Combined Starter Culture Research Articles

Temperature-Dependent Metabolic Interactions Between Streptococcus thermophilus and Lactobacillus delbrueckii ssp. bulgaricus in Milk Fermentation: Insights from GC-IMS Metabolomics

Streptococcus (S.) thermophilus and Lactobacillus (L.) delbrueckii ssp. bulgaricus are widely used as a combined starter culture for milk fermentation, often at temperatures of 37°C and 42°C. To investigate the metabolic interplay between these 2 species during the fermentation process, this study examined the growth and fermentation characteristics of different S. thermophilus strains cocultured with L. delbrueckii ssp. bulgaricus ND02 at these 2 temperature conditions. Gas chromatography-ion mobility spectrometry (GC-IMS) metabolomics was employed to analyze changes in the milk metabolome during 3 key fermentation stages: initiation (F0, pH 6.50 ± 0.02), curdling (F1, pH 5.20 ± 0.02), and endpoint (F2, pH 4.50 ± 0.02). The results showed that 42°C fermentation promoted rapid bacterial growth, with significantly reduced fermentation time compared with 37°C. Interestingly, 37°C fermentation favored the enrichment of volatile fatty acids like 2-methylpropanoic acid, 3-methylbutanoic acid, and ethyl acetate. In contrast, 42°C fermentation led to increased levels of ketones such as acetone, 2-hexanone, 2-pentanone, and 2-heptanone. Sensory evaluation indicated that the 42°C fermented milk had higher overall scores. Discriminatory flavor metabolites were more abundant during the later fermentation stage (F1 to F2), while the underlying metabolic pathways became increasingly active. These findings provide insights into the dynamic changes in volatile metabolite profiles of fermented milk produced under different temperature and time conditions using varied starter culture combinations. The results are valuable for optimizing dairy fermentation processes and product quality.

Oral Administration of Fermented Milk from Co-Starter Containing Lactobacillus plantarum Y44 Shows an Ameliorating Effect on Hypertension in Spontaneously Hypertensive Rats.

Fermented dairy foods such as yogurt exhibit some beneficial effects on consumers, including relieving the symptoms of hypertension. This study aims to obtain fermented dairy products from a co-starter that have a great flavor and the auxiliary function of reducing blood pressure after longtime consumption. Commercial starter cultures composed of Lactobacillus delbrueckii subsp. bulgaricus CICC 6047 and Streptococcus thermophilus CICC 6038 were combined with Lactobacillus plantarum strains Y44, Y12, and Y16, respectively, as a combined starter culture to ferment the mixed milk of skim milk and soybean milk. The fermented milk produced using the combined starter culture mixed with L. plantarum Y44 showed an angiotensin-converting-enzyme (ACE) inhibitory activity (53.56 ± 0.69%). Some peptides that regulate blood pressure were released in the fermented milk, such as AMKPWIQPK, GPVRGPFPII, LNVPGEIVE, NIPPLTQTPV, and YQEPVL. In spontaneously hypertensive rat (SHR) oral-administration experiments compared with the gavage unfermented milk group, the gavage feeding of SHRs with the fermented milk produced using the combined starter culture mixed with L. plantarum Y44 significantly reduced the blood pressure of the SHRs after long-term intragastric administration, shown with the systolic blood pressure (SBP) and diastolic blood pressure (DBP) decreasing by 23.67 ± 2.49 mmHg and 15.22 ± 2.62 mmHg, respectively. Moreover, the abundance of short-chain fatty acids (SCFA), bacterial diversity in the gut microbiota, and SCFA levels including acetic acid, propionic acid, and butyric acid in the feces of the SHRs were increased via oral administration of the fermented milk produced using the combined starter culture containing L. plantarum Y44. Furthermore, the ACE-angiotensin II (Ang II)-angiotensin type 1 (AT 1) axis was downregulated, the angiotensin-converting-enzyme 2 (ACE 2)-angiotensin(1-7) (Ang1-7)-Mas receptor axis of the SHRs was upregulated, and then the RAS signal was rebalanced. The fermented milk obtained from the combined starter culture shows the potential to be a functional food with antihypertension properties.

