Effects of hybrid, kernel maturity, and storage period on the bacterial community in high-moisture and rehydrated corn grain silages.

This study evaluated changes in the bacterial community in high-moisture and rehydrated corn grain silage, and their correlation with fermentation quality attributes in distinct corn hybrids, the storage period, and kernel maturity at plant harvest. Most silages achieved good fermentation (pH<4.2). Rehydrated corn had a higher pH across all storage periods evaluated and increased dry matter losses. Leuconostoc and Lactococcus were the dominant genera in fresh material, while Lactobacillus and Acetobacter were prevalent in silages. Clostridium and Enterococcus prevailed in rehydrated corn after 120 days storage, and Clostridium was highly and positively correlated with acetone, butyric acid, and 2,3-butanediol contents. The storage period and kernel maturity were the most important factors responsible for changes in the bacterial community of silages. Results confirmed the existence of a specific bacterial microbiome that was unique for each maturity and storage time. Variations in these factors also affected the fermentation quality through influencing the bacterial community.

Metabolic cross-feeding in imbalanced diets allows gut microbes to improve reproduction and alter host behaviour.

The impact of commensal bacteria on the host arises from complex microbial-diet-host interactions. Mapping metabolic interactions in gut microbial communities is therefore key to understand how the microbiome influences the host. Here we use an interdisciplinary approach including isotope-resolved metabolomics to show that in Drosophila melanogaster, Acetobacter pomorum (Ap) and Lactobacillus plantarum (Lp) a syntrophic relationship is established to overcome detrimental host diets and identify Ap as the bacterium altering the host's feeding decisions. Specifically, we show that Ap uses the lactate produced by Lp to supply amino acids that are essential to Lp, allowing it to grow in imbalanced diets. Lactate is also necessary and sufficient for Ap to alter the fly's protein appetite. Our data show that gut bacterial communities use metabolic interactions to become resilient to detrimental host diets. These interactions also ensure the constant flow of metabolites used by the microbiome to alter reproduction and host behaviour.

Optimized culture conditions for bacterial cellulose production by Acetobacter senegalensis MA1.

BACKGROUND: Cellulose, the most versatile biomolecule on earth, is available in large quantities from plants. However, cellulose in plants is accompanied by other polymers like hemicellulose, lignin, and pectin. On the other hand, pure cellulose can be produced by some microorganisms, with the most active producer being Acetobacter xylinum. A. senengalensis is a gram-negative, obligate aerobic, motile coccus, isolated from Mango fruits in Senegal, capable of utilizing a variety of sugars and produce cellulose. Besides, the production is also influenced by other culture conditions. Previously, we isolated and identified A. senengalensis MA1, and characterized the bacterial cellulose (BC) produced. RESULTS: The maximum cellulose production by A. senengalensis MA1 was pre-optimized for different parameters like carbon, nitrogen, precursor, polymer additive, pH, temperature, inoculum concentration, and incubation time. Further, the pre-optimized parameters were pooled, and the best combination was analyzed by using Central Composite Design (CCD) of Response Surface Methodology (RSM). Maximum BC production was achieved with glycerol, yeast extract, and PEG 6000 as the best carbon and nitrogen sources, and polymer additive, respectively, at 4.5 pH and an incubation temperature of 33.5 °C. Around 20% of inoculum concentration gave a high yield after 30 days of inoculation. The interactions between culture conditions optimized by CCD included alterations in the composition of the HS medium with 50 mL L- 1 of glycerol, 7.50 g L- 1 of yeast extract at pH 6.0 by incubating at a temperature of 33.5 °C along with 7.76 g L- 1 of PEG 6000. This gave a BC yield of wet weight as 469.83 g L- 1. CONCLUSION: The optimized conditions of growth medium resulted in enhanced production of bacterial cellulose by A. senegalensis MA1, which is around 20 times higher than that produced using an unoptimized HS medium. Further, the cellulose produced can be used in food and pharmaceuticals, for producing high-quality paper, wound dressing material, and nanocomposite films for food packaging.

Analysis of the influence of flame sterilization included in sampling operations on shake-flask cultures of microorganisms.

