Effect of exopolysaccharides (EPSs) produced by Lactobacillus delbrueckii subsp. bulgaricus strains to bacteriophage and nisin sensitivity of the bacteria

Lactobacillus strains used in this study were isolated from village-type yogurt and raw milk. The isolates were identified as Lactobacillus delbrueckii subsp. bulgaricus by 16 s rDNA sequence analysis and API 50 CHL identification systems. The exopolysaccharide (EPS) production of the strains growth in skim milk were investigated. In addition sensitivity and insensitivity of these strains against domestic bacteriophages and nisin were examined. It was deduced that those strains which had relatively high EPS-producing capacity were insensitive against phages and nisin. Linear relationships were determined between EPS production of the bacteria and bacteriophage and nisin insensitivity of the bacteria.There was a negative correlation between EPS production quantity and phage and nisin sensitivity of the bacteria. Of all the strains, L. delbrueckii subsp bulgaricus B3 produced the highest EPS quantity, and it was insensitive against phages and nisin. Based on these results, it is suggested that L. delbrueckii subsp bulgaricus B3 can be used with the starter culture in dairy industry for stable and high-quality yogurt production.     

Diversity of Lactic Acid Bacteria Isolated from Brazilian Water Buffalo Mozzarella Cheese

The water buffalo mozzarella cheese is a typical Italian cheese which has been introduced in the thriving Brazilian market in the last 10 y, with good acceptance by its consumers. Lactic acid bacteria (LAB) play an important role in the technological and sensory quality of mozzarella cheese. In this study, the aim was to evaluate the diversity of the autochthones viable LAB isolated from water buffalo mozzarella cheese under storage. Samples were collected in 3 independent trials in a dairy industry located in the southeast region of Brazil, on the 28th day of storage, at 4 ºC. The LAB were characterized by Gram staining, catalase test, capacity to assimilate citrate, and production of CO₂ from glucose. The diversity of LAB was evaluated by RAPD‐PCR (randomly amplified polymorphic DNA‐polymerase chain reaction), 16S rRNA gene sequencing, and by Vitek 2 system. Twenty LAB strains were isolated and clustered into 12 different clusters, and identified as Streptococcus thermophilus, Enterococcus faecium, Enterococcus durans, Leuconostoc mesenteroides subsp. mesenteroides, Lactobacillus fermentum, Lactobacillus casei, Lactobacillus delbrueckii subsp. bulgaricus, and Lactobacillus helveticus. Enterococcus species were dominant and citrate‐positive. Only the strains of L. mesenteroides subsp. mesenteroides and L. fermentum produced CO₂ from glucose and were citrate‐positive, while L. casei was only citrate positive. This is the first report which elucidates the LAB diversity involved in Brazilian water buffalo mozzarella cheese. Furthermore, the results show that despite the absence of natural whey cultures as starters in production, the LAB species identified are the ones typically found in mozzarella cheese.     

Rheological and physical characteristics of non-fat set yogurt prepared with EPS-producing Streptococcus thermophilus and an H+-ATPase-defective mutant Lactobacillus delbrueckii subsp. bulgaricus

The rheological and physical characteristics of non-fat set yogurt produced using co-cultures of ropy EPS-producing Streptococcus thermophilus S3.3 and Lactobacillus delbrueckii subsp. bulgaricus LTM and of Streptococcus thermophilus S3.3 and Lactobacillus delbrueckii subsp. bulgaricus H+-ATPase-defective mutant L6 under different conditions were studied. Yogurts produced with mutant L6 had a higher pH and water-holding capacity and better textural properties compared to those produced with the LTM strain. The highest storage modulus (G′) and thixotropic loops were found for yogurt made with L6 at 42°C on day 21 of cold storage. This yogurt contained a network of thick, continuous protein aggregates and large void spaces.     

