Alterations in cell arrangements of group B streptococcus due to virulence factor expression can bias estimates of bacterial populations based on colony count measures.

Group B streptococcus (GBS) is a chain-forming commensal bacterium and opportunistic pathogen that resides in the gastrointestinal and genitourinary tract of healthy adults. GBS can cause various infections and related complications in pregnant and nonpregnant women, adults, and newborns. Investigations of the mechanisms by which GBS causes disease pathogenesis often utilize colony count assays to estimate bacterial population size in experimental models. In other streptococci, such as group A streptococcus and pneumococcus, variation in the chain length of the bacteria that can occur naturally or due to mutation can affect facets of pathogenesis, such as adherence to or colonization of a host. No studies have reported a relationship between GBS chain length and pathogenicity. Here, we used GBS strain 874391 and several derivative strains displaying longer chain-forming phenotypes (874391pgapC, 874391ΔcovR, 874391Δstp1) to assess the impact of chain length on bacterial population estimates based on the colony-forming unit (c.f.u.) assay. Disruption of GBS chains via bead beating or sonication in conjunction with fluorescence microscopy was used to compare chaining phenotypes pre- and post-disruption to detect long- and short-chain forms, respectively. We used a murine model of GBS colonization of the female reproductive tract to assess whether chaining may affect bacterial colonization dynamics in the host during chronic infection in vivo. Overall, we found that GBS exhibiting long-chain form can significantly affect population size estimates based on the colony count assay. Additionally, we found that the length of chaining of GBS can affect virulence in the reproductive tract colonization model. Collectively, these findings have implications for studies of GBS that utilize colony count assays to measure GBS populations and establish that chain length can affect infection dynamics and disease pathogenesis for this important opportunistic pathogen.

Fluorescent carbon dots for labeling of bacteria: mechanism and prospects-a review.

The search for bacteria-labeling agents that are more efficient and less toxic compared to existing staining dyes is ongoing. Fluorescent quantum dots and carbon dots (CDs) have been extensively researched for various bioimaging applications. Priority is given to CDs due to several advantages, including lower toxicity, versatility in tuning their properties, and better photostability compared to metal-based quantum dots. Although significant progress is still needed to replace existing dyes with CDs for bacteria labeling, they offer promising potential for further improvement in efficiency. Surface charges and functional groups have been reported as decisive factors for bacterial discrimination and live/dead assays; however, a complete guideline for preparing CDs with optimum properties for efficient staining and predicting their labeling performance is lacking. In this review, we discuss the application of fluorescent CDs for bacterial labeling and the underlying mechanisms and principles. We primarily focus on the application and mechanism of CDs for Gram differentiation, live imaging, live/dead bacteria differentiation, bacterial viability testing, biofilm imaging, and the challenges associated with application of CDs. Based on proposed mechanisms of bacterial labeling and ambiguous results reported, we provide our view and guidelines for the researchers in this field to overcome the challenges associated with bacteria labeling using fluorescent CDs.

A framework for leveraging network course-based undergraduate research experience (CURE) faculty to develop, validate, and administer an assessment instrument.

Over the last several years, nationally disseminated course-based undergraduate research experiences (CUREs) have emerged as an alternative to developing a novel CURE from scratch, but objective assessment of these multi-institution (network) CUREs across institutions is challenging due to differences in student populations, instructors, and fidelity of implementation. The time, money, and skills required to develop and validate a CURE-specific assessment instrument can be prohibitive. Here, we describe a co-design process for assessing a network CURE [the Prevalence of Antibiotic Resistance in the Environment (PARE)] that did not require support through external funding, was a relatively low time commitment for participating instructors, and resulted in a validated instrument that is usable across diverse PARE network institution types and implementation styles. Data collection efforts have involved over two dozen unique institutions, 42 course offerings, and over 1,300 pre-/post-matched assessment record data points. We demonstrated significant student learning gains but with small effect size in both content and science process skills after participation in the two laboratory sessions associated with the core PARE module. These results show promise for the efficacy of short-duration CUREs, an educational research area ripe for further investigation, and may support efforts to lower barriers for instructor adoption by leveraging a CURE network for developing and validating assessment tools.

