Isolation and identification of probiotic bacteria from Iranian traditional cheese produced by lamb or kid abomasum microorganisms

Different strains of lactic acid bacteria are the most common micro-organisms known as probiotics. Two major kinds of probiotic bacteria are members of the genera Lactobacillus and Bifidobacterium, that are generally associated with the genus Lactobacillus which play a vital role in the body health, as well as the ability to produce antimicrobials and vitamins in the gastrointestinal tract. The purpose of the study was the isolation of probiotics in traditional cheeses that are produced from the natural flora of the newborn mammalian digestive tract. Bacteria from lamb and kid abomasum are added to milk. The curd is slaughtered from the abomasum of a lamb or kid that is suckling and has now no longer began out to devour forage, after which its belly is dried, that is referred to as curd. In this study, sixteen samples of cheeses local to Markazi province of Iran have been prepared. The isolated bacteria have been examined for morphological, biochemical, probiotic properties, and molecular identity. Out of sixteen traditional cheese samples, five kinds of Lactobacillus have been diagnosed through PCR. L. casei strain J026 strain was identified in the traditional cheese samples as the most genera. Using probiotic starters from local sources is a beneficial strategy for producing traditional cheese, which the native strains are more compatible with the humans’ intestinal flora and therefore may also higher play their probiotic’s characteristic.

KARGI TULUM PEYNİRİNİN BAZI KİMYASAL, TEKSTÜREL VE MİKROBİYOLOJİK ÖZELLİKLERİNİN BELİRLENMESİ

Kargı Tulum cheese is a type of cheese produced in the highlands in summer seasons from sheep, goat, cow and buffalo milk and their mixtures in certain proportions in the Kargı district of Çorum and put on the market in autumn. It is particularly consumed in Çorum, Kastamonu, Samsun and Ankara provinces. For the marketing of cheeses, tulums made of sheep and lamb skins are cleaned and cut into small pieces and sewn. Cheese curds, after the whey draining, are pressed into the prepared tulums by being compressed so that there is no airgap in them. Traditional production and sales methods are stil used in Kargı Tulum cheese, therefore standardized products cannot be obtained in every production. For this reason, the stages involved in the traditional cheese production and the packaging material used for ripening affect the chemical and textural properties, microbial load and therefore, quality of the product. There are few number of studies on Kargı Tulum cheese and no extensive study examined the texture and rheological characteristics of it. Cheese texture is influenced by many factors, such as composition, microbiological load, ripening conditions and proteolysis; therefore texture properties should be evaluated together with the chemical and hygenic parameters. In this study, some chemical (pH, titrationacidity, total solids, fat), textural (TPA) and microbiological characteristics (total mesophilic count, mold and yeast counts, coliform) of tulum cheese samples produced according to the tradional method with three replicates were determined. Moisture content of the Kargı Tulum cheese samples were 43.45%, fat content was found % 25.5, pH was 4.55, acidity (lacticacid %) was 2.55. According to TS 3001 Tulum Cheese Standard our Kargı Tulum cheese samples were classified as full fat cheese and our chemical composition results were in accordance with the standard. We found statistically significant differences between cheese samples for all texture parameters measured (firmness, adhessiveness, springiness, gumminess, resilience) except for chewiness. Average total mesophilic aerobic bacteria countwas 6.40 log cfu.g-1 mean mold and yeast counts were found to be 1.07 log cfu.g-1. Coliform group bacteria were not found in the analyzed samples. When microbiological results were compared statistically, it was determined that there was no significant difference between cheese samples.

An in-Depth Investigation of the Safety of Wooden Shelves Used for Traditional Cheese Ripening

