Phenotypic and phylogenetic characterization of Lactobacillus species isolated from traditional Lighvan cheese

Lighvan cheese (Lighvan panir) is among the most famous traditional cheese in Iran for its desired aroma and flavor. Undoubtedly, the lactic acid bacteria especially the genus Lactobacillus are the critical factors in developing the aroma, flavor, and texture in Lighvan cheese. In this study, the Lactobacillus population of the main Lighvan cheese was investigated. The Lactobacillus of the main Lighvan cheese was isolated using specific culture methods according to previously published Guidelines. Then, the phylogenetic features were investigated and the phenotypic characteristics were examined using specific culture methods. Twenty-eight Gram-positive bacterial species were identified belonged to the genus Lactobacillus. According to the same sequences as each other, three groups (A, B, and C) of isolates were categorized with a high degree of similarity to L. fermentum (100%) and L. casei group (L. casei, L. paracasei, and L. rhamnosus) (99.0 to 100%). Random amplified polymorphic DNA (RAPD) fingerprint analysis manifested the presence of three clusters that were dominant in traditional Lighvan cheese. Cluster І was divided into 4 sub-clusters. By the result of carbohydrate fermentation pattern and 16S rRNA sequencing, isolates were identified as L. rhamnosus. The isolates in clusters II and III represented L. paracasei and L. fermentum, respectively as they were identified by 16S rRNA sequencing and fermented carbohydrate patterns. Our result indicated that the specific aroma and flavor of traditional Lighvan cheese can be related to its Lactobacillus population including L. fermentum, L. casei, L. paracasei, and L. rhamnosus. Graphical abstract

Study of the Effect of Echinophora platyloba Ethanolic Extract on Survival of Listeria monocytogenes in Lighvan Cheese During Ripening Time

Listeria monocytogenes is one of the most frequent foodborne bacteria that can be transmitted through dairy products. The demand for replacing chemical preservatives with natural compounds has increased recently. The aim of this study was to investigate the effect of Echinophora platyloba DC ethanolic extract on the survival of L. monocytogenes in Lighvan cheese during ripening time. Three concentrations of E. platyloba ethanolic extract (0, 0.5, 1 and 1.5%) were added to raw milk at the beginning of Lighvan cheese manufacture, and the population of L. monocytogenes was counted on days 15, 30, 60 and 90. Furthermore, the pH and salt concentration of Lighvan cheese were evaluated in these periods. The results showed that an increase in the concentration of ethanolic extract, as well as the ripening time of Lighvan cheese, resulted in a significant decrease (p<0.05) in the levels of L. monocytogenes (cfu/g). Moreover, the logarithm of the L. monocytogenes population (log cfu/g) was significantly decreased (p<0.05) as the concentration of the added ethanolic extract was enhanced over the ripening time, but no significant changes in pH and salt concentration were observed in Lighvan cheese (p>0.05). It was found that the optimal concentration of E. platyloba ethanolic extract for the complete inhibition of L. monocytogenes was 1-1.5% following 90 days of the ripening.

Antimicrobial Effects of Zataria multiflora and Ocimum basilicum on Escherichia coli O157:H7 During Ripening of Traditional Lighvan Cheese

Background: Escherichia coli O157:H7 is one of the most common causes of contamination in Lighvan cheese processing. Using from natural antimicrobial essential oils is applied method to decrease the rate of microbial contamination of dairy products. The present investigation was done to study the antimicrobial effects of Z. multiflora and O. basilicum essential oils on survival of E. coli O157:H7 during ripening of traditional Lighvan cheese. Methods: Leaves of the Z. multiflora and O. basilicum plants were subjected to the Clevenger apparatus. Concentrations of 0, 100 and 200 ppm of the Z. multiflora and 0, 50 and 100 ppm of O. basilicum essential oils and also 103 and 105 cfu/ml numbers of E. coli O157:H7 were used. The numbers of the E. coli O157:H7 bacteria were analyzed during the days 0, 30, 60 and 90 of the ripening period. Results: Z. multiflora and O. basilicum essential oils had considerable antimicrobial effects against E. coli O157:H7. Using the essential oils caused decrease in the numbers of E. coli O157:H7 bacteria in 90th days of ripening (P <0.05). Using from Z. multiflora at concentration of 200 ppm can reduce the survival of E. coli O157:H7 in Lighvan cheese. Conclusion: Using Z. multiflora and O. basilicum essential oils as good antimicrobial agents can reduce the risk of foodborne bacteria and especially E. coli O157:H7 in food products.

Biochemical changes of Iranian probiotic Lighvan cheese

Lighvan cheese from ewe and goat milk was produced according to a traditional protocol, and with the addition of 9 log₁₀ cfu/g fresh cells of Bifidobacterium lactis subsp. animalis. Probiotic Lighvan cheese was studied to determine the survival of B. lactis subsp. animalis and biochemical changes during 60 days of ripening of probiotic Lighvan cheese. Lipolysis level and organoleptic assessments were analysed. B. lactis subsp. animalis cells survived in cheese samples at concentrations up to 6.84 log₁₀ cfu/g for at least 60 days of storage time. The lipolysis level increased continuously until the end of the ripening period.The ripening stage was the main factor affecting the cheese sensory properties.