Traditional national fermented products and cheeses are a source for the search for species and strains of lactic acid bacteria (LAB) which are not within the range of bacterial agents used in the dairy industry. Classical and modern genetic-molecular methods are used to identify LAB isolated from such products. The purpose of our work was isolation and identification of LAB from traditional Carpathian cheeses made from ewe's milk and the study of their technological properties. Three samples of cheese were selected for our research – one sample of brine cheese bryndza and one sample of budz (bryndza before salting), produced in the highlands of the Carpathians and one sample of buts, produced in the foothills zone. 106 cultures were isolated from the samples of cheese. Genus and species identification was completed using classical microbiological and molecular genetics methods. Based on the complex of tinctorial, cultural, physiological and biochemical indices, the LAB isolated were assigned to the following genera and species: Lactococcus spp. (26 cultures), including L. lactis (13 cultures) and L. garvieae (13 cultures); Lactobacillus spp. – L. plantarum (31 cultures); Enterococcus spp. – E. faecium (25 cultures); Leuconostoc spp. – L. mesenteroides (24 cultures). These results were confirmed by molecular genetics methods. The largest range of species was found in a sample of bryndza from the Carpathian highlands. The isolated cultures were studied according to technological properties – milk-coagulation activity, acid-forming ability and resistance to different concentrations of kitchen salt. Most strains of L. lactis ssp. lactis, L. plantarum and L. mesenteroides were active acid-forming agents and coagulated milk in 3–9 hours, while L. garvieae and E. faecium coagulated milk after more than 24 hours. More than 80% of cultures showed resistance to 4% of kitchen salt solution, E. faecium was observed to have the highest salt tolerance. The results of RAPD typing showed significant intra-species heterogeneity, which indicates the need for further research on identification of individual strains. In all samples of cheese, L. lactis, L. garvieae, E. faecium were detected, which shows that they are typical representatives within the traditional Carpathian bryndza. Particular attention was paid to E. faecium, as many researchers have indicated probiotic properties of individual strains, as well as the ability to synthesize volatile substances that enrich the flavor bouquet of cheeses. Today strains of E. faecium are involved in the bacterial composition of starter cultures for cheeses.

No abstract

The creation of new types of functional food products is an actual direction of the development of the food industry at the present time. The purpose of the work was to develop the technology, to investigate the properties and quality indicators of kefir using celery puree. Experimental studies of organoleptic, physicochemical and microbiological indicators of kefir were conducted in the laboratory of the department of technology of milk and dairy products. For the production of kefir we used a fermentation culture directly applied by DVS company “CHR Hansen” (Denmark) Kefir-1, which includes Debaryomyces hansenii yeast and meso- and thermophilic lactococci: Lactococcus lactis subsp. cremoris, Lactococcus lactis subsp. lactis biovar. diacetylactis, Lactococcus lactis subsp. lactis, Leuconostoc, Streptococcus thermophilus. Stalks of celery were used to prepare puree in laboratory conditions. Stalks celery were washed, cut into small cubes. Chopped celery cubes were placed in a vessel, a small amount of water was added, brought to a boil. After that, the water was drained, and the celery cubes were carefully ground with a blender to a smooth puree-like consistency. At the first stage, kefir recipes were developed using different amounts of celery puree. We produced 3 samples of kefir: sample 1 – use of celery puree in the amount of 20 % to the weight of the normalized mixture; sample 2 – 30 %; sample 3 – 35 %. Kefir was made by the tank method. The cooled milk was sent for normalization. After normalization, the milk mixture was sent for pasteurization at a temperature of (95 ± 1) °C with a holding time of 5 minutes. Fermentation culture Kefir-1 was added to the milk mixture cooled to a temperature of (30 ± 1) ºС. To evenly distribute the culture, mixing was used for 10-15 minutes. The mixture was subjected to fermentation at a temperature of (30 ± 1) ºС. After fermentation, celery puree was added to the milk mixture in the amount of 20, 30 and 35 % to the product weight. The mixture was thoroughly mixed and the product was immediately cooled to a temperature of 8 °C and sent to maturation for 12 hours. Organoleptic, physical and chemical and microbiological indicators were studied in the finished product in accordance with DSTU 4417:2005 “Kefir. Specifications”. The titrated acidity was determined according to GOST 3624-92 “Milk and dairy products. Titrometric methods of determining acidity”. Active acidity was measured using an electronic pH meter “Muttler Toledo MP220”. The total number of lactic acid cultures was determined by parallel inoculation of dilutions of yogurt samples in Petri dishes on Lactobacagar medium followed by incubation in a thermostat at a temperature of (37 ± 1) ºС for 3 days under anaerobic conditions. According to organoleptic parameters, in particular, better appearance and consistency, it is recommended to use celery puree in the amount of 35 % of the product weight in kefir technology. The physical and chemical parameters of the milk-vegetable product after maturation are as follows: titrated acidity 75–80 ºT, active acidity 4.8–4.7 units pH, mass fraction of fat 2.5 %. During storage for 14 days, acidity changed the least in milk-vegetable kefir with celery puree in a sample with 35 % puree, which is caused, in our opinion, by the presence of essential oils in celery puree, which suppress the development of lactic acid microflora. The results of determining the number of viable lactic acid bacteria in kefir during storage at a temperature of (4 ± 2) ºС for 14 days indicate the intensification of the growth of lactic acid bacteria. Their development was more active in the sample using the smallest amount of celery puree.