Immunomodulatory effects of polysaccharides produced by Lactobacillus delbrueckii ssp. bulgaricus OLL1073R-1

The extracellular polysaccharides (EPS) produced by lactic acid bacteria (LAB) are associated with the rheology, texture, and mouthfeel of fermented milk products, including yogurt. This study investigated the immunomodulatory effects of EPS purified from the culture supernatant of Lactobacillus delbrueckii ssp. bulgaricus (L. bulgaricus) OLL1073R-1. The crude EPS were prepared from the culture supernatant of L. bulgaricus OLL1073R-1 by standard chromatographic methods, and were fractionated into neutral EPS and acidic EPS (APS). Acidic EPS were further fractionated into high molecular weight APS (H-APS) and low molecular weight APS (L-APS). High molecular weight APS were shown to be phosphopolysaccharides containing D-glucose, D-galactose, and phosphorus. Stimulation of mouse splenocytes by H-APS significantly increased interferon-γ production, and, moreover, orally administered H-APS augmented natural killer cell activity. Oral administration of yogurt fermented with L. bulgaricus OLL1073R-1 and Streptococcus thermophilus OLS3059 to mice showed a similar level of immunomodulation as H-APS. However, these effects were not detected following administration of yogurt fermented with the starter combination of L. bulgaricus OLL1256 and S. thermophilus OLS3295. We conclude from these findings that yogurt fermented with L. bulgaricus OLL1073R-1, containing immunostimulative EPS, would have an immunomodulatory effect on the human body.     

Productions and monomer compositions of exopolysaccharides by Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophilus strains isolated from traditional home-made yoghurts and raw milk

In this study, a total of forty‐five strains of lactobacilli and streptococci were determined exopolysaccharide (EPS) production in skim milk and Man Rogosa and Sharpe (MRS)/M17 medium, viscosity and proteolytic activity. The exopolysaccharide production by lactobacilli strains during growth in MRS medium was twenty‐one to 211 mg L^?1, while in skim milk was to thirty‐six to 315 mg L^?1. The EPS production by streptococci strains during growth in M17 medium was sixteen to 114 mg L^?1, while in skim milk was to twenty‐four to 140 mg L^?1. The EPS production of strains was lower in MRS/M17 medium than skim milk. Results showed that it was not clear correlation between the viscosity and EPS production of some strains. All strains were shown proteolytic activity. Positive correlations between exopolysaccharide production and proteolytic activity in skim milk were found some strains of Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophilus. These results indicated that the high exocellular protease‐producing strains can produce high EPS in skim milk. The monomer compositions of the EPSs formed by selected five strains were analysed. Mannose dominated (99–100%) on the EPS produced by L. delbrueckii subsp. bulgaricus and S. thermophilusstrains (except L. delbrueckii subsp. bulgaricus 22) in skim milk and MRS/M17 medium. Besides, the EPSs of strains in skim milk contained small amount of lactose.     

Fat-free yogurt made using a galactose-positive exopolysaccharide-producing recombinant strain of Streptococcus thermophilus

To prevent textural defects in low-fat and fat-free yogurts, fat substitutes are routinely added to milk. In situ production of exopolysaccharides (EPS) by starter cultures is an acknowledged alternative to the addition of biothickeners. With the aim of increasing in situ EPS production, a recombinant galactose-positive EPS⁺Streptococcus thermophilus strain, RD-534-S1, was generated and compared with the parent galactose-negative EPS⁺ strain RD-534. The RD-534-S1 strain produced up to 84 mg/L of EPS during a single-strain milk fermentation process, which represented 1.3 times more than the EPS produced by strain RD-534. Under conditions that mimic industrial yogurt production, the starter culture consisting of RD-534-S1 and (EPS⁻) Lactobacillus bulgaricus L210R strain (RD-534-S1/L210R) led to an EPS production increase of 1.65-fold as compared with RD-534-S1 alone. However, the amount of EPS produced did not differ from that found in yogurts produced using an isogenic starter culture that included the parent S. thermophilus strain RD-534 and Lb. bulgaricus L210R (RD-534/L210R). Moreover, the gel characteristics of set-style yogurt and the rheological properties of stirred-style yogurt produced using RD-534-S1/L210R were similar to the values obtained for yogurts made with RD-534/L210R. In conclusion, it is possible to increase the production of EPS by ropy S. thermophilus strains through genetic engineering of galactose metabolism. However, when used in combination with Lb. bulgaricus for yogurt manufacture, the EPS overproduction of recombinant strain is not significant.     

