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Appl. Microbiol., Volume 5, Issue 3 (September 2025) – 24 articles

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35 pages, 1022 KiB  
Review
Mechanistic Role of Heavy Metals in Driving Antimicrobial Resistance: From Rhizosphere to Phyllosphere
by Rahul Kumar, Tanja P. Vasić, Sanja P. Živković, Periyasamy Panneerselvam, Gustavo Santoyo, Sergio de los Santos Villalobos, Adeyemi Nurudeen Olatunbosun, Aditi Pandit, Leonard Koolman, Debasis Mitra and Pankaj Gautam
Appl. Microbiol. 2025, 5(3), 79; https://doi.org/10.3390/applmicrobiol5030079 (registering DOI) - 4 Aug 2025
Abstract
Heavy metal pollution represents a pervasive environmental challenge that significantly exacerbates the ever-increasing crisis of antimicrobial resistance and the capacity of microorganisms to endure and proliferate despite antibiotic interventions. This review examines the intricate relationship between heavy metals and AMR, with an emphasis [...] Read more.
Heavy metal pollution represents a pervasive environmental challenge that significantly exacerbates the ever-increasing crisis of antimicrobial resistance and the capacity of microorganisms to endure and proliferate despite antibiotic interventions. This review examines the intricate relationship between heavy metals and AMR, with an emphasis on the underlying molecular mechanisms and ecological ramifications. Common environmental metals, including arsenic, mercury, cadmium, and lead, exert substantial selective pressures on microbial communities. These induce oxidative stress and DNA damage, potentially leading to mutations that enhance antibiotic resistance. Key microbial responses include the overexpression of efflux pumps that expel both metals and antibiotics, production of detoxifying enzymes, and formation of protective biofilms, all of which contribute to the emergence of multidrug-resistant strains. In the soil environment, particularly the rhizosphere, heavy metals disrupt plant–microbe interactions by inhibiting beneficial organisms, such as rhizobacteria, mycorrhizal fungi, and actinomycetes, thereby impairing nutrient cycling and plant health. Nonetheless, certain microbial consortia can tolerate and detoxify heavy metals through sequestration and biotransformation, rendering them valuable for bioremediation. Advances in biotechnology, including gene editing and the development of engineered metal-resistant microbes, offer promising solutions for mitigating the spread of metal-driven AMR and restoring ecological balance. By understanding the interplay between metal pollution and microbial resistance, we can more effectively devise strategies for environmental protection and public health. Full article
50 pages, 3314 KiB  
Review
Applied Microbiology for Sustainable Agricultural Development
by Barbara Sawicka, Piotr Barbaś, Viola Vambol, Dominika Skiba, Piotr Pszczółkowski, Parwiz Niazi and Bernadetta Bienia
Appl. Microbiol. 2025, 5(3), 78; https://doi.org/10.3390/applmicrobiol5030078 (registering DOI) - 1 Aug 2025
Viewed by 72
Abstract
Background: Developments in biology, genetics, soil science, plant breeding, engineering, and agricultural microbiology are driving advances in soil microbiology and microbial biotechnology. Material and methods: The literature for this review was collected by searching leading scientific databases such as Embase, Medline/PubMed, Scopus, and [...] Read more.
Background: Developments in biology, genetics, soil science, plant breeding, engineering, and agricultural microbiology are driving advances in soil microbiology and microbial biotechnology. Material and methods: The literature for this review was collected by searching leading scientific databases such as Embase, Medline/PubMed, Scopus, and Web of Science. Results: Recent advances in soil microbiology and biotechnology are discussed, emphasizing the role of microorganisms in sustainable agriculture. It has been shown that soil and plant microbiomes significantly contribute to improving soil fertility and plant and soil health. Microbes promote plant growth through various mechanisms, including potassium, phosphorus, and zinc solubilization, biological nitrogen fixation, production of ammonia, HCN, siderophores, and other secondary metabolites with antagonistic effects. The diversity of microbiomes related to crops, plant protection, and the environment is analyzed, as well as their role in improving food quality, especially under stress conditions. Particular attention was paid to the diversity of microbiomes and their mechanisms supporting plant growth and soil fertility. Conclusions: The key role of soil microorganisms in sustainable agriculture was highlighted. They can support the production of natural substances used as plant protection products, as well as biopesticides, bioregulators, or biofertilizers. Microbial biotechnology also offers potential in the production of sustainable chemicals, such as biofuels or biodegradable plastics (PHA) from plant sugars, and in the production of pharmaceuticals, including antibiotics, hormones, or enzymes. Full article
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14 pages, 990 KiB  
Article
Comparative Analysis of the Biomass Production and Nutritional Profiles of Two Wild-Type Strains of Yarrowia lipolytica
by David Torres-Añorve and Georgina Sandoval
Appl. Microbiol. 2025, 5(3), 77; https://doi.org/10.3390/applmicrobiol5030077 (registering DOI) - 1 Aug 2025
Viewed by 86
Abstract
Sustainability represents a significant global challenge, requiring a balance between environmental impact and the use of natural resources. White biotechnology, which uses microorganisms and enzymes for environmentally friendly products and processes, offers promising solutions to support a growing population. Within this context, the [...] Read more.
