Applied Microbiology

Anorexia nervosa (AN) is a psychiatric illness with harmful physical consequences. Studies have observed differences in the faecal microbiota of patients with AN compared to healthy controls. Diet has an impact on the gut microbiota, facilitating an altered community, such changes could impact the gut–brain axis. In this study, a three-stage gut model system that mimics the luminal microbiology of the large intestine was conducted to identify relationships between diet and gut microbiota. A microbial medium was developed to provide nutrients more appropriate to restricting subtype AN (R-AN). The model was inoculated with faeces and samples were taken to compare differences in the microbiota and end products following the fermentation of healthy control medium (HC) compared to R-AN medium. Then, 16S amplicon sequencing along with flow cytometry–fluorescence in situ hybridisation were used to ascertain changes in the microbiota. Gas chromatography (GC) was used to assess changes in microbial metabolites. There were reduced levels of SCFA following the fermentation of R-AN medium. The fermentation of R-AN media led to fewer total bacteria numbers, along with less bifidobacteria and Rumincoccus proximally, but more Clostridium and Enterobacteriaceae. Nutrient-deficient medium resulted in reduced neurotransmitter-producing bacteria, reduced butyrate-producing bacteria, and increased protein-utilising bacteria, all of which could be maintaining factors in AN. The model system provides a novel tool for exploring how extreme dietary changes impact the microbiota and could therefore could be useful for assessing appropriate gut–brain targeted treatments. Full article

With the increase in mercury pollution around the world, several bacteria have been identified that are capable of resisting mercury toxicity. With this in mind, the aim of this review was to determine which genes are involved in mercury resistance, which bacterial genera exhibit this resistance, and which bacterial isolation sources have been most reported. To answer these questions, the PICO method (population, intervention, comparison, and outcome) was used, three databases were searched, and 17 relevant articles were included. As a result, resistance is due to a set of mer genes that transcribe mer proteins. The most important genes identified were merA and merR, and their proteins confer resistance by reducing Hg to Hg⁺² or Hg⁰. Among the bacteria studied, those of the genera Pseudomonas, Escherichia, and the phylum Cyanobacteria stand out, the most important being Escherichia coli and Synechocystis sp., which are highly efficient and fast at reducing Hg. Based on the results, Escherichia coli and Synechocystis sp. are promising candidates for reducing environmental Hg, especially in aquatic environments. However, there is a lack of studies on the mechanism of bioremediation carried out by cyanobacteria and the influence of abiotic factors on the presence and/or expression of mer genes. Full article

The global food security crisis is emphasized by the alarming amount of food waste, where about one-third of the world’s food production, roughly 1.3 billion metric tons, is lost annually. Pathogens, such as Botrytis cinerea, contribute significantly to this loss by attacking stored agricultural produce. These attacks typically start when pathogens infiltrate small fruit wounds, remain dormant, and then switch to an aggressive necrotrophic stage upon ripening, causing significant postharvest food losses. In response to this challenge, this study presents an innovative application of Reverse Transcriptase Loop-Mediated Isothermal Amplification (RT-LAMP). This method is increasingly recognized for its simplicity and effectiveness, distinguishing itself from more complex molecular diagnostic techniques. This study focuses on developing a heat-dry RT-LAMP desiccation method designed to be simple, robust, rapid, sensitive, and specific in detecting Botrytis cinerea. This method lies in its utilization of a desiccation process, where heat is utilized to preserve crucial components such as primers and enzymes in the presence of trehalose. A 5% trehalose with an amplification time of 1 h and 40 min was optimal for the assay detection of latent Botrytis cinerea. This method exhibited a sensitivity of 10 femtograms and was tailored specifically to the Botrytis cinerea PLF marker. Validation was performed using RNA extracted from an infected tomato, establishing a detection threshold of 1 ng/µL, approximately 500 pg of synthesized DNA target marker. This discovery holds significant implications, suggesting the potential for developing dry RT-LAMP kits that are adaptable for both laboratory and field usage. Furthermore, this method shows promise as a diagnostic tool for other neglected pathogenic diseases, representing a substantial advancement in agricultural pathology and supporting endeavors to enhance food security. Full article

