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Introduction The natural behavior of horses is to spend the most of their time foraging. The feeding time of stabled horses is often far from enough to fulfill their natural eating behavior. Hay-net technique is a method aiming to promote natural foraging behavior, slowing ingestion and increasing the length of feeding time. This study was conducted to investigate the effects of using a slow-feed (hay-net) on feeding time, forage intake rate, apparent digestibility, volatile fatty acids (VFA) concentration, bacterial population, and the activity of cellulolytic and amylolytic enzymes in the colon of horses. Materials and Methods Eight Arabian horses (body weight= 396± 45 kg, age= 9±3 years) were used in a cross-over design. The length of the experimental period was eight weeks, which included two periods of four weeks in rotation. In each period, feed was provided for four horses freely and for other four horses using a hay net. Fecal samples were collected on days 21 to 28 of each period. Results and Discussion The results showed that using hay net significantly increased feeding time from 257 to 659 minutes per day and decreased forage intake rate from 1592 and 610 g/h (P<0.05). The hay-net did not affect the total VFA and relative VFAs concentrations (P< 0.05). Lactobacillus populations in horses with hay-net increased compared to those without hay-net (P<0.05). The populations of cellulolytic, amylolytic, Escherichia coli, and coliforms bacteria were unchanged by hay-net. The activity of carboxy methyl cellulase enzyme significantly increased (P<0.05), while the activity of microcrystalline cellulase, amylase, and filter paper decomposition activity was not affected by hay-net. Conclusion In comparison with the more traditional methods, slow-feed hay net devices successfully limited the forage intake rate, expanded feeding time, and changed the activity of some fiber-decomposing enzymes in the large intestine of horses; however, the recommendation of hay-nets for nutritional management in horses needs further investigations. [ABSTRACT FROM AUTHOR]

Introduction: Anorexia Nervosa (AN) stands as a severe mental disorder characterized by the highest mortality rate within the realm of psychiatric conditions. Its etiology encompasses genetic, neurobiological, environmental, and developmental factors. Recent investigations have shed light on the potential impact of the gut microbiota on the genesis and progression of AN. Aim of the Study: The aim of this paper is to provide a comprehensive review of the existing findings on relationship between gut dysbiosis and AN. The intention is to present new perspectives that contribute to a more detailed understanding of the multifaceted nature of this serious disorder. Description of the State of Knowledge: The gut microbiota takes a central role in modulating various physiological processes. Dysbiosis, which refers to an imbalance in gut microbiota composition, has recently drawn attention because of its association with several mental disorders. In the context of AN, studies have shown reduced microbial diversity and notable changes in specific bacterial populations. The influence of the gut microbiota on AN includes disturbances in the digestive system, changes in eating behaviours, and associations with related conditions such as anxiety and depression. Mechanisms such as the gut-brain axis, hormonal regulation and molecular mimicry contribute to these associations. Conclusions: Recognition of the pivotal role played by the gut microbiota in AN opens up avenues for potential therapeutic interventions. Pro- and prebiotics, along with fecal transplantation, emerge as promising modalities in the treatment of AN. Ongoing research is essential to further elucidate this evolving field, ultimately facilitating the development of targeted interventions for individuals with AN. [ABSTRACT FROM AUTHOR]

Mathematical models of cancer and bacterial evolution have generally stemmed from a gene-centric framework, assuming clonal evolution via acquisition of resistance-conferring mutations and selection of their corresponding subpopulations. More recently, the role of phenotypic plasticity has been recognized and models accounting for phenotypic switching between discrete cell states (e.g., epithelial and mesenchymal) have been developed. However, seldom do models incorporate both plasticity and mutationally driven resistance, particularly when the state space is continuous and resistance evolves in a continuous fashion. In this paper, we develop a framework to model plastic and mutational mechanisms of acquiring resistance in a continuous gradual fashion. We use this framework to examine ways in which cancer and bacterial populations can respond to stress and consider implications for therapeutic strategies. Although we primarily discuss our framework in the context of cancer and bacteria, it applies broadly to any system capable of evolving via plasticity and genetic evolution. [ABSTRACT FROM AUTHOR]

Bacterial genomes often harbor integrated viruses (prophages), which provide novel functions but also lyse cells under stressful conditions. A new paper combines mathematical models with experimental evolution to determine how prophages are maintained in bacterial populations despite their fitness costs. [ABSTRACT FROM AUTHOR]

In combination therapy, bacteria are challenged with two or more antibiotics simultaneously. Ideally, separate mutations are required to adapt to each of them, which is a priori expected to hinder the evolution of full resistance. Yet, the success of this strategy ultimately depends on how well the combination controls the growth of bacteria with and without resistance mutations. To design a combination treatment, we need to choose drugs and their doses and decide how many drugs get mixed. Which combinations are good? To answer this question, we set up a stochastic pharmacodynamic model and determine the probability to successfully eradicate a bacterial population. We consider bacteriostatic and two types of bactericidal drugs—those that kill independent of replication and those that kill during replication. To establish results for a null model, we consider non‐interacting drugs and implement the two most common models for drug independence—Loewe additivity and Bliss independence. Our results show that combination therapy is almost always better in limiting the evolution of resistance than administering just one drug, even though we keep the total drug dose constant for a 'fair' comparison. Yet, exceptions exist for drugs with steep dose–response curves. Combining a bacteriostatic and a bactericidal drug which can kill non‐replicating cells is particularly beneficial. Our results suggest that a 50:50 drug ratio—even if not always optimal—is usually a good and safe choice. Applying three or four drugs is beneficial for treatment of strains with large mutation rates but adding more drugs otherwise only provides a marginal benefit or even a disadvantage. By systematically addressing key elements of treatment design, our study provides a basis for future models which take further factors into account. It also highlights conceptual challenges with translating the traditional concepts of drug independence to the single‐cell level. [ABSTRACT FROM AUTHOR]

Background: While the urogenital microbiota has recently been characterized in healthy male and female dogs, the influence of sex hormones on the urogenital microbiome of bitches is still unknown. A deeper understanding of the cyclic changes in urinary and vaginal microbiota would allow us to compare the bacterial populations in healthy dogs and assess the impact of the microbiome on various urogenital diseases. Therefore, the aim of this study was to characterize and compare the urogenital microbiota during different phases of the estrous cycle in healthy female dogs. DNA extraction, 16 S rDNA library preparation, sequencing and informatic analysis were performed to determine the vaginal and urinary microbiota in 10 healthy beagle dogs at each phase of the estrous cycle. Results: There were no significant differences in alpha and beta diversity of the urinary microbiota across the different cycle phases. Similarly, alpha diversity, richness and evenness of vaginal bacterial populations were not significantly different across the cycle phases. However, there were significant differences in vaginal beta diversity between the different cycle phases, except for between anestrus and diestrus. Conclusion: This study strongly suggests that estrogen influences the abundance of the vaginal microbiota in healthy female dogs, but does not appear to affect the urinary microbiome. Furthermore, our data facilitate a deeper understanding of the native urinary and vaginal microbiota in healthy female dogs. [ABSTRACT FROM AUTHOR]

