Journal of microbiological methods最新文献

{"title":"Development of a multiplex PCR assay for rapid detection of Clostridium perfringens, Clostridium septicum and Paeniclostridium sordellii in necrotising soft tissue infections","authors":"Anjali Anil ,&nbsp;Tanvi Vashist ,&nbsp;Divya Nair ,&nbsp;Kamlesh Bisht ,&nbsp;Pallab Ray ,&nbsp;Cherring Tandup ,&nbsp;Karthick Rangasamy ,&nbsp;Archana Angrup","doi":"10.1016/j.mimet.2026.107420","DOIUrl":"10.1016/j.mimet.2026.107420","url":null,"abstract":"<div><h3><u>Background</u></h3><div>Necrotising soft tissue infections (NSTIs) are fulminant infections requiring rapid diagnosis and intervention. Conventional methods, such as anaerobic culture, are slow and insensitive for clostridial pathogens.</div></div><div><h3>Objectives</h3><div>To develop and evaluate a multiplex PCR assay targeting three clinically relevant clostridia directly in NSTI samples.</div></div><div><h3>Methods</h3><div>A multiplex conventional PCR targeting the <em>cpa</em> (<em>Clostridium perfringens</em>), <em>csa</em> (<em>Clostridium septicum</em>), and 16S rRNA (<em>Paeniclostridium sordellii</em>) genes was optimised and applied to tissue/pus samples from 87 clinically diagnosed NSTI patients. Anaerobic culture served as the reference. Sensitivity, specificity, and diagnostic turnaround times were compared.</div></div><div><h3>Results</h3><div>Seven patients (8.0%, 95% CI 3.3–16.2) were positive for clostridia by PCR, compared with four (4.6%, 95% CI 1.3–11.4) by culture. PCR identified three additional culture-negative cases. While McNemar's test did not show statistical significance (<em>p</em> = 0.25), PCR achieved a sensitivity of 100% (95% CI 39.8–100.0), specificity of 96.4% (95% CI 89.8–99.2), and a negative predictive value of 100%. Turnaround time was 3–4 h, compared with 48–96 h for culture. Both patients with monomicrobial <em>C. septicum</em> NSTI succumbed rapidly, highlighting its prognostic importance.</div></div><div><h3>Conclusion</h3><div>This is the first report of a multiplex PCR assay for direct detection of <em>C. perfringens, C. septicum,</em> and <em>P. sordellii</em> in NSTIs. The assay enhances detection, reduces turnaround time, and provides clinically relevant species-level identification. Multiplex PCR may serve as a valuable adjunct to culture in early NSTI diagnostics and could be expanded to include additional anaerobic pathogens.</div></div>","PeriodicalId":16409,"journal":{"name":"Journal of microbiological methods","volume":"243 ","pages":"Article 107420"},"PeriodicalIF":1.9,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146157375","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Determination of bacteria-specific odor signatures using low-cost metal oxide gas sensors","authors":"Emre Yavuzer ,&nbsp;Mahmut Yılmaz","doi":"10.1016/j.mimet.2026.107429","DOIUrl":"10.1016/j.mimet.2026.107429","url":null,"abstract":"<div><div>In this study, a feasibility assessment was conducted to evaluate the potential of commercially available low-cost gas sensors for differentiating bacterial species inoculated into fish muscles. The signals were obtained by determining how low-cost gas sensors detected the volatile organic compound (VOC) patterns produced by each bacterial species (<em>Enterococcus faecalis, Pseudomonas luteola, Proteus mirabilis</em>, and <em>Photobacterium damselae</em>) in trout tissue. It was observed that MQ3 and MQ4 sensors yielded high rates for all bacteria, indicating a strong increase in broad VOC load consistent with the known cross-sensitivity range of MQ sensors as a result of bacterial metabolism. In addition to cumulative VOC patterns, time-dependent derivative analysis (dVOC/dt) was applied to reveal bacterium-specific metabolic differences in more detail. Thus, it was determined that <em>Photobacterium damselae</em>, unlike other bacteria, exhibited suppressed VOC production kinetics in the early stages, demonstrating that dynamic VOC fingerprints are a powerful tool for bacterial differentiation. The results demonstrate the feasibility of dynamic VOC fingerprinting using minimal sensor configurations for exploratory bacterial discrimination.