Diversity of lactic acid bacteria associated with traditional fermented dairy products in Mongolia

Spontaneous milk fermentation has a long history in Mongolia, and beneficial microorganisms have been handed down from one generation to the next for use in fermented dairy products. The objective of this study was to investigate the diversity of lactic acid bacteria (LAB) communities in fermented yak, mare, goat, and cow milk products by analyzing 189 samples collected from 13 different regions in Mongolia. The LAB counts in these samples varied from 3.41 to 9.03 log cfu/mL. Fermented yak and mare milks had almost identical mean numbers of LAB, which were significantly higher than those in fermented goat milk but slightly lower than those in fermented cow milk. In total, 668 isolates were obtained from these samples using de Man, Rogosa, and Sharpe agar and M17 agar. Each isolate was considered to be presumptive LAB based on gram-positive and catalase-negative properties, and was identified at the species level by 16S rRNA gene sequencing, multiplex PCR assay, and restriction fragment length polymorphism analysis. All isolates from Mongolian dairy products were accurately identified as Enterococcus faecalis (1 strain), Enterococcus durans (3 strains), Lactobacillus brevis (3 strains), Lactobacillus buchneri (2 strains), Lactobacillus casei (16 strains), Lactobacillus delbrueckii ssp. bulgaricus (142 strains), Lactobacillus diolivorans (17 strains), Lactobacillus fermentum (42 strains), Lactobacillus helveticus (183 strains), Lactobacillus kefiri (6 strains), Lactobacillus plantarum ssp. plantarum (7 strains), Lactococcus lactis ssp. lactis (7 strains), Leuconostoc lactis (22 strains), Leuconostoc mesenteroides (21 strains), Streptococcus thermophilus (195 strains), and Weissella cibaria (1 strain). The predominant LAB were Strep. thermophilus and Lb. helveticus, which were isolated from all sampling sites. The results demonstrate that traditional fermented dairy products from different regions of Mongolia have complex compositions of LAB species. Such diversity of LAB provides useful information for further studies of probiotic strain selection and starter culture design, with regard to the industrial production of traditional fermented milk.     

Comparison of Associative Growth and Proteolytic Activity of Yogurt Starters in Whole Milk from Camels and Cows

Growth and proteolytic activities were studied using yogurt starter cultures incubated in pasteurized whole milk from camels and cows at 42°C as single and mixed cultures. In general, the growth of four strains of Streptococcus thermophilus and three strains of Lactobacillus delbrueckii ssp. bulgaricus was higher in cow milk than in camel milk. However, proteolysis was higher in camel milk than in cow milk. Lactobacillus delbrueckii ssp. bulgaricus LB12 in combination with streptococci had lowered pH more than did the other lactobacilli. Mixed cultures released the same amount of free amino groups as the corresponding single cultures, except for L. delbrueckii ssp. bulgaricus LB12.     

