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Appl. Microbiol., Volume 6, Issue 1 (January 2026) – 20 articles

Cover Story (view full-size image): Conventional and organic agriculture can both cause soil microbial community structure (SMCS) destruction, infertility, and abandonment. The application of soil productivity-improving biofertilizers is a sustainable practice that requires holistic knowledge, including complex biointeractions, diverse microbial metabolism, and culture requirements, the last of which rely on methodology and technology. In this study, a holistic culture-based and meta-analysis approach was employed to explore pristine and domesticated soils for presumptive plant growth-promoting (PGP) bacteria. Various soil samples were logistically acquired and processed using enrichment and heat alternatives. Morphologically diverse isolates were streak-purified and analyzed for 16S rRNA bacterial identification. Meta-analysis of PGP bacteria in domesticated environments was conducted using Google Search and NCBI PubMed. View this paper
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26 pages, 4076 KB  
Article
Genetic Determinants Associated with Persistence of Listeria Species and Background Microflora from a Dairy Processing Environment
by Vaishali Poswal, Sanjeev Anand, Jose L. Gonzalez-Hernandez and Brian Kraus
Appl. Microbiol. 2026, 6(1), 20; https://doi.org/10.3390/applmicrobiol6010020 - 21 Jan 2026
Cited by 2 | Viewed by 1057
Abstract
Listeria monocytogenes is a persistent foodborne pathogen capable of surviving in food processing environments, often in association with diverse environmental microflora. This study examines genomic determinants of persistence, specifically stress adaptation and biofilm-associated traits, in environmental Listeria species and other environmental microflora from [...] Read more.
Listeria monocytogenes is a persistent foodborne pathogen capable of surviving in food processing environments, often in association with diverse environmental microflora. This study examines genomic determinants of persistence, specifically stress adaptation and biofilm-associated traits, in environmental Listeria species and other environmental microflora from a dairy processing facility by analyzing whole-genome sequences of 6 environmental Listeria isolates, 4 ATCC reference strains, and 22 air and floor swab cultures, annotated using the RAST platform. Subsystem analysis revealed that Listeria isolates carried a defined set of genes linked to biofilm formation, antimicrobial resistance, and stress response, though in lower abundance than environmental cultures. Listeria exhibited fewer flagellar genes but greater consistency in core stress-related genes, including those for disinfectant and osmotic stress resistance, with SigB operon and RpoN genes highlighting strong stress tolerance. In contrast, environmental cultures exhibited broader transcriptional regulators (RpoE, RpoH) and greater diversity in acid and heat shock response genes, indicating distinct survival strategies. All examined Listeria species harbor biofilm and stress-resistance genes enabling independent survival, while environmental microbiota show greater genetic diversity that may promote persistence and multispecies biofilm formation. This study underscores the complex genetic landscape that may contribute to the persistence of Listeria and environmental microbiota in dairy processing environments, providing foundational insights for environmental cross contamination control strategies. Full article
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20 pages, 923 KB  
Review
Practical Insights and Emerging Trends for Strategic Cloning of Large Biosynthetic Gene Clusters from Bacteria
by Louise Davison, Zoë Alice Bell and Hong Gao
Appl. Microbiol. 2026, 6(1), 19; https://doi.org/10.3390/applmicrobiol6010019 - 21 Jan 2026
Viewed by 1772
Abstract
Cloning large biosynthetic gene clusters (BGCs) is fundamental to unlocking microbial natural product potential for drug discovery and biotechnology. These clusters encode diverse bioactive compounds, but their size, high GC content, and complex architecture pose significant technical challenges. This review scrutinises recent advances [...] Read more.
Cloning large biosynthetic gene clusters (BGCs) is fundamental to unlocking microbial natural product potential for drug discovery and biotechnology. These clusters encode diverse bioactive compounds, but their size, high GC content, and complex architecture pose significant technical challenges. This review scrutinises recent advances in BGC cloning strategies, categorising them into three major groups: (1) direct release-and-capture methods, (2) genome-integrated preconditioning systems, and (3) CRISPR-assisted hybrid platforms. This review compares the strengths, limitations, and reported efficiencies of BGC cloning strategies, highlighting trade-offs in precision, scalability, and workflow complexity. Emerging trends, such as AI-driven genome mining, modular synthetic biology toolkits, and high-throughput automation, are reshaping the cloning landscape, enabling predictive design and streamlined assembly of clusters exceeding 100 kb. By integrating comparative analysis with future perspectives, this review provides outlines on how next-generation strategies will accelerate heterologous expression, natural product discovery, and sustainable biomanufacturing. Full article
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21 pages, 4039 KB  
Article
16S rRNA Metagenomic Profiling Reveals Diet-Induced Shifts in Gut Microbial Diversity and Taxonomic Structure in Guinea Pigs
by José Cantaro Segura, Héctor Cántaro-Segura and Raul Blas
Appl. Microbiol. 2026, 6(1), 18; https://doi.org/10.3390/applmicrobiol6010018 - 20 Jan 2026
Cited by 1 | Viewed by 2035
Abstract
Diet plays a pivotal role in shaping the gut microbiota, influencing host physiology, immune function, and nutrient metabolism. In this study, we evaluated the impact of three distinct feeding systems—Forage only, Balanced feed only, and Mixed system—on the cecal microbiota of guinea pigs [...] Read more.