Shake-flask cultures of microorganisms involve flame sterilization during sampling, which produces combustion gas with high CO2 concentrations. The gaseous destination has not been deeply analyzed. Our aim was to investigate the effect of flame sterilization on the headspace of the flask and on the shake-flask culture. In this study, the headspace CO2 concentration was found to increase during flame sterilization ~0.5-2.0% over 5-20 s empirically using the Circulation Direct Monitoring and Sampling System. This CO2 accumulation was confirmed theoretically using Computational Fluid Dynamics; it was 9% topically. To evaluate the influence of CO2 accumulation without interference from other sampling factors, the flask gas phase formed by flame sterilization was reproduced by aseptically supplying 99.8% CO2 into the headspace, without sampling. We developed a unit that can be sampled in situ without interruption of shaking, movement to a clean bench, opening of the culture-plug, and flame sterilization. We observed that the growth behaviour of Escherichia coli, Pelomonas saccharophila, Acetobacter pasteurianus, and Saccharomyces cerevisiae was different depending on the CO2 aeration conditions. These results are expected to contribute to improving microbial cell culture systems.

Starter consortia for on-farm cocoa fermentation and their quality attributes.

A starter consortium of Saccharomyces cerevisiae (Y), Lactobacillus plantarum (LAB), and Acetobacter aceti (AAB) was defined to ferment the Cocoa beans (Theobroma cacao). Emphasis was laid to optimize the microbial concentration with a functional ratio of selected cultures. A central composite rotatable design (CCRD) was employed to study the effect of inoculum size (0-20% w/v) with alcohol, titrable acidity, polyphenols, anthocyanin, cut test, and sensory as response variables. The significant (p < 0.05) response surface models with high coefficients of determination values (R2) ranging from 0.82 to 0.93 were considered for the experimental data, which represented the polynomial response models for describing the constraints. Based on the design, the concentration of consortia ranged 9.03X103 CFU/g of Y, 5.9X104 CFU/g of LAB, and 7.0X104 CFU/g of AAB. The graphical optimization of superimposed contour plots fulfilled the desired metabolites; alcohol (Y1) ≤ 11 mg/g, titrable acidity (Y2) ≥ 0.25%, polyphenols (Y3) ≤ 4.0 mg/g, anthocyanin (Y4) ≤ 14 mg/g, sensory (Y5) ≥ 6.0, and cut test (Y6)≥95%. Thus, validation through a field trial was confirmed to adopt the techno-economic feasibility on-farm process with precise inoculums. The effect of starter consortia on Cocoa fermentation and quality was found to be significant.

Highly efficient asymmetric reduction of 2-octanone in biphasic system by immobilized Acetobacter sp. CCTCC M209061 cells.

Highly efficient asymmetric reduction of 2-octanone to (R)-2-octanol catalyzed by immobilized Acetobacter sp. CCTCC M209061 cells was achieved in a biphasic system. Bioreduction conducted in aqueous single phase buffer was limited due to poor solubility and toxicity towards cells cause by product accumulation. Introduction of [C4MIM]·Ac accelerated the biotransformation process, giving 99% yield, >99% product e.e. and 1.42-fold higher initial reaction rate in conversion of 10 mM 2-octanone as substrate, compared with 99% yield, 97.3% product e.e. and 1.57 µmol min-1 of initial reaction rate in a aqueous single phase buffer system containing 6 mM 2-octanone as the optimal substrate concentration. Moreover, in the [C4MIM]·Ac-containing buffer/n-tetradecane biphasic system, the optimal substrate concentration was enhanced by 83 times (500 mM) in comparison with that in aqueous single phase buffer, resulting in 53.4% yield (267 mM), 99% product e.e. with 8.9 mM g-1 h-1 space time yield.

Growth and metabolite production of a grape sour rot yeast-bacterium consortium on different carbon sources.

The present study was designed to evaluate possible sugar-based trophic interactions between acetic acid bacteria (AAB) and non-Saccharomyces yeasts (NSY) involved in table grape sour rot, a disease in which berries spoilage is caused by the accumulation of several microbial metabolites. Acetobacter syzygii LMG 21419 (As) and Candida zemplinina CBS 9494 (Cz), a simplified AAB-NSY association responsible for table grape sour rot, grew differently in a minimal medium (YP) supplemented with glucose, ethanol, acetic and gluconic acid under monoculture conditions. In As -Cz co-culture media, after 24 h of incubation, As showed high relative abundance in YP-ethanol, whereas Cz was the dominant strain in YP-glucose medium. Co-culture in YP-glucose showed that glucose was converted into ethanol by Cz that, in turn, promoted the growth of As population. Gluconic acid was the main bacterial metabolite from glucose in monoculture, whereas acetic acid putatively derived from ethanol oxidation was found only in co-culture. However, gluconic acid showed inhibitory effect against As whereas acetic acid mainly inhibited Cz. Negative effects of both metabolites were mitigated in the glucose-supplemented medium. The results suggest a possible metabolic- based temporal succession between AAB and NSY during grape sour rot development. At the begin of sour rot, low glucose concentration promotes NSY producing ethanol, then, the AAB could take advantage from the oxidation of ethanol into acetic acid, becoming the dominant microbial sour rot population during the late stages of the process.