Transcriptional heterogeneity in isogenic bacterial populations can play various roles in bacterial evolution, but its detection remains technically challenging. Here, we use microbial split-pool ligation transcriptomics to study the relationship between bacterial subpopulation formation and plasmid-host interactions at the single-cell level. We find that single-cell transcript abundances are influenced by bacterial growth state and plasmid carriage. Moreover, plasmid carriage constrains the formation of bacterial subpopulations. Plasmid genes, including those with core functions such as replication and maintenance, exhibit transcriptional heterogeneity associated with cell activity. Notably, we identify a cell subpopulation that does not transcribe conjugal plasmid transfer genes, which may help reduce plasmid burden on a subset of cells. Our study advances the understanding of plasmid-mediated subpopulation dynamics and provides insights into the plasmid-bacteria interplay. Transcriptional heterogeneity in isogenic bacterial populations can play various roles in bacterial evolution, but its detection remains technically challenging. Here, Cyriaque et al. use microbial split-pool ligation transcriptomics to study the relationship between bacterial subpopulation formation and plasmid-host interactions at the single-cell level, providing insights into plasmid-bacteria dynamics. [ABSTRACT FROM AUTHOR]

Small intestinal bacterial overgrowth (SIBO) is characterized by an increase in the bacterial population of the small intestine due to an imbalance between the amount of bacteria and the intestinal barrier. Pediatric SIBO presents with a wide spectrum of symptoms, ranging from mild gastrointestinal complaints to malabsorption or malnutrition. Breath tests are commonly used as noninvasive diagnostic tools for SIBO, but a standardized methodology is currently unavailable. Intestinal flora produces methane which slows intestinal transit and increases the contractile activity of small intestine. Emerging literature suggests a correlation between overgrowth of methanogenic bacteria in the intestines and constipation. Treatment of SIBO involves administration of antibacterial therapy in addition to management of underlying conditions and optimal dietary adjustments. However, research on antibiotic treatment for pediatric patients with constipation and SIBO is limited and has yielded conflicting results. In the current review, we summarize the state-of-the-art of the field and discuss previous treatment attempts and currently used regimens for SIBO patients with constipation, with a focus on pediatric populations. [ABSTRACT FROM AUTHOR]

The Internet of Things (IoT) is made up of billions of interconnected devices that can transmit and receive data over the Internet. IoT devices have many vulnerabilities that attackers could use to compromise their security because of the heterogeneity of device connectivity. Distributed denial-of-service (DDoS) attacks against those applications become more common as IoT applications continue to expand and devolve. Identifying DDoS attacks is a difficult process due to the variety of IoT devices connected. The present article proposed a new method to detect DDoS attacks using an optimized Elman recurrent neural network (ERNN) based on chaotic bacterial colony optimization (CBCO) called CBCO-ERNN. The proposed method uses CBCO for obtaining optimal parameters (weights and biases) and structure (number of hidden neurons) of ERNN architecture. The chaos theory is applied to improve BCO's exploration and exploitation capabilities by initializing the bacterial population and selecting the appropriate chemotaxis step size value. The CBCO approach is used to train the ERNN model to avoid local optima and enhance the convergence rate. The performance of the CBCO-ERNN is tested and evaluated using four benchmark attack datasets such as the BoT-IoT, CIC-IDS2017, CIC-DDoS2019, and IoTID20 datasets, and five performance metrics are considered: accuracy, sensitivity, specificity, precision, and F-Score. According to the experimental results, the CBCO-ERNN method provides a high detection and a faster convergence rate when compared to earlier algorithms. [ABSTRACT FROM AUTHOR]

Introduction: Symbiotic microbial have a significant impact on the growth and metabolism of medicinal plants. Schisandra chinensis is a very functionally rich medicinal herb; however, its microbial composition and diversity have been poorly studied. Methods: In the present study, the core microbiomes associated with the rhizospheric soil, roots, stems, leaves, and fruits of S. chinensis from six geographic locations were analyzed by a macro-genomics approach. Results: Alpha and beta diversity analyses showed that the diversity of microbial composition of S. chinensis fruits did not differ significantly among the geographic locations as compared to that in different plant compartments. Principal coordinate analysis showed that the microbial communities of S. chinensis fruits from the different ecological locations were both similar and independent. In all S. chinensis samples, Proteobacteria was the most dominant bacterial phylum, and Ascomycota and Basidiomycota were the most dominant fungal phyla. Nitrospira, Bradyrhizobium, Sphingomonas, and Pseudomonas were the marker bacterial populations in rhizospheric soils, roots, stems and leaves, and fruits, respectively, and Penicillium, Golubevia, and Cladosporium were the marker fungal populations in the rhizospheric soil and roots, stems and leaves, and fruits, respectively. Functional analyses showed a high abundance of the microbiota mainly in biosynthesis. Discussion: The present study determined the fungal structure of the symbiotic microbiome of S. chinensis, which is crucial for improving the yield and quality of S. chinensis. [ABSTRACT FROM AUTHOR]

The interactions between a virus and its host vary in space and time and are affected by the presence of molecules that alter the physiology of either the host or the virus. Determining the molecular mechanisms at the basis of these interactions is paramount for predicting the fate of bacterial and phage populations and for designing rational phage-antibiotic therapies. We study the interactions between stationary phase Burkholderia thailandensis and the phage ΦBp-AMP1. Although heterogeneous genetic resistance to phage rapidly emerges in B. thailandensis, the presence of phage enhances the efficacy of three major antibiotic classes, the quinolones, the beta-lactams and the tetracyclines, but antagonizes tetrahydrofolate synthesis inhibitors. We discovered that enhanced antibiotic efficacy is facilitated by reduced antibiotic efflux in the presence of phage. This new phage-antibiotic therapy allows for eradication of stationary phase bacteria, whilst requiring reduced antibiotic concentrations, which is crucial for treating infections in sites where it is difficult to achieve high antibiotic concentrations. Author summary: Bacteriophages are viruses that infect and kill bacteria and therefore are considered to be a very valuable alternative to the antibiotic molecules that are currently employed in the clinic but towards which bacteria are becoming increasingly resistant. Here we show that when infected with a bacteriophage termed ΦBp-AMP1, the bacterium Burkholderia thailandensis becomes more susceptible to three major antibiotic classes routinely employed to treat infections caused by Burkholderia. We discovered that enhanced antibiotic efficacy in the presence of this bacteriophage is due to a decreased capability of the bacterium to expel antibiotics when it is infected with this bacteriophage. Understanding how bacteriophages and antibiotic molecules interact with each other is essential for optimizing bacteriophage-antibiotic therapy against bacterial infections. [ABSTRACT FROM AUTHOR]