Diet plays a pivotal role in shaping the gut microbiota, influencing host physiology, immune function, and nutrient metabolism. In this study, we evaluated the impact of three distinct feeding systems—Forage only, Balanced feed only, and Mixed system—on the cecal microbiota of guinea pigs (Cavia porcellus) using 16S rRNA gene amplicon sequencing in a randomized allocation of 18 males across the three diets (n = 6 per group) over 7 weeks. A total of 2,135,852 high-quality reads were obtained, with rarefaction curves and Good’s coverage confirming sufficient sequencing depth. Alpha diversity indices revealed significantly higher microbial richness and evenness in the mixed group, while beta diversity analyses demonstrated distinct microbial community structures across diets. Taxonomic profiling showed that forage-based diets enriched fiber-degrading genera such as Fibrobacter and Treponema, whereas the Balanced feed group favored mucin- and protein-degrading bacteria like Akkermansia and Bacteroides. LEfSe and t-test analyses identified several biomarkers and diet-specific genera, suggesting functional divergence in microbial metabolism. Forage-fed animals showed microbiota associated with short-chain fatty acid production and enhanced fiber utilization, while the Balanced feed group showed microbial traits linked to mucin degradation and potential gut barrier disruption. These findings highlight the strong influence of dietary composition on gut microbial ecology and suggest that fiber-rich diets promote a more diverse and functionally beneficial cecal microbiome in guinea pigs. Full article
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43 pages, 1794 KB  
Review
Microbial Fermentation: A Sustainable Strategy for Producing High-Value Bioactive Compounds for Agriculture, Animal Feed, and Human Health
by Victor Eduardo Zamudio-Sosa, Luis Angel Cabanillas-Bojórquez, Evangelina García-Armenta, Marilyn Shomara Criollo-Mendoza, José Andrés Medrano-Felix, Alma Haydee Astorga-Gaxiola, José Basilio Heredia, Laura Aracely Contreras-Angulo and Erick Paul Gutiérrez-Grijalva
Appl. Microbiol. 2026, 6(1), 17; https://doi.org/10.3390/applmicrobiol6010017 - 18 Jan 2026
Cited by 3 | Viewed by 4002
Abstract
Microbial fermentation is a key biotechnological tool for producing bioactive metabolites such as alkaloids, carotenoids, essential oils, and phenolic compounds, among others, with applications in human health, agriculture, and food industries. This review comprehensively reviews recent information on the synthesis of valuable compounds [...] Read more.
Microbial fermentation is a key biotechnological tool for producing bioactive metabolites such as alkaloids, carotenoids, essential oils, and phenolic compounds, among others, with applications in human health, agriculture, and food industries. This review comprehensively reviews recent information on the synthesis of valuable compounds and enzymes through fermentation processes. Here, we discuss the advantages of the different types of fermentation, such as submerged and solid-state fermentation, in optimizing metabolite production by bacteria, fungi, and yeast. The role of microbial metabolism, enzymatic activity, and fermentation conditions in enhancing the bioavailability and functionality of these compounds is discussed. Integrating fermentation with emerging biotechnologies, including metabolic engineering, further enhances yields and specificity. The potential of microbial-derived bioactive compounds in developing functional foods, pharmaceuticals, and eco-friendly agricultural solutions positions fermentation as a pivotal strategy for future biotechnological advancements. Therefore, microbial fermentation is a sustainable tool to obtain high-quality metabolites from different sources that can be used in agriculture, animal, and human health. Full article
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22 pages, 956 KB  
Article
Growth of Listeria monocytogenes in Goat’s Pasteurized Milk Cheese During Maturation: Its Prediction from a Milk Model Medium
by Yara Loforte, Mariem Zanzan, André Martinho de Almeida, Vasco Cadavez and Ursula Gonzales-Barron
Appl. Microbiol. 2026, 6(1), 16; https://doi.org/10.3390/applmicrobiol6010016 - 16 Jan 2026
Viewed by 1271
Abstract
Previous research showed that a strain of Leuconostoc mesenteroides, isolated from goat’s raw milk cheese, was effective in slowing down the growth and reducing the maximum concentration of L. monocytogenes when evaluated in a milk model; furthermore, the extent of inhibition was [...] Read more.
Previous research showed that a strain of Leuconostoc mesenteroides, isolated from goat’s raw milk cheese, was effective in slowing down the growth and reducing the maximum concentration of L. monocytogenes when evaluated in a milk model; furthermore, the extent of inhibition was dependent on the milk’s initial pH. The objectives of this study were as follows: (1) to determine whether the growth of L. monocytogenes in goat’s pasteurized milk cheese during maturation could be approximated from growth data obtained in the milk model medium, either in monoculture or in coculture with L. mesenteroides, and if so, (2) to model a milk-to-cheese conversion factor (Cf) for L. monocytogenes growth rate. Challenge tests were conducted by inoculating L. monocytogenes in monoculture and in coculture with L. mesenteroides in goat’s pasteurized milk adjusted at initial pH levels of 5.5, 6.0, and 6.5. The process of cheesemaking continued, and cheeses were ripened at 12 °C for 12 days. Each experimental growth curve was adjusted to a pH-driven dynamic model where the microbial maximum growth rate is a function of pH. As observed in the milk model medium, in coculture with L. mesenteroides, the optimum growth rate (μopt) of L. monocytogenes in maturing cheese was affected by the initial pH of milk: the lowest rate of 0.863 ± 0.042 day−1 was obtained at the initial pH 5.5, in comparison to 1.239 ± 0.208 and 1.038 ± 0.308 day−1 at pH 6.0 and 6.5, respectively. Regardless of the milk’s initial pH, L. mesenteroides did not reduce the maximum load of L. monocytogenes in maturing cheeses, as it did in the milk medium. On the contrary, at the milk’s initial pH of 5.5, 6.0, and 6.5, L. mesenteroides was able to decrease, on average, 2.2-fold, 1.5-fold, and 1.9-fold the μopt of L. monocytogenes in both milk medium and cheese, without significant differences between matrices. Following such validation in goat’s cheese, the square root of milk-to-cheese Cf for L. monocytogenes was estimated as 0.751 (SE = 0.0108), and the type of culture (monoculture and coculture) was not found to affect Cf (p = 0.320). In conclusion, this work validated the pre-acidification of milk as an efficient strategy that, when combined with the use of a protective culture, can synergically enhance the control of L. monocytogenes in cheese. Full article
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14 pages, 1715 KB  
Article
Using Phytoplankton as Bioindicators of Tourism Impact and Seasonal Eutrophication in the Andaman Sea (Koh Yaa, Thailand)
by Tassnapa Wongsnansilp, Manoch Khamcharoen, Jaran Boonrong and Wipawee Dejtisakdi
Appl. Microbiol. 2026, 6(1), 15; https://doi.org/10.3390/applmicrobiol6010015 - 13 Jan 2026
Cited by 1 | Viewed by 1040
Abstract
This study focuses on the diversity of phytoplankton in the Koh Yaa region of Thailand and their relationship with environmental variables, aiming to assess whether human activities (primarily tourism) pose potential threats to the marine ecosystem and provide scientific support for eco-sustainable tourism [...] Read more.