Forcing fermentation: Profiling proteins, peptides and polyphenols in lab-scale cocoa bean fermentation.

This study encompassed the lab-scale fermentation of cocoa beans in 300-g heaps under controlled laboratory conditions, in order to replicate the microbial dynamics and metabolomic changes that usually occur in large-scale spontaneous fermentations. Growth profiles of yeast and acetic acid bacteria (AAB) with the native assortment of microbes as well as with the use of a starter culture were very similar to those observed in literature. Greater production of acetic acid by AAB not only led to more acidic-tasting liquor but also contributed to bitterness, due to polyphenol preservation. It also brought about a drastic drop in pH leading to greater proteolytic activity. Peptides generated through proteolysis also showed incredible similarity to those reported in literature, in particular, those speculated to be involved in cocoa-specific flavour. A closer look at the naturally occurring peptide repertoires of our fermentation trials, generated by the breakdown of cocoa storage protein, pointed to a potential peptide responsible for cocoa-specific aroma.

Producing Acetic Acid of Acetobacter pasteurianus by Fermentation Characteristics and Metabolic Flux Analysis.

The acetic acid bacterium Acetobacter pasteurianus plays an important role in acetic acid fermentation, which involves oxidation of ethanol to acetic acid through the ethanol respiratory chain under specific conditions. In order to obtain more suitable bacteria for the acetic acid industry, A. pasteurianus JST-S screened in this laboratory was compared with A. pasteurianus CICC 20001, a current industrial strain in China, to determine optimal fermentation parameters under different environmental stresses. The maximum total acid content of A. pasteurianus JST-S was 57.14 ± 1.09 g/L, whereas that of A. pasteurianus CICC 20001 reached 48.24 ± 1.15 g/L in a 15-L stir stank. Metabolic flux analysis was also performed to compare the reaction byproducts. Our findings revealed the potential value of the strain in improvement of industrial vinegar fermentation.

Impacts of bioprocess engineering on product formation by Acetobacter pasteurianus.

Aerobic Acetobacter pasteurianus is one of the most widely used bacterial species for acetic acid and vinegar production. The acetic acid condition is the primary challenge to the industrial application of A. pasteurianus. Thus, numerous endeavors, including strain improvement and process control, have been performed to improve the product formation and acetic acid tolerance of A. pasteurianus. The metabolic features of A. pasteurianus have been gradually elucidated through omic techniques, such as genomics and proteomics. In this mini review, we summarized bioprocess engineering methods that improved product formation of A. pasteurianus by exploiting its metabolic features. Moreover, given that A. pasteurianus is an important functional microorganism in traditional vinegar production, we discuss its metabolism when cocultured with other microorganisms in traditional vinegar production.

Spatiotemporally Heterogeneous Population Dynamics of Gut Bacteria Inferred from Fecal Time Series Data.

A priority in gut microbiome research is to develop methods to investigate ecological processes shaping microbial populations in the host from readily accessible data, such as fecal samples. Here, we demonstrate that these processes can be inferred from the proportion of ingested microorganisms that is egested and their egestion time distribution, by using general mathematical models that link within-host processes to statistics from fecal time series. We apply this framework to Drosophila melanogaster and its gut bacterium Acetobacter tropicalis Specifically, we investigate changes in their interactions following ingestion of a food bolus containing bacteria in a set of treatments varying the following key parameters: the density of exogenous bacteria ingested by the flies (low/high) and the association status of the host (axenic or monoassociated with A. tropicalis). At 5 h post-ingestion, ~35% of the intact bacterial cells have transited through the gut with the food bolus and ~10% are retained in a viable and culturable state, leaving ~55% that have likely been lysed in the gut. Our models imply that lysis and retention occur over a short spatial range within the gut when the bacteria are ingested from a low density, but more broadly in the host gut when ingested from a high density, by both gnotobiotic and axenic hosts. Our study illustrates how time series data complement the analysis of static abundance patterns to infer ecological processes as bacteria traverse the host. Our approach can be extended to investigate how different bacterial species interact within the host to understand the processes shaping microbial community assembly.IMPORTANCE A major challenge to our understanding of the gut microbiome in animals is that it is profoundly difficult to investigate the fate of ingested microbial cells as they travel through the gut. Here, we created mathematical tools to analyze microbial dynamics in the gut from the temporal pattern of their abundance in fecal samples, i.e., without direct observation of the dynamics, and validated them with Drosophila fruit flies. Our analyses revealed that over 5 h after ingestion, most bacteria have likely died in the host or have been egested as intact cells, while some living cells have been retained in the host. Bacterial lysis or retention occurred across a larger area of the gut when flies ingest bacteria from high densities than when flies ingest bacteria from low densities. Our mathematical tools can be applied to other systems, including the dynamics of gut microbial populations and communities in humans.

