Applied Microbiology

The growing threat of antibiotic-resistant Gram-negative bacteria highlights the urgent need for innovative, non-bactericidal therapeutic strategies. Quorum-sensing (QS) inhibition has emerged as a promising approach to attenuate bacterial virulence without exerting selective pressure. This study evaluated the antimicrobial, anti-QS, and antibiofilm properties of aqueous extracts from five medicinal plants native to northeastern Mexico: Gymnosperma glutinosum, Ibervillea sonorae, Larrea tridentata, Olea europaea, and Tecoma stans. Disk diffusion and violacein quantification assays using Chromobacterium violaceum demonstrated significant QS inhibition by G. glutinosum and T. stans, with violacein reductions of 60.02% and 52.72%, respectively, at 40 mg/mL. While L. tridentata and O. europaea exhibited antibacterial activity, I. sonorae showed no growth or pigment inhibition but achieved the highest biofilm disruption (89.89%) against Salmonella typhimurium. UPLC-MS analysis identified chlorogenic acid, kaempferol, and D-(−)-quinic acid as major constituents, compounds previously associated with QS modulation. These findings highlight the potential of traditional Mexican plant species as sources of QS inhibitors and bio-film-disrupting agents, supporting their further development as alternatives to conventional antibiotics. Full article

The whole-genome sequencing of lactic acid bacteria provides a valuable resource for identifying the genetic determinants underlying molecular mechanisms related to their probiotic properties. Analysis of draft genome sequences relies on bioinformatics tools for genetic data processing and in silico analytical methods to pinpoint the genetic determinants encoding biologically active compounds. The aim of this study was to perform the phenotypic determination of the antibiotic sensitivity and bioinformatics analyses on whole-genome assemblies from LAB from the human oral microbiome, and determine the presence of acquired antibiotic resistance genes, peptidases, adhesion proteins, and bacteriocins. Bioinformatics processing was performed in order to establish the molecular mechanisms responsible for the previously observed probiotic properties. The tested LAB strains exhibited a broad spectrum of antibiotic multiresistance, but did not possess acquired antibiotic resistance genes. The detected genes for peptidase activity were from the Pep family of hydrolase enzymes. Genetic determinants for adhesion proteins contained LPxTG, YSIRK, KxYKxGKxW, and SEC 10/PgrA domains, as well as MucBP domains. Lectins were found for five of the strains with the presence of WxL domains from the CscC protein family and L-type lectin domains. The in silico analyses show that some of the tested strains possessed mechanisms for bacteriocin production. Full article

Background: Clinical features of asthma are associated with differences in the lower airway microbiome. However, knowledge is limited on whether airway microbiota composition differs between individuals residing in different geographic regions and if asthma-associated differences in lower airway microbiota are similar between distinct populations. Methods: Existing 16S rRNA gene sequence data, generated from sputum collected from adults with or without asthma (n = 74) from two single-center cohort studies in the U.S. and United Arab Emirates, were re-processed for merged computational analysis using standard available tools. Potential differences between study sites, asthma status and specific clinical factors (inhaled corticosteroid use, ICS; obesity) were examined. Results: Differences in sputum bacterial composition, assessed by alpha- and beta-diversity measures, were associated with study site. Despite this, asthma-related differences were discerned in both cohorts. Specifically, sputum microbiota of asthmatic patients on ICS treatment displayed reduced bacterial phylogenetic diversity, compared to those not on ICS treatment (p = 0.006). Sputum bacterial composition also differed by obesity status (unweighted Unifrac distance PERMANOVA, p = 0.004). Specific genera were identified in both cohorts that were differentially enriched between obese vs. non-obese subjects, including Rothia and Veillonella (obesity-associated) and Campylobacter (non-obesity-associated). Conclusions: Our findings suggest clinical factors associated with differences in the lower airway microbiome in asthma may transcend variation related to geographic area of residence. Full article