This study focuses on the diversity of phytoplankton in the Koh Yaa region of Thailand and their relationship with environmental variables, aiming to assess whether human activities (primarily tourism) pose potential threats to the marine ecosystem and provide scientific support for eco-sustainable tourism management decisions in the region. In April, August, and December 2024, corresponding to peak season, off-season, and shoulder season, a total of 156 discrete samples were collected from four coastal sites to analyze water quality parameters such as temperature, pH, total nitrogen (TN), and total phosphorus (TP), along with plankton diversity and abundance. Statistical analyses including two-way ANOVA with Duncan’s Multiple Range Test (DMRT), Pearson correlation analysis, and principal component analysis (PCA) were applied. The results showed a declining trend in plankton abundance over time, peaking at 1009 × 106 cells/m3 in April and dropping to 281 × 106 cells/m3 by December. A total of 15 types of phytoplankton were identified across four phyla: Bacillariophyta, Cyanobacteria, Dinoflagellata, and Chlorophyta. Notably, Chaetoceros from Bacillariophyta accounted for 47% of phytoplankton, while Oscillatoria from Cyanobacteria made up 29.6%. The diversity index and evenness index improved from 1.34 and 0.46 in April to 1.88 and 0.64 in December, respectively. Environmental factors like pH, temperature, and TP significantly affected phytoplankton abundance (p < 0.01), with TP levels ranging from 0.27 to 0.69 mg/L. These results indicate possible pollution in this region, and changes in phytoplankton abundance were linked to seasonal climate variations—especially during peak tourist seasons—which may exacerbate eutrophication affecting community structures. Full article
(This article belongs to the Topic Environmental Bioengineering and Geomicrobiology)
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18 pages, 1082 KB  
Article
Lactic Acid Bacteria from Traditional Fermented Milk: Antimicrobial Potential Against Foodborne Pathogens
by Kaoutar Boussif, Ahmed Elidrissi, Abdelkhaleq Elmoslih, Youssef Ezzaky, Mariem Zanzan and Fouad Achemchem
Appl. Microbiol. 2026, 6(1), 14; https://doi.org/10.3390/applmicrobiol6010014 - 13 Jan 2026
Cited by 3 | Viewed by 1563
Abstract
Lactic acid bacteria (LAB) are increasingly recognized for their role in food biopreservation due to their ability to synthesize antimicrobial compounds. Milk naturally harbors a wide variety of LAB, offering a promising source for identifying strains with biopreservative potential. This study investigated the [...] Read more.
Lactic acid bacteria (LAB) are increasingly recognized for their role in food biopreservation due to their ability to synthesize antimicrobial compounds. Milk naturally harbors a wide variety of LAB, offering a promising source for identifying strains with biopreservative potential. This study investigated the antagonistic effects, safety characteristics, and technological properties of LAB strains isolated from traditionally fermented milk. Thirty-two dairy samples were analyzed, and the resulting LAB isolates were screened for inhibitory activity against Listeria monocytogenes CECT 4032 and Staphylococcus aureus CECT 976 using agar spot and well diffusion assays. All tested strains exhibited strong antimicrobial effects, with particularly notable inhibition of L. monocytogenes. After phenotypic screening, five representative isolates were selected for molecular identification and further assessment of safety-related attributes, functional capabilities, auto- and co-aggregation properties. 16S rRNA gene sequencing revealed that four strains belonged to the genus Enterococcus, specifically, one E. faecium and three E. durans, while one was classified as a Lactococcus species. Moreover, none of the strains showed proteolytic or lipolytic activities which highlights their potential use in dairy fermentation processes. Full article
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22 pages, 3544 KB  
Article
Advancing Sustainable Wheat Production in the Andes Through Biofertilization with AzospirillumTrichoderma and Fermented Anchovy-Based Under Rainfed Conditions
by Edwin Villegas, Fernando Escobal, Toribio Tejada, Peter Piña, Hector Cántaro-Segura, Luis Diaz-Morales and Daniel Matsusaka
Appl. Microbiol. 2026, 6(1), 13; https://doi.org/10.3390/applmicrobiol6010013 - 13 Jan 2026
Viewed by 1175
Abstract
Wheat (Triticum aestivum L.) sustains global caloric intake, but its productivity in Andean highlands is constrained by soil fertility and input reliance. This study represents one of the first field-based evaluations of biofertilizers under high-altitude, rainfed Andean conditions, addressing a major knowledge [...] Read more.