Strong responses of Drosophila melanogaster microbiota to developmental temperature.

Physiological responses to changes in environmental conditions such as temperature may partly arise from the resident microbial community that integrates a wide range of bio-physiological aspects of the host. In the present study, we assessed the effect of developmental temperature on the thermal tolerance and microbial community of Drosophila melanogaster. We also developed a bacterial transplantation protocol in order to examine the possibility of reshaping the host bacterial composition and assessed its influence on the thermotolerance phenotype. We found that the temperature during development affected thermal tolerance and the microbial composition of male D. melanogaster. Flies that developed at low temperature (13°C) were the most cold resistant and showed the highest abundance of Wolbachia, while flies that developed at high temperature (31°C) were the most heat tolerant and had the highest abundance of Acetobacter. In addition, feeding newly eclosed flies with bacterial suspensions from intestines of flies developed at low temperatures changed the heat tolerance of recipient flies. However, we were not able to link this directly to a change in the host bacterial composition.

Effect of aspartic acid and glutamate on metabolism and acid stress resistance of Acetobacter pasteurianus.

BACKGROUND: Acetic acid bacteria (AAB) are widely applied in food, bioengineering and medicine fields. However, the acid stress at low pH conditions limits acetic acid fermentation efficiency and high concentration of vinegar production with AAB. Therefore, how to enhance resistance ability of the AAB remains as the major challenge. Amino acids play an important role in cell growth and cell survival under severe environment. However, until now the effects of amino acids on acetic fermentation and acid stress resistance of AAB have not been fully studied. RESULTS: In the present work the effects of amino acids on metabolism and acid stress resistance of Acetobacter pasteurianus were investigated. Cell growth, culturable cell counts, acetic acid production, acetic acid production rate and specific production rate of acetic acid of A. pasteurianus revealed an increase of 1.04, 5.43, 1.45, 3.30 and 0.79-folds by adding aspartic acid (Asp), and cell growth, culturable cell counts, acetic acid production and acetic acid production rate revealed an increase of 0.51, 0.72, 0.60 and 0.94-folds by adding glutamate (Glu), respectively. For a fully understanding of the biological mechanism, proteomic technology was carried out. The results showed that the strengthening mechanism mainly came from the following four aspects: (1) Enhancing the generation of pentose phosphates and NADPH for the synthesis of nucleic acid, fatty acids and glutathione (GSH) throughout pentose phosphate pathway. And GSH could protect bacteria from low pH, halide, oxidative stress and osmotic stress by maintaining the viability of cells through intracellular redox equilibrium; (2) Reinforcing deamination of amino acids to increase intracellular ammonia concentration to maintain stability of intracellular pH; (3) Enhancing nucleic acid synthesis and reparation of impaired DNA caused by acid stress damage; (4) Promoting unsaturated fatty acids synthesis and lipid transport, which resulted in the improvement of cytomembrane fluidity, stability and integrity. CONCLUSIONS: The present work is the study to show the effectiveness of Asp and Glu on metabolism and acid stress resistance of A. pasteurianus as well as their working mechanism. The research results will be helpful for development of nutrient salts, the optimization and regulation of high concentration of cider vinegar production process.

Efficacy of gaseous ozone to counteract postharvest table grape sour rot.