E. coli attaches to, and forms biofilms on various surfaces, including latex and polystyrene, contributing to nosocomial spread. E. coli responds to both exogenous and endogenous insulin, which induces behavioral changes. Human insulin, a quorum signal surrogate for microbial insulin, may affect the ability of E. coli to interact with latex and polystyrene in the presence of various sugars. E. coli ATCC 25923 was grown in peptone (1%) yeast nitrogen base broth to either the logarithmic or stationary growth phase. Adherence to latex was determined using 6 × 6 mm latex squares placed in a suspension of washed cells (10³ CFU/mL; 30 min; 37 °C) in buffer containing insulin at 2, 20, and 200 µU/mL (Humulin^® R; Lilly) with and without mannose, galactose, fructose, sorbose, arabinose, xylose, lactose, maltose, melibiose, glucose-6-phosphate, glucose-1-phosphate, and glucosamine at concentrations reported to affect behavioral response. Attachment levels to latex were determined by the press plate method. Biofilm levels were measured in a similar fashion but with overnight cultures in flat bottom uncoated polystyrene plates. Controls were media, insulin, sugar, or buffer alone. Glucose served as the positive control. Overall, the stationary phase cells’ adherence to latex was greater, regardless of the test condition, than was measured for the logarithmic phase cells. The effect of insulin on adherence to latex was insulin and sugar concentration dependent. The addition of insulin (200 µU/mL) resulted in a significantly (p < 0.05) increased adherence to latex and biofilm formation on polystyrene compared with sugar alone for 12 of the 13 sugars tested with stationary phase bacteria and 10 of the 13 sugars tested with logarithmic phase bacteria. Adherence in response to sorbose was the only sugar tested that was unaffected by insulin. These findings show that insulin enhances E. coli’s association with materials in common usage in medical environments in a nutrition-dependent manner. Full article

Overuse of antibiotics is a concern in ‘culture-negative sepsis’ but it is unclear whether this is due to infection with viruses, fungi or other microbes that are not easily cultured, or whether it results from inflammatory processes. In a prospective study, we enrolled 50 preterm neonates with culture-positive sepsis (CP), culture-negative sepsis (CN), and asymptomatic preterm controls (CO). The microbiome of stool, skin, and blood, including bacterial, viral and fungal components and serum cytokine profiles were evaluated. The microbiome alpha or beta diversity did not differ between CN and CO groups. A MaAsLin analysis revealed increased relative abundances of specific bacterial and fungal genera in stool and skin samples in the CN group compared to CO. The virome analysis identified 24 viruses from skin samples, but they were not statistically different among the three groups. The cytokine and chemokine biomarker profiles were elevated in the CP group but were not statistically different between the CN and CO groups. Although the CN group had a longer hospital stay and higher BPD rates than the controls in unadjusted analyses, these differences were not significant after adjusting for gestational age and birth weight. The CN infants demonstrated microbial shifts without systemic immune activation or significantly worse clinical outcomes, supporting the rationale for discontinuing antibiotics in the absence of positive cultures. Full article

The rumen microbiome impacts beef cattle feed efficiency, a key economic factor in production systems. This study investigated the rumen microbiome of Charolais bulls with divergent residual feed intake-expected progeny difference (RFI-EPD) values to identify microbial taxa associated with feed efficiency. Forty Charolais bulls were evaluated for feed intake and growth over 60 days, and RFI values were determined. The 10 most efficient (NegRFI) and 10 least efficient (PosRFI) bulls were selected for microbiome analysis. Rumen fluid samples were collected and analyzed via 16S rRNA gene sequencing. Microbial analysis revealed no significant differences in alpha or beta diversity between groups, but differential abundance analysis identified 20 operational taxonomic units (OTUs) as more prevalent in NegRFI bulls, while 15 OTUs were more abundant in PosRFI bulls. Two OTUs from the key genus Prevotella showed different relative abundances in the two RFI-EPD groups. NegRFI bulls had a higher relative abundance of Prevotella OTU 109358, while PosRFI bulls had more Prevotella OTU 626329. Additionally, OTUs from Ruminococcus, a genus involved in fiber degradation and volatile fatty acid (VFA) production, were more abundant in NegRFI bulls. In contrast, PosRFI bulls had a higher abundance of OTUs from Oscillospira and F16, both linked to butyrate production. The results of this study support the need for further exploration into the role of microbial taxa associated with feed efficiency. A deeper understanding of the functional profile of the microbiota could aid in the development of microbiome-informed strategies to enhance nutrient utilization and performance in beef cattle. Full article