Nutritional and sensory quality of Ofada rice sourdough bread made with selected lactic acid bacteria strains

Celiac disease ranks highest among immunological disorders attributed to gluten consumption thus, posing great demands on gluten-free products. Rice is a gluten-free cereal with promising dietary applications. Ofada rice, a native southwestern Nigerian variety, can be improved with sourdough technology to develop functional aglutenic bread. Ofada rice sourdough bread (ORSB) was made with Fructilactobacillus fructivorans RY1, Weissella viridescens RY9 and Lactobacillus acidophilus RY10 as individual and combined starter cultures. Physical qualities, proximate contents, sensorial attributes and shelf-life of the ORSBs were evaluated. Sourdough bread with the highest (3.10 cm3/g) and lowest specific volumes (2.02 cm3/g) were the sample made single culture of W. viridescens and F. fructivorans, respectively. The highest crude protein (8.48 %) was found in ORSB with only F. fructivorans and ORSB with L. acidophilus singly had the highest content (0.44 %) of crude fibre. Nevertheless, the least crude protein (5.25 %) and crude fibre (0.28 %) were observed in samples containing F. fructivorans, W. viridescens, and L. acidophilus as combined starters. The sourdough bread with F. fructivorans as a starter scored best for texture and overall acceptability. The ORSBs had shelf-life ranging from three to four days. It was revealed in the study that using the specific LAB starters could improve the physicochemical attributes and acceptability of bread from Ofada rice flour.

Investigation of the biochemical activity of starter compositions enriched with propionic acid bacteria on various raw materials

Currently, there is a need to develop an assortment of low-lactose or lactose-free dairy products and plant-based products due to an increase in the number of consumers with lactase deficiency, allergic to cow's milk components, cholesterol problems, etc. For the development of plant and lactose-free bioproducts, it is possible to use various starter cultures enriched with probiotics. Various starter cultures were used in the work, propionic acid bacteria were used as a probiotic. The objects were lactose-free milk and vegetable bases: rice milk, oat milk and coconut milk; the control was selected milk. A comparative analysis of the fermentation process of various compositions of starter cultures enriched with propionic acid bacteria on various raw materials was carried out. The duration of fermentation of vegetable raw materials is 8-10 hours, the acid-forming activity of combined starter cultures is quite high. The number of viable cells of lactic acid and propionic acid bacteria in the obtained bioproducts has a high value and amounts to 107-108 K.O.E. /cm3. This confirms that the nutrient medium for the cultivation of selected starter compositions is favorable, the microflora of the starter composition develops harmoniously. Lactic acid microorganisms and propionic acid bacteria make it possible to obtain fermented biological products with a variety of taste characteristics inherent in a certain type of raw material. According to the results of the research, the developed starter compositions meet the requirements for starter cultures, have valuable technological properties that allow expanding the range of plant and lactose-free biological products.

A Comparative Study of Skimmed Milk and Cassava Flour on the Viability of Freeze-Dried Lactic Acid Bacteria as Starter Cultures for Yogurt Fermentation.

The purpose of this study was to evaluate the survival rates and fermentation performance of three freeze-dried lactic acid bacterial cultures previously isolated from Ghanaian traditional fermented milk. LAB cultures, i.e., Lactobacillus delbrueckii, Lactococcus lactis and Leuconostoc mesenteroides, were frozen in the chamber of a Telstar (Lyoquest) laboratory freeze dryer for 10 h at -55 °C (as single and combined cultures) using skimmed milk and cassava flour as cryoprotectants held in plastic or glass cryovials. For viability during storage, freeze-dried LAB cultures were stored in a refrigerator (4 °C) and at room temperature (25 °C) for 4 weeks. The survival of freeze-dried cultures was determined by growth kinetics at 600 nm (OD600). The performance of freeze-dried LAB cultures after 4 weeks of storage was determined by their growth, acidification of milk during yogurt fermentation and consumer sensory evaluation of fermented milk using a nine-point hedonic scale. The survival rates for LAB ranged between 60.11% and 95.4% following freeze-drying. For single cultures, the highest survival was recorded for Lactobacillus delbrueckii (L12), whereas for combined cultures, the highest survival was observed for Lactococcus lactis (L3) combined with Lactobacillus delbrueckii (L12). The consumer acceptability results showed that yogurts produced from a combined starter culture of Lactococcus lactis and Lactobacillus delbrueckii or from a single culture of Lactococcus lactis were the most preferred products with Lactococcus lactis and Lactobacillus delbrueckii possessing high survival rates and high consumer acceptability in yogurt production. These findings are crucial and can be adopted for large-scale production and commercialization of yogurt.