Extending the Potency and Lifespan of Antibiotics: Inhibitors of Gram-Negative Bacterial Efflux Pumps.

Efflux is a natural process found in all prokaryotic and eukaryotic cells that removes a diverse range of substrates from inside to outside. Many antibiotics are substrates of bacterial efflux pumps, and modifications to the structure or overexpression of efflux pumps are an important resistance mechanism utilized by many multidrug-resistant bacteria. Therefore, chemical inhibition of bacterial efflux to revitalize existing antibiotics has been considered a promising approach for antimicrobial chemotherapy over two decades, and various strategies have been employed. In this review, we provide an overview of bacterial multidrug resistance (MDR) efflux pumps, of which the resistance nodulation division (RND) efflux pumps are considered the most clinically relevant in Gram-negative bacteria, and describe over 50 efflux inhibitors that target such systems. Although numerous efflux inhibitors have been identified to date, none have progressed into clinical use because of formulation, toxicity, and pharmacokinetic issues or a narrow spectrum of inhibition. For these reasons, the development of efflux inhibitors has been considered a difficult and complex area of research, and few active preclinical studies on efflux inhibitors are in progress. However, recently developed tools, including but not limited to computational tools including molecular docking models, offer hope that further research on efflux inhibitors can be a platform for research and development of new bacterial efflux inhibitors.

RamanCluster: A deep clustering-based framework for unsupervised Raman spectral identification of pathogenic bacteria.

Raman spectroscopy serves as a powerful and reliable tool for the characterization of pathogenic bacteria. The integration of Raman spectroscopy with artificial intelligence techniques to rapidly identify pathogenic bacteria has become paramount for expediting disease diagnosis. However, the development of prevailing supervised artificial intelligence algorithms is still constrained by costly and limited well-annotated Raman spectroscopy datasets. Furthermore, tackling various high-dimensional and intricate Raman spectra of pathogenic bacteria in the absence of annotations remains a formidable challenge. In this paper, we propose a concise and efficient deep clustering-based framework (RamanCluster) to achieve accurate and robust unsupervised Raman spectral identification of pathogenic bacteria without the need for any annotated data. RamanCluster is composed of a novel representation learning module and a machine learning-based clustering module, systematically enabling the extraction of robust discriminative representations and unsupervised Raman spectral identification of pathogenic bacteria. The extensive experimental results show that RamanCluster has achieved high accuracy on both Bacteria-4 and Bacteria-6, with ACC values of 77 % and 74.1 %, NMI values of 75 % and 73 %, as well as AMI values of 74.6 % and 72.6 %, respectively. Furthermore, compared with other state-of-the-art methods, RamanCluster exhibits the superior accuracy on handling various complicated pathogenic bacterial Raman spectroscopy datasets, including situations with strong noise and a wide variety of pathogenic bacterial species. Additionally, RamanCluster also demonstrates commendable robustness in these challenging scenarios. In short, RamanCluster has a promising prospect in accelerating the development of low-cost and widely applicable disease diagnosis in clinical medicine.

Gut bacterium Burkholderia cepacia (BsNLG8) and immune gene Defensin A contribute to the resistance against Nicotine-induced stress in Nilaparvata lugens (Stål).