Sustainability represents a significant global challenge, requiring a balance between environmental impact and the use of natural resources. White biotechnology, which uses microorganisms and enzymes for environmentally friendly products and processes, offers promising solutions to support a growing population. Within this context, the yeast Yarrowia lipolytica stands out, so we investigated the generation of biomass from two wild strains (ATCC 9773 and NRRL Y-50997) using different carbon sources. Additionally, protein content and amino acid profiles were assessed via standardized analytical methods to evaluate their potential as nutritional yeasts. Both strains demonstrated potential as nutritional yeasts, with biomass productivities of up to 35.5 g/L and 42 g/L, respectively. The protein content was high, with 58.8% for ATCC 9773 and 58.2% for NRRL Y-50997. Furthermore, the strains presented essential amino acid contents of 62.6% and 41.5%, with lysine being the most abundant amino acid. These findings underscore the versatility and productivity of Y. lipolytica, highlighting its potential for sustainable biotechnological applications such as single-cell protein production. Full article
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18 pages, 1685 KiB  
Article
Influence of Isolation Source on the Probiotic Properties and Health Benefits of Yeasts: Insights from Metabarcoding and Cultivation Approaches
by Kanyarat Kanyakam and Cheunjit Prakitchaiwattana
Appl. Microbiol. 2025, 5(3), 76; https://doi.org/10.3390/applmicrobiol5030076 - 30 Jul 2025
Viewed by 125
Abstract
The study aimed to identify potential sources of novel probiotic yeasts exhibiting health-promoting properties. A combination of metabarcoding analysis and cultural methods was employed to investigate and isolate yeasts from various sources, including rice wine, palm wine, fermented shrimp paste at different stages [...] Read more.
The study aimed to identify potential sources of novel probiotic yeasts exhibiting health-promoting properties. A combination of metabarcoding analysis and cultural methods was employed to investigate and isolate yeasts from various sources, including rice wine, palm wine, fermented shrimp paste at different stages of natural fermentation, and lychee peels. The two analytical methods revealed distinct yeast profiles, and each source exhibited a unique composition of yeast species. Through metabarcoding and cultural methods, it was demonstrated that lychee peels harbored a greater diversity of genera compared to other sources. The evaluation of the probiotic properties of yeasts revealed that lychee peel yielded the highest proportion of isolates with potential probiotic activity (53.33%), followed by palm wine (25%), fermented shrimp paste (10%), and rice wine (9.09%). Moreover, yeast isolates with health-promoting properties as evaluated in this study, including Starmerella meliponinorum L12 and Pichia terricola L9 from lychee peels, demonstrated notable antioxidant activity and cholesterol-reducing properties, respectively. These findings represent the first report providing initial insights into the influence of yeast sources and serve as a guideline for the targeted selection of yeasts with specific probiotic and health-promoting attributes. Full article
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16 pages, 1196 KiB  
Article
Sustainable Bioconversion of Cashew Apple Bagasse Hemicellulosic Hydrolysate into Xylose Reductase and Xylitol by Candida tropicalis ATCC 750: Impact of Aeration and Fluid Dynamics
by Juliana de França Serpa, Franciandro Dantas dos Santos, Carlos Eduardo Alves Soares, Benevides Costa Pessela and Maria Valderez Ponte Rocha
Appl. Microbiol. 2025, 5(3), 75; https://doi.org/10.3390/applmicrobiol5030075 (registering DOI) - 30 Jul 2025
Viewed by 162
Abstract
This study aimed to evaluate the production of xylose reductase (XR), an enzyme responsible for converting xylose into xylitol, by Candida tropicalis ATCC 750 using hemicellulosic hydrolysate from cashew apple bagasse (CABHM) as a low-cost carbon source. The effects of temperature, aeration, and [...] Read more.
This study aimed to evaluate the production of xylose reductase (XR), an enzyme responsible for converting xylose into xylitol, by Candida tropicalis ATCC 750 using hemicellulosic hydrolysate from cashew apple bagasse (CABHM) as a low-cost carbon source. The effects of temperature, aeration, and fluid dynamics on XR biosynthesis were also investigated. The highest XR production (1.53 U mL−1) was achieved at 30 °C, with 8.3 g·L−1 of xylitol produced by the yeast under microaerobic conditions, demonstrating that aeration and fluid dynamics are important factors in this process. Cellular metabolism and enzyme production decreased at temperatures above 35 °C. The maximum enzymatic activity was observed at pH 7.0 and 50 °C. XR is a heterodimeric protein with a molecular mass of approximately 30 kDa. These results indicate that CABHM is a promising substrate for XR production by C. tropicalis, contributing to the development of enzymatic bioprocesses for xylitol production from lignocellulosic biomass. This study also demonstrates the potential of agro-industrial residues as sustainable feedstocks in biorefineries, aligning with the principles of a circular bioeconomy. Full article
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36 pages, 1502 KiB  
Review
A Critical Review on the Role of Lactic Acid Bacteria in Sourdough Nutritional Quality: Mechanisms, Potential, and Challenges
by Youssef Mimoune Reffai and Taoufiq Fechtali
Appl. Microbiol. 2025, 5(3), 74; https://doi.org/10.3390/applmicrobiol5030074 - 29 Jul 2025
Viewed by 259
Abstract
Sourdough fermentation, driven by the biochemical activity of lactic acid bacteria (LAB), presents a scientifically promising approach to addressing nutritional limitations in cereal-based staples. This review critically examines both the underlying mechanisms by which LAB enhance the nutritional profile of sourdough and the [...] Read more.
