaržija, D., Bogovič Matijašić, B. (2012). Phenotypic and genotypic characterization of indigenous Lactobacillus community from traditional Istrian ewe's cheese. Food Technology and Biotechnology, 50(3), 362–370.
Smit, G., Smit, B.A., Engel, W.J.M. (2005). Flavour formation by lactic acid bacteria and biochemical flavour profiling of cheese products. FEMS Microbiology Reviews, 29, 591–610.
Terzaghi, B.E., Sandine, W.E. (1975). Improved medium for lactic streptococci and their bacteriophages. Applied Microbiology, 29, 807-813.
Thierry, A., Maillard, M.B., Richoux, R., Lortal, S. (2006). Ethyl ester formation is enhanced by ethanol addition in mini Swiss cheese with and without added propionibacteria. Journal of Agricultural and Food Chemistry, 54(18), 6819−6824.