Antimicrobial tolerance is a significant concern in the food industry, as it poses risks to food safety and public health. To overcome this challenge, synergistic combinations of antimicrobials have emerged as a potential solution. In this study, the combinations of two essential oil constituents (EOCs), namely carvacrol (CAR) and eugenol (EUG), with the quaternary ammonium compounds (QACs) benzalkonium chloride (BAC) and didecyldimethylammonium chloride (DDAC) were evaluated for their antimicrobial effects against Escherichia coli and Bacillus cereus, two common foodborne bacteria. The checkerboard assay was employed to determine the fractional inhibitory concentration index (FICI) and the fractional bactericidal concentration index (FBCI), indicating the presence of bactericidal, but not bacteriostatic, synergy in all QAC–EOC combinations. Bactericidal synergism was clearly supported by Bliss independence analysis. The bactericidal activity of the promising synergistic combinations was further validated by time–kill curves, achieving a >4-log10 reduction of initial bacterial load, which is significant compared to typical industry standards. The combinations containing DDAC showed the highest efficiency, resulting in the eradication of bacterial population in less than 2–4 h. These findings emphasize the importance of considering both bacteriostatic and bactericidal effects when evaluating antimicrobial combinations and the potential of EOC–QAC combinations for sanitization and disinfection in the food industry. [ABSTRACT FROM AUTHOR]

Modern lifestyle greatly influences human well-being. Indeed, nowadays people are centered in the cities and this trend is growing with the everincreasing population. The main habitat for modern humans is defined as the built environment (BE). The modulation of life quality in the BE is primarily mediated by a biodiversity of microbes. They derive from different sources, such as soil, water, air, pets, and humans. Humans are the main source and vector of bacterial diversity in the BE leaving a characteristic microbial fingerprint on the surfaces and spaces. This review, focusing on articles published from the early 2000s, delves into bacterial populations present in indoor and outdoor urban environments, exploring the characteristics of primary bacterial niches in the BE and their native habitats. It elucidates bacterial interconnections within this context and among themselves, shedding light on pathways for adaptation and survival across diverse environmental conditions. Given the limitations of culture-based methods, emphasis is placed on culture-independent approaches, particularly high-throughput techniques to elucidate the genetic and -omic features of BE bacteria. By elucidating these microbiota profiles, the review aims to contribute to understanding the implications for human health and the assessment of urban environmental quality in modern cities. [ABSTRACT FROM AUTHOR]

Bacteria employ quorum sensing as a remarkable mechanism for coordinating behaviors and communicating within their communities. In this study, we introduce a MATLAB Graphical User Interface (GUI) that offers a versatile platform for exploring the dynamics of quorum sensing. Our computational framework allows for the assessment of quorum sensing, the investigation of parameter dependencies, and the prediction of minimum biofilm thickness required for its initiation. A pivotal observation from our simulations underscores the pivotal role of the diffusion coefficient in quorum sensing, surpassing the influence of bacterial cell dimensions. Varying the diffusion coefficient reveals significant fluctuations in autoinducer concentration, highlighting its centrality in shaping bacterial communication. Additionally, our GUI facilitates the prediction of the minimum biofilm thickness necessary to trigger quorum sensing, a parameter contingent on the diffusion coefficient. This feature provides valuable insights into spatial constraints governing quorum sensing initiation. The interplay between production rates and cell concentrations emerges as another critical facet of our study. We observe that higher production rates or cell concentrations expedite quorum sensing, underscoring the intricate relationship between cell communication and population dynamics in bacterial communities. While our simulations align with mathematical models reported in the literature, we acknowledge the complexity of living organisms, emphasizing the value of our GUI for standardizing results and facilitating early assessments of quorum sensing. This computational approach offers a window into the environmental conditions conducive to quorum sensing initiation, encompassing parameters such as the diffusion coefficient, cell concentration, and biofilm thickness. In conclusion, our MATLAB GUI serves as a versatile tool for understanding the diverse aspects of quorum sensing especially for non-biologists. The insights gained from this computational framework advance our understanding of bacterial communication, providing researchers with the means to explore diverse ecological contexts where quorum sensing plays a pivotal role. [ABSTRACT FROM AUTHOR]

Selenium (Se) and zinc (Zn) are essential animal microminerals. Combining Se and Zn (Se-Zn) as a feed additive in its influence on rumen fermentation patterns is still very limited, so further investigation is needed. The present study explored the supplementation impact of combined Se-Zn from organic sources on rumen enzyme activity and relative abundance of several bacterial species through an in vitro method. Five treatments, each with six replicates were used in the study. The first group treated without Se and Zn supplementation (T0, control), the second group treated with 0.3 ppm Se + 60 ppm Zn (T1), the third group treated with 0.45 ppm Se + 60 ppm Zn (T2), the fourth group treated with 0.3 ppm Se + 90 ppm Zn (T3), and the fifth group treated with 0.45 ppm Se + 90 ppm Zn (T4). The parameters observed included rumen microbial enzyme activities (carboxyl methyl cellulase, amylase, protease) and the relative abundance of rumen microbes (Ruminococcus sp., Ruminococcus flavefaciens, Ruminococcus albus, Streptococcus sp., Prevotella ruminicola, and Eubacterium ruminantium). Results indicated that carboxyl methyl cellulase (CMC-ase) and amylase activities raised in T2, T3, and T4 in comparison to T1 and T0 treatments. Protease activity and protein enzyme content increased in T2 compared to all treatments. The relative abundance of Ruminococcus sp. and Ruminococcus albus was higher in T2 and T3 compared to T0 treatment. Furthermore, an elevated Ruminococcus flavefaciens was indicated in T2 compared to other treatments. The T2, T3, and T4 led to higher abundances of Eubacterium ruminantium, Prevotella ruminicola, and Ruminococcus albus compared to T0 and T1. It is concluded that organic Se and Zn enhanced the relative abundance of several bacterial species and the activity of enzymes in the rumen; optimal results are recommended when combining 0.45 ppm Se + 60 ppm Zn. [ABSTRACT FROM AUTHOR]

The coleopteran pests of banana (Musa species), pseudo stem weevil (Odoiporus longicollis Olivier) and rhizome weevil (Cosmopolites sordidus Germer) are not only the most destructive and resilient but also intriguing pests due to their ability to survive for long periods in the most oxygen-deficient conditions like inside the banana stem/rhizome that has limited nutrition. In the recent past, the vast and varied roles of the gut microbiome in insects that aid in metabolic activities like nutrient assimilation, insecticide detoxification, abiotic stress tolerance and copulation have been studied extensively. Very little is known about the gut microbiome of banana weevils and the present study aims at exploration of gut microbiomes of both the pseudostem and rhizome weevils at larval (SWL and RWL respectively) and adult stages (SWA and RWA and respectively). The results revealed that RWL harbored the highest culture-dependent bacterial population as compared to others. Culture-independent bacteriome analyses confirmed that the abundance of observed species (OTUs) found in all four samples were in the order: RWL (616) > SWL (506) > SWA (463) > RWA (413). Diversity analyses showed that RWL exhibited the highest Chao index (593.44), while SWL exhibited the highest Shannon and Simpson indices of 4.22 and 0.89, respectively. Further, it was observed that Proteobacteria, Firmicutes and Bacteroidetes were the major phyla in banana weevils and they sheltered important bacteria that have numerous metabolic functions. [ABSTRACT FROM AUTHOR]