Sourdough fermentation, driven by the biochemical activity of lactic acid bacteria (LAB), presents a scientifically promising approach to addressing nutritional limitations in cereal-based staples. This review critically examines both the underlying mechanisms by which LAB enhance the nutritional profile of sourdough and the translational challenges in realizing these benefits. Key improvements explored include enhanced mineral bioavailability (e.g., up to 90% phytate reduction), improved protein digestibility, an attenuated glycemic response (GI ≈ 54 vs. ≈75 for conventional bread), and the generation of bioactive compounds. While in vitro and animal studies extensively demonstrate LAB’s potential to reshape nutrient profiles (e.g., phytate hydrolysis improving iron absorption, proteolysis releasing bioactive peptides), translating these effects into consistent human health outcomes proves complex. Significant challenges hinder this transition from laboratory to diet, including the limited bioavailability of LAB-derived metabolites, high strain variability, and sensitivity to fermentation conditions. Furthermore, interactions with the food matrix and host-specific factors, such as gut microbiota composition, contribute to inconsistent findings. This review highlights methodological gaps, particularly reliance on in vitro or animal models, and the lack of long-term, effective human trials. Although LAB hold significant promise for nutritional improvements in sourdough, translating these findings to validated human benefits necessitates continued efforts in mechanism-driven strain optimization, the standardization of fermentation processes, and rigorous human studies. Full article
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20 pages, 1766 KiB  
Review
Recent Development of Exploring Ferroptosis-Inspired Effect of Iron as a Feasible Strategy for Combating Multidrug Resistant Bacterial Infections
by Nalin Abeydeera
Appl. Microbiol. 2025, 5(3), 73; https://doi.org/10.3390/applmicrobiol5030073 - 28 Jul 2025
Viewed by 818
Abstract
The increasing threat of antimicrobial resistance (AMR), along with the limited availability of new lead compounds in the drug development pipeline, highlights the urgent need to discover antimicrobial agents with innovative mechanisms of action. In this regard, metal complexes offer a unique opportunity [...] Read more.
The increasing threat of antimicrobial resistance (AMR), along with the limited availability of new lead compounds in the drug development pipeline, highlights the urgent need to discover antimicrobial agents with innovative mechanisms of action. In this regard, metal complexes offer a unique opportunity to access mechanisms distinct from those of conventional antibiotics. Although iron (Fe) is an essential element for all forms of life, including pathogenic bacteria, it also poses a serious risk of cytotoxicity due to its redox activity, which can trigger the production of reactive oxygen species (ROS) via the Fenton reaction. This review highlights recent advances in the development of iron-based antimicrobial agents that harness the toxicity resulting from dysregulated iron uptake, thereby inducing bacterial cell death through oxidative stress. These findings may guide the development of effective treatments for pathogenic infections and offer new perspectives on leveraging redox chemistry of iron to combat the growing threat of global bacterial resistance. Full article
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22 pages, 2596 KiB  
Article
Cardio-Protective Effects of Microencapsulated Probiotic and Synbiotic Supplements on a Myocardial Infarction Model Through the Gut–Heart Axis
by Doha A. Mohamed, Hoda B. Mabrok, Hoda S. El-Sayed, Sherein Abdelgayed and Shaimaa E. Mohammed
Appl. Microbiol. 2025, 5(3), 72; https://doi.org/10.3390/applmicrobiol5030072 - 27 Jul 2025
Viewed by 292
Abstract
Myocardial infarction (MI) is an inflammatory disease responsible for approximately 75% of sudden cardiac deaths. In this study, we aimed to evaluate the cardio-protective influence of microencapsulated probiotic and synbiotic dietary supplements in vivo and in molecular docking studies. MI was induced in [...] Read more.