Wheat (Triticum aestivum L.) sustains global caloric intake, but its productivity in Andean highlands is constrained by soil fertility and input reliance. This study represents one of the first field-based evaluations of biofertilizers under high-altitude, rainfed Andean conditions, addressing a major knowledge gap in low-input mountain agroecosystems. This study evaluated three seed-applied biofertilizers—Azospirillum brasilense, Trichoderma viride (Trichomax), and an anchovy (Engraulis ringens) based liquid biofertilizer, compared with an untreated control and a soil-test mineral fertilization benchmark in rainfed wheat (Triticum aestivum L.) cv. INIA 405 in the central Andes of Peru. A 5 × 5 Latin square design (25 plots) was established under farmer-realistic conditions. At physiological maturity (Zadoks 9.5), plant height, spike length, grains per spike, thousand-grain weight, test weight, root dry mass, and grain yield were recorded. Mineral fertilization achieved the highest yield (1.20 ± 0.79 t ha−1), nearly doubling the control (0.60 ± 0.47 t ha−1). Notably, A. brasilense delivered an intermediate yield of 0.90 ± 0.64 t ha−1, representing a 50% increase over the control—accompanied by a marked rise in root dry mass. T. viride and the anchovy-based input yielded 0.85 ± 0.59 and 0.81 ± 0.59 t ha−1, respectively. Grain physical quality remained stable across treatments (thousand-grain weight ≈ 42 g; test weight 68–75 kg hL−1). Trait responses were complementary: root dry mass increased with mineral fertilization and A. brasilense, whereas spike length increased with mineral fertilization and the anchovy-based input. Overall, the evidence supports biofertilizers, particularly A. brasilense, as effective complements that enable partial fertilizer substitution within integrated nutrient-management strategies for sustainable wheat production in Andean rainfed systems. Full article
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32 pages, 9835 KB  
Article
Microbial Synergy Between Azospirillum brasilense and Glomus iranicum Promotes Root Biomass and Grain Yield in Andean Quinoa Cultivars
by Miriam Gutierrez, Eugenia Quispe-Medina, Cayo García-Blásquez Morote, José Antonio Quispe-Tenorio, Héctor Cántaro-Segura, Luis Díaz-Morales and Daniel Matsusaka
Appl. Microbiol. 2026, 6(1), 12; https://doi.org/10.3390/applmicrobiol6010012 - 13 Jan 2026
Cited by 1 | Viewed by 1453
Abstract
Quinoa (Chenopodium quinoa Willd.) is a strategic crop for climate-smart agriculture in the Andes, yet yield gains are constrained by soil degradation and low-input systems. We tested whether synergistic bioinoculation with a plant growth-promoting rhizobacterium (Azospirillum brasilense) and an arbuscular [...] Read more.
Quinoa (Chenopodium quinoa Willd.) is a strategic crop for climate-smart agriculture in the Andes, yet yield gains are constrained by soil degradation and low-input systems. We tested whether synergistic bioinoculation with a plant growth-promoting rhizobacterium (Azospirillum brasilense) and an arbuscular mycorrhizal fungus (Glomus iranicum var. tenuihypharum) enhances root function and grain productivity under field conditions. A split-plot RCBD was conducted in Ayacucho, Peru (2735 m a.s.l.) using four cultivars, Blanca de Junín (BJ), INIA 441 Señor del Huerto (SH), INIA 415 Pasankalla (RP) and INIA 420 Negra Collana (NC) and four treatments: uninoculated control, Azospirillum, Glomus and co-inoculation. Vegetative, root and yield traits were quantified; ANOVA, Tukey/Dunnett contrasts, correlations and PCA were applied. Co-inoculation consistently outperformed single inoculants, increasing root diameter, length, branching, dry weight and volume dry weight, while also enlarging panicle dimensions and raising grain weight per panicle and thousand-seed weight. Grain yield reached 4.94 ± 0.59 t ha−1 under co-inoculation, almost triple that of the control (1.71 ± 0.28 t ha−1) and about 1.5 times higher than single inoculations. Genotypic effects were pronounced; BJ and SH combined superior root biomass with higher yield, RP maximized grain size and hectoliter weight, whereas NC responded weakly. Significant genotype × treatment interactions indicated cultivar-dependent microbiome benefits. Correlation and PCA linked root biomass and stem/panicle architecture to yield formation, positioning co-inoculation along trait vectors associated with belowground vigor and productivity. These results demonstrate a robust microbial synergy that translates root gains into yield, supporting co-inoculation as a scalable, low-input strategy for sustainable intensification of quinoa in highland agroecosystems. Full article
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25 pages, 1850 KB  
Article
Recovery, Identification, and Presumptive Agricultural Application of Soil Bacteria
by Guadalupe Steele, Andrew K. Rindsberg and Hung King Tiong
Appl. Microbiol. 2026, 6(1), 11; https://doi.org/10.3390/applmicrobiol6010011 - 9 Jan 2026
Viewed by 1263
Abstract
Conventional and organic agriculture can both cause soil microbial community structure (SMCS) destruction, infertility, and abandonment. The application of soil productivity-improving biofertilizers is a sustainable practice that requires holistic knowledge, including complex biointeractions, diverse microbial metabolism, and culture requirements, the last of which [...] Read more.