Abstract. In the article a comparative analysis of the use of the bacterial preparation Herobacterin and the starter RSF-742 (Chr. Hansen, Denmark) in the technology of brine cheese was conducted. Herobacterin is a bacterial preparation created using bacteria Lactococcus lactis, Lactobacillus plantarum, Enterococcus faecium, Leuconostoc mesenteroides and Lactococcus garvieae, isolated from traditional Carpathian brine cheese brynza and identified using classical microbiological and modern molecular genetic methods (RAPD-PCR, RFLP-PCR, sequencing of the 16S rRNA gene). Анотація. Показано результати порівняльного аналізу використання створеного бакконцентрату Геробактерин із бактеріальним препаратом RSF-742 (Chr. Hansen, Данія) у технології розсільного сиру бринза. Геробактерин − бак-теріальний препарат, створений на міжвидовій, багатоштамовій основі з використанням бактерій Lactococcus lactis, Lactobacillus plantarum, Enterococcus faecium, Leuconostoc mesenteroides і Lactococcus garvieae, виділених із традицій-ної карпатської бринзи та ідентифікованих із використанням класичних мікробіологічних і сучасних молекулярно-генетичних методів (RAPD-PCR, RFLP-PCR, секвенування гену 16S рРНК). Наведено результати досліджень органо-лептичних, фізико-хімічних, синеретичних та мікробіологічних показників сиру бринза із використанням бакконцент-рату Геробактерин у порівнянні із препаратом прямого внесення RSF-742, до складу якого входять такі культури: Lactococcus lactis subsp. сremoris, Lactococcus lactis subsp. lactis, Streptococcus thermophilus, Lactobacillus helveticus. Використання препарату Геробактерин здійснює позитивний вплив на органолептичні, фізико-хімічні та мікробіоло-гічні показники бринзи, усі параметри відповідали вимогам ДСТУ 7065:2009. Рівень виживання молочнокислих бак-терій у бринзі протягом визрівання і зберігання є високим, що підтверджує правильність відбору штамів до бакконце-нтрату Геробактерин, які продемонстрували добру пристосованість до складу і властивостей овечого молока.Ключові слова: бринза, технологія, бакконцентрат Геробактерин, препарат мікробіальний RSF-742, органоле-птичні показники, фізико-хімічні показники, кількість життєздатних клітин.

A promising area for improving probiotics is the search for new sources of strains and the development of complex preparations which would include different types of bacterial cultures that complement each other. Sources of selection may be traditional dairy products, in particular, cheeses made from raw milk. Wild strains can be endowed with antibacterial properties. The antagonistic action of lactic acid bacteria (LAB) has long attracted the attention of researchers and scientists. The aim of the study was to investigate the antagonistic activity against pathogenic and opportunistic microorganisms of LAB strains isolated from traditional Carpathian cheese. Three samples of cheese were selected for the research – one sample of brynza and budz (brynza before salting), made in the highlands of the Carpathians, and one sample of budz, made in the foothills. LAB were identified using classic microbiological and modern molecular genetic methods (RAPD-PCR, RFLP-PCR, 16S rRNA gene sequencing). The objects of our studies were five strains of LAB: Lactococcus lactis IMAU32258, L. garvieae JB2826472, Enterococcus durans FMA8, E. faecium L3-23, E. faecium IMAU9421. Technological parameters such as acid-forming activity of milk fermentation, resistance to high concentrations of NaCl and temperature optimums of cultivation were taken as the main criteria for assessing the suitability of LAB for inclusion in fermentation preparations. Antagonistic activity was determined by agar diffusion (agar well method) and optic density of test cultures using a Multiscan FC microplate reader (Thermo scientifiс, USA) at the wave of 620 nm. There were four reference strains of pathogenic and opportunistic microorganisms were test cultures: Listeria monocytogenes PCM 2191, Staphylococcus aureus PCM 458, Escherichia coli PCM 2208, Salmonella typhimurium PCM 2182. Strains of the test cultures were received from the collection of microorganisms of the Institute of Biology and Biotechnology the (University of Rzeszów, Poland). According to the ability of LAB strains to form lactic acid, L. lactis IMAU32258 was the best acid-forming agent with an acid-forming energy of 94 °T. E. faecium was characterized by moderate levels of active and titratable acidity. Less pronounced acid-forming ability was determined for the species E. durans and L. garvieae. Cultures of the genus E. faecium, L. garvieae and E. durans were the most resistant to high concentrations of NaCl (6.5%). Regarding temperature optimums, we found that strains of E. faecium and E. durans species grew both at temperatures of 10, 15 and 45 °C, whereas no growth of L. lactis IMAU32258 and L. garvieae JB282647 2 was observed at 45 °C. Among the studied bacteria, the strains of E. durans FMA8 and E. faecium L3-23 were characterized by the highest antagonistic activity in producing the largest zones of growth inhibition and optic density of pathogenic and opportunistic microorganisms. The strain L. garvieae JB282647 2 exhibited the lowest level of antagonistic activity against pathogenic and opportunistic microorganisms.