Host genetics and environmental factors have been associated with effects on the mouse fecal microbiome; however, the commercial source of mice remains the dominant factor. Increasing evidence indicates that supplier-specific microbiomes confer differences in disease susceptibility in models of inflammatory conditions, as well as baseline behavior and body morphology. However, current knowledge regarding the compositional differences between suppliers is based on targeted-amplicon sequencing data, and functional differences between these communities remain poorly defined. We applied a multi-omic (metagenomic and metatranscriptomic) approach to biomolecules extracted from murine feces representative of two U.S. suppliers of research mice, which differ in composition, and influence baseline physiology and behavior as well as disease severity in models of intestinal disease. We reconstructed high-quality metagenome-assembled genomes, frequently containing genomic content unique to each supplier. Transcriptional activity and pathway analyses revealed key functional differences between the metagenomes associated with each supplier including carbohydrate, fatty acid, and sulfite metabolism. These data provide a detailed characterization of the baseline differences in the fecal metagenome of mice from two U.S. commercial suppliers, suggesting that these functional differences are influenced by differences in the initial inoculum of colony founders, as well as additional taxa gained during growth of the production colony. Full article

On commercial banana (Musa spp.) plantations, soils are often supplemented with phosphorus (P) fertiliser to optimise production. Such additions may influence the diversity and function of soil microbial communities, which play important roles in P cycling and affect plant fitness. Here, we characterised the effects of P addition on the diversity and function of banana-associated microbial communities. P addition was associated with significant increases in soil P and the activities of alpha-glucosidase, chitinase, arylsulphatase, and acid phosphatase, but not beta-glucosidase or xylosidase. P addition also expedited bunch emergence and harvest, but did not influence fruit yield, plant height, or foliar P. There were no significant effects of P addition on the alpha or beta diversity of bacterial, fungal, and nematode communities, including members of the core microbiome. The only exceptions to this was an increase in the relative abundance of a Fusarium population in roots. These results indicate that phosphorus application to banana soils may stimulate microbial enzyme activities with minor or negligible effects on microbial diversity. Full article

Staphylococcus aureus is a leading cause of bloodstream infection (SAB), with up to 30% mortality. Despite treatment with standard antibiotics, one in three patients develops a persistent infection, which portends a five-fold increase in the risk of death. Persistent SAB has been attributed in part to the inability of antistaphylococcal antibiotics to eradicate intracellular S. aureus surviving inside macrophages. (-)- Epigallocatechin gallate (EGCG) is a catechin found in green tea that has been widely studied for its broad biological activities, ranging from anticancer to antibacterial activity. However, EGCG is greatly limited by its poor drug-like properties in terms of stability, membrane permeability, and bioavailability. In this study, we established through a series of in vitro experiments that structural modifications of EGCG enhanced drug-like properties while maintaining or improving its antistaphylococcal activity. Our lead EGCG analogs (MCC-1 and MCC-2) showed improved biochemical properties along with increased potency against extracellular S. aureus and restored susceptibility of β-lactam agents to methicillin-resistant S. aureus (MRSA). Importantly, the lead analogs but not EGCG potentiated macrophage- and antibiotic-mediated clearance of intracellular bacteria. Overall, EGCG analogs showed promise for further development as adjunctive therapy candidates for the treatment of SAB. Full article

The rhizosphere, a narrow region of soil surrounding plant roots, is an environment rich in microbial diversity that profoundly influences plants’ health, growth, and agricultural productivity. This microbial community, known as the rhizosphere microbiome, consists of a complex array of bacteria, fungi, archaea, and other microorganisms that engage in complex interactions with plant roots. These microorganisms contribute to nutrient cycling, mineral uptake facilitation, and protection against soil-borne pathogens, thereby promoting plant growth and resilience towards biotic and abiotic stresses. Additionally, microbial signaling molecules, including phytohormones such as auxins, cytokinin, gibberellins, ethylene, and abscisic acid, play a pivotal role in regulating these interactions by modulating plants’ responses to environmental stressors. Recent advancements in microbiomics have enabled a deeper understanding of the rhizosphere’s diversity, composition, and functions, paving the way for more sustainable agricultural practices. By harnessing the potential of the rhizosphere microbiome, innovative strategies can be developed to reduce dependency on synthetic agrochemicals, enhance soil fertility, and increase crop yields. This review discusses the diversity and mechanisms of plant–microbe interactions, focusing on the role of microbial signaling molecules, and explores their applications in promoting agricultural sustainability. The insights gained from microbiomics studies can revolutionize farming practices by reducing dependency on chemical inputs, enhancing crop productivity, and nurturing soil health and environmental sustainability. Full article