Nicotine, a naturally occurring alkaloid found in tobacco, is a potent neurotoxin extensively used to control Nilaparvata lugens (Stål), a destructive insect pest of rice crops. The insect gut harbors a wide array of resident microorganisms that profoundly influence several biological processes, including host immunity. Maintaining an optimal gut microbiota and immune homeostasis requires a complex network of reciprocal regulatory interactions. However, the underlying molecular mechanisms driving these symbiotic exchanges, particularly between specific gut microbe and immunity, remain largely unknown in insects. Our previous investigations identified and isolated a nicotine-degrading Burkholderia cepacia strain (BsNLG8) with antifungal properties. Building on those findings, we found that nicotine intake significantly increased the abundance of a symbiotic bacteria BsNLG8, induced a stronger bacteriostatic effect in hemolymph, and enhanced the nicotine tolerance of N. lugens. Additionally, nicotine-induced antimicrobial peptides (AMPs) exhibited significant antibacterial effects against Staphylococcus aureus. We adopted RNA-seq to explore the underlying immunological mechanisms in nicotine-stressed N. lugens. Bioinformatic analyses identified numerous differentially expressed immune genes, including recognition/immune activation (GRPs and Toll) and AMPs (i.e., Defensin, Lugensin, lysozyme). Temporal expression profiling (12, 24, and 48 hours) of immune genes revealed pattern recognition proteins and immune effectors as primary responders to nicotine-induced stress. Defensin A, a broad-spectrum immunomodulatory cationic peptide, exhibited significantly high expression. RNA interference-mediated silencing of Defensin A reduced the survival, enhanced nicotine sensitivity of N. lugens to nicotine, and decreased the abundance of BsNLG8. The reintroduction of BsNLG8 improved the expression of immune genes, aiding nicotine resistance of N. lugens. Our findings indicate a potential reciprocal immunomodulatory interaction between Defensin A and BsNLG8 under nicotine stress. Moreover, this study offers novel and valuable insights for future research into enhancing nicotine-based pest management programs and developing alternative biocontrol methods involving the implication of insect symbionts.

Combining deep learning and droplet microfluidics for rapid and label-free antimicrobial susceptibility testing of colistin.

Efficient tools for rapid antibiotic susceptibility testing (AST) are crucial for appropriate use of antibiotics, especially colistin, which is now often considered a last resort therapy with extremely drug resistant Gram-negative bacteria. Here, we developed a rapid, easy and miniaturized colistin susceptibility assay based on microfluidics, which allows for culture and high-throughput analysis of bacterial samples. Specifically, a simple microfluidic platform that can easily be operated was designed to encapsulate bacteria in nanoliter droplets and perform a fast and automated bacterial growth detection in 2 h, using standardized samples. Direct bright-field imaging of compartmentalized samples proved to be a faster and more accurate detection method as compared to fluorescence-based analysis. A deep learning powered approach was implemented for the sensitive detection of the growth of several strains in droplets. The DropDeepL AST method (Droplet and Deep learning-based method for AST) developed here allowed the determination of the colistin susceptibility profiles of 21 fast-growing Enterobacterales (E. coli and K. pneumoniae), including clinical isolates with different resistance mechanisms, showing 100 % categorical agreement with the reference broth microdilution (BMD) method performed simultaneously. Direct AST of bacteria in urine samples on chip also provided accurate results in 2 h, without the need of complex sample preparation procedures. This method can easily be implemented in clinical microbiology laboratories, and has the potential to be adapted to a variety of antibiotics, especially for last-line antibiotics to optimize treatment of patients infected with multi-drug resistant strains.

NIR-driven multifunctional PEC biosensor based on aptamer-modified PDA/MnO2 photoelectrode for bacterial detection and inactivation.