</div></div>","PeriodicalId":16409,"journal":{"name":"Journal of microbiological methods","volume":"243 ","pages":"Article 107429"},"PeriodicalIF":1.9,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146147466","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"KguS/KguR two-component system as a molecular marker for E. coli strains associated with chronic urinary tract infections (cUTI)","authors":"Ricardo E. Ahumada-Cota ,&nbsp;Ulises Hernández-Chiñas ,&nbsp;Brenda Recillas-Farfán ,&nbsp;Armando Navarro-Ocaña ,&nbsp;José Molina-López ,&nbsp;María G. Balbuena-Alonso ,&nbsp;María E. Chávez-Berrocal ,&nbsp;Carlos A. Eslava-Campos","doi":"10.1016/j.mimet.2026.107421","DOIUrl":"10.1016/j.mimet.2026.107421","url":null,"abstract":"<div><div>The identification of key genes responsible for the colonization and infection of <em>E. coli</em> strains associated with diarrhea (DEC) has made significant progress. Unfortunately, for uropathogenic <em>E. coli</em> (UPEC) strains, there are currently no defined genes that contribute to the identification of strains associated with urinary tract infections. This paper presents data regarding genes associated with the virulence and adaptability of UPEC strains isolated from urinary tract infections (UTI), aiming to evaluate the possibility of identifying a molecular marker for this pathotype. We analyzed 166 <em>E. coli</em> strains (90 recovered from urine and 76 isolated from feces) characterized by serotyping, phylogenetic analysis, and PCR detection of <em>fimH</em>, <em>iutA</em>, <em>fyuA</em>, <em>feoB</em>, <em>ompT</em>, <em>kguS</em> and <em>kguR</em>, genes associated with the virulence and metabolism of UPEC. Herein, results showed a higher prevalence of <em>kguS/kguR</em> combination in strains isolated from persistent UTI (peUTI), which correlated with strains of classic UPEC serogroups, a higher virulence gene load, and inclusion in pathogenic phylogroups (B2 and D). The presence of the genes in strains isolated from feces showed that the <em>kguS/kguR</em> presence was minimal (14%), while the presence of <em>kguR</em> was almost inexistent (1%), contrary to <em>kguS</em> (39%). The results obtained suggest that the presence of the <em>kguS/kguR</em> pair encoding the homonymous two-component system (TCS) plays an important role in the adaptation and colonization of the urinary tract by <em>E. coli</em> strains that cause UTI. In conclusion, the combination of <em>kguS/kguR</em> could be used as a molecular marker to identify pathogenic strains with a high capacity for adaptation to the urinary environment.</div></div>","PeriodicalId":16409,"journal":{"name":"Journal of microbiological methods","volume":"243 ","pages":"Article 107421"},"PeriodicalIF":1.9,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146142707","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Production of semi-dry wine using cape gooseberry under very high gravity conditions with preserved antioxidant properties","authors":"Somil Kumar ,&nbsp;Naveen Malik ,&nbsp;Akhilesh Kumar ,&nbsp;Sunil Kumar ,&nbsp;Vikram Kumar ,&nbsp;Umesh kumar Dwivedi ,&nbsp;Rajesh Sharma ,&nbsp;Ajay Bhardwaj ,&nbsp;Sudarshan Singh Lakhawat ,&nbsp;Pushpender Kumar Sharma","doi":"10.1016/j.mimet.2026.107418","DOIUrl":"10.1016/j.mimet.2026.107418","url":null,"abstract":"<div><div>In this study, we have reported production of a semi-dry fruit wine using <em>Cape Gooseberry</em> for the first time using very high gravity conditions at laboratory scale. Employing industrial strain <em>Saccharomyces cerevisiae,</em> the batch fermentation was carried out at room temperature in 1-l flasks in triplicates. The moisture content of fruit was calculated to be 80% after 24 days. The °Bx of raw fruit juice was found to be 4.9 and was raised to 25°Bx on addition of sucrose, the yeast assimilable nitrogen content of cape goose berry juice was calculated to be 280 mg N/L. Average fermentation time calculated based on cessation of bubbling and attainment of final °Bx of 3.8 ± 0.3 was found to be 21 days. The initial pH of raw juice was observed to be 4.51 ± 0.04 before fermentation, while post fermentation it was calculated to be 3.63 ± 0.03. The ethanol estimated using HPLC, GC-FID and refractometric analysis showed concentration of 13.29 ± 0.26, 13.15 ± 0.03 and 13.63 ± 0.374% respectively. The antioxidant activity of raw juice vs processed cape gooseberry wine was calculated to be 65.06% and 39.64% respectively, while it was estimated to be 53.86% in one-year-old preserved cape gooseberry wine sample. The titratable acidity and volatile acidity of wine was determined to be 6 g/l and 0.15 g/l respectively and was found to be well within the desired range. Our results provide a clear demonstration of feasibility of very high gravity fermentation for producing semi-dry Cape Gooseberry wine with moderate antioxidant preservation.