The main goal of this research was to characterize the bacterial diversity of wooden boards used for aging traditional Sicilian cheeses and to evaluate whether pathogenic bacteria are associated with these surfaces. Eighteen cheese dairy factories producing three traditional cheese typologies (PDO Pecorino Siciliano, PDO Piacentinu Ennese and Caciocavallo Palermitano) were selected within Sicily region. The wooden shelf surfaces were sampled by a destructive method to detach wood splinters as well as by a non-destructive brushing to collect microbial cells. Scanning electron microscopy showed the presence of almost continuous bacterial formations on the majority of the shelves analysed. Yeasts and fungal hyphae were also visualized, indicating a complexity of the plank communities. The amplicon library of the 16S rRNA gene V3-V4 region was pair-end sequenced using the Illumina MiSeq system allowing the identification of 14 phyla, 32 classes, 52 orders, 93 families and 137 genera. Staphylococcus equorum was identified from all wooden surfaces with a maximum abundance of 64.75%. Among cheese surface ripening bacteria, Brevibacterium and Corynebacterium were detected in almost all samples. Several halophilic ( Halomonas , Tetragenococcus halophilus , Chromohalobacter, Salimicrobium , Marinococcus , Salegentibacter , Haererehalobacter , Marinobacter and Idiomarinaceae) and moderately halophilic ( Salinicoccus , Psychrobacter and Salinisphaera ) bacteria were frequently identified. Lactic acid bacteria (LAB) were present at low percentages with the genera Leuconostoc , Lactococcus , Lactobacillus , Pediococcus and Streptococcus . The levels of wooden shelf viable microorganisms ranged between 2.4 and 7.8 log CFU/cm 2 . In some cases, LAB were counted at very high levels (8.2 log CFU/cm 2 ). Members of Enterobacteriaceae family were detected in a viable state only for six samples. Coagulase positive staphylococci, Salmonella spp. and Listeria monocytogenes were not detected. Seventy-five strains belonged to Leuconostoc , Lactococcus , Pediococcus , Enterococcus , Lactobacillus and Weissella genera. IMPORTANCE This study provides evidence for the lack of pathogenic bacteria on the wooden shelves used to ripen internal bacterially ripened semi-hard and hard cheeses produced in Sicily. These three cheeses are non-inoculated on their surfaces and surface ripening is not considered to occur or, at least, not at the same extent of surface inoculated smear cheeses. Several bacterial groups identified from the wooden shelves are typically associated with smear cheeses strongly suggesting that PDO Pecorino Siciliano, PDO Piacentinu Ennese and Caciocavallo Palermitano cheese rind contributes to their final organoleptic profiles.

ASSESSMENT OF PROBIOTIC PROPERTIES AND SAFETY OF LACTIC ACID BACTERIA ISOLATED FROM SOUTH SULAWESI ETHNIC CHEESE

Probiotic bacteria have been used widely as a functional food and health supplements. The functionality and safety of probiotics are the prerequisites given by WHO (World Health Organization) and FAO (Food Agricultural Organization) before utilizing probiotics. This study focuses on assessing probiotic properties and the safety of the lactic acid bacteria (LAB) isolated from dangke—traditional cheese of South Sulawesi. In the current study, the assessment of probiotic properties was carried by assessing its tolerance against low pH and bile salts. Safety assessments were divided into two assays viz., susceptibility testing and hemolytic activity. LAB from dangke demonstrated tolerance against low pH, bile salt and susceptibility against four types of antibiotics (ampicillin, cefotaxime, erythromycin, and tetracycline). Unfortunately, resistance towards gentamycin and an -hemolytic activity was observed. This bacterium met the functional criteria from probiotics but failed to meet the safety criteria for probiotic safety.

Proteomics Analysis in Dairy Products: Cheese, a Review

Cheese is a worldwide produced and consumed commodity. There are many varieties of cheese from soft to hard, white to yellow, and fresh to aged after ripening. Especially, each category has its own producing technology. Many countries have labeled their most traditional cheese as Protective Designation of Origin (PDO). Moreover, several studies using advanced technologies, such as proteomics, have been performed to enhance labeling. In this review, broadly diffused and marketed, as well as Mediterranean countries’ special interest in Mediterranean diet-related PDO cheeses have been chosen as a reference. The aim of this work was to highlight the use of proteomics methods to examine how cheese proteins and peptides rearrange after ripening and use of starters. Further, we aimed to examine what kind of proteins are produced. Finally, we focused on bioactive molecules in cheeses and distinction of the original product from its counterfeit.

Tobacco BY2 cells expressing recombinant cardosin B as an alternative for production of active milk clotting enzymes

Cynara cardunculus L. or cardoon is a plant that is used as a source of milk clotting enzymes during traditional cheese manufacturing. This clotting activity is due to aspartic proteases (APs) found in the cardoon flower, named cyprosins and cardosins. APs from cardoon flowers display a great degree of heterogeneity, resulting in variable milk clotting activities and directly influencing the final product. Producing these APs using alternative platforms such as bacteria or yeast has proven challenging, which is hampering their implementation on an industrial scale. We have developed tobacco BY2 cell lines as an alternative plant-based platform for the production of cardosin B. These cultures successfully produced active cardosin B and a purification pipeline was developed to obtain isolated cardosin B. The enzyme displayed proteolytic activity towards milk caseins and milk clotting activity under standard cheese manufacturing conditions. We also identified an unprocessed form of cardosin B and further investigated its activation process. The use of protease-specific inhibitors suggested a possible role for a cysteine protease in cardosin B processing. Mass spectrometry analysis identified three cysteine proteases containing a granulin-domain as candidates for cardosin B processing. These findings suggest an interaction between these two groups of proteases and contribute to an understanding of the mechanisms behind the regulation and processing of plant APs. This work also paves the way for the use of tobacco BY2 cells as an alternative production system for active cardosins and represents an important advancement towards the industrial production of cardoon APs.