Conventional and organic agriculture can both cause soil microbial community structure (SMCS) destruction, infertility, and abandonment. The application of soil productivity-improving biofertilizers is a sustainable practice that requires holistic knowledge, including complex biointeractions, diverse microbial metabolism, and culture requirements, the last of which rely on methodology and technology. In this study, a holistic culture-based and meta-analysis approach was employed to explore pristine and domesticated soils for presumptive plant growth-promoting (PGP) bacteria. Various soil samples were logistically acquired and processed using enrichment and heat alternatives. Morphologically diverse isolates were streak-purified and analyzed for 16S rRNA bacterial identification. Meta-analysis of PGP bacteria in domesticated environments was conducted using Google Search and NCBI PubMed. Soil fertility was analyzed for the pH and nitrogen/phosphorus/potassium (NPK) contents using biochemical tests. Notably, 7 genera and 15 species were differentially recovered, with Bacillus being the most prevalent and diverse in species. Conversely, Aeromonas, Lactobacillus, Lelliottia, Pseudomonas, and Staphylococcus were found only in pristine soil. While soil pH was consistent in all pristine soil samples, NPK contents ranged widely across the pristine (i.e., P/K) and domesticated samples (i.e., N/P/K). These findings could enhance biofertilizer SMCS, function, and effectiveness in the agricultural productivity needed to feed the expanding population. Full article
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22 pages, 1078 KB  
Review
In Vitro Models for Emerging Infectious Disease Detection and Host–Pathogen Interaction Studies
by Said Ezrari, Abdessamad Ikken, Oussama Grari, Mohamed Ou-zine, Mohammed Lahmer, Abderrazak Saddari and Adil Maleb
Appl. Microbiol. 2026, 6(1), 10; https://doi.org/10.3390/applmicrobiol6010010 - 7 Jan 2026
Viewed by 2258
Abstract
Many emerging and re-emerging infectious diseases have been observed over the last few decades around the globe due to population growth, international travel, environmental changes, and microbial adaptation and evolution, despite advances in the medical field. The spread of these diseases is related [...] Read more.
Many emerging and re-emerging infectious diseases have been observed over the last few decades around the globe due to population growth, international travel, environmental changes, and microbial adaptation and evolution, despite advances in the medical field. The spread of these diseases is related to complex interactions between pathogens and their hosts. Accordingly, this review summarises current knowledge on infection development and discusses methods used for detection and modeling. Recent studies have revealed the limitations of two-dimensional models and increasingly rely on 3D systems, including spheroids, organoids, and organ-on-a-chip systems, that offer more realistic tissue environments, allowing researchers to more effectively study host–pathogen interactions. Overall, the integration of complementary approaches and the development of 3D models are crucial for enhancing diagnosis, developing new therapeutic approaches, and strengthening control strategies of emerging outbreaks. Full article
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16 pages, 1688 KB  
Article
Effect of Trichoderma atroviride Application on Tea Yield and Its Impact on the Soil Microbiome in a New Zealand Tea Plantation
by Prashansani M. D. Silva, Travis R. Glare, John Graham Hampton, Diwakar R. W. Kandula and Josefina Narciso
Appl. Microbiol. 2026, 6(1), 9; https://doi.org/10.3390/applmicrobiol6010009 - 4 Jan 2026
Viewed by 1403
Abstract
New Zealand’s only tea (Camellia sinensis) plantation supplies a niche market for organically produced high value tea but faces challenges from climatic conditions and the decision to use only organic production methods. Fungi from the genus Trichoderma have been commercialised in [...] Read more.
New Zealand’s only tea (Camellia sinensis) plantation supplies a niche market for organically produced high value tea but faces challenges from climatic conditions and the decision to use only organic production methods. Fungi from the genus Trichoderma have been commercialised in New Zealand and elsewhere as disease-suppressing and plant growth-promoting agents. However, the potential benefits of using Trichoderma as a microbial biostimulant for tea cultivation have not been investigated in New Zealand. The ability of T. atroviride application to stimulate tea plant growth at a tea plantation was investigated over one year of production. The study involved foliar application of the biostimulant either once, twice or three times, one month apart, using 12 g of a commercially formulated spore mixture of four strains of T. atroviride per 5 m2 of experimental plots. Treatment with T. atroviride significantly increased tea yield by between 17% and 28% compared to the control over the harvesting season, but there were no statistically significant yield differences among the number of applications. The foliar applied T. atroviride was not detected in the soil or root samples six months after application, in either a soil metabarcoding analysis or on re-isolation media. This was likely due to the dense tea foliage and ground cover under the tea plants which impeded its movement to the soil. While the specific nature of T. atroviride interaction with perennial crops like tea is not known, in this trial it appeared to have remained on the phyllosphere and provided biostimulation without reaching the soil. Full article
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17 pages, 1157 KB  
Article
Isolation and Characterization of Phosphate-Solubilizing Rhizobacteria from Solanum tuberosum with Plant Growth-Promoting Activity
by Michel Leiva-Mora, Pamela Elizabeth Mera Guzmán, Rafael Isaías Mera-Andrade, Alicia Monserrath Zabala Haro, Luis Rodrigo Saa, Paúl Loján, Catherine Lizzeth Silva Agurto, Luis Fabián Salazar-Garcés, Betty Beatriz González Osorio, Dariel Cabrera Mederos and Orelvis Portal
Appl. Microbiol. 2026, 6(1), 8; https://doi.org/10.3390/applmicrobiol6010008 - 3 Jan 2026
Cited by 5 | Viewed by 2513
Abstract
Phosphate-solubilizing rhizobacteria associated with the Solanum tuberosum L. cultivar ‘Superchola’ were isolated and characterized to improve our understanding of plant growth promotion in agricultural systems. Bacteria were isolated by serial dilutions, and the morphology of the colonies was characterized on nutrient agar culture [...] Read more.
Phosphate-solubilizing rhizobacteria associated with the Solanum tuberosum L. cultivar ‘Superchola’ were isolated and characterized to improve our understanding of plant growth promotion in agricultural systems. Bacteria were isolated by serial dilutions, and the morphology of the colonies was characterized on nutrient agar culture medium. In addition, morphological identification was achieved by Gram staining. The ability to solubilize phosphate was assessed in Pikovskaya agar culture medium, while molecular identification involved the amplification of the partial 16S rRNA gene using the polymerase chain reaction. In the Píllaro canton, the highest number of colony-forming units per gram of soil was recorded at 9.72 × 109. Among the isolated strains, 62% exhibited circular morphology, 92% had a smooth texture, and 85% displayed entire margins. Notably, 83% of the isolates were Gram-negative, with 50% exhibiting a bacillary form. The most effective phosphate solubilizers were from the Mocha canton, particularly the isolate CC-FCAGP-BSF6, which showed superior solubilization capacity. Molecular identification revealed bacterial isolates from four genera, i.e., Bacillus, Pseudomonas, Lysinibacillus, and Paenibacillus. These strains exhibited significant phosphate solubilization in vitro and resulted in increased leaf area (0.21–0.49, p = 0.038), fresh mass (0.46–0.87, p = 0.014), dry mass (0.092–0.096, p = 0.047), and leaf area index (0.14–0.33, p = 0.026) in the S. tuberosum cultivar ‘Superchola’ in vitro plants. This study identifies bacterial species associated with the rhizosphere of S. tuberosum in Ecuador and highlights their potential for promoting plant growth and solubilizing phosphates. Full article
(This article belongs to the Special Issue Microorganisms: A Way Forward for Sustainable Development?)