Probiotic Strains as Starter Cultures Improve Angiotensin-converting Enzyme Inhibitory Activity in Soy Yogurt

Suitability of soy yogurt as a system for delivering probiotics and other bioactive compounds was assessed by fermenting soy milk using starter culture containing Lactobacillus delbrueckii ssp. bulgaricus Lb1466, Streptococcus thermophilus St1342, and probiotic organisms (Lactobacillus acidophilus LAFTI® L10, Bifidobacterium lactis LAFTI® B94, and Lactobacillus paracasei LAFTI® L26). Fermentations were terminated at different pH of 4.50, 4.55, and 4.60 and metabolic patterns of cultures (viability, proteolytic activity, organic acids production, angiotensin‐converting enzyme (ACE) inhibitory activity) were investigated during 28 d of storage at 4 °C. The presence of probiotics enhanced the growth of L. delbrueckii ssp. bulgaricus Lb1466 and S. thermophilus St134 in soy yogurt in comparison to the control produced by sole yogurt culture. In general, different termination pH had no effect (P > 0.05) on the viability of probiotic organisms that maintained good viability in soy yogurt during cold storage. Higher levels of essential growth factors in the form of peptides and amino acids in soy yogurts may have promoted the growth of L. acidophilus LAFTI® L10, B. lactis LAFTI® B94, and L. paracasei LAFTI® L26. The use of probiotic strains as a part of starter culture in soy yogurt resulted in a substantial increase in in vitro ACE inhibitory activity compared with the control produced by yogurt culture only. This improvement of ACE inhibition in soy yogurt is partly due to higher proteolytic activity of probiotics.     

Screening selected strains of probiotic lactic acid bacteria for their ability to produce biogenic amines (histamine and tyramine)

The aim of this study was to investigate the production of biogenic amines (BA), histamine and tyramine by some probiotic lactic acid bacteria (LAB). Fifteen strains representing six LAB species were screened qualitatively by growing them in a decarboxylase medium. Quantitative analysis was carried out by HPLC analysis with direct derivatization of acid extracts. Lactobacillus casei (TISTR 389) and Lactobacillus delbrueckii subsp. bulgaricus (TISTR 895) were found to produce BA. The highest levels of histamine (1820.9 ± 3.5 mg L^?1) and tyramine (5486.99 ± 47.6 mg L^?1) formation were observed for the TISTR 389 strain, while TISTR 895 produced only histamine (459.1 ± 0.63 mg L^?1) in the decarboxylase broth. Biogenic amine potential was not observed for the Lactobacillus acidophilus, Lactobacillus lactis subsp. lactis, Lactococcus lactis subsp. lactis, and Lactobacillus plantarum strains studied. This study confirmed that BA formation is strain dependent and not related to the species. Therefore, careful screening for amino acid decarboxylase activity is recommended before selecting LAB as appropriate starter or probiotic strains in food and dairy industry.     

Development of a pentaplex PCR assay for the simultaneous detection of Streptococcus thermophilus, Lactobacillus delbrueckii subsp. bulgaricus, L. delbrueckii subsp. lactis, L. helveticus, L. fermentum in whey starter for Grana Padano cheese

A pentaplex PCR assay for the rapid, selective and simultaneous detection of Lactobacillus helveticus, L. delbrueckii subsp. lactis, L. delbrueckii subsp. bulgaricus, Streptococcus thermophilus, and L. fermentum, was developed. The target sequences were a group of genes coding for beta-galactosidase production (S. thermophilus and L. delbrueckii subsp. bulgaricus), for cell-enveloped associated proteinase synthesis (L. helveticus), for dipeptide transport system production (L. delbrueckii subsp. lactis) and for arginine-ornithine antiporter protein production (L. fermentum). The analytical specificity of the assay was evaluated with 5 reference strains and 140 lactic acid bacterial strains derived from raw milk cheeses and belonging to the Lactobacillus, Streptococcus, Lactococcus and Enterococcus genera. The identification limit for each target strain was 10³ CFU/ml. This new molecular assay was used to investigate the LAB population by direct extraction of DNA from the 12 whey cultures for Grana Padano. The pentaplex PCR assay revealed a good correspondence with microbiological analyses and allowed to identify even minor LAB community members which, can be out-competed in vitro by numerically more abundant microbial species.     