Myocardial infarction (MI) is an inflammatory disease responsible for approximately 75% of sudden cardiac deaths. In this study, we aimed to evaluate the cardio-protective influence of microencapsulated probiotic and synbiotic dietary supplements in vivo and in molecular docking studies. MI was induced in rats with the injection of isoproterenol (i.p. 67 mg/kg). Plasma lipid profiles and the levels of oxidative stress markers, inflammatory markers, and cardiac enzymes were determined. The expression levels of MMP-7 and IL-1β in the heart muscle were measured. The impact of dietary supplements on fecal bacterial counts was evaluated across all rat groups. A histopathological examination of cardiac tissue was performed. The cardio-protective potential of cyanidin 3-diglucoside 5-glucoside and arabinoxylan was studied using molecular docking. The results demonstrate that all tested dietary supplements induced an improvement in all the biochemical parameters in association with an improvement in myocardial muscle tissue. The mRNA expression levels of MMP-7 and IL-1β were significantly downregulated by all dietary supplements. All dietary supplements increased the fecal counts of probiotic strains. In the molecular docking analysis, cyanidin 3-diglucoside 5-glucoside exhibited binding affinity values of −8.8 and −10 for lactate dehydrogenase (LDH) and Paraoxonase 1 (PON1), respectively. Arabinoxylan showed similar binding affinity (−8.8) for both LDH and PON1. Conclusion: Microencapsulated probiotic and synbiotic dietary supplements demonstrated notable cardio-protective influence in vivo and in molecular docking studies. These supplements may serve as promising candidates for the prevention of myocardial infarction. Full article
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21 pages, 6746 KiB  
Article
Harnessing Wild Jackfruit Extract for Chitosan Production by Aspergillus versicolor AD07: Application in Antibacterial Biodegradable Sheets
by Adhithya Sankar Santhosh and Mridul Umesh
Appl. Microbiol. 2025, 5(3), 71; https://doi.org/10.3390/applmicrobiol5030071 - 20 Jul 2025
Viewed by 393
Abstract
A fungal strain with comparably high chitosan yield was isolated from the Shivaganga hills and identified as Aspergillus versicolor AD07 through molecular characterization. Later, the strain was cultivated on Sabouraud Dextrose Broth (SDB) and wild jackfruit-based media to evaluate its potential for chitosan [...] Read more.
A fungal strain with comparably high chitosan yield was isolated from the Shivaganga hills and identified as Aspergillus versicolor AD07 through molecular characterization. Later, the strain was cultivated on Sabouraud Dextrose Broth (SDB) and wild jackfruit-based media to evaluate its potential for chitosan production. Among the various media formulations, the highest chitosan yield (178.40 ± 1.76 mg/L) was obtained from the jackfruit extract medium with added peptone and dextrose. The extracted chitosan was characterized through FTIR, XRD (reported a crystallinity index of 55%), TGA/DTG, and DSC analysis, confirming the presence of key functional groups and high thermal resistance. The extracted chitosan was fabricated into a sheet incorporated with 1% lemongrass oil; the sheet exhibited strong antibacterial activity against Escherichia coli (30 mm) and Bacillus megaterium (48 mm). The biodegradation studies reported a weight loss of 38.93 ± 0.51% after 50 days of soil burial. Further, the chitosan film was tested as a packaging material for paneer, demonstrating better preservation by maintaining nutritional quality and reducing microbial load over a 14-day storage period. These findings highlight the potential of A. versicolor AD07-derived chitosan, cultivated on a waste substrate medium, as a sustainable biopolymer for food packaging applications. Full article
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16 pages, 709 KiB  
Perspective
The Gut–Brain Axis in Schizophrenia: A Systems-Level Understanding of Psychiatric Illness
by Austin Mardon, Haadiya Chaudhry, Jonathan Harline, Catherine Mardon, Jenna Banks, Eric Hodgson and Jean-Luc Leong-Sit
Appl. Microbiol. 2025, 5(3), 70; https://doi.org/10.3390/applmicrobiol5030070 - 18 Jul 2025
Viewed by 389
Abstract
Schizophrenia is a complex psychiatric disorder traditionally linked to neurotransmitter dysregulation, particularly within dopamine and glutamate pathways. However, recent evidence implicates the gut–brain axis as a potential contributor to its pathophysiology. This perspective article proposes a systems-level understanding of schizophrenia that incorporates the [...] Read more.
Schizophrenia is a complex psychiatric disorder traditionally linked to neurotransmitter dysregulation, particularly within dopamine and glutamate pathways. However, recent evidence implicates the gut–brain axis as a potential contributor to its pathophysiology. This perspective article proposes a systems-level understanding of schizophrenia that incorporates the role of gut microbial dysbiosis specifically, reductions in short-chain fatty acid (SCFA)-producing taxa, and elevations in pro-inflammatory microbes. These imbalances may compromise gut barrier integrity, stimulate systemic inflammation, and disrupt neurochemical signaling in the brain. We synthesize findings from animal models, clinical cohorts, and microbial intervention trials, highlighting mechanisms such as SCFA regulation, altered tryptophan–kynurenine metabolism, and microbial impacts on neurotransmitters. We also explore microbiome-targeted interventions like probiotics, prebiotics, dietary strategies, and fecal microbiota transplantation (FMT) and their potential as adjunctive therapies. While challenges remain in causality and translation, integrating gut–brain axis insights may support more personalized and biologically informed models of schizophrenia care. Full article
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23 pages, 3262 KiB  
Article
An Exploratory Study on the Growth Dynamics of Alkalihalophilus marmarensis Using a Model-Based Approach
by Yağmur Atakav, Eldin Kurpejović, Dilek Kazan and Nihat Alpagu Sayar
Appl. Microbiol. 2025, 5(3), 69; https://doi.org/10.3390/applmicrobiol5030069 - 17 Jul 2025
Viewed by 199
Abstract
Alkalihalophilus marmarensis is an obligate alkaliphile with exceptional tolerance to high-pH environments, making it a promising candidate for industrial bioprocesses that require contamination-resistant and extremophilic production platforms. However, its practical deployment is hindered by limited biomass formation under extreme conditions, which constrains overall [...] Read more.
