Tina wooden vat biofilm: A safe and highly efficient lactic acid bacteria delivering

Sylvie Lortal a,c,*, Annalisa Di Blasi b, Marie-Noëlle Madec a,c, Concetta Pediliggieri b, Laura Tuminello b, Gaëlle Tanguy a,c, Jacques Fauquant a,c, Yohan Lecuona a,c, Patrizia Campo b, Stefania Carpino b, Giuseppe Licitra b

a INRA, UMR1253, Science et Technologie du Lait et de l’Oeuf, F-35 Rennes, France
b CoRFiLaC, 25 km 5, S.P. Ragusa Mare, 97100 Ragusa, Sicily, Italy
c AGROCAMPUS OUEST, UMR1253, Science et Technologie du Lait et de l’Oeuf, F-35 Rennes, France


In the Sicilian PDO Ragusano cheese making, raw milk is placed in a wooden vat called a Tina. As no starter is added, lactic acid is produced by milk flora and flora released from the Tina biofilm. The aim of this work was to assess the safety and efficiency of this natural inoculation system. From 15 Tinas’ biofilms, bacteria total counts varied from 103 to 106 CFU/cm2, with the predominance of thermophilic lactic acid bacteria. Low counts of yeasts and moulds were found in a few Tinas. Salmonella, Listeria monocytogenes, Escherichia coli O157:H7 were totally absent, as assessed by conventional plating and the Bax detection system after enrichment, highlighting the safety of the system. From four Tinas out of the 15, micropieces of wood were observed by confocal and scanning electron microscopy. The biofilm entrapped in a matrix covered almost entirely the surface of the wood. Polysaccharides were detected in the four Tinas. In three of the latter, cocci were predominant in the ecosystem whereas in the other one, cocci, bacilli, yeasts and moulds were observed. Fifty litres of microfiltrated milk (b10 CFU/mL) were poured in the four Tinas for 10 min of contact. Enumeration of lactic acid bacteria, yeasts and enterococci were performed in the milk after contact. Depending on the Tina, from 5·104 to 106 CFU/mL of Streptococcus thermophilus were released into the milk, and from 104 to 105 CFU/mL of thermophilic lactobacilli. Spontaneous acidification after contact confirmed the high efficiency of biofilm lactic acid bacteria delivery.

Keywords: Wooden vat, Cheese, Biofilm, Ragusano cheese, Lactic acid bacteria, Food safety

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Sylvie Lortal