Characterization of volatile compounds in fermented milk using solid-phase microextraction methods coupled with gas chromatography-mass spectrometry

Lactic acid bacteria (LAB) are industrially important bacteria that are widely used in the fermented food industry, especially in the manufacture of yogurt. Characteristic flavors are produced by LAB during fermentation and storage that affect the quality and acceptability of fermented milk products. In this study, the volatile compounds in milk fermented by Streptococcus thermophilus IMAU80842 alone, Lactobacillus delbrueckii ssp. bulgaricus IMAU20401 alone, or both species together were identified using solid-phase microextraction methods coupled with gas chromatography-mass spectrometry. A total of 53, 43, and 32 volatile compounds were identified in milk fermented by S. thermophilus alone, L. delbrueckii ssp. bulgaricus alone, or both species together, respectively. The presence of some important flavor compounds was confirmed: acetic acid, acetaldehyde, acetoin, 2,3-butanedione, ethanol, and 1-heptanol. Our results demonstrate that the composition of the volatile compounds in fermented milk depends on the species of LAB used and whether they are used alone or in combination. This is important for the selection of appropriate starter cultures for the production of different types of fermented milk product with particular flavors.     

Characterization of functional, biochemical and textural properties of synbiotic low-fat yogurts during refrigerated storage

This study examined the influence of exopolysaccharides (EPS) produced in situ on the viability of Streptococcus thermophilus, Lactobacillus delbrueckii ssp. bulgaricus and probiotics, the proteolytic and ACE-inhibitory activities, and textural and rheological properties of inulin-containing probiotic yogurt during refrigerated storage. Two types of yogurt were prepared using strains of S. thermophilus not producing EPS (NEPY) and producing EPS (EPY). The yield of crude EPS increased (by 2.4 times) until day 21 of storage. Presence of EPS showed a protective effect on the survival of L. delbrueckii ssp. bulgaricus and Lactobacillus acidophilus but not on S. thermophilus, Lactobacillus casei and Bifidobacterium longum. No changes in post-acidification, lactic acid content or the ACE-inhibition activity of the two types of yogurt were observed. Overall, EPS containing yogurts exhibited higher proteolysis in the presence of inulin and probiotics (0.698 units) than the corresponding control (0.563 units). The storage and loss moduli (Pa), yield stress (Pa), consistency index (Pa s) and thixotrophic behaviour (Pa s⁻¹) of both samples were similar at day 1 and the influence of EPS was observable only after day 7. Such a variation of the effect of EPS on the textural and rheological properties of low-fat yogurt appears to be partially due to the presence of probiotics.     

The effect of wild lactic acid bacteria on the production of goat’s milk soft cheese

A blend of four wild strains of lactic acid bacteria, Lactobacillus delbrueckii subsp. bulgaricus, Streptococcus thermophilus, Lactobacillus paracasei subsp. paracasei and Enterococcus faecalis, was used as starter to produce a goat’s milk soft cheese. The cheese was analysed microbiologically, physicochemically and organoleptically through a 30‐day period of ripening. Counts of the starter cultures increased in the first 24 h and then remained stable during ripening. Yeasts and coliforms were found at low numbers initially and gradually decreased to undetectable levels. Micrococcal counts were high throughout ripening. The end product was a soft cheese characterised by a mild, aromatic taste as well as a smooth structure.     

Effect of lactose hydrolysis on the milk-fermenting properties of Lactobacillus delbrueckii ssp. bulgaricus 2038 and Streptococcus thermophilus 1131