Alkalihalophilus marmarensis is an obligate alkaliphile with exceptional tolerance to high-pH environments, making it a promising candidate for industrial bioprocesses that require contamination-resistant and extremophilic production platforms. However, its practical deployment is hindered by limited biomass formation under extreme conditions, which constrains overall productivity. This study presents a model-driven investigation of how pH (8.8 and 10.5), culture duration (24 and 48 h), and nitrogen source composition (peptone and meat extract) affect cell dry mass, lactate, and protease synthesis. Using the response surface methodology and multi-objective optimization, we established predictive models (R2 up to 0.92) and uncovered key trade-offs in biomass and metabolite yields. Our findings reveal that peptone concentration critically shapes the metabolic output, with low levels inhibiting growth and high levels suppressing protease activity. Maximum cell dry mass (4.5 g/L), lactate (19.3 g/L), and protease activity (43.5 U/mL) were achieved under distinct conditions, highlighting the potential for targeted process tuning. While the model validation confirmed predictions for lactate, deviations in cell dry mass and protease outputs underscore the complexity of growth–product interdependencies under nutrient-limited regimes. This work delivers a foundational framework for developing fermentations with A. marmarensis and advancing its application in sustainable, high-pH industrial bioprocesses. The insights gained here can be further leveraged through synthetic biology and bioprocess engineering to fully exploit the metabolic potential of obligate alkaliphiles like A. marmarensis. Full article
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21 pages, 1929 KiB  
Review
Antimicrobial Compounds from Anaerobic Microorganisms: A Review of an Untapped Reservoir
by Mamta Mishra, Upasana Sharma, Manisha Rawat, Harshvardhan, Shelley Sardul Singh and Suresh Korpole
Appl. Microbiol. 2025, 5(3), 68; https://doi.org/10.3390/applmicrobiol5030068 - 15 Jul 2025
Viewed by 375
Abstract
Anaerobes, the oldest evolutionary life forms, have been unexplored for their potential to produce secondary metabolites due to the difficulties observed in their cultivation. Antimicrobials derived from anaerobic bacteria are an emerging and valuable source of novel therapeutic agents. The urgent need for [...] Read more.
Anaerobes, the oldest evolutionary life forms, have been unexplored for their potential to produce secondary metabolites due to the difficulties observed in their cultivation. Antimicrobials derived from anaerobic bacteria are an emerging and valuable source of novel therapeutic agents. The urgent need for new antimicrobial agents due to rising antibiotic resistance has prompted an investigation into anaerobic bacteria. The conventional method of antimicrobial discovery is based on cultivation and extraction methods. Antibacterial and antifungal substances are produced by anaerobic bacteria, but reports are limited due to oxygen-deficient growth requirements. The genome mining approach revealed the presence of biosynthetic gene clusters involved in various antimicrobial compound synthesis. Thus, the current review is focused on antimicrobials derived from anaerobes to unravel the potential of anaerobic bacteria as an emerging valuable source of therapeutic agents. These substances frequently consist of peptides, lipopeptides, and other secondary metabolites. Many of these antimicrobials have distinct modes of action that may be able to go around established resistance pathways. To this effect, we discuss diverse antimicrobial compounds produced by anaerobic bacteria, their biosynthesis, heterologous production, and activity. The findings suggest that anaerobic bacteria harbor significant biosynthetic potential, warranting further exploration through recombinant production for developing new antibiotics. Full article
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19 pages, 1439 KiB  
Article
Applied Metagenomic Profiling of Domestic Cat Feces from Cali, Colombia: An Exploratory Approach
by Monica Pimienta, Hernan Florez-Rios, Angie Patiño-Montoya, Anyelo Florez, Lizeth Mejia, Raul Sedano and Andres Castillo
Appl. Microbiol. 2025, 5(3), 67; https://doi.org/10.3390/applmicrobiol5030067 - 8 Jul 2025
Viewed by 400
Abstract
This exploratory study presents the first metagenomic assessment of the gut microbiome in domestic cats from Cali, Colombia. Fecal samples were collected from 10 healthy, sterilized domestic cats, aged 8 months to over 2 years, with variation in sex (7 females, 3 males), [...] Read more.
