Comparison of Methods for the Detection of Biofilm Formation in Yeast and Lactic Acid Bacteria Species Isolated from Dairy Products

Lactic acid bacteria (LAB) and some yeast species are common microorganisms found in dairy products and most of them are responsible for the fermentation of foods. Such cultures are isolated and used as a starter culture in the food industry because of providing standardisation of the final product during the food processing. Choice of starter culture is the most important step for the production of fermented food. Isolated LAB and yeast cultures which have the ability to create a biofilm layer can be preferred as a starter in the food industry. The biofilm formation could be beneficial to extend the period of usage time of microorganisms as a starter. On the other hand, it is an undesirable property in pathogens, since biofilm structure allows a microorganism become more resistant to stress conditions such as antibiotic presence. It is thought that the resistance mechanism could be turned into an advantage by promoting the effective microorganisms which are used in the food industry as starter culture and also which have potential to stimulate the gastrointestinal system. Development of the biofilm layer is observed in some LAB and yeast strains. The resistance could make LAB and yeast strains dominant microflora in the human gastrointestinal system; thus, competition against pathogen microorganisms can be provided more easily. Based on this circumstance, in the study, 10 LAB and 10 yeast strains were isolated from various dairy products, such as cheese, yoghurt, kefir, and cream. Samples were obtained from farmer markets and bazaars in Bursa, Turkey. As a part of this research, all isolated strains were identified and their ability of biofilm formation was detected with two different methods and compared with each other. The first goal of this research was to determine whether isolates have the potential for biofilm production, and the second was to compare the validity of two different methods, which are known as “Tube method” and “96-well plate-based method”. This study may offer an insight into developing a point of view about biofilm formation and its beneficial properties in LAB and yeast cultures used as a starter in the food industry.

Screening of Antagonistic/Synergistic Effect between Lactic Acid Bacteria (LAB) and Yeast Strains Isolated from Kefir

Kefir is a traditional fermented refreshing beverage which is known for its valuable and beneficial properties for human health. Mainly yeast species, lactic acid bacteria (LAB) strains and fewer acetic acid bacteria strains live together in a natural matrix named “kefir grain”, which is formed from various proteins and polysaccharides. Different microbial species live together in slimy kefir grain and it has been thought that synergetic effect could take place between microorganisms, which belong to different genera and species. In this research, yeast and LAB were isolated from kefir samples obtained from Uludag University Food Engineering Department. The cell morphology of isolates was screened by microscopic examination. Gram reactions of bacteria isolates were determined by Gram staining method, and as well catalase activity was examined. After observing the microscopic/morphological and physical, enzymatic properties of all isolates, they were divided into the groups as LAB and/or yeast according to their physicochemical responses to the applied examinations. As part of this research, the antagonistic/synergistic efficacy of the identified five LAB and five yeast strains to each other were determined individually by disk diffusion method. The antagonistic or synergistic effect is one of the most important properties in a co-culture system that different microorganisms are living together. The synergistic effect should be promoted, whereas the antagonistic effect is prevented to provide effective culture for fermentation of kefir. The aim of this study was to determine microbial interactions between identified yeast and LAB strains, and whether their effect is antagonistic or synergistic. Thus, if there is a strain which inhibits or retards the growth of other strains found in Kefir microflora, this circumstance shows the presence of antagonistic effect in the medium. Such negative influence should be prevented, whereas the microorganisms which have synergistic effect on each other should be promoted by combining them in kefir grain. Standardisation is the most desired property for industrial production. Each microorganism found in the microbial flora of a kefir grain should be identified individually. The members of the microbial community found in the glue-like kefir grain may be redesigned as a starter culture regarding efficacy of each microorganism to another in kefir processing. The main aim of this research was to shed light on more effective production of kefir grain and to contribute a standardisation of kefir processing in the food industry.

Preliminary Study of Antimicrobial Activity against Escherichia coli and Probiotic Properties of Lactic Acid Bacteria Isolated from Thailand Fermented Foods

The lactic acid bacteria (LAB) were isolated from 10 samples of fermented foods (Sa-tor-dong and Bodo) in South locality of Thailand. The 23 isolates of lactic acid bacteria were selected, which were exhibited a clear zone and growth on MRS agar supplemented with CaCO3. All of lactic acid bacteria were tested on morphological and biochemical. The result showed that all isolates were Gram’s positive, non-spore forming but only 10 isolates displayed catalase negative. The 10 isolates including BD1 .1, BD 1.2, BD 2.1, BD2.2, BD 2.3, BD 3.1, BD 4.1, BD 5.2, ST 4.1 and ST 5.2 were selected for inhibition activity determination. Only 2 strains (ST 4.1 and BD 2.3) showed inhibition zone on agar, when using Escherichia coli sp. as target strain. The ST 4.1 showed highest inhibition zone on agar, which was selected for probiotic property testing. The ST4.1 isolate could grow in MRS broth containing a high concentration of sodium chloride 6%, bile salts 7%, pH 4-10 and vary temperature at 15-45°C.