A notable shift in understanding the human microbiome's influence on cardiovascular disease (CVD) is underway, although the causal association remains elusive. A systematic review and meta-analysis were conducted to synthesise current knowledge on microbial taxonomy and metabolite variations between healthy controls (HCs) and those with CVD. An extensive search encompassing three databases identified 67 relevant studies (2012–2023) covering CVD pathologies from 4707 reports. Metagenomic and metabolomic data, both qualitative and quantitative, were obtained. Analysis revealed substantial variability in microbial alpha and beta diversities. Moreover, specific changes in bacterial populations were shown, including increased Streptococcus and Proteobacteria and decreased Faecalibacterium in patients with CVD compared with HC. Additionally, elevated trimethylamine N-oxide levels were reported in CVD cases. Biochemical parameter analysis indicated increased fasting glucose and triglycerides and decreased total cholesterol and low- and high-density lipoprotein cholesterol levels in diseased individuals. This study revealed a significant relationship between certain bacterial species and CVD. Additionally, it has become clear that there are substantial inconsistencies in the methodologies employed and the reporting standards adhered to in various studies. Undoubtedly, standardising research methodologies and developing extensive guidelines for microbiome studies are crucial for advancing the field. [ABSTRACT FROM AUTHOR]

Effective disinfection methods are crucial in the cold chain transportation process of food due to the specificity of temperature and the diversity of contaminated flora. The objective of this study was to investigate the sanitizing effect of different disinfectants on various fungi at – 20 °C to achieve accurate disinfection of diverse bacterial populations. Peracetic acid, hydrogen peroxide, and potassium bisulfate were selected as low-temperature disinfectants and were combined with antifreeze. The sanitizing effect of these cryogenic disinfectants on pathogens such as Bacillus subtilis black variant spores (ATCC9372), Staphylococcus aureus (ATCC 6538), Candida albicans (ATCC 10231), Escherichia coli (8099), and poliovirus (PV-1) was sequentially verified by bactericidal and virus inactivation experiments. After a specified time of disinfection, a neutralizing agent was used to halt the sanitizing process. The study demonstrates that different disinfectants exhibit selective effects during the low-temperature disinfection process. Peracetic acid, hydrogen peroxide, and potassium monopersulfate are suitable for the low-temperature environmental disinfection of bacterial propagules, viruses, and fungal contaminants. However, for microorganisms with strong resistance to spores, a low-temperature disinfectant based on peracetic acid should be chosen for effective disinfection treatment. Our results provide a valuable reference for selecting appropriate disinfectants to sanitize various potential pathogens in the future. [ABSTRACT FROM AUTHOR]

Simple Summary: Sorghum is a nutritious crop that thrives in diverse climates and can withstand harsh growing conditions, making it an attractive option for feeding farmed fish in various regions around the world. In this study, the effects of different sorghum-containing diets on carp were tested, revealing a reduction in growth rates, notable changes in flesh protein content, and significant effects on intestinal microbiota and oxidative stress. This indicates sorghum's potential health and metabolic impacts on Cyprinus carpio. This study investigates the impact of including sorghum in the diet of the common carp (Cyprinus carpio) on its growth, blood parameters, meat composition, intestinal microbiota, and oxidative stress. Experimental diets with varying sorghum content (0%—V0 or control, 10%—V1, 20%—V2, and 30%—V3) were administered to carp weighing 43 g initially. Notably, in the 30% variant, sorghum entirely replaced corn and barley in the diet. Chemical analysis of sorghum unveiled a protein content of 14% and a fat content of 3.9%. Sorghum inclusion led to a decline in final body weight and weight gain, particularly notable in the V3 group with 30% sorghum. However, other physiological parameters, such as feed conversion ratio, specific growth rate, and organ indices, remained unaffected. Protein and salt content in carp flesh increased with higher sorghum inclusion levels, while hematological parameters showed minimal variations. Analysis of the intestinal microbiota revealed increases in both aerobic and anaerobic bacterial populations with sorghum inclusion. Furthermore, sorghum concentration inversely correlated with glutathione levels and positively correlated with malondialdehyde content, indicating a disruption of antioxidant defense mechanisms and elevated oxidative stress. [ABSTRACT FROM AUTHOR]

Exposure to environmental stressors, including certain antibiotics, induces stress responses in bacteria. Some of these responses increase mutagenesis and thus potentially accelerate resistance evolution. Many studies report increased mutation rates under stress, often using the standard experimental approach of fluctuation assays. However, single-cell studies have revealed that many stress responses are heterogeneously expressed in bacterial populations, which existing estimation methods have not yet addressed. We develop a population dynamic model that considers heterogeneous stress responses (subpopulations of cells with the response off or on) that impact both mutation rate and cell division rate, inspired by the DNA-damage response in Escherichia coli (SOS response). We derive the mutant count distribution arising in fluctuation assays under this model and then implement maximum likelihood estimation of the mutation-rate increase specifically associated with the expression of the stress response. Using simulated mutant count data, we show that our inference method allows for accurate and precise estimation of the mutation-rate increase, provided that this increase is sufficiently large and the induction of the response also reduces the division rate. Moreover, we find that in many cases, either heterogeneity in stress responses or mutant fitness costs could explain similar patterns in fluctuation assay data, suggesting that separate experiments would be required to identify the true underlying process. In cases where stress responses and mutation rates are heterogeneous, current methods still correctly infer the effective increase in population mean mutation rate, but we provide a novel method to infer distinct stress-induced mutation rates, which could be important for parameterising evolutionary models. Author summary: How does environmental stress, especially from antibiotics, affect mutation rates in bacteria? This question has often been examined by estimating mutation rates using fluctuation assays, an experiment dating back to Luria and Delbrück in the 1940s. In this study, we consider variation in stress responses within bacterial populations, as revealed by recent single-cell studies, which is neglected in currently available mutation-rate estimation methods. Our approach involves a population dynamic model inspired by the DNA-damage response in E. coli (SOS response). It accounts for a subpopulation with high expression of the stress response, which increases the mutation rate and decreases the division rate of a cell. We use computer simulations to generate synthetic fluctuation assay data. Notably, we find that over a wide range of scenarios, existing models and our heterogeneous-response model cannot be distinguished using fluctuation assay data alone. This emphasises the need for separate experiments to uncover the true underlying processes. Nevertheless, when stress responses are known to be heterogeneous, our study offers a novel method for accurately estimating mutation rates specifically associated with the high expression of the stress response. Uncovering the heterogeneity in stress-induced mutation rates could be important for predicting the evolution of antibiotic resistance. [ABSTRACT FROM AUTHOR]