Sensitive detection and effective inactivation of bacteria are essential in preventing foodborne bacterial infection that poses a significant threat to human health. Herein, a near-infrared (NIR)-driven multifunctional photoelectrochemical (PEC) biosensor was constructed for detection and inactivation of S. aureus. Based on the covalent bonding between amine and carboxyl groups, carboxyl-functionalized SA31 aptamer was immobilized on the PDA/MnO2 photoelectrode. In the presence of S. aureus, SA31 aptamer can specifically capture S. aureus, causing the decrease of photocurrent signal owing to steric hindrance effect. Leveraging photocurrent-off signal, there existed a satisfied linear relationship between the photocurrent variation and the logarithm of S. aureus concentration, achieving a wide linear range from 10 to 107 CFU/mL with a low detection limit of 2.0 CFU/mL. Notably, PDA/MnO2 with peroxidase-like activity facilitated the catalytic oxidation of S. aureus with assistance of hydrogen peroxide (H2O2) to cause the inactivation of S. aureus. Desorption of inactivated S. aureus from the photoelectrode led to a recovery of photocurrent signal, enabling a "signal on" switch. Simultaneously, the excellent photothermal performance of the PDA/MnO2 converted light energy into heat energy under the irradiation of NIR light (808 nm, 1.5 W/cm2), triggering the synergistic antibacterial effect against S. aureus (97.36%). This work provides a novel strategy for fabricating the detection and inactivation of bacteria in practical applications.

Detecting Contamination Events during Robotic Total Joint Arthroplasty.

BACKGROUND: Robot-assisted total joint arthroplasty (robotic-TJA) has become more widespread over the last 20 years due to higher patient satisfaction and reduced complications. However, robotic-TJA may have longer operative times and increased operating room traffic, which are known risk factors for contamination events. Contamination of surgical instruments may be contact- or airborne-related with documented scalpel blade contamination rates up to 9%. The robot arm is a novel instrument that comes in and out of the surgical field, so our objective was to assess whether the robot-arm is a source of contamination when used in robotic-TJA compared to other surgical instruments. METHODS: This was a prospective, single-institution, single-surgeon pilot study involving 103 robotic TJAs. The robot-arm was swabbed prior to incision and after closure. Pre- and postoperative control swabs were also collected from the suction tip and scalpel blade. Swabs were incubated for 24 hours on tryptic soy agar followed by inspection for growth of any contaminating bacteria. RESULTS: A contamination event was detected in 10 cases (10%). The scalpel blade was the most common site of contamination (8%) followed by the robot-arm (2%) and suction tip (0%). DISCUSSION: Contamination of the robot-arm during robotic-TJA is minimal when compared to contamination of the scalpel blade.

Microbiological etiology and current resistance patterns in acute calculous cholecystitis.

INTRODUCTION: The current treatment for acute calculous cholecystitis (ACC) is early laparoscopic cholecystectomy, in association with appropriate empiric antibiotic therapy. In our country, the evolution of the prevalence of the germs involved and their resistance patterns have been scarcely described. The aim of the study was to analyze the bacterial etiology and the antibiotic resistance patterns in ACC. METHODS: We conducted a single-center, retrospective, observational study of consecutive patients diagnosed with ACC between 01/2012 and 09/2019. Patients with a concomitant diagnosis of pancreatitis, cholangitis, postoperative cholecystitis, histology of chronic cholecystitis or carcinoma were excluded. Demographic, clinical, therapeutic and microbiological variables were collected, including preoperative blood cultures, bile and peritoneal fluid cultures. RESULTS: A total of 1104 ACC were identified, and samples were taken from 830 patients: bile in 89%, peritoneal fluid and/or blood cultures in 25%. Half of the bile cultures and less than one-third of the blood and/or peritoneum samples were positive. Escherichia coli (36%), Enterococcus spp (25%), Klebsiella spp (21%), Streptococcus spp (17%), Enterobacter spp (14%) and Citrobacter spp (7%) were isolated. Anaerobes were identified in 7% of patients and Candida spp in 1%. Nearly 37% of patients received inadequate empirical antibiotic therapy. Resistance patterns were scrutinized for each bacterial species. The main causes of inappropriateness were extended-spectrum beta-lactamase-producing bacteria (34%) and Enterococcus spp (45%), especially in patients older than 80 years. CONCLUSIONS: Updated knowledge of microbiology and resistance patterns in our setting is essential to readjust empirical antibiotic therapy and ACC treatment protocols.