Effect of Lactic Acid Fermentation on Phytochemical Content, Antioxidant Capacity, Sensory Acceptability and Microbial Safety of African Black Nightshade and African Spider Plant Vegetables

Traditional preparation of African indigenous vegetables (AIVs) such as African black nightshade (Solanum nigrum) and African spiderplant (Cleome gynandra) involves either boiling and discarding the first water or lengthy boiling. Fermentation is considered a better alternative processing technique due to the enhanced retention of phytochemical contents and sensory properties. However, little is known about the impact of lactic acid fermentation on the phytochemical content, antioxidant capacity, sensory acceptability and microbial safety of the African black nightshade and African spiderplant. This study aimed to ferment AIVs using combined starter cultures (Lactobacillus fermentum and Lactococcus lactis) and further determine their effect on the phytochemical content (phenolic compounds and flavonoids), antioxidant capacity, sensory acceptability and microbial safety of the vegetables. There was a marked increase in phenol and flavonoid contents in all fermented vegetables (p < 0.05). The highest phenol content was 228.8 mg/g GAE (gallic acid equivalent) in the starter-culture-inoculated African black nightshade, while flavonoid content was 10.6 mg/g QE (quercetin equivalent) in the same. Starter-culture-inoculated AIVs presented significantly higher antioxidant capacity with a 60–80% radical scavenging activity compared to levels in uninoculated batches (p < 0.05). Fermented vegetables were more liked than the boiled vegetables and were microbiologically safe. In conclusion, lactic fermentation of AIVs increased phytochemical contents (phenolic compounds and flavonoids), maintained antioxidant capacity and improved product safety and sensory acceptability. Therefore, fermentation and consumption of the African indigenous vegetables are to be encouraged.

Milk fermented by combined starter cultures comprising three Lactobacillus strains exerts an alleviating effect on loperamide-induced constipation in BALB/c mice.

Fermented dairy food, such as yogurt, exhibits some beneficial effects on consumers, including alleviating constipation. In this study, Lactobacillus delbrueckii subsp. bulgaricus DPUL-36, Lactobacillus paracasei DPUL-40 and Lactobacillus paracasei DPUL-44 were used as combined starter cultures at a bacterial cell ratio of 1 : 1 : 1 for reconstituted skim milk fermentation. The milk fermented with the combined starter culture showed good sensory properties. During the storage period, the yogurt showed high lactic acid bacteria vitality and quality stability. Constipated BALB/c mice induced by loperamide (Lop) were orally administered with the combined starter culture fermented milk for 14 days. The results clearly showed that oral administration of the fermented milk relieved Lop-induced constipation in the mice, as evidenced by the significantly increased fecal water content, reduced first black stool time, improved gastrointestinal transmission rate, recovered colon tissue damage, increased level of excitatory neurotransmitters (motilin, gastrin, and substance P) and reduced level of inhibitory neurotransmitters (vasoactive intestinal peptide, somatostatin and endothelin-1) of the mice. Compared with the mice in the Lop group, oral administration of the fermented milk significantly increased the concentrations of acetic acid, propionic acid, butyric acid, isovaleric acid and valeric acid in the feces of the mice, and furthermore, exerted a regulatory effect on the gut microbiota of the mice by up-regulating the abundance of Lactobacillus and Bacteroides, and decreasing the abundance of Helicobacter, Pseudomonas and Porphyromonas. Our results indicated that the combined starter culture fermented milk can effectively alleviate Lop-induced-constipation in BALB/c mice. The relationship between the nutrient profiles and the health promoting function of the yogurt should be further illustrated.