Phytoplankton blooms provoke bacterioplankton blooms, from which bacterial biomass (necromass) is released via increased zooplankton grazing and viral lysis. While bacterial consumption of algal biomass during blooms is well-studied, little is known about the concurrent recycling of these substantial amounts of bacterial necromass. We demonstrate that bacterial biomass, such as bacterial alpha-glucan storage polysaccharides, generated from the consumption of algal organic matter, is reused and thus itself a major bacterial carbon source in vitro and during a diatom-dominated bloom. We highlight conserved enzymes and binding proteins of dominant bloom-responder clades that are presumably involved in the recycling of bacterial alpha-glucan by members of the bacterial community. We furthermore demonstrate that the corresponding protein machineries can be specifically induced by extracted alpha-glucan-rich bacterial polysaccharide extracts. This recycling of bacterial necromass likely constitutes a large-scale intra-population energy conservation mechanism that keeps substantial amounts of carbon in a dedicated part of the microbial loop. Phytoplankton blooms provoke bacterioplankton blooms, from which bacterial biomass (necromass) is released via zooplankton grazing and viral lysis. Here, Beidler et al. show that the bacterial biomass, including alpha-glucan polysaccharides generated from the consumption of algal organic matter, is reused by microbes in vitro and during a diatom-dominated bloom. [ABSTRACT FROM AUTHOR]

Heteroresistance is a medically relevant phenotype where small antibiotic-resistant subpopulations coexist within predominantly susceptible bacterial populations. Heteroresistance reduces treatment efficacy across diverse bacterial species and antibiotic classes, yet its genetic and physiological mechanisms remain poorly understood. Here, we investigated a multi-resistant Klebsiella pneumoniae isolate and identified three primary drivers of gene dosage-dependent heteroresistance for several antibiotic classes: tandem amplification, increased plasmid copy number, and transposition of resistance genes onto cryptic plasmids. All three mechanisms imposed fitness costs and were genetically unstable, leading to fast reversion to susceptibility in the absence of antibiotics. We used a mouse gut colonization model to show that heteroresistance due to elevated resistance-gene dosage can result in antibiotic treatment failures. Importantly, we observed that the three mechanisms are prevalent among Escherichia coli bloodstream isolates. Our findings underscore the necessity for treatment strategies that address the complex interplay between plasmids, resistance cassettes, and transposons in bacterial populations. Bacterial heteroresistance is a medically relevant phenotype where small antibiotic-resistant subpopulations coexist within predominantly susceptible bacterial populations. Here, Nicoloff et al. describe how three different mechanisms that increase the copy number of resistance genes can lead to unstable and transient heteroresistance. [ABSTRACT FROM AUTHOR]

Mangroves are important coastal wetlands along tropical and subtropical regions. Pneumatophore, a kind of aerial root, is among the prominent components of a mangrove ecosystem, which provides microhabitats for a range of prokaryotic (bacteria and cyanobacteria) microbial assemblages, whose role in the maintenance of mangrove ecology often remains neglected. Very few studies are available on pneumatophore-associated prokaryotic microorganisms (PAPMs). The majority of them are related to the microscopic identification of cyanobacteria, with very limited research on the bacterial population, even though they demand more attention. Also, very scarce information is available on biotic and abiotic factors shaping the PAPMs. The objective of this review is to highlight the structural and functional importance of prokaryotic organisms associated with pneumatophores. This review begins with a brief introduction of what mangrove pneumatophores are, then focuses on the PAPMs, accentuating the breadth and depth of information gained from previous research. We further discuss how a combination of a traditional cultivable approach and a newly developed omics approach can be efficaciously employed to untangle PAPMs. This review provides updated information on PAPMs, which will intensify the visibility and necessity of pneumatophore-associated microbial community research. [ABSTRACT FROM AUTHOR]

Antimicrobial resistance (AMR) in bacteria is a major public health threat and conjugative plasmids play a key role in the dissemination of AMR genes among bacterial pathogens. Interestingly, the association between AMR plasmids and pathogens is not random and certain associations spread successfully at a global scale. The burst of genome sequencing has increased the resolution of epidemiological programs, broadening our understanding of plasmid distribution in bacterial populations. Despite the immense value of these studies, our ability to predict future plasmid-bacteria associations remains limited. Numerous empirical studies have recently reported systematic patterns in genetic interactions that enable predictability, in a phenomenon known as global epistasis. In this perspective, we argue that global epistasis patterns hold the potential to predict interactions between plasmids and bacterial genomes, thereby facilitating the prediction of future successful associations. To assess the validity of this idea, we use previously published data to identify global epistasis patterns in clinically relevant plasmid-bacteria associations. Furthermore, using simple mechanistic models of antibiotic resistance, we illustrate how global epistasis patterns may allow us to generate new hypotheses on the mechanisms associated with successful plasmid-bacteria associations. Collectively, we aim at illustrating the relevance of exploring global epistasis in the context of plasmid biology. Antimicrobial resistance (AMR) in bacteria is a major public health threat and plasmids play a key role in AMR gene dissemination. This Review discusses the relevance of global epistasis patterns for predicting AMR evolution and generating hypotheses about the mechanisms. [ABSTRACT FROM AUTHOR]

In the present study, biological indicators were used to assess the impact of applying spent mushroom substrate and manure on the soil environment. The use of spent mushroom substrate had a varied effect on the microorganisms. Stimulation was recorded in the abundance of copiotrophic bacteria and fungi, but only in the first year of the study. In the case of cellulolytic bacteria, this effect was visible only in single plots. Similar observations were also noted regarding the relative DNA content (in relation to the control), which increased for both bacteria and fungi after applying spent mushroom substrate. In the soil fertilized with spent mushroom substrate, a decrease in DNA concentration was observed, but only in the first and second year. For enzymatic activity, the use of spent mushroom substrate alone proved to be more favorable, but this effect was again observed only in the first year of the study. The application of manure caused similar changes as observed with the use of spent mushroom substrate. These observations indicate a similar impact of spent mushroom substrate and manure on the parameters tested. The research presented suggests the use of both classical methods and methods based on the analysis of DNA extracted from soil to study the impact of spent mushroom substrate on the activity of soil microbial populations. [ABSTRACT FROM AUTHOR]