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17 pages, 724 KB  
Article
The Use of Confidence Intervals in Differential Abundance Analysis of Microbiome Data
by Elizaveta Vinogradova, Almagul Kushugulova, Samat Kozhakhmetov and Maxim Baltin
Appl. Microbiol. 2026, 6(1), 7; https://doi.org/10.3390/applmicrobiol6010007 - 2 Jan 2026
Viewed by 1550
Abstract
Differential abundance analysis (DAA) is a critical task in microbiome research aimed at identifying microbial signatures that reliably characterize groups. Research suggests that microbiome systems are relatively stable and resilient, yet even small changes under certain conditions can trigger dysbiosis. The high dimensionality [...] Read more.
Differential abundance analysis (DAA) is a critical task in microbiome research aimed at identifying microbial signatures that reliably characterize groups. Research suggests that microbiome systems are relatively stable and resilient, yet even small changes under certain conditions can trigger dysbiosis. The high dimensionality of microbiome datasets exacerbates the challenge of detecting such changes by posing a multiple comparison problem that requires hypothesis filtration. Standard filtration using multiple comparison correction procedures is designed for scenarios with a high number of true positives and is often too conservative for microbiome data, where the proportion of true signals can be very low. Therefore, there is a substantial need for hypothesis filtration methods tailored to microbiome data. Confidence intervals (CIs) for between-group differences offer a powerful alternative to p-value filtration, as their range simultaneously conveys information about the significance, potential magnitude, and direction of the effect, as well as the certainty of the estimate itself. Microbial data can be adequately modeled using a negative binomial (NB) distribution, and its location parameter can be robustly estimated with the Hodges–Lehmann estimator (HLE). Using synthetic and experimental data, we demonstrate that hypothesis filtration based on CIs for the two-sample HLE is a robust method for comparing microbial data. Our analysis demonstrates that the HLE-CI approach provides the same level of precision as filtration using multiple-adjustment methods while achieving significantly higher recall in microbiome DAA. The results of this study suggest that HLE-CI-based filtration can be an effective step in the search for microbiome biomarkers. Full article
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14 pages, 279 KB  
Article
Evaluation of the Capacity of Purple Nonsulfur Bacteria from In-Dyke Alluvial Soil to Solubilize Mica-Derived Potassium and Promote Hybrid Maize Growth
by Tran Ngoc Han, Nguyen Thanh Toan, Nguyen Thi Tuyet Hue, Le Thi My Thu, Phung Thi Hang, Nguyen Duc Trong, Tran Trong Khoi Nguyen, Le Thanh Quang, Ly Ngoc Thanh Xuan, Ngo Thanh Phong and Nguyen Quoc Khuong
Appl. Microbiol. 2026, 6(1), 6; https://doi.org/10.3390/applmicrobiol6010006 - 30 Dec 2025
Cited by 2 | Viewed by 771
Abstract
Potassium (K) is a vital macronutrient for plant growth and yield, yet most soil K occurs in insoluble mineral forms, limiting availability to crops. Reliance on chemical K fertilizers is unsustainable due to cost and environmental concerns. Microbial solubilization of mineral K, particularly [...] Read more.
Potassium (K) is a vital macronutrient for plant growth and yield, yet most soil K occurs in insoluble mineral forms, limiting availability to crops. Reliance on chemical K fertilizers is unsustainable due to cost and environmental concerns. Microbial solubilization of mineral K, particularly by purple nonsulfur bacteria (PNSB), offers an eco-friendly alternative. This study focused on isolating mica-potassium-solubilizing purple nonsulfur bacteria (MK-PNSB) from in-dyke alluvial soil and assessing their effects on hybrid maize germination and seedling growth. Among the isolates, the results showed that strain M-Wa-19 released the highest amount of soluble K under microaerobic light conditions (27.4 mg∙L−1). Under aerobic dark conditions, M-Wa-24 and M-Wa-26 released 20.1–21.0 mg∙L−1 of soluble K. Strains M-Wa-21, M-Wa-25, and M-Sl-13 solubilized K in the range of 14.3–25.1 mg∙L−1 and 12.9–24.4 mg∙L−1 under both incubation conditions. The selected strains were identified by 16S rRNA as Rhodopseudomonas palustris strain M-Sl-13 (PX588604), Rhodoplanes pokkaliisoli strain M-Wa-19 (PX588605), Afifella marina strain M-Wa-21 (PX588606), Rhodocista pekingensis strain M-Wa-24 (PX588607), Rhodocista pekingensis strain M-Wa-25 (PX588608), and Rhodocista pekingensis strain M-Wa-26 (PX588609). None exhibited toxicity to maize seeds; instead, all enhanced seed vigor indices by up to 99.7% and improved plant height and root biomass by 19.0–26.2% and 14.4–22.9%, respectively, under static hydroponic conditions. At a 1:1000 (bacteria and distilled water) dilution rate, strains M-Wa-26, M-Wa-25, M-Sl-13, M-Wa-24, M-Wa-19, and M-Wa-21, along with the six-strain mixture, improved seed vigor index by 3.96–7.91%. These findings suggest that MK-PNSB, individually or in mixtures, hold promise as biofertilizer candidates for sustainable K management in crop production. Full article
16 pages, 2728 KB  
Article
Extensive Molecular Characterization of a New Bacterial Cellulose Hydrogel (BCH) Producing Gluconacetobacter Isolate from Nigeria-Sourced Agro-Residue
by Samuel Charles Olabode, Barde Yelwa Luka, Oluwafemi Abiodun Adepoju, Ibrahim Zubairu Waziri, Lockta Joel, Rabiu Bukar Mohammed, Mohammed Nasir Shuaibu, Abdullahi Balarabe Sallau, Tahir Turaki Mohammed, Sa’adiya Halima Mahmud, Abdullahi Abdulmalik Salman, Ghulam Jeelani, Yakubu Kokori Enevehe Ibrahim and Emmanuel Oluwadare Balogun
Appl. Microbiol. 2026, 6(1), 5; https://doi.org/10.3390/applmicrobiol6010005 - 28 Dec 2025
Viewed by 1242
Abstract
Bacterial cellulose hydrogels (BCHs) are characterized as exopolysaccharides of glucose polymers consisting of β–1–4–glycosidic linkage with various degrees of polymerization which are synthesized by bacteria. There is a paucity of information on the isolation and characterization of a BCH producer isolate from Nigeria. [...] Read more.