Application of ginseng powder and combined starter culture for improving the oxidative stability, microbial safety and quality characteristics of sausages

This work aimed to investigate the effects of ginseng powder and combined starter culture (the mass ratio of Lactobacillus plantarum to Lactobacillus rhamnosus was 1:1, Lp-Lr) on oxidative stability, microbial growth and quality characteristics of sausages. Compared with natural fermentation, the combination of ginseng powder and Lp-Lr in sausages reduced the oxidation degree of lipid and protein, improved the DPPH and ABTS radical scavenging capacities, and increased the variety and content of identified volatile flavor compounds. High-throughput sequencing analysis revealed the combination of ginseng powder and Lp-Lr improved the abundance of lactic acid bacteria, and inhibited the growth of harmful bacteria (i.e., Acinetobacter, Halovibrio and Alkalibacillus) in sausages after 30 days. During storage, the pH values and nitrite contents of all sausages were within the appropriate ranges, and the addition of ginseng powder and Lp-Lr in sausages displayed a smaller variable range of color and texture than natural fermented sausages. These findings suggest that the ginseng powder and Lp-Lr had positive effects on quality, oxidative stability, flavor profiles and microbial safety in sausages.

The fermented soy whey produced by a combined lactic acid bacteria starter shows improved flavor and the function in alleviating dextran sulphate sodium induced colitis in mice

ABSTRACT In this study, Lactiplantibacillus plantarum BC299, Lactobacillus delbrueckii subsp. bulgaricus 717 and Streptococcus thermophilus 176 were used as combined starter cultures to produce a fermented soy whey beverage. The results showed that the fermentation of the combined starter cultures increased the content of lactic acid, acetic acid, succinic acid, and malic acid, while decreased the content of citric acid, fumaric acid and propionic acid. Meanwhile, the fermentation also effectively changed the proportion of volatile flavor substances. The higher sensory score of the fermented soy whey also confirmed the flavor improvement effect of the combined starter cultures. In addition, the in vivo results of animal experiments showed that the fermented soy whey could significantly improve the disease activity index (DAI) and colon morphology in dextran sulfate sodium (DSS) induced colitis mice. This study showed that the combined lactic acid bacteria starter cultures had important value-added relevance on the effective reuse of the soy whey by-product.

Application of starter cultures on milk and milk-containing mixtures

Milk-containing products occupy a large place in the range of popular products. Replacing milk fat or protein with vegetable fat can reduce the biological value of the product and its organoleptic properties. the effect of direct application starter cultures on raw materials that use functionally necessary ingredients (structure-forming agents, salt, sugar, etc.), food additives (thickeners, stabilizers, preservatives, etc.) and a variety of non-dairy components (non-dairy fats, non-dairy proteins, meat products, fruits, vegetables, spices, herbs, nuts, etc.) is ambiguous. The paper deals with the use of homogeneous and combined starter cultures in the production of fermented milk products from natural and normalized milk and milk-containing mixtures with a substitute for milk fat. The introduction of components of plant origin into the product leads to a decrease in the content of milk protein in the mixtures, which in turn leads to the formation of clots with a non-uniform consistency. Slow acid formation in milk-containing mixtures indicates a decrease in milk sugar in them with this type of normalization in fat, relative to the normalized milk according to the classical technology. This pattern was revealed both when using starter cultures on lactic acid thermophilic streptococci, and on combined starter cultures. In this paper, it is proposed to use Streptococcus salivarius subsp in ferments for milk-containing mixtures. thermophilus strain ST-95. The introduction of the ST-95 strain into the starter culture made it possible to obtain from the milk-containing mixture a fermented milk product with well-pronounced organoleptic properties, a stable clot structure and sufficiently high taste properties. The greatest positive effect was obtained on combined ferments.