The gut microbiota plays an essential role in its host’s nutrition, development and behavior. Although crickets are becoming major ecosystemic model systems and have important societal applications, such as alternative animal proteins or biocatalysts, little is known about their gut microbiome acquisition and how environmental factors shape this community. Therefore, in this study, we exposed sand field crickets to soils with different characteristics and microbial communities to test the influence of these on gut microbial community composition. We used 16S/18S rRNA gene Illumina sequencing to analyze different soil and gut communities, targeting the three domains of life, Archaea, Bacteria, and Eukaryotes. Our results showed a dominance of Mucoromycota fungi and Bacteroidota in the gut microbiota. We were unable to retrieve sufficient read numbers for the Archaea. Most of the microbial taxa that were identified can degrade soil-derived complex organic matter, likely helping the host digest its food. The soil characteristics had a significant impact on the gut microbial community structure, supporting our assumption that the environment plays an essential role in gut microbiota acquisition. Host sex also had an impact on the gut community, possibly because the female guts were bigger in mass, leading to differences in oxygen concentrations. Full article

Salmonella has had a long and intimate relationship with humans and continues to raise concerns for human health, but this close bond also provides opportunities for new therapeutics and treatments. Although Salmonella enterica serovar Typhi is the principal organism that comes to mind in terms of death and morbidity, it is the non-typhoidal Salmonellae that have the most health and economic implications. The developed world has had a challenging relationship with Salmonella, particularly in the UK and the EC/EU, experiencing significant Salmonella outbreaks in the 1980s and 1990s. As a consequence, the research focus was on understanding the nature of infection in food animals and on developing ways and means of controlling zoonotic infections. This led to the development of numerous model systems for the study of Salmonella both in vitro and in vivo. The introduction of vaccination has all but eliminated Salmonella in eggs and reduced perceived risk held by the general public. At the same time as Salmonella in eggs was being brought under control in the UK and EU, the danger posed by antibiotic resistance was beginning to emerge. In the past, with the efficacy of antibiotics against Gram-negative bacteria being unchallenged, there was limited focus on the threat posed by antibiotic resistance in non-typhoidal Salmonella. However, the identification of Salmonella as the first ‘multidrug-resistant’ organism, the presence of invasive non-typhoidal Salmonella in North Africa and the emergence of monophasic Salmonella enterica serovar Typhimurium across Europe, Asia and the Americas have prompted renewed interest in Salmonella research, particularly in the context of non-infectious disease, biofilm studies and antibiotic resistance. At the same time, research has continued to develop ways of taking advantage of what Salmonella offers in the way of pathogenic factors and the therapeutic and treatment applications in areas such as vaccine development, cancer therapeutics and drug delivery and the role of Salmonella in non-infectious diseases supported by developments in molecular and genomic methods. Full article

This review presents an approach to the incorporation of cyanobacteria and microalgae in yogurts and explores their impact on the nutritional, rheological, sensory, and antioxidant qualities of these products. First, the yogurt market context and its relationship with nutritional quality are outlined, emphasizing the quest for functional foods that meet consumer demands for healthy and nutritious products. A discussion of the incorporation of cyanobacteria and microalgae, especially Spirulina platensis, in foods, particularly yogurt, is then presented, highlighting the nutritional and functional benefits that this type of biomass can provide to the final product. The fermentation process and the quantity of algae to be incorporated are discussed to understand their fundamental role in the characteristics of the final product. In addition, this article considers some challenges such as sensory and rheological changes in the product resulting from the interaction of milk, algal biomass, and the fermentation process. Addressing these challenges involves delineating how these interactions contribute to changes in the traditionally consumed product, while obtaining a pro- and prebiotic product is crucial for creating an innovative dairy product that diversifies the market for derived dairy products with increased functional properties. Full article

The emergence of antimicrobial resistance (AMR) in pathogens transmitted through food poses a significant threat to global public health, complicating infection treatment and increasing mortality rates. This review explores the role of resistome mapping as a crucial tool for understanding the transmission dynamics of antimicrobial resistance genes (ARGs) in foodborne pathogens, such as Salmonella, Escherichia coli, Listeria monocytogenes, and Campylobacter spp., as well as various techniques for resistome mapping, such as metagenomic sequencing, PCR-based methods, and whole-genome sequencing (WGS), highlighting the significance of horizontal gene transfer (HGT) as a key mechanism for ARG dissemination in foodborne pathogens. Additionally, we investigated the influence of agricultural practices and environmental factors on AMR development, highlighting the critical need for improved surveillance, antibiotic stewardship, and global collaboration to mitigate the spread of resistant ARGs through the food chain. The perceptions gained from resistome mapping play an essential role in developing effective approaches to address AMR and to ensure food safety. Full article

