Microbial screening of animal skin bags used in traditional cheesemaking.

Bouhezza is a traditional Algerian cheese produced and ripened in goatskin bags called Djeld. The aim of this study was to characterize the microbial ecosystem from Djeld (fresh and dried Djeld for making Bouhezza cheese) and the changes introduced by Lben microflora during its preparation and to identify its role in cheesemaking and its safety. Two replicates of fresh and dried skin bags (FS and DS) were sampled and analyzed before and after contact with Lben. The microbiological results showed no pathogens. Skins observed before the addition of Lben were less populated 2.86 and 3.20 log CFU cm-2 than skins examined after the addition of Lben (approximately 6.0 log CFU cm-2), suggesting a potential role of Lben in releasing some microorganisms into the skin during its time in the Djeld. However, an increase in mesophilic lactic acid bacteria and yeasts was observed in Lben after different periods of interaction with the skin. PCR-TTGE revealed the predominance of lactic acid bacteria (Lactiplantibacillus plantarum, Limosilactobacillus fermentum, Staphylococcus equorum subsp. linens, Lactococcus cremoris, Streptococcus thermophilus) and a few high-GC-content bacteria (Lacticaseibacillus paracasei, Brevibacterium casei). Transfer of several microbial species was observed between the goatskin bag biofilm and Lben during the overnight interaction. Bands corresponding to Lacticaseibacillus paracasei, Brevibacterium casei, and Lactobacillus delbrueckii subsp. lactis were detected in the fresh skin profile and in Lben after contact with the fresh skin. Lacticaseibacillus paracasei was found in dried skin and Lben after contact with dry skin. Lactobacillus helveticus and Enterococcus faecalis appeared in the Lben profile and persisted in Lben and the biofilm-covered dry skin after interaction. These results demonstrate an exchange of specific microbial populations between goatskin bag biofilm and Lben during the traditional preparation method, suggesting that the diversity of goatskin biofilm contributes to the microbial diversity of Lben used in the production of Bouhezza cheese.

Uncommonly thorough hydrolysis of peptides during ripening of Ragusano cheese revealed by tandem mass spectrometry.

Ragusano is a pasta filata cheese produced from raw milk in Sicily. The proteolysis was extensively analyzed after stretching (day 0), at 4 and 7 months of ripening through soluble nitrogen, urea-PAGE, and peptide identification by tandem mass spectrometry. After stretching, 123 peptides were identified: 72 arising from ß-casein, 34 from α(s1)-casein, and 17 from α(s2)-casein. The main protein splitting corresponded to the action of plasmin, chymosin, cathepsin D, cell envelope proteinase, and peptidase activities of lactic acid bacteria. Unlike other types of cheeses, <10% residual ß- and α(s)-caseins remained intact at 7 months, indicating original network organization based on large casein fragments. The number of identified soluble peptides also dramatically decreased after 4 and 7 months of ripening, to 47 and 25, respectively. Among them, bioactive peptides were found, that is, mineral carrier, antihypertensive, and immunomodulating peptides and phosphopeptides.

Tina wooden vat biofilm: a safe and highly efficient lactic acid bacteria delivering system in PDO Ragusano cheese making.

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 10(3) to 10(6) CFU/cm(2), 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 (<10 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 5.10(4) to 10(6) CFU/mL of Streptococcus thermophilus were released into the milk, and from 10(4) to 10(5) CFU/mL of thermophilic lactobacilli. Spontaneous acidification after contact confirmed the high efficiency of biofilm lactic acid bacteria delivery.