The influence of starter cultures on the lactic acid bacteria microbiota of Petrovac sausage

Petrovac sausage (Petrovská klobása) is a high-quality fermented dry sausage produced traditionally in the municipality of Ba?ki Petrovac (Vojvodina, Serbia). The product is characterised by specific and recognised texture, aroma and colour, produced without additives or preservatives. Lactic acid bacteria (LAB) microbiota plays an important role in production of the sausage. The aim of the paper is to monitor the changes in LAB during the production of Petrovac sausage. Samples of sausages were prepared without and with the addition of starter culture Staphylococcus xylosus as well as combined starter culture Lactiplantibacillus plantarum and S. xylosus, and produced at two different temperature ranges. A total number of 495 strains were isolated from 33 samples of Petrovac sausage during 120 days of production process. Characterisation of the isolates was performed by phenotypic tests, while molecular identification of the representative strains was done by 16S ribosomal DNA sequencing. The total number of LAB was about 8 log (Colony Forming Unit (CFU))/g in all samples, while the number of staphylococci was about 4 log CFU/g. Molecular identification confirmed that all isolates belonged to the following species: Levilactobacillus brevis, Leuconostoc mesenteroides, Lactiplantibacillus plantarum and Pediococcus pentosaceus. Lactobacilli and Leuconostoc spp. dominate the total LAB strains, while P. pentosaceus was isolated at the lowest frequency.

Sour milk beverage preparation technology

In order to expand the range of dairy products for the elderly, a technology has been developed for preparing a fermented milk drink with stevia, using a combined starter culture from kefir fungi, bulgarian bacillus, thermophilic streptococcus and bifidobacteria. During the research, modern standard methods were used. The research was carried out in the laboratory of the Research Institute of Agroecology of the Gorsk State Agrarian University, in the laboratory of food products of the commodity-technological faculty of the Gorsky State Agrarian University, in the Scientific Laboratory of Feed and Metabolism of the Stavropol State Agrarian University. In laboratory conditions, the quality of milk and fermented milk drink was determined. The organoleptic, physicochemical, microbiological indicators of the quality of milk and milk drink, the amino acid composition of proteins, the content of toxic elements, and nutritional value have been investigated. During our research, we found that the quality of the used pasteurized cow's milk meets the requirements of the standard and TR CU. During laboratory studies, it was found that the quality indicators of the fermented milk drink are within the requirements of the current NTD. According to economic calculations, the production of this product is profitable.

Development of fermented milk product based on mare milk and lactic microorganisms association

Since ancient times, mare's milk has been widely used by many peoples. Thanks to its unique composition, it is now used in the nutrition of people with allergies to cow's milk, in the technologies of foods for special dietary uses, etc. To expand the range of products with useful properties, it is advisable to develop fermented milk products based on mare's milk. Numerous studies indicate that the use of fermented milk products provides various health benefits. And the use of probiotic cultures in the composition of starter cultures for fermented milk products allows you to add a number of functional properties to the product. However, the only fermented milk product available on the market from mare's milk is koumiss. Therefore, the development of new fermented milk products based on mare's milk is relevant and in demand. The aim of the study was to develop a fermented milk product based on mare's milk using an association consisting of yogurt starter culture and a probiotic strain of L. rhamnosus F. Material and methods. For the development of a fermented milk product based on mare's milk, the yogurt starter STBp (S. thermophilus and L. delbrueckii subsp bulgaricus) and the probiotic culture L. rhamnosus F (GenBank MN994629) from the collection of lactic acid and probiotic microorganisms of VNIMI were chosen. As a basis for the fermented milk product, mare's milk was used with the addition of dry mare's or dry cow's milk, pre-pasteurized at a temperature of 65±1 °C with a holding time of 30 min and cooled to a fermentation temperature of 37±1 °C. In the course of the work, the activity of acid formation during product fermentation, the duration of fermentation and the dynamics of the number of lactic acid bacteria and L. rhamnosus F probiotic strain during fermentation were studied, depending on the ratio of cultures in the ferment, the dose of the ferment introduced and the base for fermentation. The antimicrobial activity of the product was determined by agar well diffusion assay. Results. It was found that L. rhamnosus F has a low acid-forming activity, so the use of a combined starter culture (association) was proposed. It was determined that the addition of cow's milk powder to mare's milk positively affects the activity of acid formation; after 6 hours of fermentation, the pH value varied in the range of 4.6-4.83, and after 8 hours pH was 4.44-4.65. When dry mare's milk was added, the pH value after 8 hours of fermentation was in the range of 4.71-4.98 pH units. The influence of the amount of combined starter culture (association) and its cultures ratio, the duration of fermentation on the content of lactic acid bacteria, including the probiotic strain L. rhamnosus F in the fermented milk product based on mare's milk, was established. The largest amount of L. rhamnosus F was contained in the product fermented by 7% association with the ratio of cultures 1/4 and 1/6 and after 8 h it amounted to 7-9.5×108 CFU/cm3. Conclusion. Summarizing the results of the research, the technology of a fermented milk product based on mare's milk was developed: the amount of starter culture (associations) - 7%, the ratio of cultures - 1 part of the starter culture for yogurt and 6 parts of L. rhamnosus F (1/6), the fermentation temperature - 37±1 °C, the duration of fermentation - 6 hours with the addition of cow's milk powder and 8 hours with the addition of mare's milk powder. The developed fermented milk product has antimicrobial activity against opportunistic and pathogenic microorganisms E. coli ATCC 25922, S. aureus ATCC 6538, S. typhimurium ATCC 14028.