Significant heavy metals contamination is often caused by rapid industrialization, which is devastating to both public health and the environment. Conventional processes of metal removal also result in the accumulation of secondary waste. This work proposes the use of a novel fungal biomass (microwave heat dried) from Arthrinium malaysianum for the biosorption of toxic chromium. We have meticulously explored and investigated the interactions of hexavalent chromium with dried biomass using several cutting-edge techniques like FTIR for studying the involvement of functional groups on the biomass surface, XRD for the surface architecture changes after metal binding, XPS to unravel the reduction of hexavalent chromium into its non-toxic form, and FESEM-EDX for the visualization of the ultra-structure of fungal cell surface. The Langmuir isotherm demonstrates that the maximum removal capacity Q_max of Cr(VI) is 102.310 mgg⁻¹, at a pH of 3.5 with 100% removal of Cr(VI). There were substantial changes in the surface architecture during adsorption, confirmed by FESEM and AFM studies. FTIR and XPS data analysis indicated that carbonyl, hydroxyl, phosphate, and amine groups were responsible for the conversion of Cr(VI) (toxic) to Cr(III) (non-toxic). The IR spectra of biomass treated with Cr showed a decreased C-O stretching intensity and slight shriveling of the -OH band, and the bands in the FTIR spectra at 1642 cm⁻¹ to 1635 cm⁻¹ and at 1549 cm⁻¹ to 1547 cm⁻¹ shifted and appeared quite distinct. XRD revealed that the chromium-treated biomass had greater crystalline features and also the appearance of a wide peak where 2θ = 20°, approximately, indicating an amorphous nature at 576.0 eV and in highly loaded chromium (500 mg/L) biomass, with the Cr2p level displaying a slight shift, eventually terminating in a (576.0 eV) Cr₂O₃ to Cr(III) peak. Since the FTIR and XPS data obtained revealed that Cr(VI) reduces to Cr(III), this fungal biomass can also be used for generating metallic nanoparticles during biosorption. Thus, we suggest that the above-mentioned fungal biomass could be a very useful biomaterial for future translational research. We are in the process of fabricating beads with powdered biomass for further studies. Full article

In this study, pigment-producing yeast (Rhodotorula alborubescens) was isolated from the soil sample, demonstrating the potential for carotenoid biosynthesis. Physiological, morphological, biochemical, and molecular characterization confirmed the identity of the isolate. Optimization of the physical parameters for carotenoid production was achieved through batch shake flask experiments. The optimal conditions were determined to be 84 h of incubation at pH 6.0 and 28 °C under white light irradiation, utilizing the Yeast Peptone Dextrose (YPD) medium composed of 10 g/L yeast extract, 5 g/L of peptone, and 5 g/L dextrose, resulting in maximum carotenoid content. Further, the presence of β-carotene was confirmed using High-Performance Liquid Chromatography and Fourier Transform Infrared Spectroscopy. These findings highlight the potential of the isolated soil yeast (R. alborubescens) as a potential source of carotenoids, offering natural alternatives for various industrial applications. Full article

Microbially induced calcite precipitation (MICP) has attracted attention as an environmentally friendly soil stabilization method, with Sporosarcina pasteurii being a key ureolytic bacterium in this process. However, its behavior in oxygen-limited environments remains poorly understood, limiting the predictability of MICP outcomes in natural soils. This study investigated the population dynamics of Sporosarcina in compacted soil reactors operated under aerobic and anaerobic conditions, including saturated environments. Quantitative PCR and 16S rRNA gene sequencing revealed that Sporosarcina thrived and became dominant under aerobic, unsaturated conditions, but failed to maintain a high abundance under anaerobic or saturated conditions. These findings indicate that gas-phase oxygen—not merely its presence in the overlying atmosphere—is essential for effective Sporosarcina-driven MICP. The results highlight a critical environmental constraint that limits the application of biostimulation strategies relying on indigenous Sporosarcina in oxygen-poor soils. This study provides the first in situ evidence linking oxygen availability and microbial dominance in MICP systems, with implications for optimizing microbial soil stabilization in real-world conditions. Full article

The traditional fermentation of table olives is a complex and dynamic, process, carried out by a consortium of microorganisms that interact with each other and contribute to the uniqueness and attractiveness of the final product. Fermentation is conducted by yeasts and lactic acid bacteria (LAB) that coexist in olive fruits. The succession of one microbial population to the detriment of others depends on internal and external factors that affect the process, e.g., the maturation degree of fruits, cultivar, endophytic, or epiphytic state of microorganisms, pH, water activity, temperature, and salt concentration. Thus, studying microbiota evolution and their identification in natural table olive fermentations is paramount. This review aims to provide an overview of the knowledge on the natural fermentation of table olives, namely regarding microbial dynamics, as to report the main species involved in the fermentation process, highlight the influence of the olive oil ecosystem on the origin of the microbiota and consequently on the obtaining of the final product. The results report a total of 97 yeast species and 45 LAB species described in olives and brine over the last few decades. Full article