Bacterial cellulose hydrogels (BCHs) are characterized as exopolysaccharides of glucose polymers consisting of β–1–4–glycosidic linkage with various degrees of polymerization which are synthesized by bacteria. There is a paucity of information on the isolation and characterization of a BCH producer isolate from Nigeria. The study, therefore, aimed to characterize a new Acetobacter species that had previously been confirmed to produce BCH. The BCH-producing isolate was characterized by PCR amplification of the full-length 16S rRNA gene, as well as whole-genome sequencing analysis. The whole-genome sequence of the isolate was determined using the Illumina next-generation sequencing (NGS) platform, with downstream analysis of genomic reads through the metaWRAP pipeline. The BCH producer isolate was identified to be Acetobacter orientalis strain Zaria-B1, based on sequence identity with the reference Acetobacter orientalis strain VVS. Based on its annotated genome, the isolate had an approximate genomic size of 3.1 Mbp, 45 total RNAs, a GC content of 52.5%, 3046 total protein-encoding genes, an N50 of 253,774 bp, and an L50 of 4, as well as 30 contigs. The nucleotide BLAST of the cellulose synthase gene sequence confirmed the bin to be Acetobacter orientalis. The whole-genome characterization alongside the 16S rRNA genotyping confirmed the BCH-producing isolate to be Acetobacter orientalis. Full article
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11 pages, 1658 KB  
Article
Bioactive Potential of Soft Coral-Associated Bacteria Collected from the Red Sea, Egypt
by Doaa S. Aboelwafa, Abdel-Hamied M. Rasmey, Akram A. Aboseidah, Noura S. Dosoky and Samar M. Abdelrahman
Appl. Microbiol. 2026, 6(1), 4; https://doi.org/10.3390/applmicrobiol6010004 - 25 Dec 2025
Viewed by 1213
Abstract
In this study, we used a culture-dependent approach to explore the biochemical potential of bacteria associated with two genera of soft corals collected from the Red Sea (phylum Cnidaria, class Anthozoa, subclass Octocorallia, order Alcyonaceae, and family Alcyoniidae). The soft corals were identified [...] Read more.
In this study, we used a culture-dependent approach to explore the biochemical potential of bacteria associated with two genera of soft corals collected from the Red Sea (phylum Cnidaria, class Anthozoa, subclass Octocorallia, order Alcyonaceae, and family Alcyoniidae). The soft corals were identified as Cladiella sp. and Paralemnalia sp. The associated bacteria were isolated on marine agar, nutrient agar, starch casein agar, ISP2 Agar, and M1 agar. The highest proportion of strains was recovered using marine agar, followed by nutrient agar and M1. We focused on Gram-positive bacteria and evaluated their cytotoxicity and antimicrobial activity. About 24% of the bacterial samples demonstrated promising cytotoxicity against Ehrlich ascites carcinoma (EAC). Out of 12 bioactive isolated strains, two bacterial isolates showed strong cytotoxicity, with IC50 values of 134.47 and 148.5 µg/mL, respectively. Nine isolates displayed significant antimicrobial activity against two tested pathogens. Based on the 16S rRNA gene sequence, two bioactive bacterial isolates were identified as Bacillus subtilis and Microbacterium sp. These findings indicate that bacteria associated with soft corals could be a valuable source of new bioactive compounds with potential uses in drug development. Furthermore, our data add important insights to the understudied field of host-microbiome relationships in soft corals. Full article
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29 pages, 3252 KB  
Article
Metagenomic and Proxy Monitoring of Surfactant Degradation by Microbial Consortia from Oil-Contaminated Soil
by Morena India Mokoena, Rosina Nkuna and Tonderayi Sylvester Matambo
Appl. Microbiol. 2026, 6(1), 3; https://doi.org/10.3390/applmicrobiol6010003 - 24 Dec 2025
Cited by 1 | Viewed by 1454
Abstract
Surfactants are harmful, persistent pollutants that are often found in contaminated soils, wastewater, and industrial effluents in complex mixes. Due to their chemical diversity and persistence, they present a bioremediation challenge. Using long-read shotgun metagenomics, 16S rRNA amplicon sequencing, PICRUSt2 functional prediction, and [...] Read more.