Microbial communities associated with seagrass beds play a crucial role in maintaining the balance of seagrass ecosystems. However, the driving mechanisms behind the structure and functional succession of seagrass microbial communities are still unclear despite the close interaction between seagrass and surrounding microorganisms. To enhance our knowledge of the diversity and functional characteristics of microbial communities in seagrass beds, we employed 16S rRNA gene amplicon sequencing to investigate bacterial communities in seagrass leaves, roots, seawater, and sediments in Caofeidian Zostera marina seagrass beds of Hebei Province, Northern China. Our results highlighted that specific types of bacteria were enriched in different sample compartments, indicating the importance of habitat in influencing microbial diversity and community structure in seagrass bed ecosystems. Notably, the microbial community structure of seagrass leaves and roots showed more similarity to that found in seawater and sediments. Among all the samples, the phylum Pseudomonadota exhibited the highest relative abundance, particularly in sediment samples where they accounted for over 95% of the total bacterial population. In addition, the enrichment of Vibrio, an opportunistic pathogen in several plant samples, alerted us to seagrass and its surrounding marine environments. Finally, functional predictions of microbial communities using PICRUSt2 revealed variations in microbial functions, indicating specific metabolic preferences of microbial communities in different natural environments. The present research sheds light on the mechanisms underlying microbial community succession and their ecological function in seagrass beds. [ABSTRACT FROM AUTHOR]

In the dairy industry bacteriophage (phage) contamination significantly impairs the production and quality of products like yogurt and cheese. To combat this issue, the strains of bacteria used as starter cultures possess mechanisms that make them resistant to phage infection, such as envelope resistance, or processes that render them immune to phage infection, such as restriction-modification and CRISPR-Cas. Lactococcus lactis, used to manufacture cheese and other dairy products, can also block the reproduction of infecting phages by abortive infection (Abi), a process in which phage-infected cells die before the phage replicate. We employ mathematical-computer simulation models and experiments with two Lactococcus lactis strains and two lytic phages to investigate the conditions under which Abi can limit the proliferation of phages in L. lactis populations and prevent the extinction of their populations by these viruses. According to our model, if Abi is almost perfect and there are no other populations of bacteria capable of supporting the replication of the L. lactis phages, Abi can protect bacterial populations from succumbing to infections with these viruses. This prediction is supported by the results of our experiment, which indicate that Abi can help protect L. lactis populations from extinction by lytic phage infections. However, our results also predict abortive infection is only one element of L. lactis defenses against phage infection. Mutant phages that can circumvent the Abi systems of these bacteria emerge. The survival of L. lactis populations then depends on the evolution of envelope mutants that are resistant to the evolved host-range phage. [ABSTRACT FROM AUTHOR]

Heterotrophs may live in a wider range of environmental conditions and have long been used in probiotic treatments in aquaculture. For detritivores like shrimp, tilapia and carp, heterotrophic bacteria are essential constituents of their diet. In the present study, aimed to isolate and identify heterotrophic bacteria from fermentation sample of freshwater and brackish water shrimp farms. Liquid enrichment medium was used to isolate the bacteria. Bacteria were identified using DNA barcoding analysis of the 16S rRNA gene sequence using primer 533 F, 1100 Rand phylogenetic analysis with the MEGA X Software. The sequences obtained from the samples were aligned and edited using MEGA-X software. The results revealed that up to four bacteria were isolated from Shrimp ponds. These bacteria were EW1, SP1, SP2 and FD2. Based on the 16S rRNA gene sequence barcoding analysis, EW1 and SP1 was identified as Priestia flexa, SP2 identified as Acinetobacter venetianus and FD2 identified as Bacillus pumilus. These heterotrophic bacteria could be potential candidates to improve health of Shrimps. [ABSTRACT FROM AUTHOR]

The efficiency of replication error repair is a critical factor governing the emergence of mutations. However, it has so far been impossible to study this efficiency at the level of individual cells and to investigate if it varies within isogenic cell populations. In addition, why some errors escape repair remains unknown. Here we apply a combination of fluorescent labelling of the Escherichia coli Mismatch Repair (MMR) complex, microfluidics, and time-lapse microscopy, to monitor in real-time the fate of >20000 replication errors. We show that i) many mutations result from errors that are detected by MMR but inefficiently repaired ii) this limited repair efficiency is due to a temporal constraint imposed by the transient nature of the DNA strand discrimination signal, a constraint that is likely conserved across organisms, and iii) repair capacity varies from cell to cell, resulting in a subpopulation of cells with higher mutation rate. Such variations could influence the fitness and adaptability of populations, accelerating for instance the emergence of antibiotic resistance. An interdisciplinary approach following replication errors in Escherichia coli unveils that many spontaneous mutations originate from inefficient repair, and that repair capacity is variable between single cells within a bacterial population. [ABSTRACT FROM AUTHOR]

This study explores the possibility of utilising electrochemically assisted anaerobic digestion supplemented with carbon-based materials to stimulate methanogenesis. Two different carbonaceous materials—commercial activated carbon (AC), and pyrolysed argan (PA, derived from argan shells)—were employed as supplements, with cheese whey (CW) being used as the substrate. Methane production slightly increased in the electrochemically assisted digesters, potentially translating into a 2–4% increase in the output of industrial digesters. In addition, reactors supplemented with PA also exhibited better production rates (496–508 L·kgVS−1), although there was no observed improvement in the quantity of biogas at the end of the biodegradability experiment. In contrast, when commercial AC was used as the supplement, the start-up phase was accelerated (5 days), although methane productivity decreased (273–352 L·kgVS−1). These observations were supported by microbiological analyses, demonstrating that the reactors with the poorest performance (those supplemented with AC) experienced the most significant decrease in both archaeal and bacterial populations. [ABSTRACT FROM AUTHOR]

Dietary supplementation with triglyceride tributyrin (TBT), a butyrate precursor, has been associated with beneficial effects on fish health and improvements in the ability of carnivorous fish to tolerate higher levels of plant-based protein. In this study, we aimed to investigate the effects of a plant-based diet supplemented with TBT on the structural diversity and putative function of the digesta-associated bacterial communities of rainbow trout (Oncorhynchus mykiss). In addition to this, we also assessed the response of fish gut digestive enzyme activities and chyme metabolic profile in response to TBT supplementation. Our results indicated that TBT had no significant effects on the overall fish gut bacterial communities, digestive enzyme activities or metabolic profile when compared with non-supplemented controls. However, a more in-depth analysis into the most abundant taxa showed that diets at the highest TBT concentrations (0.2% and 0.4%) selectively inhibited members of the Enterobacterales order and reduced the relative abundance of a bacterial population related to Klebsiella pneumoniae, a potential fish pathogen. Furthermore, the predicted functional analysis of the bacterial communities indicated that increased levels of TBT were associated with depleted KEGG pathways related to pathogenesis. The specific effects of TBT on gut bacterial communities observed here are intriguing and encourage further studies to investigate the potential of this triglyceride to promote pathogen suppression in the fish gut environment, namely in the context of aquaculture. [ABSTRACT FROM AUTHOR]