The aim of the work was the study of keeping probiotic properties of sour-milk butter with inclusion of Lactobacillus acidophilus La-5 (La-5) monoculture. Flora Danica mesophile culture independently (FD); in combination with La-5 and La-5 independently were used for fermenting cream. The output consistence of culture in cream was 1×106 CFU/cm3. In autumn-winter and spring-summer period of the year four butter groups were prepared, they differed by temperature of cream fermentation: I group – (30±1) ºС; II – (37±1) ºС; III – stage regimes of combination of fermentation and physical maturing; IV group – introduction of cultures into oil kernel; the output concentration – 1·108 CFU/cm3. As to the features of summer and winter periods, in summer one cream fermentation is more active that is indicated by more number of cells of both microbial cultures. The best parameters of viable cells keeping were typical to the samples at FD+La-5 use and temperature of cream fermentation (30±1) ºС. Storage life of sour-cream butter with probiotic properties is 35 days at temperature 0…-5 ºС.

No abstract

The article presents the results of determining the ability of enterococci extracted from traditional Carpathian cheese bryndza to produce biologically active substances, in particular, amino acids, B vitamins and cations (ammonium, potassium, sodium, magnesium, calcium). It was found that the studied strains of enterococci in different quantities synthesized both essential and essential amino acids. Thus, the essential amino acid lysine was found in the cultivation of strains of E. durans SB18, E. durans SB20, in particular, its concentration was significantly increased by 15.6 and 10.4 %, respectively (P < 0.05) compared to the control. A probable increase in the essential amino acid histidine by 20 and 53.3 % (P < 0.05) was detected in the cultivation of only E. faecium SB12 and E. durans SB18. In addition, it was found a probable increase in threonine synthesis by enterococci E. durans SB6 and E. durans SB18, respectively – 33.3 and 39.6 % (P < 0.05). The replacement amino acid serine was able to synthesize strains of E. faecium SB12, E. durans SB18 and E. durans SB20, while its concentration increased by 40.0 (P < 0.001), 30.0 and 35.0 %, respectively < 0.01), and strains of E. durans, SB6, and E. durans SB18 synthesized glycine, the concentration of which increased by – 10.2 and 16.2 %, respectively (P < 0.01). In addition, it was found that the studied strains in small quantities synthesized B vitamins, or not synthesized at all. In all experimental samples the most vitamin B1 was detected, its concentration increased from 8.5 to 10.0 times (P < 0.001). Riboflavin was synthesized by three strains of enterococci – E. durans SB6, E. durans SB18, E. durans SB20, so the concentration of vitamin B2 probably increased, respectively, 4.1, 2.0 and 2.0 times (P < 0.05). Enterococci E. durans SB6, E. faecium SB12, E. durans SB18 and E. durans SB20 synthesized in significant quantities only vitamin B3, in particular, its concentration probably increased by 1.5, 1.5 (P < 0.05), respectively, 1.5 (P < 0.01) and 1.6 (P < 0.001) times, and vitamin B5 was produced by E. faecium SB12, E. durans SB18 and E. durans SB20, the concentration of nicotinic acid increased, respectively, 2.9 (P < 0.05), 8.4 and 9.5 (P < 0.001) times. Analysis of the macroelement composition of the supernatant of enterococci showed that strains of E. durans, SB6, E. faecium SB12, E. durans SB18 and E. durans SB20 are able to produce only Calcium, in particular, found a probable increase, respectively, in 1.8, 2.4, 1.6 and 1.4 times (P < 0.05).