Intensification of kvas biotechnology using non-traditional raw materials

In the production of beverages of incomplete alcoholic and lactic acid fermentation, it is important to use non-traditional raw materials that have advantages over well-known analogues. Of particular interest in the production of fermented beverages is the use of malt extract prepared from triticalic malt. Serum and products derived from it are biologically complete, have dietary and medicinal properties. To achieve this goal accomplished objectives: to assess the quality and applicability of triticale malt extract for kvass production; explore the possibility of using whey as a processing aid in the production of kvass on the basis of triticale malt extract; to choose a combination of microorganisms to obtain a mixed leaven with the best biotechnological properties; conduct a comparative analysis of the duration of fermentation samples of kvas wort. The most intensive decrease in the dry matter content was observed in the experimental sample of leavened wort prepared on the basis of malt triticalic extract with the use of whey and combined sourdough from dry baking yeast of the brand "SAF-Moment" of the species Saccharomyces cerevisiae and lactic acid bacteria L. lactis, L. cremoris, L. diacetylactis, Str. Thermophilus. The duration of fermentation of leavened wort in the test sample was 10 hours, which is 6–8 hours less than the traditional technology. As a result of this work, the possibility of using triticalic malt extract for the production of kvass was proved. According to the main physical and chemical parameters, triticale malt extract is not inferior to traditional raw materials. The possibility of using whey as a technological additive for fermentation of leavened wort based on triticalic malt extract is investigated. The composition of leavened wort and a combination of microorganisms were selected to produce a combined starter culture with the best biotechnological properties. A comparative analysis of the duration of fermentation of samples of leavened wort is carried out.

Cовершенствование технологии кисломолочного напитка «Быырпах»

В настоящее время на отечественном рынке стремительно развивается ассортимент молочных продуктов с пробиотическими свойствами. В сложившейся конкурентной среде при разработке новых технологий кисломолочных продуктов необходимо помимо благоприятного воздействия на микрофлору человека учитывать потребительскую привлекательность органолептических показателей. В работе представлены исследования по совершенствованию технологии популярного в Якутии напитка Быырпах. Это кисломолочный напиток, производимый из нормализованного до массовой доли жира 1% пастеризованного охлажденного молока, заквашенного лактобактериями и дрожжами до кислотности 70-90 oТ. Напиток серийно производится молокоперерабатывающими предприятиями в Якутии по ТУ9220-001-067027-02. Молоко заквашивают пересадочным способом комбинированной закваской на чистых культурах термофильного стрептококка, болгарской палочки и дрожжей, сбраживающих лактозу. Для активизации спиртового брожения добавляют сахар. В связи с этим для повышения биологической ценности и снижения санитарно-гигиенических рисков производства напитка в работе предлагается использование бактериального концентрата микробного консорциума. Бактериальный концентрат содержит микробный консорциум, сформированный длительным совместным культивированием микрофлоры кефирной грибковой закваски и термофильных лактобактерий Lactobacillus acidophilus, Lactobacillus bulgaricus. Полученный микробный консорциум по своему составу близок естественной популяции микроорганизмов курунги и кумыса. Доказана ее высокая антибиотическая и адгезивная активность. В результате исследований по выбору оптимальной дозы вносимого бактериального концентрата установлено, что 0,01% позволяет в течение 10 часов культивирования получить продукт с титруемой кислотностью 90 oТ. Так же установлено, что для активизации дрожжевой микрофлоры в технологии нужно предусмотреть созревание кисломолочного продукта при 16 oС в течение 8 часов. Готовый продукт обладает антибиотической активностью по отношению к патогенной и гнилостной микрофлоре. Изучение органолептических показателей свидетельствует, что применения бактериального концентрата микробного консорциума позволяет получить продукт, соответствующий по своим показателям прохладительному напитку Быырпах.