Beer lees (BL), a by-product of beer production, consist mainly of dead yeast cells with potential nutritional value. On the other hand, yeast extract (YE), obtained through the lysis of yeast cells, is commonly used as a nutrient-rich supplement for the growth of fastidious microorganisms such as lactic acid bacteria (LAB). However, YE is a high-cost ingredient. Therefore, the aim of this study was to optimize the use of BL as a low-cost alternative source of YE through different lysis treatments, evaluating its suitability to support the growth of UNQLpc 10 and UNQLp 11 strains in a whey permeate (WP)-based medium. Growth kinetics and cell viability were compared with those obtained in MRS broth. The best results were observed with sonicated BL, up to 10 logarithmic units, which supported LAB growth comparable to MRS. Although autolyzed BL promoted lower bacterial growth than sonicated BL, it showed greater cell disruption and higher levels of nitrogen, proteins, and amino acids (5.32%, 26.0%, and 277 nM, respectively). Additionally, autolyzed BL exhibited lower concentrations of reducing sugars and a higher presence of Maillard reaction products, as indicated by colorimetric analysis. These changes, which may be related to the formation of Maillard reaction products during the autolysis process, could have negatively affected the nutritional quality of the extract and, thus, reduced its effectiveness as a bacterial growth promoter. Full article

Sialidases are gradually entering various areas of human practice—in medicine and pharmacy, as antiviral, antitumor, diagnostic, and vaccine preparations; for the chemoenzymatic synthesis of regioselective sialoglycoconjugates; and for the structural analysis of sialoglycoproteins. Optimizing the synthesis conditions of these commercially important enzymes would be beneficial for enhancing their production and expanding potential applications. Since sialidase producers are often pathogenic microorganisms, the use of saprophytic bacteria could be an alternative to reduce the health risk when working with them. So far, the topic has not been widely discussed. By a single-factor optimization method, the most suitable fermentation conditions for achieving maximum sialidase production by the non-model strain Oerskovia paurometabola O129 were established. The dynamics of enzyme accumulation during the growth phases and the optimal physicochemical parameters for cultivation were determined (30 °C, pH 8.0, agitation at 200 rpm, for 28 h). The addition of various inducers and surfactants to improve enzyme yield was also investigated. The effect of surfactants on bacterial sialidase production was tested for the first time. Maximum enzyme production (98.3 U/mL), representing about a three-fold increase compared to non-optimized conditions, was obtained by culturing the strain under optimal conditions and by the synergistic action of glucomacropeptide and Tween 80. A new, simple, and cost-effective laboratory model for optimizing sialidase production by the saprophytic strain O. paurometabola O129 in submerged fermentation was proposed. Future work may involve scaling up the process and exploring genetic or metabolic enhancements for targeted biomedical and industrial applications. Full article

The study of bacteriocins has significantly enhanced our understanding of microbial interactions, notably within the genus Streptococcus. Among the most functionally diverse and clinically relevant bacteriocins are those belonging to the lantibiotic class, which exhibit potent antimicrobial properties and are central to the competitive dynamics of streptococcal species. This review focuses on the discovery and characterization of bacteriocins produced by Streptococcus pyogenes and Streptococcus salivarius, emphasizing their biological significance within their exclusive human host. A cornerstone of these studies has been the development and application of the pioneer agar culture-based bacteriocin detection methodology, known as streptococcal bacteriocin fingerprinting. This approach has proven invaluable for the initial detection and differentiation of a wide array of bacteriocin-like inhibitory substances (BLIS) in streptococcal populations. A central theme of this review is the diverse biological roles of lantibiotics in S. pyogenes and S. salivarius, particularly in relation to microbial competition, colonization dynamics, and host interactions. The expression of lantibiotic determinants provides distinct advantages to the producing strain, including enhanced niche establishment and the ability to suppress competing microbes. Furthermore, the presence of specific lantibiotic immunity mechanisms safeguards the producer from self-inhibition and potential antagonism from closely related competitors. In S. pyogenes, lantibiotic production has been implicated in virulence modulation, raising important questions about its role in pathogenicity and host immune evasion. Conversely, S. salivarius, a prominent commensal and probiotic candidate species, utilizes its lantibiotic arsenal to confer colonization benefits and mediate beneficial interactions, especially within the oral and upper respiratory tract microbiomes. The implications of in situ lantibiotic expression extend beyond microbial ecology, presenting opportunities for innovative probiotic and therapeutic applications. The potential for harnessing bacteriocin-producing streptococci in antimicrobial interventions, particularly in combating antibiotic-resistant pathogens, underscores the translational relevance of these findings. This review integrates historical and contemporary perspectives on streptococcal bacteriocin research, providing insights into future avenues for leveraging these bioactive peptides in clinical and biotechnological contexts. Full article