Yogurt is a well-known nutritious and probiotic food and is traditionally fermented from milk using the symbiotic starter culture of Streptococcus thermophilus and Lactobacillus delbrueckii ssp. bulgaricus. However, yogurt consumption may cause health problems in lactose-intolerant individuals, and the demand for lactose-free yogurt has been increasing. The standard method to prepare lactose-free yogurt is to hydrolyze milk by lactase; however, this process has been reported to influence the fermentation properties of starter strains. This study aimed to investigate the fermentation properties of an industrial starter culture of L. bulgaricus 2038 and S. thermophilus 1131 in lactose-hydrolyzed milk and to examine the metabolic changes induced by glucose utilization. We found that the cell number of L. bulgaricus 2038, exopolysaccharide concentration, and viscosity in the coculture of L. bulgaricus 2038 and S. thermophilus 1131 was significantly increased in lactose-hydrolyzed milk compared with that in unhydrolyzed milk. Although the cell number of S. thermophilus 1131 showed no difference, production of formic acid and reduction of dissolved oxygen were enhanced in lactose-hydrolyzed milk. Further, in lactose-hydrolyzed milk, S. thermophilus 1131 was found to have increased the expression of NADH oxidase, which is responsible for oxygen reduction. These results indicated that glucose utilization promoted S. thermophilus 1131 to rapidly reduce the dissolved oxygen amount and produce a high concentration of formic acid, presumably resulting in the increased cell number of L. bulgaricus 2038 in the coculture. Our study provides basic information on the metabolic changes in starter strains in lactose-hydrolyzed milk, and demonstrates that lactose-free yogurt with increased cell number of L. bulgaricus can be prepared without delay in fermentation and decrease in the cell number of S. thermophilus.     

Microbial diversity and dynamics of Pecorino di Filiano PDO,a traditional cheese of Basilicata region (Southern Italy)

The evolution of microbial populations of ‘Pecorino di Filiano’ (PF) cheese was investigated during ripening in natural cave and storeroom. 62.5% of isolates grow at 45 and 15 °C and 77.7% showed high salt concentrations tolerance. Brevibacterium linens was dominant in surface samples. Lactobacillus delbrueckii subsp. bulgaricus and Lactobacillus paracasei subsp. paracasei were more frequently isolated both surface and core samples, while Leuconostoc lactis and Leuconostoc mesenteroides subsp. mesenteroides prevailed among Leuconostoc isolates. Our results suggest the importance of the ripening environment of cheeses and how a biological ecosystem affects and produces the typical features of artisanal products.     

The effect of using an exopolysaccharide‐producing culture on the physicochemical properties of low‐fat and reduced‐fat Kasar cheeses

A mixed starter culture containing exopolysaccharide (EPS)‐producing strains of Streptococcus thermophilus and Lactobacillus delbrueckii subsp. bulgaricus was combined with Lactobacillus helveticus LH301 and used in the manufacture of low‐fat and reduced‐fat Kasar cheeses. For comparison, low‐fat (C10) and reduced‐fat (C20) cheeses were made using EPS‐producing (EPS⁺) starter strain and EPS‐non‐producing (EPS^?) starter strain. The physicochemical properties of the cheeses were assessed in terms of chemical composition, texture, microstructure and microbial content over 90 days. Cheeses made with EPS‐producing culture (EPS10 and EPS20) had lower protein contents than control cheeses with 10% and 20% fat in dry basis (C10 and C20). Scanning electron microscopy images showed that using EPS‐producing culture resulted in a less compact protein matrix and sponge‐like structure in the cheese samples. In general, cheeses made using EPS‐producing culture had lower total viable counts. This could be related to the reduced survivability of EPS‐producing cells in the cheese matrix during ripening due to autolysis ability.     