Effect of Co-Inoculation of Candida zemplinina, Saccharomyces cerevisiae and Lactobacillus plantarum for the Industrial Production of Negroamaro Wine in Apulia (Southern Italy).

The employment of multi-species starter cultures has growing importance in modern winemaking for improving the complexity and wine attributes. The assessment of compatibility for selected species/strains at the industrial-scale is crucial to assure the quality and the safety associated with fermentations. An aspect particularly relevant when the species belong to non-Saccharomyces, Saccharomyces spp. and malolactic bacteria, three categories with different biological characteristics and oenological significance. To the best of our knowledge, the present report is the first study regarding the utilization of a combined starter culture composed of three strains of non-Saccharomyces, Saccharomyces cerevisiae and Lactobacillus plantarum for production of wine at the industrial scale. More in-depth, this work investigated the oenological potential of the autochthonous characterized strains from the Apulian region (Southern Italy), Candida zemplinina (syn. Starmerella bacillaris) 35NC1, S. cerevisiae (NP103), and L. plantarum (LP44), in co-inoculation following a complete scale-up scheme. Microbial dynamics, fermentative profiles and production of volatile secondary compounds were assessed in lab-scale micro-vinification tests and then the performances of the mixed starter formulation were further evaluated by pilot-scale wine production. The above results were finally validated by performing an industrial-scale vinification on 100HL of Negroamaro cultivar grape must. The multi-starter formulation was able to rule the different stages of the fermentation processes effectively, and the different microbial combinations enhanced the organoleptic wine features to different extents. The findings indicated that the simultaneous inoculation of the three species affect the quality and quantity of several volatile compounds, confirming that the complexity of the wine can reflect the complexity of the starter cultures. Moreover, the results underlined that the same mixed culture could differently influence wine quality when tested at the lab-, pilot- and industrial-scale. Finally, we highlighted the significance of employment non-Saccharomyces and L. plantarum, together with S. cerevisiae, autochthonous strains in the design of custom-made starter culture formulation for typical regional wine production with pronounced unique quality.

Lactic and Propionic Acid Bacteria: the Formation of a Community for the Production of Functional Products with Bifidogenic and Hypotensitive Properties

The work presents the results of studies on the creation of a combined starter culture based on lactic and propionic acid bacteria and the functional properties of products based on it. It was shown that the fermented product obtained from a combined starter based on Lactococcus lactis ssp. cremoris CR201, Lactobacillus rhamnosus F and Propionibacterium shermanii E2 strains in a ratio of 2 : 1 : 6 (vol/vol), respectively, had a pronounced antagonistic effect on Escherichia coli and Staphylococcus aureus and causative agents of food spoilage Penicillium brevicompactum and Torulaspora delbrueckii, as well as ACE inhibitory activity in vitro. Bifidogenic and moderate hypotensive effects were demonstrated with in-vivo models. The addition of whey-protein hydrolysates to skimmed milk also leads to an increase in the growth of propionic acid bacteria and an increase in the number of viable cells in the finished fermented beverage, which increased its antimicrobial activity in vitro and bifidogenic effect in vivo.