Nanobubble applications in aquaculture industry for improving harvest yield, wastewater treatment, and disease control.

The ever-growing demand for aquaculture has led the industry to seek novel approaches for more sustainable practices. These attempts aim to increase aquaculture yield by increasing energy efficiency and decreasing footprint and chemical demand without compromising animal health. For this, emerging nanobubbles (NBs) aeration technology gained attention. NBs are gas-filled pockets suspended as sphere-like cavities (bulk NBs) or attached to surfaces (surface NBs) with diameters of <1 µm. Compared to macro bubbles and microbubbles, NBs have demonstrated unique characteristics such as long residence time in water, higher gas mass transfer efficiency and hydroxyl radical production. This paper focuses on reviewing NB technology in aquaculture systems by summarizing and discussing uses and implications. Three focus areas were targeted to review the applicability and effects of NBs in aquaculture: (i) NBs aeration to improve the aquaculture harvest yield and subsequent wastewater treatment; (ii) NB application for inactivation of harmful microorganisms; and (iii) NBs for reducing oxidative stress and improving animal health. Thus, this study reviews the research studies published in the last 10 years in which air, oxygen, ozone, and hydrogen NBs were tested to improve gas mass transfer, wastewater treatment and control of pathogenic microorganisms. The experimental results indicated that air and oxygen NBs yield significantly higher productivity, growth rate, total harvest, survival rate, and less oxygen consumption in fish and shrimp farming. Secondly, the application of air and ozone NBs demonstrated the ability of efficient pollutant degradation. Third, NB application demonstrated effective control of infectious bacteria and viruses, and thus increased fish survival, as well as different gene expression patterns that induce immune responses to infections. Reviewed studies lack robust comparative analyses of the efficacy of macro- and microbubble treatments. Also, potential health and safety implications, as well as economic feasibility through factors such as changes in capital infrastructure, routine maintenance and energy consumption need to be considered and evaluated in parallel to applicability. Therefore, even with a promising future, further studies are needed to confirm the benefits of NB treatment versus conventional aquaculture practices.

Biochar pH reduction using elemental sulfur and biological activation using compost or vermicompost.

This study aimed to improve biochar's quality for arid land applications by using elemental sulfur as a pH reducer agent co-applied with compost or vermicompost as biological activators. Biochar pH was decreased by the addition of elemental sulfur, with the highest reduction from 8.1 to 7.2 occurring when co-amended with vermicompost. Elemental sulfur increased the water-soluble concentrations of calcium, magnesium, and many other elements, and stimulated substrate-induced respiration, especially when co-amended with vermicompost. The bacterial diversity community structure were significantly affected by all treatments. The Shannon index significantly increased in response to compost and sulfur treatments, while the vermicompost treatments showed higher microbial evenness and equitability diversity indices. Multivariate analyses indicated that elemental sulfur oxidation was associated with specific sulfur-oxidizing bacterial clusters. Integrating biochar with sulfur and (vermi)compost was found to be a promising sustainable technology for managing excessive biochar alkalinity, increasing its fertility and potential for application in aridlands.

Broad-Spectrum Activity of Membranolytic Cationic Macrocyclic Peptides Against Multi-Drug Resistant Bacteria and Fungi.