Surfactants are harmful, persistent pollutants that are often found in contaminated soils, wastewater, and industrial effluents in complex mixes. Due to their chemical diversity and persistence, they present a bioremediation challenge. Using long-read shotgun metagenomics, 16S rRNA amplicon sequencing, PICRUSt2 functional prediction, and physicochemical proxies (total organic carbon, dissolved oxygen, chemical oxygen demand, foaming activity, etc.), this study investigated the aerobic biodegradation of SDS, SLS, rhamnolipids, Triton X-100, and CTAB (individually/mixed, 4% w/v) by microbial consortia enriched from oil-contaminated soil for 14 days. Pseudomonadota was dominant (85–90%), with Pseudomonas (60%) driving SLS and SDS degradation, while Paraburkholderia and Bordetella were dominant in recalcitrant surfactant degradation. Among the surfactants, SLS, rhamnolipids, and the combination of all surfactants demonstrated higher degradation by virtue of total organic carbon reductions of 50%, 56%, and 50%, respectively, and a foaming activity decline of 45–64%. The combination of surfactants with CTAB showed a 21% reduction in TOC, most likely due to CTAB’s known bactericidal effects. PICRUSt2 showed differential enrichment in alkyl oxidation, sulfate ester hydrolysis, aromatic ring cleavage, and fatty acid/sulfur genes and pathways. This study establishes inexpensive, scalable proxy indicators for monitoring surfactant bioremediation when direct metabolite analysis is impractical. Full article
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27 pages, 3698 KB  
Article
Biocontrol and Plant Growth-Promoting Potential of Bacillus and Actinomycetes Isolated from the Rhizosphere and Phyllosphere of Potato (Solanum tuberosum L.) from Different Agroecological Zones of Peru
by Lizbeth Mamani-Rojas, Raihil Rengifo, Leslie Velarde-Apaza, Max Ramírez-Rojas and Hector Cántaro-Segura
Appl. Microbiol. 2026, 6(1), 2; https://doi.org/10.3390/applmicrobiol6010002 - 23 Dec 2025
Cited by 3 | Viewed by 1876
Abstract
Potato (Solanum tuberosum L.) is a key staple crop in the Peruvian Andes, but its productivity is threatened by fungal pathogens such as Rhizoctonia solani and Alternaria alternata. In this study, 71 native bacterial strains (39 from phyllosphere and 32 from [...] Read more.
Potato (Solanum tuberosum L.) is a key staple crop in the Peruvian Andes, but its productivity is threatened by fungal pathogens such as Rhizoctonia solani and Alternaria alternata. In this study, 71 native bacterial strains (39 from phyllosphere and 32 from rhizosphere) were isolated from potato plants across five agroecological zones of Peru and characterized for their plant growth-promoting (PGPR) and antagonistic traits. Actinomycetes demonstrated broader enzymatic profiles, with 2ACPP4 and 2ACPP8 showing high proteolytic (68.4%, 63.4%), lipolytic (59.5%, 60.6%), chitinolytic (32.7%, 35.5%) and amylolytic activity (76.3%, 71.5%). Strain 5ACPP5 (Streptomyces decoyicus) produced 42.8% chitinase and solubilized both dicalcium (120.6%) and tricalcium phosphate (122.3%). The highest IAA production was recorded in Bacillus strain 2BPP8 (95.4 µg/mL), while 5ACPP6 was the highest among Actinomycetes (83.4 µg/mL). Siderophore production was highest in 5ACPP5 (412.4%) and 2ACPP4 (406.8%). In vitro antagonism assays showed that 5ACPP5 inhibited R. solani and A. alternata by 86.4% and 68.9%, respectively, while Bacillus strain BPP4 reached 51.0% inhibition against A. alternata. In greenhouse trials, strain 4BPP8 significantly increased fresh tuber weight (11.91 g), while 5ACPP5 enhanced root biomass and reduced stem canker severity. Molecular identification confirmed BPP4 as Bacillus halotolerans and 5ACPP5 as Streptomyces decoyicus. These strains represent promising candidates for the development of bioinoculants for sustainable potato cultivation in Andean systems. Full article
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18 pages, 2234 KB  
Article
Validation of L-Lactic Acid Production Using Companilactobacillus farciminis KUJ 25-S for Sustainable Bio-Polylactic Acid Manufacturing
by Kangsadan Boonprab, Vichien Kitpreechavanich and Mingkwan Nipitwattanaphon
Appl. Microbiol. 2026, 6(1), 1; https://doi.org/10.3390/applmicrobiol6010001 - 19 Dec 2025
Viewed by 1393
Abstract
Companilactobacillus farciminis KUJ 25-S was isolated from fermented fish and identified using 16S rRNA gene sequencing with 30.0 g/L of L-LA (L-lactic acid), with 97% LA per sum of DL-LA. The characteristics of LA and its stereoisomers were confirmed using TLC, chiral-HPLC, and [...] Read more.
Companilactobacillus farciminis KUJ 25-S was isolated from fermented fish and identified using 16S rRNA gene sequencing with 30.0 g/L of L-LA (L-lactic acid), with 97% LA per sum of DL-LA. The characteristics of LA and its stereoisomers were confirmed using TLC, chiral-HPLC, and enzymatic techniques. Based on various conditions using liquid MRS broth (static condition, glucose 10%, NaCl 5%, 37 °C for 48 h), the highest growth and LA formation of the culture were at a low temperature (25 °C) and decreased at 37, 45, and 55 °C, respectively. The broth could grow and produce acid at an initial pH in the range 4–11, with a low initial pH of 4 promoting the highest LA formation. LA formation and growth were inversely proportional to the NaCl concentration in the 0.5–30% range. High glucose concentrations suppressed LA formation. The growth-promotion effect varied with glucose concentration (5–40%), with the optimum concentration for LA production being 20% glucose. On the other hand, if used in microoxic conditions, the absence of NaCl was more favorable to acidification than the addition of NaCl (5% NaCl). C. farciminis KUJ 25-S was proposed as a suitable method to produce L-LA based on using the appropriate line for further industrial use. Full article
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