</div></div>","PeriodicalId":16409,"journal":{"name":"Journal of microbiological methods","volume":"243 ","pages":"Article 107418"},"PeriodicalIF":1.9,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146142713","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Standardization and Comparative Evaluation of quick Inoculation Techniques for Screening Pigeonpea genotype Resistance to Phytophthora cajani","authors":"Pragati Nema ,&nbsp;Raj K. Mishra ,&nbsp;Sonika Pandey ,&nbsp;Rajani Sasode","doi":"10.1016/j.mimet.2026.107419","DOIUrl":"10.1016/j.mimet.2026.107419","url":null,"abstract":"<div><div>Phytophthora stem blight (PSB), caused by <em>Phytophthora cajani</em>, is a destructive disease of pigeonpea (<em>Cajanus cajan</em> L.) that can lead to complete crop loss under favorable conditions. Effective resistance breeding is constrained by the lack of reliable and reproducible screening methods. The present study aimed to standardize and compare artificial inoculation techniques for consistent induction of Phytophthora blight under controlled conditions. Ten inoculation methods targeting different infection pathways were evaluated in greenhouse pot experiments over two consecutive seasons (2023–24 and 2024–25) using susceptible (UPAS-120, ICP-7119, ICP-2376) and moderately resistant (KPBR-80-2-1, IPAC-79, IPAB-7-2-1-7) pigeonpea cultivars. Disease incidence was recorded to assess the efficiency and reproducibility of each technique. All methods successfully established infection; however, disease severity varied significantly among techniques and genotypes. The leaf-lamina inoculation method consistently produced the highest and most uniform disease incidence, recording 97.7% and 96.6% in the susceptible cultivar UPAS-120 and 83.3% and 86.6% in the moderately resistant cultivar KPBR-80-2-1 across the two seasons. Node inoculation emerged as the second most reliable method. The study identifies most efficient and quick, reproducible inoculation techniques that enable rapid and high-throughput resistance screening, providing a robust methodological framework to support pigeonpea breeding and Phytophthora blight management.</div></div>","PeriodicalId":16409,"journal":{"name":"Journal of microbiological methods","volume":"243 ","pages":"Article 107419"},"PeriodicalIF":1.9,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146142710","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Combining isolation chips with velvet transfer for high-throughput primary screening of antibiotic-producing soil-dwelling bacteria","authors":"Sabina D. Hidat ,&nbsp;Joan Masatu ,&nbsp;Nelson E. Masota ,&nbsp;Rogers Mwakalukwa ,&nbsp;Paul Malaba ,&nbsp;Joseph Sempombe ,&nbsp;Doreen Mloka","doi":"10.1016/j.mimet.2026.107397","DOIUrl":"10.1016/j.mimet.2026.107397","url":null,"abstract":"<div><div>Antimicrobial resistance (AMR) remains a significant threat to global health, requiring various mitigation strategies. Soil-dwelling microbes have long been a valuable source of novel antibiotics; however, their low cultivability using standard methods has often led to the discovery of known antibiotics. This bottleneck has been partially overcome by using isolation chips (iChips), which enhance the cultivability (both in terms of number and diversity) of soil bacteria. This has created a need to find new screening methods that allow high-throughput screening, are less resource-intensive, and can screen multiple test organisms.</div><div>To address this, soil samples were collected from three different regions of Tanzania, and numerous bacterial soil species colonies were grown in iChips and then transferred to master plates. Replica plates were created using a velvet transfer method and utilized for parallel agar overlay assays against <em>Escherichia coli</em>, <em>Klebsiella pneumoniae</em>, and <em>Staphylococcus aureus</em>. Master plates with 33–62 colonies showing variation in size, morphology, and spatial arrangement were produced by cultivating agar plugs from iChips. Three to four replicates, which preserved these characteristics at the transfer efficiencies of 91.3–100% were obtained without contamination. Zones of inhibition -to-colony diameter ratios ranging 1.15–3.27 were obtained following parallel primary screening against test strains, hence enabling the identification of potential antibiotic producers.