This work aims at studying the efficacy of low doses of gaseous ozone in postharvest control of the table grape sour rot, a disease generally attributed to a consortium of non-Saccharomyces yeasts (NSY) and acetic acid bacteria (AAB). Sour rot incidence of wounded berries, inoculated with 8 NSYstrains, or 7 AAB, or 56 yeast-bacterium associations, was monitored at 25 °C up to six days. Sour rot incidence in wounded berries inoculated with yeast-bacterium associations resulted higher than in berries inoculated with one single NSY or AAB strain. Among all NSY-AAB associations, the yeast-bacterium association composed of Candida zemplinina CBS 9494 (Cz) and Acetobacter syzygii LMG 21419 (As) showed the highest prevalence of sour rot; thus, after preliminary in vitro assays, this simplified As-Cz microbial consortium was inoculated in wounded berries that were stored at 4 °C for ten days under ozone (2.14 mg m-3) or in air. At the end of cold storage, no berries showed sour-rot symptoms although ozonation mainly affected As viable cell count. After additional 12 days at 25 °C, the sour rot index of inoculated As-Cz berries previously cold-stored under ozone or in air accounted for 22.6 ± 3.7% and 66.7 ± 4.5%, respectively. Molecular analyses of dominant AAB and NSY populations of both sound and rotten berries during post-refrigeration period revealed the appearance of new strains mainly belonging to Gluconobacter albidus and Hanseniaspora uvarum species, respectively. Cold ozonation resulted an effective approach to extend the shelf-life of table grapes also after cold storage.

Commensal bacteria and essential amino acids control food choice behavior and reproduction.

Choosing the right nutrients to consume is essential to health and wellbeing across species. However, the factors that influence these decisions are poorly understood. This is particularly true for dietary proteins, which are important determinants of lifespan and reproduction. We show that in Drosophila melanogaster, essential amino acids (eAAs) and the concerted action of the commensal bacteria Acetobacter pomorum and Lactobacilli are critical modulators of food choice. Using a chemically defined diet, we show that the absence of any single eAA from the diet is sufficient to elicit specific appetites for amino acid (AA)-rich food. Furthermore, commensal bacteria buffer the animal from the lack of dietary eAAs: both increased yeast appetite and decreased reproduction induced by eAA deprivation are rescued by the presence of commensals. Surprisingly, these effects do not seem to be due to changes in AA titers, suggesting that gut bacteria act through a different mechanism to change behavior and reproduction. Thus, eAAs and commensal bacteria are potent modulators of feeding decisions and reproductive output. This demonstrates how the interaction of specific nutrients with the microbiome can shape behavioral decisions and life history traits.

Effect of chitosan and SO2 on viability of Acetobacter strains in wine.

Wine spoilage is an important concern for winemakers to preserve the quality of their final product and avoid contamination throughout the production process. The use of sulphur dioxide (SO2) is highly recommended to prevent wine spoilage due to its antimicrobial activity. However, SO2 has a limited effect on the viability of acetic acid bacteria (AAB). Currently, the use of SO2 alternatives is favoured in order to reduce the use of chemicals and improve stabilization in winemaking. Chitosan is a biopolymer that is approved by the European authorities and the International Organization of Vine and Wine to be used as a fining agent and antimicrobial in wines. However, its effectiveness in AAB prevention has not been studied. Two strains of Acetobacter, adapted to high ethanol environments, were analysed in this study. Both chitosan and SO2 effects were compared in artificially contaminated wines. Both molecules reduced the metabolic activity of both AAB strains. Although AAB populations were detected by culture independent techniques, their numbers were reduced with time, and their viability decreased following the application of both products, especially with chitosan.

Improving fermented quality of cider vinegar via rational nutrient feeding strategy.

This work aimed to find a rational nutrient feeding strategy for cider vinegar fermentation based on adequate information on the nutritional requirement of acetic acid bacteria. Through single nutrient lack experiment assay, necessary nutrient recipe for Acetobacter pasteurianus CICIM B7003 in acetous fermentation was confirmed. Compounds from the essential nutrient recipe were tested further to find out the key substrates significantly influencing cider vinegar fermentation. The findings showed that aspartate, glutamate, proline and tryptophan should be considered in detail for optimizing nutritional composition of cider. Finally, a nutrient feeding strategy that simultaneously adds proline, glutamate, aspartate and tryptophan to form final concentrations of 0.02g/L, 0.03g/L, 0.01g/L and 0.005g/L in cider was achieved by orthogonal experiment design. Comparing to the original fermentation, the yield of acetic acid from alcohol reached 93.3% and the concentration of most volatile flavor compounds increased with the rational nutrient feeding strategy.