A conventional, group-specific PCR method was developed to identify each of the four previously defined major taxa (Myoviridae, Siphoviridae, Podoviridae and Microviridae) of somatic coliphages and used to classify isolates from sewage. Somatic coliphage infectivity detection, occurrence and survival in primary human sewage effluent was observed over time to further understand the presence and behavior of the groups of somatic coliphages at two environmental temperatures (4 and 25 °C). Over time, the taxonomic composition of the somatic coliphage population in sewage changed, with the Microviridae family becoming the most prevalent family in the sewage population after several weeks. Based on their persistence and prevalence in environmental waters, phages belonging to the Microviridae family provide supporting information on sewage contamination and possibly of human enteric viruses in sewage-contaminated water. Full article

This study explores the integration of genome sequencing and digital droplet polymerase chain reaction (ddPCR)-based methods for tracking the diversity of COVID-19 variants in wastewater. The research focuses on monitoring various Omicron subvariants during a period of significant viral evolution. Genome sequencing, particularly using Oxford Nanopore Technology (ONT), provides a detailed view of emerging variants, surpassing the limitations of PCR-based detection kits that rely on known sequences. Of the 43 samples analyzed, 39.5% showed matching results between the GT Molecular ddPCR kits and sequencing, though only 4% were exact matches. Some mismatches occurred due to newer subvariants like XBB and BQ.1, which the ddPCR kits could not detect. This emphasized the limitations of ddPCR kits, which rely on known variant sequences, while sequencing provides real-time data on emerging variants, offering a more comprehensive view of circulating strains. This study highlights the effectiveness of combining these methodologies to enhance early detection and inform public health strategies, especially in regions with limited clinical sequencing capabilities. Full article

V. lutrae is an emerging human pathogen attributed to increasing hospitalization cases in humans; however, its biology and epidemiology are under-explored. The present study explored V. lutrae recovery, prevalence, and biology. A two-step enrichment method (i.e., step 1, nourishment; step 2, heat, 80 °C, 20 min) and thiosulfate–citrate–bile salts–sucrose (TCBS) agar were employed for recovering V. lutrae in raw seafood. Bacterial colonies were streaked for purification before 16S rRNA bacterial identification. Confirmed V. lutrae isolates were analyzed for their culture-challenged turbidity and virulence. Of 41 bacterial isolates, 9 confirmed V. lutrae, including regular (33%; nourished 24 h) and heat-resistant (67%; nourished 48 h plus heating) isolates, were exclusively from yellow colonies (i.e., TCBS) and were exclusively recovered from nourished shrimp (78%) and crab (22%) only. The culture and virulence biology revealed that they could diversely tolerate salinity (i.e., 0–17.5% additional NaCl), pasteurization (63 °C, 8 h), oxygen availability, and antibiotic sensitivity (i.e., erythromycin, gentamicin, and vancomycin). Further, this pathogen exhibited no visible hemolytic and alkalization activities. Emerging foodborne pathogens could readily evade the established food safety regime. The present study reveals systematic investigation and diverse phenotypes of V. lutrae to enhance its detection and contribute to public health initiatives. Full article

With the anticipated population growth by 2050, the demand for high-quality protein for human consumption is set to rise. To enhance the sustainability of U.S. cattle production, producers and researchers have traditionally concentrated on improving the feed efficiency of steers through advancements in genetics, nutrition, and microbiome tools, resulting in a more marketable beef product. However, without successful pregnancies, there would be no marketable animals to feed. Despite extensive research on hormonal impacts on cattle physiology, including nutrition and reproduction, there is limited knowledge about how the rumen microbial environment is impacted by pregnancy and feed efficiency in female beef cattle. Understanding the rumen microbiome’s role in feed efficiency and its response to hormonal changes during pregnancy is crucial for advancing sustainable beef cattle production. Therefore, this review highlights the importance of understanding the microbial dynamics in the rumen of pregnant beef females. Although progress has been made, gaps remain in understanding how varying nutritional requirements throughout pregnancy affect the rumen microbiome, highlighting the need for continued research. Addressing these areas will lead to more efficient and sustainable cattle production practices, benefiting beef production and contributing to global food security. Full article