This exploratory study presents the first metagenomic assessment of the gut microbiome in domestic cats from Cali, Colombia. Fecal samples were collected from 10 healthy, sterilized domestic cats, aged 8 months to over 2 years, with variation in sex (7 females, 3 males), diet (processed or raw), and outdoor access (5 with, 5 without). Using 16S rRNA gene metabarcoding and pooled shotgun metagenomic sequencing, the study characterized the taxonomic composition and functional potential of the feline gut microbiome. Dominant phyla included Bacillota and Bacteroidota, with substantial inter-individual variation. Peptoclostridium was the most consistently abundant genus, while Megamonas and Megasphaera showed higher variability. Shotgun analysis detected antibiotic resistance genes (ErmG, ErmQ) and virulence factors (pfoA, plc, colA, nanJ, nagI) in Clostridium perfringens, highlighting potential zoonotic risk. The composition of the gut microbiota was influenced primarily by diet and outdoor access, while age and gender had more moderate effects. The study concludes that lifestyle and environmental factors play a key role in shaping the gut microbiome of domestic cats. We recommend further longitudinal and larger-scale studies to better understand the dynamics of feline microbiota and their implications for animal and public health within a One Health framework. Full article
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18 pages, 457 KiB  
Article
Application of Trichoderma spp. to Control Colletotrichum sp. and Pseudopestalotiopsis spp., Causing Agents of Fruit Rot in Pomelo (Citrus maxima (Burm.) Merr.)
by Nguyen Quoc Khuong, Le Ba Duy, Vo Minh Thuan, Nguyen Thanh Ngan, Phan Chan Hiep, Le Thanh Quang, Nguyen Duc Trong, Ha Ngoc Thu, Do Thi Xuan, Le Thi My Thu, Tran Trong Khoi Nguyen, Ly Ngoc Thanh Xuan and Ngo Thanh Phong
Appl. Microbiol. 2025, 5(3), 66; https://doi.org/10.3390/applmicrobiol5030066 - 6 Jul 2025
Viewed by 358
Abstract
Fruit rot seriously damages pomelo production. Given concerns regarding the safety of chemical agents, biological alternatives are becoming more preferable. Therefore, the experiment aimed to (i) identify the pathogens causing pomelo fruit rot disease and (ii) select Trichoderma spp. strains controlling the determined [...] Read more.
Fruit rot seriously damages pomelo production. Given concerns regarding the safety of chemical agents, biological alternatives are becoming more preferable. Therefore, the experiment aimed to (i) identify the pathogens causing pomelo fruit rot disease and (ii) select Trichoderma spp. strains controlling the determined pathogens in Ben Tre, Vietnam. Three pathogenic fungal strains isolated from diseased pomelo fruits were selected. The three pathogenic fungal strains were randomly injected into 9 healthy pomelo fruits. The strain PCP-B02-A2 led to a completely rotten fruit on day 17 after infection, while strains PCP-B02-B2 and PCP-B03-A1 had infected spots whose lengths were 17.5 and 28.1 mm, became larger, and eventually led to the whole fruit rot. The pathogens were identified by the internal transcribed spacer (ITS) technique as Colletotrichum gloeosporioides PCP-B02-A2, Pseudopestalotiopsis camelliae sinensis PCP-B03-A1, and P. chinensis PCP-B02-B2. Twenty-five Trichoderma spp. strains were isolated. The ITS technique identified four strains, including Trichoderma asperellum TP-B01, T. harzianum TP-B08, T. harzianum TP-B09, and T. asperellum TP-C25. The PCP-B02-A2 strain had antagonism at 66.7–68.7%, while those of PCP-B02-B2 and PCP-B03-A1 were 64.2–71.1% and 55.7–57.4%, respectively. Full article
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20 pages, 1509 KiB  
Article
Studying the Population Dynamics of NSLAB and Their Influence on Spores During Cheese Ripening
by Rakesh Kaushik and Sanjeev Anand
Appl. Microbiol. 2025, 5(3), 65; https://doi.org/10.3390/applmicrobiol5030065 - 3 Jul 2025
Viewed by 314
Abstract
Cheese ripening involves microbial changes, with starter lactic acid bacteria (SLAB) initiating fermentation and nonstarter lactic acid bacteria (NSLAB) driving flavor and texture development. However, heat-resistant spores of Clostridium and Bacillus can survive pasteurization and cause spoilage during ripening. This study evaluated NSLAB [...] Read more.
Cheese ripening involves microbial changes, with starter lactic acid bacteria (SLAB) initiating fermentation and nonstarter lactic acid bacteria (NSLAB) driving flavor and texture development. However, heat-resistant spores of Clostridium and Bacillus can survive pasteurization and cause spoilage during ripening. This study evaluated NSLAB dynamics in the presence of spores during cheese ripening. Cheddar cheese samples at pilot-scale level (110 L) with four treatments, namely control, with spores of B. licheniformis (T1), with spores of Cl. tyrobutyricum (T2), and both spores (T3) at 2.0 Log10 CFU/mL, were ripened at 7 °C for six months. SLAB declined from 8.0 to 0.2 Log10 CFU/g, while NSLAB increased from 2.0 to 8.5 Log10 CFU/g by month three and maintained their counts up to six months, unaffected by spore presence. Spore counts were ≤1.45 Log10 CFU/g in controls but reached 2.94 ± 0.02 (T2) and 2.48 ± 0.03 (T3), correlating with spoilage signs after five months. MALDI-TOF identified L. rhamnosus (up to 37%) and L. paracasei (up to 25%) as dominant NSLAB across treatments. Physicochemical parameters were not significantly affected by higher spore levels. While NSLAB dominated, they were inadequate to prevent spoilage in spore-inoculated samples exceeding 2.0 logs during cheese ripening. Full article
(This article belongs to the Special Issue Applied Microbiology of Foods, 3rd Edition)
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30 pages, 2010 KiB  
Review
Functional Versatility of Vibrio cholerae Outer Membrane Proteins
by Annabelle Mathieu-Denoncourt and Marylise Duperthuy
Appl. Microbiol. 2025, 5(3), 64; https://doi.org/10.3390/applmicrobiol5030064 - 3 Jul 2025
Viewed by 931
Abstract
A key feature that differentiates Gram-positive and Gram-negative bacteria is the outer membrane, an asymmetric membrane composed of lipopolysaccharides, phospholipids, lipoproteins and integral proteins, including the outer-membrane proteins (OMPs). By being in direct contact with the extracellular milieu, the outer membrane and OMPs [...] Read more.