Effect of ammonium and amino acids on the growth of selected strains of Gluconobacter and Acetobacter.

Acetic acid bacteria (AAB) are a group of microorganisms highly used in the food industry. However, its use can be limited by the insufficient information known about the nutritional requirements of AAB for optimal growth. The aim of this work was to study the effects of different concentrations and sources of nitrogen on the growth of selected AAB strains and to establish which nitrogen source best encouraged their growth. Two strains of three species of AAB, Gluconobacter japonicus, Gluconobacter oxydans and Acetobacter malorum, were grown in three different media with diverse nitrogen concentrations (25, 50, 100, and 300mgN/L and 1gN/L) as a complete solution of amino acids and ammonium. With this experiment, the most favourable medium and the lowest nitrogen concentration beneficial for the growth of each strain was selected. Subsequently, under these conditions, single amino acids or ammonium were added to media individually to determine the best nitrogen sources for each AAB strain. The results showed that nitrogen requirements are highly dependent on the nitrogen source, the medium and the AAB strain. Gluconobacter strains were able to grow in the lowest nitrogen concentration tested (25mgN/L); however, one of the G. oxydans strains and both A. malorum strains required a higher concentration of nitrogen (100-300mgN/L) for optimal growth. In general, single nitrogen sources were not able to support the growth of these AAB strains as well as the complete solution of amino acids and ammonium.

Diversity of the microbiota involved in wine and organic apple cider submerged vinegar production as revealed by DHPLC analysis and next-generation sequencing.

Unfiltered vinegar samples collected from three oxidation cycles of the submerged industrial production of each, red wine and organic apple cider vinegars, were sampled in a Slovene vinegar producing company. The samples were systematically collected from the beginning to the end of an oxidation cycle and used for culture-independent microbial analyses carried out by denaturing high pressure liquid chromatography (DHPLC) and Illumina MiSeq sequencing of 16S rRNA gene variable regions. Both approaches showed a very homogeneous bacterial structure during wine vinegar production but more heterogeneous during organic apple cider vinegar production. In all wine vinegar samples Komagataeibacter oboediens (formerly Gluconacetobacter oboediens) was a predominating species. In apple cider vinegar the acetic acid and lactic acid bacteria were two major groups of bacteria. The acetic acid bacterial consortium was composed of Acetobacter and Komagataeibacter with the Komagataeibacter genus outcompeting the Acetobacter in all apple cider vinegar samples at the end of oxidation cycle. Among the lactic acid bacterial consortium two dominating genera were identified, Lactobacillus and Oenococcus, with Oenococcus prevailing with increasing concentration of acetic acid in vinegars. Unexpectedly, a minor genus of the acetic acid bacterial consortium in organic apple cider vinegar was Gluconobacter, suggesting a possible development of the Gluconobacter population with a tolerance against ethanol and acetic acid. Among the accompanying bacteria of the wine vinegar, the genus Rhodococcus was detected, but it decreased substantially by the end of oxidation cycles.

Microbial community, and biochemical and physiological properties of Korean traditional black raspberry (Robus coreanus Miquel) vinegar.

BACKGROUND: The aim of this study was to elucidate the changes in microbial community and biochemical and physiological properties of traditional Muju black raspberry (Robus coreanus Miquel) vinegar (TMBV) during fermentation by culture-independent methods. RESULTS: During vinegar fermentation, ethanol produced up to 120 g L(-1) until day 35, with continuously increasing yeast concentration to a total of log 7.6 CFU mL(-1) . After day 35, acetic acid bacteria (AAB) concentrations rose to log 5.8 CFU mL(-1) until day 144. Denaturing gradient gel electrophoresis analysis showed that Saccharomyces cerevisiae was detected until day 87 of the fermentation, at which point Acetobacter pasteurianus gradually took over as the dominant species. Total sugar was reduced to 6.6 °Brix and total acidity produced up to 44 g L(-1) . CONCLUSION: In this study, we established the physicochemical analysis and growth dynamics of yeast and AAB during alcoholic and acetic acid fermentation of black raspberry by a traditional method. Overall, S. cerevisiae and A. pasteurianus species appeared to dominate the TMBV fermentation. In conclusion, this study demonstrated a suitable fermentation system for TMBV by the static surface method. © 2015 Society of Chemical Industry.