Isolation and Probiotic Characterization of Arsenic-Resistant Lactic Acid Bacteria for Uptaking Arsenic

The growing health hazardous impact of arsenic (As) contamination in environment is the impetus of the present investigation. Application of lactic acid bacteria (LAB) for the removal of toxic and heavy metals from water has been reported. This study was performed in order to isolate and characterize the Asresistant LAB from mud and sludge samples for using as efficient As uptaking probiotic. Isolation of As-resistant LAB colonies was performed by spread plate technique using bromocresol purple impregnated-MRS (BP-MRS) agar media provided with As @ 50 μg/ml. Isolated LAB were employed for probiotic characterization process, acid and bile tolerance, lactic acid production, antibacterial activity and antibiotic tolerance assays. After As-resistant and removal characterizations, the LAB were identified using 16S rDNA sequencing. A total of 103 isolates were identified as As-resistant strains of LAB. The survival of 6 strains (As99-1, As100-2, As101-3, As102-4, As105-7, and As112-9) was found after passing through the sequential probiotic characterizations. Resistant pattern pronounced hollow zones at As concentration >2000 μg/ml in As99-1, As100-2, and As101-3 LAB strains, whereas it was found at ~1000 μg/ml in rest 3 strains. Among 6 strains, the As uptake efficiency of As102-4 (0.006 μg/h/mg wet weight of cell) was higher (17 – 209%) compared to remaining LAB. 16S rDNA sequencing data of 3 (As99- 1, As100-2, and As101-3) and 3 (As102-4, As105-7, and As112-9) LAB strains clearly showed 97 to 99% (340 bp) homology to Pediococcus dextrinicus and Pediococcus acidilactici, respectively. Though, there was no correlation between the metal resistant and removal efficiency of LAB examined but identified elevated As removing LAB would probably be a potential As uptaking probiotic agent. Since present experiment concerned with only As removal from pure water, As removal and removal mechanism in natural condition of intestinal milieu should be assessed in future studies.

Preliminary Study of Fermented Pickle of Tabah Bamboo Shoot (Gigantochloa nigrociliata (Buese) Kurz)

Processing tabah bamboo shoot as fermented pickle is one of the way to increase the shelf life of this bamboo shoot. The advantage of this shoot is low concentration of hydro cyanic acid (HCN) make it potential for functional food product. This study aimed to determine the characteristic of tabah bamboo shoot pickle such as total of lactic acid bacteria (LAB), pH, total acidity, and hydro cyanic acid (HCN) content, and also find the LAB’s type involved during fermentation, and organic acids’ profiles. The pickle was made by natural fermentation with 6% salt concentration and fermentation conducted for 13 days. The result showed during the fermentation time, in the 4th day LAB’s number was highest as much as 72 x 107 CFU/ml and the lowest pH was 3.09. We also found decreasing in HCN from 37.8 ppm at the beginning to 20.52 ppm at the end of fermentation process. The organic acids detected during the fermentation were lactic acid with the highest concentration was 0.0546 g/100 g and small amount of acetic acid. By using PCR method, the 18 of LABs which had rod shape were detected as member of Lactobacillus spp., in which 17 strains detected as L. plantarum.

Antibacterial Activity of Lactic Acid Bacteria Isolated from Table Olives against Skin Pathogens

The aim of this study was to assess the effect of LAB isolated from Iranian native olives on the opportunistic skin pathogens, Pseudomonas aeruginosa and Staphylococcus aureus. Lactic Acid Bacteria were isolated from the brine of each sample in the prior of time. The samples were spread on MRS agar for isolation of lactobacillus and for lactococcus. 28 strains of labs were isolated. The labs were centrifuged, the supernatant was strewed and pellet was used to inoculation in wells or at blank disks. 20μl of each pellet was inoculated to blank disks and 40μl of each pellet was inoculated to each well. The result of disk and well diffusion agar against these pathogens were confirmed each other. The size of inhibition zone was different according to the type of bacteria, the method and the concentrations of labs.