</div><div>This is the first report of the combined iChip and velvet transfer workflow that offers a high-throughput, resource-efficient, and scalable method for primary screening of various soil-derived bacteria for antibacterial Natural Products (NPs), while also maintaining the master plates for downstream investigations.</div></div>","PeriodicalId":16409,"journal":{"name":"Journal of microbiological methods","volume":"242 ","pages":"Article 107397"},"PeriodicalIF":1.9,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146003742","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Microbial biofilms and nanoparticles synergistic interaction: A cutting-edge nano-bioremediation method for environmental cleanup and site restoration","authors":"Manoj Kumar ,&nbsp;Meera Goswami ,&nbsp;Deepak Pant ,&nbsp;Kulamani Parida ,&nbsp;Anand Giri","doi":"10.1016/j.mimet.2026.107406","DOIUrl":"10.1016/j.mimet.2026.107406","url":null,"abstract":"<div><div>In nature, bacteria have two different growth modes: a unicellular life stage, in which the cells are free-moving (planktonic), and a multicellular life stage wherein the cells are immobile and reside within a biofilm structure, composed of a complex network of extracellular polymeric substances (EPS). The EPS contribute to the distinctive characteristics of the biofilm's lifestyle, environmental degradation, as well as virulence and antibiotic resistance. The present review critically analyses the functional significance of extracellular polymeric substances (EPS), characteristics of biofilms and nanoparticle-biofilm interactions for the removal of emerging environmental contaminants. Nanoparticles can also function as emulsifiers, enhancing bioavailability by providing droplet surfaces that facilitate microbial attachment and promote microbial proliferation. The enhancement of certain biofilms through positive interactions with nanoparticles will enable the design of systems that facilitate the development of cost-effective alternatives of interest, including enhanced bioremediation, biodegradation, oil spill mitigation, and improved microbial fuel cell (MFC) performance. Nano-bioremediation is effective across a wide range of emerging environmental contaminants such as PAHs, chlorinated compounds, pHthalates, plastic heavy metals, etc. and provides a sustainable alternative to conventional remediation approaches.</div></div>","PeriodicalId":16409,"journal":{"name":"Journal of microbiological methods","volume":"242 ","pages":"Article 107406"},"PeriodicalIF":1.9,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146044304","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Strain-level typing of Candida albicans and Saccharomyces cerevisiae using Fourier transform infrared spectroscopy and whole-genome sequencing","authors":"Asuka Kashiwaba ,&nbsp;Asako Mitani ,&nbsp;Takumi Sonoda ,&nbsp;Naofumi Shigemune ,&nbsp;Hiroki Takahashi","doi":"10.1016/j.mimet.2026.107405","DOIUrl":"10.1016/j.mimet.2026.107405","url":null,"abstract":"<div><div>Rapid, low-cost strain-level typing is essential for source attribution and microbiological control in clinical and industrial settings. Fourier-transform infrared (FT-IR) spectroscopy using IR Biotyper method was previously optimized to achieve an identification level equivalent to that of whole-genome sequencing (WGS) discrimination of <em>Wickerhamomyces anomalus</em>. However, the species of yeast studied were limited; further, studies that have used the results of WGS as a reference for strain identification in yeast remain limited. Therefore, here, the generalizability of this approach to <em>Candida albicans</em> and <em>Saccharomyces cerevisiae</em> was assessed using publicly available whole-genome datasets as references. Single-nucleotide polymorphisms from 29<em>C. albicans</em> and 27 <em>S. cerevisiae</em> genomes were analyzed to build phylograms and select 5 and 4 mutually distant strains for FT-IR. FT-IR spectra were obtained after a standardized 24-h rotational culture in SD liquid medium, with three independent passages per strain. The same and similar strains clustered tightly, and different strains formed non-overlapping clusters; branches for the same and similar strains were &lt; 0.1 on IR dendrograms, whereas branches for distinct strains were &gt; 0.1. The discriminatory ability of FT-IR fully agreed with the WGS-based same/different classifications. Standardized liquid culture likely reduced cell-state heterogeneity, improving reproducibility. These results extend FT-IR strain typing beyond <em>W. anomalus</em>, providing a rapid, low-cost tool for contamination source attribution.