A key feature that differentiates Gram-positive and Gram-negative bacteria is the outer membrane, an asymmetric membrane composed of lipopolysaccharides, phospholipids, lipoproteins and integral proteins, including the outer-membrane proteins (OMPs). By being in direct contact with the extracellular milieu, the outer membrane and OMPs participate in multiple functions in Gram-negative bacteria, including controlling nutrient and molecule access to the cytoplasm, membrane vesicle formation and resistance to environmental stresses. OMPs have a characteristic barrel shape formed by antiparallel β-strands, with or without channels that allow diffusion of substrates through the outer membrane. The marine bacterium Vibrio cholerae is responsible for non-invasive gastroenteritis and cholera disease by consumption of contaminated water or food. Its OMPs, besides having a porin function, contribute to resistance to osmotic pressure and antimicrobial agents, intracellular signaling, adhesion to host cells and biofilm formation, amongst other functions. In this review, in addition to quickly reviewing the general structure of the outer membrane, the OMPs and how they reach the outer membrane, the functions attributed to these proteins are compiled. The mechanisms used by each of the described OMP to accomplish these functions in the marine pathogenic bacterium V. cholerae are discussed. Potential clinical and bioengineering applications of OMPs, such as diagnostic tools, vaccine development, and targeted antimicrobial or anti-virulence strategies are presented. What is known about the OMPs of V. cholerae is presented below. Full article
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37 pages, 7888 KiB  
Article
Comprehensive Analysis of E. coli, Enterococcus spp., Salmonella enterica, and Antimicrobial Resistance Determinants in Fugitive Bioaerosols from Cattle Feedyards
by Ingrid M. Leon, Brent W. Auvermann, K. Jack Bush, Kenneth D. Casey, William E. Pinchak, Gizem Levent, Javier Vinasco, Sara D. Lawhon, Jason K. Smith, H. Morgan Scott and Keri N. Norman
Appl. Microbiol. 2025, 5(3), 63; https://doi.org/10.3390/applmicrobiol5030063 - 2 Jul 2025
Viewed by 629
Abstract
Antimicrobial use in food animals selects for antimicrobial-resistant (AMR) bacteria, which most commonly reach humans via the food chain. However, AMR bacteria can also escape the feedyard via agricultural runoff, manure used as crop fertilizer, and even dust. A study published in 2015 [...] Read more.
Antimicrobial use in food animals selects for antimicrobial-resistant (AMR) bacteria, which most commonly reach humans via the food chain. However, AMR bacteria can also escape the feedyard via agricultural runoff, manure used as crop fertilizer, and even dust. A study published in 2015 reported AMR genes in dust from cattle feedyards; however, one of the study’s major limitations was the failure to investigate gene presence in viable bacteria, or more importantly, viable bacteria of importance to human health. Our main objective was to investigate the presence and quantity of viable bacteria and antimicrobial-resistant (AMR) determinants in fugitive bioaerosols from cattle feedyards in the downwind environment. Six bioaerosol sampling campaigns were conducted at three commercial beef cattle feedyards to assess variability in viable bacteria and AMR determinants associated with geographic location, meteorological conditions, and season. Dust samples were collected using four different sampling methods, and spiral plated in triplicate on both non-selective and antibiotic-selective media. Colonies of total aerobic bacteria, Enterococcus spp., Salmonella enterica, and Escherichia coli were enumerated. Viable bacteria, including AMR bacteria, were identified in dust from cattle feedyards. Bacteria and antimicrobial resistance genes (ARGs via qPCR) were mainly found in downwind samples. Total suspended particles (TSPs) and impinger samples yielded the highest bacterial counts. Genes encoding beta-lactam resistance (blaCMY-2 and blaCTX-M) were detected while the most common ARG was tet(M). The predominant Salmonella serovar identified was Lubbock. Further research is needed to assess how far viable AMR bacteria can travel in the ambient environment downwind from cattle feedyards, to model potential public health risks. Full article
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28 pages, 1842 KiB  
Article
Association Between Adherence Levels to the EAT-Lancet Diet in Habitual Intake and Selected Gut Bacteria in a Mexican Subpopulation
by Mariana Lares-Michel, Rafael Vázquez-Solórzano, Zyanya Reyes-Castillo, Leilani Clarissa Salaiza-Ambriz, Salvador Ramírez-Guerrero, Fatima Ezzahra Housni, Avilene Rodríguez-Lara and Jesús R. Huertas
Appl. Microbiol. 2025, 5(3), 62; https://doi.org/10.3390/applmicrobiol5030062 - 30 Jun 2025
Viewed by 304
Abstract
The EAT-Lancet diet is an outstanding model of a healthy, environmentally sustainable diet. However, its effects on the gut microbiota remain poorly explored. This study assessed the asso-ciation between adherence to the EAT-Lancet diet in habitual intake and the relative abundance of selected [...] Read more.