The Effect of Different Starter Cultures of Lactobacillus paracasei M104 and Pediococcus pentosaceus M103 on the Physicochemical and Microbial Qualities of Fermented Goat Milk

The utilization of starter cultures is vital for dairy fermentation industries. Lactobacillus paracasei M104 and Pediococcus pentosaceus M103 have been isolated from goat milk and were demonstrated to acidify goat milk during fermentation. This study was conducted to compare the physicochemical and microbial qualities of goat milk fermented using these starter cultures either as a single starter or combined in goat milk. The treatments were L. paracasei M104 (Lp) and P. pentosaceus M103 (Pp) as single starter culture fermentation and combined (Lp/Pp) fermentation at a ratio of 1:1 at 6% inoculation (v/v). For control treatment, goat milk was fermented using yogurt starter, i.e., a combination culture of Streptococcus thermophilus and L. bulgaricus (St/Lb) at a ratio of 1:1 at 6% (v/v). The fermentation was performed by incubating the inoculated goat milk at 37°C for 18 h. The pH and degree of acidity were measured every 2 h during the fermentation process, whereas the physicochemical (pH, degree of acidity, lactose, protein, and viscosity) and microbial (total lactic acid bacteria/LAB) qualities were measured after the fermentation process. Data analysis revealed that the different starter cultures had no significant effect (P > 0.05) on the pH and degree of acidity, lactose and protein content, viscosity, and total LAB content. The goat milk fermented in this study had an average pH of 4.41 ± 0.11 and a degree of acidity of 1.26 ± 0.05%, a lactose content of 5.47± 0.80%, a protein content of 6.69± 1.52%, a viscosity of 1217 ± 395 cP, and a total LAB content of 9.57 ± 1.49 log CFU/mL. In conclusion, goat milk fermented using both single and combined starter cultures of L. paracasei M104 and P. pentosaceus M103 exhibited similar physicochemical and microbial qualities and required longer fermentation time compared with the control.

Effects of different starter cultures on the biogenic amine concentrations, mutagenicity, oxidative stress, and neuroprotective activity of fermented sausages and their relationships

Five fermented sausage groups were produced (fermented sausages (FS) produced without starter culture (natural fermentation), control; FS produced with commercial starter culture Almi7™ (Staphylococcus carnosus and Lactobacillus sakei), T1; FS produced with Enterococcus thailandicus, T2; FS produced with Enterococcus faecalis, T3; FS produced with E. thailandicus and E. faecalis, T4), and they were analyzed for biogenic amine concentrations, mutagenicity, oxidative stress, and neuroprotective activity. Among the biogenic amines, only cadaverine was present in all fermented sausages, and the combined starter culture of E. thailandicus and E. faecalis (T4) produced the lowest biogenic amine concentrations. Sausages fermented by E. faecalis (T3) alone showed the lowest mutagenicity. Sausages fermented by E. thailandicus and E. faecalis (T4) showed the lowest oxidative stress and highest neuroprotective activity. Finally, the combined starter culture of E. thailandicus and E. faecalis would be optimum for producing fermented sausage regarding biogenic amine concentrations, oxidative stress, and neuroprotective activity.

Behavior and effect of combined starter cultures on microbiological and physicochemical characteristics of dry-cured ham.

The behaviour of two combined starter cultures and their influence on the microbiological and physicochemical characteristics of dry-cured ham have been evaluated. Three lots of dry-cured hams were tested during processing (0, 9, 48, 74, 112, 142, 166 and 211days). Lot 1 had no added starter culture. Lot 2 contained a starter culture of Penicillium chrysogenum, Penicillium digitatum, Penicillium nalgiovense, Debaryomyces hansenii, Lactobacillus plantarum, Lactobacillus acidophilus, Pediococcus pentosaceus and Micrococcus varians was and lot 3 had L. plantarum, L. acidophilus, P. pentosaceus and M. varians. The use of a selected starter culture based on a combination of lactic acid bacteria (LAB) and fungal strains with a demonstrated proteolytic activity such as P. chrysogenum and D. hansenii (lot 2) did not affect the main characteristics of dry-cured ham processing, even enhancing some desirable aspects, like its non-protein nitrogen contents. LAB strains were not significantly affected by combining them with fungal starter, and better counts were found with respect to control. A higher thiobarbituric acid reactive substances content was described in lot inoculated only with LAB (lot 3). Potentially pathogenic microorganisms were not detected in any of the lots studied. The starter culture used in lot 2 showed a potential interest for use in dry-cured ham production.