The emergence of multidrug-resistant (MDR) strains causes severe problems in the treatment of microbial infections owing to limited treatment options. Antimicrobial peptides (AMPs) are drawing considerable attention as promising antibiotic alternative candidates to combat MDR bacterial and fungal infections. Herein, we present a series of small amphiphilic membrane-active cyclic peptides composed, in part, of various nongenetically encoded hydrophilic and hydrophobic amino acids. Notably, lead cyclic peptides 3b and 4b showed broad-spectrum activity against drug-resistant Gram-positive (MIC = 1.5-6.2 µg/mL) and Gram-negative (MIC = 12.5-25 µg/mL) bacteria, and fungi (MIC = 3.1-12.5 µg/mL). Furthermore, lead peptides displayed substantial antibiofilm action comparable to standard antibiotics. Hemolysis (HC50 = 230 µg/mL) and cytotoxicity (>70% cell viability against four different mammalian cells at 100 µg/mL) assay results demonstrated the selective lethal action of 3b against microbes over mammalian cells. A calcein dye leakage experiment substantiated the membranolytic effect of 3b and 4b, which was further confirmed by scanning electron microscopy. The behavior of 3b and 4b in aqueous solution and interaction with phospholipid bilayers were assessed by employing nuclear magnetic resonance (NMR) spectroscopy in conjunction with molecular dynamics (MD) simulations, providing a solid structural basis for understanding their membranolytic action. Moreover, 3b exhibited stability in human blood plasma (t1/2 = 13 h) and demonstrated no signs of resistance development against antibiotic-resistant S. aureus and E. coli. These findings underscore the potential of these newly designed amphiphilic cyclic peptides as promising anti-infective agents, especially against Gram-positive bacteria.

Prevalence and Resistance Patterns of Urinary Tract Infection in Al-Madinah Al-Munawarah, Saudi Arabia: A Retrospective Study.

BACKGROUND: Urinary tract infections (UTIs) are among the most common infections and can cause numerous complications of the renal system. This study aimed to assess the prevalence of uropathogens and their antibiotic susceptibility patterns in Al-Madinah Al-Munawarah, Saudi Arabia. METHODS: Data was collected from patients with UTIs presented at King Fahad General Hospital in Al-Madinah Al-Munawarah, Saudi Arabia. In this retrospective cross-sectional study, UTI microbial-causing agents and antimicrobial resistance profiles identified using automated systems, Phoenix and VITEK2, were collected between July 2022 and June 2023. In addition, minimal demographic data, including date of collection and sex and age of patients were collected and analyzed using Chi-square test. RESULTS: The study included 1394 patients positive for UTI, comprising 50.57% males and 49.43% females (chi-square goodness-of-fit, p > 0.999). Microbial identification and antimicrobial susceptibility tests were performed on UTI-positive cultures. Among UTIs, mono-infection, caused by a single pathogen, was the most prevalent, accounting for 88.16% of cases, whereas poly-infection (caused by multiple pathogens) presented at 11.9%. The most prevalent UTIs' pathogens were E. coli (30.59%), followed by Klebsiella pneumoniae (21.40%), Enterococcus faecalis (8.46%), Pseudomonas aeruginosa (7.81%), Streptococcus agalactiae (6.35%), Enterococcus faecium (3.01%), Proteus mirabilis (3.01%), Enterobacter cloacae (2.52%), Candida sp. (2.44%), Acinetobacter calcoaceticus-baumannii (1.95%), Staphylococcus aureus (1.79%), and Enterobacter aerogenes (1.30%). The most dominant pathogens that coexisted with other uropathogens to cause UTIs were K. pneumoniae and P. mirabilis (9.32%, chi-square 5.550, p = 0.018), K. pneumoniae and P. aeruginosa (8.07%, chi-square 6.285, p = 0.012), K. pneumoniae and E. faecalis (7.45%, chi-square 5.785, p = 0.016), Candida sp. and Enterococcus faecium (4.97%, chi-square 9.176, p = 0.002, and Candida sp. and Acinetobacter calcoaceticus-baumannii (3.11%, chi-square 4.312, p=0.038)). Among the uropathogens, gram-negative pathogens showed resistance to most of the tested antimicrobials (ampicillins, cephalosporins, fluoroquinolones, trimethoprim-sulfamethoxazole, aztreonam, and nitrofurantoin). High rates of resistance were identified to cephalosporins, amoxicillin-clavulanic acid, and trimethoprim-sulfamethoxazole. CONCLUSION: This study reported UT mono-infection and poly-infection in Al-Madinah Al-Munawarah, Saudi Arabia, with a predominant representation from gram-negative bacteria, Enterobacteriaceae. Most of the UT microbial strains showed a highly resistant profile.