The EAT-Lancet diet is an outstanding model of a healthy, environmentally sustainable diet. However, its effects on the gut microbiota remain poorly explored. This study assessed the asso-ciation between adherence to the EAT-Lancet diet in habitual intake and the relative abundance of selected gut bacteria in a Mexican subpopulation. Fifty-four young adults (18–35 years) completed a validated Food Frequency Questionnaire (FFQ) and were nutritionally assessed. Participants were grouped into low, moderate, and high adherence levels to the EAT-Lancet diet. Blood samples were analysed for glucose and lipid profiles, and gDNA from faecal samples was analysed using Real-time qPCR to quantify gut bacteria. While no significant differences in bacterial abundance were observed across adherence levels, correlations emerged with increased adherence. Notably, Bifidobacterium negatively correlated with beef and lamb intake (rho −0.5, p < 0.05), and Akkermansia muciniphila negatively correlated with fish intake (rho −0.8, p < 0.05). Bilophila wadsworthia positively correlated with triglycerides, while Prevotella copri and Faecalibacterium prausnitzii negatively correlated with body fat and blood pressure, respectively. In addition, a non-significant trend toward a higher abundance of Firmicutes, Akkermansia muciniphila, and Prevotella copri was observed in the high-adherence group, whereas Lactobacillus tended to be more abundant in participants with low and moderate adherence. These findings suggest that adherence to the EAT-Lancet diet modulates gut microbiota composition. However, further controlled interventional studies are needed to confirm these effects and their implications for human health. Full article
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12 pages, 1373 KiB  
Article
Characterizing Aqueous Extracts of Native Plants in Northeastern Mexico: Prospects for Quorum-Sensing Inhibition Against Gram-Negative Bacteria
by Jose E. Quiroz-Hernandez, Gustavo Hernandez-Vidal, Orquidea Perez-Gonzalez, Uziel Castillo-Velazquez and Victor E. Aguirre-Arzola
Appl. Microbiol. 2025, 5(3), 61; https://doi.org/10.3390/applmicrobiol5030061 - 29 Jun 2025
Viewed by 392
Abstract
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 [...] Read more.
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
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15 pages, 295 KiB  
Article
In Silico Detection of Genetic Determinants for the Acquired Antibiotic Resistance and Biologically Active Compounds of Lactic Acid Bacteria from the Human Oral Microbiome
by Nikola Atanasov, Yana Evstatieva and Dilyana Nikolova
Appl. Microbiol. 2025, 5(3), 60; https://doi.org/10.3390/applmicrobiol5030060 - 29 Jun 2025
Viewed by 301
Abstract
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 [...] Read more.
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
13 pages, 1117 KiB  
Article
Respiratory Microbiota Associations with Asthma Across American and Emirati Adults: A Comparative Analysis
by Ariangela J. Kozik, Kyra Henderson, Laila Salameh, Bassam Mahboub, Mohammad T. Al Bataineh and Yvonne J. Huang
Appl. Microbiol. 2025, 5(3), 59; https://doi.org/10.3390/applmicrobiol5030059 - 29 Jun 2025
Viewed by 380
Abstract
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 [...] Read more.
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
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16 pages, 1122 KiB  
Article
Effect of r-Human Insulin (Humulin®) and Sugars on Escherichia coli K-12 Biofilm Formation
by Balbina J. Plotkin, Ira Sigar and Monika Konaklieva
Appl. Microbiol. 2025, 5(3), 58; https://doi.org/10.3390/applmicrobiol5030058 - 27 Jun 2025
Viewed by 223
Abstract
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 [...] Read more.
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
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18 pages, 1031 KiB  
Article
Microbiome Signatures and Inflammatory Biomarkers in Culture-Negative Neonatal Sepsis
by Morcos Hanna, Shixia Huang, Matthew Ross, Anaid Reyes, Dimuthu Perera, Anil Surathu, Sara Javornik Cregeen, Joseph Hagan and Mohan Pammi
Appl. Microbiol. 2025, 5(3), 57; https://doi.org/10.3390/applmicrobiol5030057 - 24 Jun 2025
Viewed by 320
Abstract
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 [...] Read more.
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
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13 pages, 487 KiB  
Article
Differential Rumen Microbial Taxa in Charolais Bulls with Divergent Residual Feed Intake
by Taylor Sidney, Emily Treon, Godstime Taiwo, Samanthia Johnson, Yarahy Leal, Peixin Fan and Ibukun M. Ogunade
Appl. Microbiol. 2025, 5(3), 56; https://doi.org/10.3390/applmicrobiol5030056 - 24 Jun 2025
Viewed by 284
Abstract
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 [...] Read more.
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
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