The biotechnological applications of chitinases are diverse. They are used in industrial sectors such as pharmaceuticals, textiles, and agriculture, including the use of recombinant chitinases for pest control, since traditional treatments affect and contaminate hive products. Bacillus licheniformis UV01 bacterium is of interest, as it expresses genes for different enzymes, including chitinase. The Chibluv01 gene was cloned into the pHTP8 vector with a His/tag for purification using affinity chromatography. It showed a specific activity of 115 U/m. The optimal pH and temperature were 7.5 and 42 °C, respectively. The choleoptericidal activity (ability to kill beetles) of the enzyme was evaluated in the larvae and adult beetles of Aethina tumida treated with immersion in a purified enzyme extract at different concentrations, and the mortality was verified at 24, 48, and 72 h. Within 24 h of application, the mortality increased by 50% in the larval stage and 56.6% in adult beetles compared to the control groups. The LC₅₀ and LC₉₀ were obtained: 104.05 U/mL and 234.36 U/mL in larvae and 92.99 U/mL and 211.14 U/mL in adults, respectively. These results indicate the potential of the application of ChiBlUV02 chitinase in pest control. Full article

This study evaluated the antimicrobial potential of a Bacillus subtilis spore-based probiotic cocktail to reduce foodborne pathogens in both nutrient-rich laboratory media and a complex food matrix (hummus). Three common foodborne pathogens—Listeria monocytogenes, Escherichia coli O157:H7, and Salmonella Typhimurium—were cultured individually in full-strength, half-strength, and quarter-strength tryptic soy broth (TSB) with or without the probiotic spores (~7 log CFU/mL). Additionally, a commercial hummus formulation was inoculated with L. monocytogenes (~3 log CFU/g) and B. subtilis spores (~7 log CFU/g) and stored at 30 °C to simulate temperature abuse. In TSB, E. coli and Salmonella grew to ~8.2 log CFU/mL in full-strength media, with no significant inhibition by the probiotics. However, L. monocytogenes showed substantial suppression: in nutrient-limited TSB, viable counts dropped below the detection limit of 1.48 log CFU/mL by 24 h in the presence of probiotics. In hummus, L. monocytogenes grew to an average of 8.22 log CFU/g in the absence of probiotics but remained significantly lower at an average of 5.03 log CFU/g when co-inoculated with B. subtilis (p < 0.05). Germination of probiotic spores was confirmed within 6 h under all conditions. These findings suggest that B. subtilis spores selectively inhibit Listeria, particularly under nutrient stress or abuse conditions. While the probiotic had limited impact on Gram-negative pathogens, its application may serve as a clean-label strategy for suppressing L. monocytogenes in ready-to-eat (RTE) foods. This dual-model approach provides insights into both mechanistic activity and practical limitations of spore probiotics in complex food matrices. Full article

Cyanobacteria are gaining significant importance as potential biocontrol agents against phytopathogenic fungi. We evaluated the inhibitory effects of a suspension of Nostoc sp. BCAC 1226 on Penicillium sp., Phytophthora sp., and Stemphylium sp. in vitro using potato dextrose agar medium. On the 7th day of incubation, Phytophthora sp. showed a reduction in colony area from 18.30 ± 1.68 to 8.55 ± 0.74 cm² (53.6% inhibition). Similarly, Penicillium sp. showed a reduction from 17.64 ± 1.46 to 8.90 ± 0.36 cm² (49.4% inhibition), and Stemphylium sp. showed a reduction from 17.76 ± 1.28 to 13.5 ± 0.73 cm² (23.7% inhibition). The inhibitory effects were more significant on the 14th day, with the growth of Phytophthora sp. further reduced to 4.9 ± 0.40 cm² (72.8% inhibition), Penicillium sp. to 5.54 ± 0.32 cm² (68.8% inhibition), and Stemphylium sp. to 8.71 ± 0.31 cm² (50.8% inhibition). These results indicate the potential antifungal activity of Nostoc sp. suspension, with the highest reduction observed in Phytophthora sp., followed by Penicillium sp. and Stemphylium sp. Future research should focus on the chemical characterization of the antifungal metabolites produced by Nostoc sp. BCAC 1226 and in vivo evaluations on economically important crops to evaluate their practical efficiency under field conditions. Full article