Selection of starters for a traditional Turkish yayik butter made from yoghurt

Butter is produced from two different materials in Turkey, cream and yoghurt. The butter produced from fresh yoghurt or ‘tulum yoghurt’ (a strained yoghurt produced from cow, goat or sheep milk) is called ‘yayik butter’ and has been traditionally produced in Turkey for centuries. In this research, we attempted to isolate and identify the natural lactic acid bacteria (LAB) of yayik butter and to select the best LAB combination for butter production. Twenty samples of yayik butter were collected from Afyon, Antalya, Isparta and Konya regions in Turkey and determined to have a mean pH of 4·78±0·33, a mean titratable acidity (lactic acid) of 0·23±0·07% and a mean NaCl of 0·55±1·22%. The mean counts of LAB (log₁₀ cfu g⁻¹) were 2·66±0·84 and 1·72±0·82 on MRS agar at 30 and 42°C, 2·44±0·93 and 1·78±0·24 on M17 agar at 30 and 42°C, and 1·64±1·196 on Sodium Azide Leuconostoc agar at 21°C, respectively. Eighty-five different LAB isolates were obtained from 20 yayik butters and identified asStreptococcus salivarius ssp. thermophilus (21·2%), Streptococcus sp. (4·7%), Lactobacillus delbrueckii ssp. bulgaricus (20%), Lactobacillus casei ssp.casei (15·3%), Lactobacillus paracasei ssp. paracasei (2·3%),Enterococcus faecium (18·8%). Leuconostoc pseudomesenteroides (Leucono-stoc mesenteroides ssp. dextranicum) (7·1%), Leuconostoc gelidum (Leuconostoc mesenteroides ssp.mesenteroides ) (4·7%) and Weissella paramesenteroides (Leuconostoc paramesenteroides) (5·9%). Combinations of S. salivarius ssp. thermophilus S51, Lb. delbrueckii ssp.bulgaricus A42, Lb. casei ssp. casei K64, Lb. paracasei ssp. paracasei A27, andLeu. pseudomesenteroides E83 were used as starter bacteria for experimental butter production from cream. Six different groups of butters were produced using different combinations of these bacteria (B, C, D and E samples), commercial culture (F sample), and without culture (A sample). Sensory evaluations showed that the experimentally produced butter sample of group B was more acceptable than the other butters. In addition, the buttermilk of sample B had lowest fact content. LAB counts of experimental butters produced with combined cultures and commercial culture were similar (6·66±1·87–6·83±0·040 and 6·81±0·13 log₁₀ cfu g⁻¹ on MRS agar, respectively).     

Identification and characterization of lactic acid bacteria isolated from traditional cone yoghurt

Cone yoghurt is a yoghurt variety produced by adding only pine cones to milk without culture in a limited area of Turkey. The present study was conducted to identify and characterize lactic flora in traditional cone yoghurt. Morphological, cultural, physiological, biochemical, and genotypic characteristics were employed to identify lactic acid bacteria isolates from cones and cone yoghurts. Streptococcus salivarius subsp. thermophilus (S. thermophilus) and Lactobacillus delbrueckii subsp. bulgaricus (L. bulgaricus) were obtained from both cones and yoghurts. Among the isolates, L. plantarum was frequently isolated except for these two bacteria (S. thermophilus and L. bulgaricus). The results indicate that the cone yoghurt has a mixed microflora contrary to the yoghurt produced by the addition of a starter culture and S. thermophilus, and L. bulgaricus in cone yoghurt originates from the pine cones.     

Characterization and Classification of Streptococcus Thermophilus and Lactobacillus Delbrueckii Subsp. Bulgaricus Strains Isolated from Traditional Greek Yogurts

ABSTRACT Streptococcus thermophilus (74 strains) and Lactobacillus delbrueckii subsp. bulgaricus (75 strains) isolates from traditional Greek yogurts were tested on the basis of their acidification kinetics, acidity, viscosity, and aroma compounds formation in milk pure cultures. Strains of Lb. bulgaricus presented greater viscosity values, higher acidification activity and produced more acetaldehyde at culture pH 4.6 than S. thermophilus strains. Acetoin was formed only by the latter strains, while diacetyl was not detected in either species. Application of cluster and discriminant analysis permitted definition of distinct groups of strains and of the most significant variables to discriminate among them. Finally, classification and regression tree analysis allowed clusters characterization by critical values of the most discriminant variables. Evaluation of results as above could be a useful tool to select, through a large number of screened strains, the suitable starters for specific purposes.