GacS/GacA is a widely distributed two-component system playing an essential role as a key global regulator, although its characterization in phytopathogenic bacteria has been deeply biased, being intensively studied in pathogens of herbaceous plants but barely investigated in pathogens of woody hosts. P. savastanoi pv. savastanoi (Psv) is characterized by inducing tumours in the stem and branches of olive trees. In this work, the model strain Psv NCPPB 3335 and a mutant derivative with a complete deletion of gene gacA were subjected to RNA-Seq analyses in a minimum medium and a medium mimicking in planta conditions, accompanied by RT-qPCR analyses of selected genes and phenotypic assays. These experiments indicated that GacA participates in the regulation of at least 2152 genes in strain NCPPB 3335, representing 37.9 % of the annotated CDSs. GacA also controls the expression of diverse rsm genes, and modulates diverse phenotypes, including motility and resistance to oxidative stresses. As occurs with other P. syringae pathovars of herbaceous plants, GacA regulates the expression of the type III secretion system and cognate effectors. In addition, GacA also regulates the expression of WHOP genes, specifically encoded in P. syringe strains isolated from woody hosts, and genes for the biosynthesis of phytohormones. A gacA mutant of NCPPB 3335 showed increased virulence, producing large immature tumours with high bacterial populations, but showed a significantly reduced competitiveness in planta. Our results further extend the role of the global regulator GacA in the virulence and fitness of a P. syringae pathogen of woody hosts. [ABSTRACT FROM AUTHOR]

Multiple Salmonella enterica serovars and strains have been reported to be able to persist inside the foliar tissue of lettuce (Lactuca sativa L.), potentially resisting washing steps and reaching the consumer. Intraspecies variation of the bacterial pathogen and of the plant host can both significantly affect the outcome of foliar colonization. However, current understanding of the mechanisms underlying this phenomenon is still very limited. In this study, we evaluated the foliar fitness of 14 genetically barcoded S. enterica isolates from 10 different serovars, collected from plant and animal sources. The S. enterica isolates were vacuum-infiltrated individually or in pools into the leaves of three- to four-week-old lettuce plants. To estimate the survival capacity of individual isolates, we enumerated the bacterial populations at 0- and 10- days post-inoculation (DPI) and calculated their net growth. The competition of isolates in the lettuce apoplast was assessed through the determination of the relative abundance change of barcode counts of each isolate within pools during the 10 DPI experimental period. Isolates exhibiting varying apoplast fitness phenotypes were used to evaluate their capacity to grow in metabolites extracted from the lettuce apoplast and to elicit the reactive oxygen species burst immune response. Our study revealed that strains of S. enterica can substantially differ in their ability to survive and compete in a co-inhabited lettuce leaf apoplast. The differential foliar fitness observed among these S. enterica isolates might be explained, in part, by their ability to utilize nutrients available in the apoplast and to evade plant immune responses in this niche. [ABSTRACT FROM AUTHOR]

Antibiotic-resistant, facultative pathogenic bacteria are commonly found in surface water; however, the factors influencing the spread and stabilization of antibiotic resistance in this habitat, particularly the role of biofilms, are not fully understood. The extent to which bacterial populations in biofilms or sediments exacerbate the problem for specific antibiotic classes or more broadly remains unanswered. In this study, we investigated the differences between the bacterial populations found in the surface water and sediment/biofilm of the Mur River and the Drava River in Austria. Samples of Escherichia coli were collected from both the water and sediment at two locations per river: upstream and downstream of urban areas that included a sewage treatment plant. The isolates were subjected to antimicrobial susceptibility testing against 21 antibiotics belonging to seven distinct classes. Additionally, isolates exhibiting either extended-spectrum beta-lactamase (ESBL) or carbapenemase phenotypes were further analyzed for specific antimicrobial resistance genes. E. coli isolates collected from all locations exhibited resistance to at least one of the tested antibiotics; on average, isolates from the Mur and Drava rivers showed 25.85% and 23.66% resistance, respectively. The most prevalent resistance observed was to ampicillin, amoxicillin–clavulanic acid, tetracycline, and nalidixic acid. Surprisingly, there was a similar proportion of resistant bacteria observed in both open water and sediment samples. The difference in resistance levels between the samples collected upstream and downstream of the cities was minimal. Out of all 831 isolates examined, 13 were identified as carrying ESBL genes, with 1 of these isolates also containing the gene for the KPC-2 carbapenemase. There were no significant differences between the biofilm (sediment) and open water samples in the occurrence of antibiotic resistance. For the E. coli populations in the examined rivers, the different factors in water and the sediment do not appear to influence the stability of resistance. No significant differences in antimicrobial resistance were observed between the bacterial populations collected from the biofilm (sediment) and open-water samples in either river. The different factors in water and the sediment do not appear to influence the stability of resistance. The minimal differences observed upstream and downstream of the cities could indicate that the river population already exhibits generalized resistance. [ABSTRACT FROM AUTHOR]

Heat flux measurement shows potential for the early detection of infectious growth. Our research is motivated by the possibility of using heat flux sensors for the early detection of infection on aortic vascular grafts by measuring the onset of bacterial growth. Applying heat flux measurement as an infectious marker on implant surfaces is yet to be experimentally explored. We have previously shown the measurement of the exponential growth curve of a bacterial population in a thermally stabilized laboratory environment. In this work, we further explore the limits of the microcalorimetric measurements via heat flux sensors in a microfluidic chip in a thermally fluctuating environment. [ABSTRACT FROM AUTHOR]

The impact of living mulches established with three officinal plants (Alchemilla vulgaris, Fragaria vesca and Mentha x piperita) on the soil bacterial microbiome and activity, the nematodes population, and the nutrient status of an organic apple orchard was assessed. The composition and diversity of the bacterial communities were differentially modified by living mulches. The activity of the bacterial microbiome associated with F. vesca was higher and utilized more C sources in comparison to other treatments. The combined analysis of the core bacterial microbiome and metabolic activity pointed to a potential effect of F. vesca on different levels of the soil's trophic network. The living mulches did not affect the overall number of nematodes, but in some cases, they modified the structure of the population: F. vesca induced the highest share of bacteria feeders and the lowest number of herbivores and fungal feeders. The living mulches modified the availability of some nutrients and the pH. Multivariate analysis of the whole dataset showed several potential inter-dependencies between the assessed parameters that are worthy of further study. In conclusion, the introduction of multifunctional living mulches based on officinal plants induced changes to the soil's genetic and functional biodiversity and chemical properties. These modifications could deliver ecosystem services particularly relevant to organic apple orchards. [ABSTRACT FROM AUTHOR]