This study explores the relationship between filamentous fungi and dental caries in isolated indigenous communities in Siltepec, Chiapas, Mexico. A total of 37 oral swabs were collected, with 22 participants harboring filamentous fungi, primarily from the genus Cladosporium. Statistical analysis using Student’s t-test and the Mann–Whitney U test revealed a significant reduction in extensive and fully cavitated caries (p < 0.0001) in individuals with fungi, while those without fungi exhibited higher rates of dental decay. Participants with fungi had a higher prevalence of healthy teeth and incipient caries. The findings suggest that traditional maize-based diets, particularly fermented beverages like pozol, may promote the growth of beneficial fungi in the oral microbiome, offering a protective effect against dental caries through microbial competition and the alteration of the oral environment. These results underline the need for further research into the long-term impact of traditional diets on oral health and the potential use of natural substances, such as probiotics and plant-based antimicrobials, to maintain oral homeostasis and prevent caries. Full article

Pseudomonas aeruginosa is a ubiquitous environmental Gram-negative bacterium and also an opportunistic pathogen for both humans and animals, causing acute or chronic infections. It has been frequently detected in healthy and diseased reptiles, more commonly in captive ones. Since most studies are primarily on clinical isolates, the pathogenic potential of strains originating from wild animals is poorly explored. We isolated the strain P. aeruginosa PM1012 from the cloacal microbiota of a common wall lizard (Podarcis muralis Laurenti, 1768) from a free-living population. The effect of temperature, pH and salinity on its growth was evaluated. Antibiotic resistance, the expression of several virulence factors as some extracellular enzymes, pyocyanin production and biofilm formation were also assessed. Apart from intrinsic resistance, the newly isolated strain P. aeruginosa PM1012 presented an antibiotic susceptibility profile with a low resistance rate limited to meropenem and intermediate to ceftazidime and aztreonam. Protease, lipase and gelatinase secretion was detected. Strong pyocyanin production was observed in the optimal range of growth conditions. An excellent biofilm-forming capacity was manifested. Full article

Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) spreads primarily through respiratory droplets, aerosols, and contaminated surfaces. While high-traffic locations like hospitals and airports have been studied extensively, detecting significant virus levels in aerosols and on environmental surfaces, campus settings remain underexplored. This study focused on two crowded buildings at the University of North Carolina at Charlotte (UNCC). From December 2021 to March 2022, we collected 16 indoor air samples and 201 samples from high-touch surfaces. During the sampling timeframe, 44.82% of surface samples from the Student Union and 28% from the University Recreational Center (UREC) tested positive for the presence of SARS-CoV-2 RNA. Median and average viral RNA copies per swab were higher in UREC (273 and 475) than in Student Union (92 and 269). However, all air samples tested negative. Surface positivity in these high-traffic campus locations was directly correlated with COVID-19 clinical cases in Mecklenburg County. The campus COVID-19 cases, driven by the Omicron wave, peaked a week before the peak detection of surface contamination. These findings underscore the importance of surface hygiene measures and highlight environmental conditions as potential contributors to COVID-19 spread on campuses. Full article

The terrestrial biota is a rich source of biologically active substances whose anti-biofilm potential is not studied enough. The aim of this review is to outline a variety of terrestrial sources of antimicrobial agents with the ability to inhibit different stages of biofilm development, expecting to give some ideas for their utilization in improved anti-biofilm treatments. It provides an update for the last 5 years on anti-biofilm plant products and derivatives, essential oils, antimicrobial peptides, biosurfactants, etc., that are promising candidates for providing novel alternative approaches to combating multidrug-resistant biofilm-associated infections. Based on the reduction in bacterial adhesion to material and cell surfaces, the anti-adhesion strategy appears interesting for the prevention of bacterial attachment in combating a broad range of mono- and multispecies bacterial biofilms. So far, few studies have been carried out in this direction. Anti-biofilm coatings made by or containing biologically active products from terrestrial biota have scarcely been studied although they are of significant interest for a reduction in infections associated with medical devices. Combination therapy with commercial antibiotics and natural products is accepted now as a promising base for future advances in anti-biofilm treatment. In vivo testing and clinical trials are necessary for clinical application. Full article