If lettuce is contaminated in the field, Shiga toxin-producing E. coli (STEC) O157:H7 can survive through the distribution chain. Prolonged cold storage during transportation may impact pathogen physiology, affecting subsequent stress survival and virulence. Greenhouse-grown Romaine lettuce, inoculated with three STEC O157:H7 strains, was harvested after 24 h and stored at 2 °C for 5 d following 4 h at harvest temperature (9 °C or 17 °C). Culturable, persister, and viable but non-culturable (VBNC) cells were quantified. Virulence was evaluated using Galleria mellonella and acid tolerance at pH 2.5 and tolerance to 20–25 ppm free chlorine were quantified. Colder harvest temperature (9 °C) before cold storage led to greater transformation of STEC O157:H7 into dormant states and decreased virulence in most cases. Increasing length of cold storage led to decreased virulence and acid tolerance of STEC O157:H7 on lettuce, while having no significant effect on chlorine tolerance. These findings highlight that entry of STEC O157:H7 into dormant states during harvest and transportation at cold temperatures leads to decreased stress tolerance and virulence with increasing cold storage. Changes in STEC O157:H7 physiology on lettuce during cold storage can be integrated into risk assessment tools for producers, which can assist in identifying practices that minimize risk of STEC O157:H7 from consumption of lettuce. Full article

The escalating threats posed by plant pathogens and the environmental repercussions of conventional agrochemicals necessitate sustainable agricultural solutions. This review focuses on plant growth-promoting microorganisms (PGPMs) such as bacteria, filamentous fungi, and yeasts, which play a pivotal role as biocontrol agents. These organisms enhance plant growth and resilience through nutrient solubilization, phytohormone production, and antagonistic activities against pathogens, offering a dual benefit of disease suppression and growth enhancement. However, the effective application of PGPMs faces challenges, including variability in field performance, survival and colonization under field conditions, and regulatory hurdles. This paper discusses these challenges and explores recent advances in utilizing these bioagents in sustainable agriculture, underscoring the importance of integrated pest management systems that reduce chemical inputs, thus promoting ecological balance and sustainable farming practices. Full article

During cocoa bean fermentation, yeasts, particularly those with pectinolytic activity, contribute to pulp degradation, facilitating and accelerating fermentation. This study aimed to identify and evaluate pectinolytic yeast strains for their suitability as starter cultures in cocoa bean fermentation. A high-throughput screening of 1139 yeasts previously isolated from cocoa bean fermentations identified three strains of Wickerhamomyces pijperi with strong pectinolytic activity. These strains also reduced the viscosity of a pectin-enriched cocoa pulp simulation medium (mCPSMpc) from 23.06 ± 0.11 mPa·s (uninoculated sample) to 4.40 ± 0.14 mPa·s, 4.22 ± 0.13 mPa·s, and 4.77 ± 0.17 mPa·s after 24 h for samples inoculated with strains H312, H403, and H404, respectively. W. pijperi H403 and H404, applied in co-culture with Limosilactobacillus fermentum 223 and Saccharomyces cerevisiae H290 in 1 kg lab-scale fermentations, significantly enhanced pulp degradation, with runoff after 23.8 h reaching 12.6–13.3%, compared to 4.7% in uninoculated controls. In 20 kg fermentations in Costa Rica, the effect was less pronounced, likely due to lower inoculation rates and environmental factors. Quantitative PCR confirmed the persistence of W. pijperi H403 in fermentations. Additionally, trial cultivations in 15 L stirred-tank bioreactors successfully demonstrated the ability to produce larger biomass quantities for upscaled applications. These findings highlight W. pijperi H403 as a promising candidate for controlled cocoa fermentation, potentially accelerating biochemical changes and improving process stability. Full article