Comparative analysis of Spodoptera frugiperda (J. E. Smith) (Lepidoptera, Noctuidae) corn and rice strains microbiota revealed minor changes across life cycle and strain endosymbiont association.

Background: Spodoptera frugiperda (FAW) is a pest that poses a significant threat to corn production worldwide, causing millions of dollars in losses. The species has evolved into two strains (corn and rice) that differ in their genetics, reproductive isolation, and resistance to insecticides and Bacillus thuringiensis endotoxins. The microbiota plays an important role in insects' physiology, nutrient acquisition, and response to chemical and biological controls. Several studies have been carried out on FAW microbiota from larvae guts using laboratory or field samples and a couple of studies have analyzed the corn strain microbiota across its life cycle. This investigation reveals the first comparison between corn strain (CS) and rice strain (RS) of FAW during different developmental insect stages and, more importantly, endosymbiont detection in both strains, highlighting the importance of studying both FAW populations and samples from different stages. Methods: The composition of microbiota during the life cycle of the FAW corn and rice strains was analyzed through high-throughput sequencing of the bacterial 16S rRNA gene using the MiSeq system. Additionally, culture-dependent techniques were used to isolate gut bacteria and the Transcribed Internal Spacer-ITS, 16S rRNA, and gyrB genes were examined to enhance bacterial identification. Results: Richness, diversity, and bacterial composition changed significantly across the life cycle of FAW. Most diversity was observed in eggs and males. Differences in gut microbiota diversity between CS and RS were minor. However, Leuconostoc, A2, Klebsiella, Lachnoclostridium, Spiroplasma, and Mucispirilum were mainly associated with RS and Colidextribacter, Pelomonas, Weissella, and Arsenophonus to CS, suggesting that FAW strains differ in several genera according to the host plant. Firmicutes and Proteobacteria were the dominant phyla during FAW metamorphosis. Illeobacterium, Ralstonia, and Burkholderia exhibited similar abundancies in both strains. Enterococcus was identified as a conserved taxon across the entire FAW life cycle. Microbiota core communities mainly consisted of Enterococcus and Illeobacterium. A positive correlation was found between Spiroplasma with RS (sampled from eggs, larvae, pupae, and adults) and Arsenophonus (sampled from eggs, larvae, and adults) with CS. Enterococcus mundtii was predominant in all developmental stages. Previous studies have suggested its importance in FAW response to B. thuringensis. Our results are relevant for the characterization of FAW corn and rice strains microbiota to develop new strategies for their control. Detection of Arsenophonus in CS and Spiroplasma in RS are promising for the improvement of this pest management, as these bacteria induce male killing and larvae fitness reduction in other Lepidoptera species.

Modern approaches for detection of volatile organic compounds in metabolic studies focusing on pathogenic bacteria: Current state of the art.

Pathogenic microorganisms produce numerous metabolites, including volatile organic compounds (VOCs). Monitoring these metabolites in biological matrices (e.g., urine, blood, or breath) can reveal the presence of specific microorganisms, enabling the early diagnosis of infections and the timely implementation of targeted therapy. However, complex matrices only contain trace levels of VOCs, and their constituent components can hinder determination of these compounds. Therefore, modern analytical techniques enabling the non-invasive identification and precise quantification of microbial VOCs are needed. In this paper, we discuss bacterial VOC analysis under in vitro conditions, in animal models and disease diagnosis in humans, including techniques for offline and online analysis in clinical settings. We also consider the advantages and limitations of novel microextraction techniques used to prepare biological samples for VOC analysis, in addition to reviewing current clinical studies on bacterial volatilomes that address inter-species interactions, the kinetics of VOC metabolism, and species- and drug-resistance specificity.

Microbiome-derived acidity protects against microbial invasion in Drosophila.