What a strain is and how many strains make up a natural bacterial population remain elusive concepts despite their apparent importance for assessing the role of intra-population diversity in disease emergence or response to environmental perturbations. To advance these concepts, we sequenced 138 randomly selected Salinibacter ruber isolates from two solar salterns and assessed these genomes against companion short-read metagenomes from the same samples. The distribution of genome-aggregate average nucleotide identity (ANI) values among these isolates revealed a bimodal distribution, with four-fold lower occurrence of values between 99.2% and 99.8% relative to ANI >99.8% or <99.2%, revealing a natural "gap" in the sequence space within species. Accordingly, we used this ANI gap to define genomovars and a higher ANI value of >99.99% and shared gene-content >99.0% to define strains. Using these thresholds and extrapolating from how many metagenomic reads each genomovar uniquely recruited, we estimated that –although our 138 isolates represented about 80% of the Sal. ruber population– the total population in one saltern pond is composed of 5,500 to 11,000 genomovars, the great majority of which appear to be rare in-situ. These data also revealed that the most frequently recovered isolate in lab media was often not the most abundant genomovar in-situ, suggesting that cultivation biases are significant, even in cases that cultivation procedures are thought to be robust. The methodology and ANI thresholds outlined here should represent a useful guide for future microdiversity surveys of additional microbial species. What a microbial strain is and how many strains make up a natural bacterial population remain elusive concepts. Here, Viver et al. analyse Salinibacter ruber isolates and metagenomes from two solar salterns, revealing gaps within the species sequence space that they use to define and quantify sub-species categories, such as genomovars and strains, that co-exist in a saltern pond. [ABSTRACT FROM AUTHOR]

Scientific drilling expeditions offer a unique opportunity to characterize microbial communities in the subsurface that have long been isolated from the surface. With subsurface microbial biomass being low in general, biological contamination from the drilling fluid, sample processing, or molecular work is a major concern. To address this, characterization of the contaminant populations in the drilling fluid and negative extraction controls are essential for assessing and evaluating such sequencing data. Here, rock cores down to 2250 m depth, groundwater-bearing fractures, and the drilling fluid were sampled for DNA to characterize the microbial communities using a broad genomic approach. However, even after removing potential contaminant populations present in the drilling fluid, notorious contaminants were abundant and mainly affiliated with the bacterial order Burkholderiales. These contaminant microorganisms likely originated from the reagents used for isolating DNA despite stringent quality standards during the molecular work. The detection of strictly anaerobic sulfate reducers such as Candidatus Desulforudis audaxviator suggested the presence of autochthonous deep biosphere taxa in the sequenced libraries, yet these clades represented only a minor fraction of the sequence counts (< 0.1 %), hindering further ecological interpretations. The described methods and findings emphasize the importance of sequencing extraction controls and can support experimental design for future microbiological studies in conjunction with continental drilling operations. [ABSTRACT FROM AUTHOR]

Laboratory evolution studies, particularly with Escherichia coli, have yielded invaluable insights into the mechanisms of antimicrobial resistance (AMR). Recent investigations have illuminated that, with repetitive antibiotic exposures, bacterial populations will adapt and eventually become tolerant and resistant to the drugs. Through intensive analyses, these inquiries have unveiled instances of convergent evolution across diverse antibiotics, the pleiotropic effects of resistance mutations, and the role played by loss-of-function mutations in the evolutionary landscape. Moreover, a quantitative analysis of multidrug combinations has shed light on collateral sensitivity, revealing specific drug combinations capable of suppressing the acquisition of resistance. This review article introduces the methodologies employed in the laboratory evolution of AMR in bacteria and presents recent discoveries concerning AMR mechanisms derived from laboratory evolution. Additionally, the review outlines the application of laboratory evolution in endeavors to formulate rational treatment strategies. [ABSTRACT FROM AUTHOR]

Simple Summary: Symbiotic bacteria play an essential role in the digestion, detoxification, and nutrient supply of bark and ambrosia beetles. They may even promote the invasiveness and adaptability of invasive beetles. However, as a newly invasive pest in China, the diversity and community structure of bacteria associated with Hylurgus ligniperda is still unclear. This hinders our ability to comprehend their invasion mechanism and ecological adaptability. Therefore, this study revealed the differences in the diversity and community composition of associated bacteria in the beetle's elytra, prothorax, and gut. Our findings revealed that the microbial population of the elytra was distinct from that of the prothorax and gut. Additionally, gender disparities existed throughout the bacterial population of elytra. The distribution of associated bacteria in different organs of males and females may indicate that they undertake different ecological functions. This study will help to understand the bacterial communities that may play important roles in Hylurgus ligniperda. Hylurgus ligniperda (Fabricius) (Curculionidae: Scolytinae) is a new invasive pest beetle in China, which colonized the Shandong province, causing devastating damage. Originating in Europe, it has spread to Oceania, Asia, North and South America. Bacterial associates have been frequently reported to play a vital role in strengthening the ecological adaptations of bark and ambrosia beetles. The environmental adaptability of H. ligniperda may be supported by their associated bacteria. Bacterial communities colonizing different body parts of insects may have different functions. However, little is known about the bacteria associated with H. ligniperda and their potential involvement in facilitating the adaptation and invasion of the beetles into new environments. In this study, we employed high-throughput sequencing technology to analyze the bacterial communities associated with male and female adults of H. ligniperda by comparing those colonizing the elytra, prothorax, and gut. Results showed that the bacterial communities of male and female adults were similar, and the elytra samples had the highest bacterial diversity and richness, followed by the gut, while the prothorax had the lowest. The dominant phyla were Proteobacteria, Firmicutes, and Actinobacteriota, while the dominant genera were Serratia, Lactococcus, Rhodococcus, unclassified Enterobacteriaceae, and Gordonia. Among these, Rhodococcus and Gordonia were the specific genera of endobacteria and ectobacteria, respectively. Differences in the distribution of associated bacteria may suggest that they have different ecological functions for H. ligniperda. The results of functional prediction showed that bacteria were enriched in terpenoid backbone biosynthesis, degradation of aromatic compounds, limonene and pinene degradation, neomycin, kanamycin and gentamicin biosynthesis, indicating that they may assist their beetles in synthesizing pheromones, degrading toxic secondary metabolites of host trees, and antagonizing pathogenic fungi. These results help us understand the interaction between H. ligniperda and bacteria and highlight possible contributions to the invasion process. [ABSTRACT FROM AUTHOR]

The European spruce bark beetle Ips typographus is a pest causing severe damage in forests dominated by Norway spruce in Europe. Microorganisms play essential roles in bark beetles, including nutrition, fitness as well as in overcoming host defenses. Here, we performed high-throughput 16S rRNA metabarcoding of I. typographus across different populations in Europe, to assess its bacterial community. We investigated four postglacial refugial areas in Europe and focused specifically on a current bark beetle hot spot in the Dolomites where we compared populations with different epidemiological phases (outbreaking vs. non-outbreaking) and across different seasons (pre-overwintering vs. overwintering). Our results show that the bacterial community structure varied among populations from the refugial areas and between different regions within the Dolomites. We found a significant difference in the bacterial community between pre-overwintering and overwintering individuals, but we did not find differences between epidemic and endemic populations. The prevalence of the genera Erwinia (which was present in all individuals) and Pseudoxanthomonas (present in almost all individuals) across all populations suggests that these taxa form the core bacterial community of I. typographus. Furthermore, several additional bacterial taxa occurred in all populations, but with variable frequencies. This study highlights a complex interaction of I. typographus and various bacterial taxa across different regions and ecological phases and provides new insights into the role of microorganisms in the biology of this important pest species. [ABSTRACT FROM AUTHOR]

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