</div></div>","PeriodicalId":16409,"journal":{"name":"Journal of microbiological methods","volume":"242 ","pages":"Article 107405"},"PeriodicalIF":1.9,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146044238","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A simple preservation method for the short-medium term storage of environmental bacteria","authors":"John E. Moore ,&nbsp;Beverley Cherie Millar","doi":"10.1016/j.mimet.2026.107409","DOIUrl":"10.1016/j.mimet.2026.107409","url":null,"abstract":"<div><div>A novel method is described for the short to medium term storage of environmental bacteria up to six months. Fifty environmental bacteria were employed in this study, which had been previously isolated from green spaces. These organisms comprised of 14 genera (10 Gram negative and 4 Gram positive) [<em>Arthrobacter, Bacillus, Buttiauxella, Carnobacterium, Citrobacter, Enterobacter, Lelliottia, Pectobacterium, Pseudomonas, Rahnella, Serratia, Sporosarcina, Stenotrophomonas, Yersinia</em>] and 16 identified species. Organisms were inoculated onto fresh plates of Standard Plate Count (SPC) agar and incubated aerobically at 30 °C for 72 h. Following this, plates with growth were stacked in metal plate racks and wrapped in plastic with a large autoclave bag and sealed with a cable tie. Plates were stored at 5 °C in the dark for six months, after which they were removed and the colonies checked for viability. All isolates were able to be recovered through passaging onto fresh SPC plates. This method is simple, inexpensive, versatile and widely adaptable and requires minimum preparation/handling/processing, utilising a small laboratory footprint for refrigerator space. Adoption and employment of this simple storage system encourages the routine archiving of environmental isolates for future research purposes.</div></div>","PeriodicalId":16409,"journal":{"name":"Journal of microbiological methods","volume":"242 ","pages":"Article 107409"},"PeriodicalIF":1.9,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146100264","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Optimization of physicochemical and antimicrobial properties of functional kombucha beverage sweetened with dried fruits using simplex centroid mixture and Plackett-Burman designs","authors":"Lamia Ayed ,&nbsp;Haifa Dhif ,&nbsp;Souhir Torjmèn ,&nbsp;Kamel Chaieb","doi":"10.1016/j.mimet.2026.107408","DOIUrl":"10.1016/j.mimet.2026.107408","url":null,"abstract":"<div><div>Traditional foods and beverages represent alternative strategies to counteract bacterial virulence. The fermentation of tea, sugar, supplemented with a symbiotic culture of bacteria and yeast (SCOBY) produces kombucha beverage, which offers several health advantages and is similar to soft drinks. This study investigates the effect of various fermentation factors on the growth levels of <em>Lactobacillus</em>, <em>Lactococcus</em>, total phenolic content and antimicrobial activities of Green Tea, Black Tea and Moringa kombucha beverage (GTBTMK) prepared with a starter SCOBY culture. Formulation of kombucha beverage was optimized using a Simplex-Centroid Mixture Design and Plackett-Burman Design. The best optimized kombucha formulation (18.98 g/L; Dried Apricot was 20 g/L; Dried plum was 10 g/L, Dried Grape was 20 g/L,11.763 g/L Green tea, 0.01238 g/L Black tea and 8.1127 g/L Moringa) contain a high phenolic content of 86.79 mg GAE/mL and exhibited a significant antimicrobial activity against <em>Bacillus cereus</em> ATCC 11778, <em>Micrococcus luteus</em> NCIMB 8166 and <em>Enterococcus faecalis</em> ATCC 29212, <em>Candida albicans</em> ATCC 90028, <em>Cryptococcus neoformans</em> ATCC 14116, <em>Aspergillus brasiliensis</em> ATCC 16404 (Inhibition zone more than 20 mm). This research provides new insights into the development of innovative, functional kombucha beverage potentially expanding the spectrum of health-promoting fermented drinks available to consumers.</div></div>","PeriodicalId":16409,"journal":{"name":"Journal of microbiological methods","volume":"242 ","pages":"Article 107408"},"PeriodicalIF":1.9,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146064144","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}