Microbial invasions underlie host-microbe interactions resulting in pathogenesis and probiotic colonization. In this study, we explore the effects of the microbiome on microbial invasion in Drosophila melanogaster. We demonstrate that gut microbes Lactiplantibacillus plantarum and Acetobacter tropicalis improve survival and lead to a reduction in microbial burden during infection. Using a microbial interaction assay, we report that L. plantarum inhibits the growth of invasive bacteria, while A. tropicalis reduces this inhibition. We further show that inhibition by L. plantarum is linked to its ability to acidify its environment via lactic acid production by lactate dehydrogenase, while A. tropicalis diminishes the inhibition by quenching acids. We propose that acid from the microbiome is a gatekeeper to microbial invasions, as only microbes capable of tolerating acidic environments can colonize the host. The methods and findings described herein will add to the growing breadth of tools to study microbe-microbe interactions in broad contexts.

Episymbiotic Saccharibacteria TM7x modulates the susceptibility of its host bacteria to phage infection and promotes their coexistence.

Bacteriophages (phages) play critical roles in modulating microbial ecology. Within the human microbiome, the factors influencing the long-term coexistence of phages and bacteria remain poorly investigated. Saccharibacteria (formerly TM7) are ubiquitous members of the human oral microbiome. These ultrasmall bacteria form episymbiotic relationships with their host bacteria and impact their physiology. Here, we showed that during surface-associated growth, a human oral Saccharibacteria isolate (named TM7x) protects its host bacterium, a Schaalia odontolytica strain (named XH001) against lytic phage LC001 predation. RNA-Sequencing analysis identified in XH001 a gene cluster with predicted functions involved in the biogenesis of cell wall polysaccharides (CWP), whose expression is significantly down-regulated when forming a symbiosis with TM7x. Through genetic work, we experimentally demonstrated the impact of the expression of this CWP gene cluster on bacterial-phage interaction by affecting phage binding. In vitro coevolution experiments further showed that the heterogeneous populations of TM7x-associated and TM7x-free XH001, which display differential susceptibility to LC001 predation, promote bacteria and phage coexistence. Our study highlights the tripartite interaction between the bacterium, episymbiont, and phage. More importantly, we present a mechanism, i.e., episymbiont-mediated modulation of gene expression in host bacteria, which impacts their susceptibility to phage predation and contributes to the formation of "source-sink" dynamics between phage and bacteria in biofilm, promoting their long-term coexistence within the human microbiome.

Microbial chassis design and engineering for production of gamma-aminobutyric acid.

Gamma-aminobutyric acid (GABA) is a non-protein amino acid which is widely applied in agriculture and pharmaceutical additive industries. GABA is synthesized from glutamate through irreversible α-decarboxylation by glutamate decarboxylase. Recently, microbial synthesis has become an inevitable trend to produce GABA due to its sustainable characteristics. Therefore, reasonable microbial platform design and metabolic engineering strategies for improving production of GABA are arousing a considerable attraction. The strategies concentrate on microbial platform optimization, fermentation process optimization, rational metabolic engineering as key metabolic pathway modification, promoter optimization, site-directed mutagenesis, modular transporter engineering, and dynamic switch systems application. In this review, the microbial producers for GABA were summarized, including lactic acid bacteria, Corynebacterium glutamicum, and Escherichia coli, as well as the efficient strategies for optimizing them to improve the production of GABA.

Therapeutic bacteria and viruses to combat cancer: double-edged sword in cancer therapy: new insights for future.

Cancer, ranked as the second leading cause of mortality worldwide, leads to the death of approximately seven million people annually, establishing itself as one of the most significant health challenges globally. The discovery and identification of new anti-cancer drugs that kill or inactivate cancer cells without harming normal and healthy cells and reduce adverse effects on the immune system is a potential challenge in medicine and a fundamental goal in Many studies. Therapeutic bacteria and viruses have become a dual-faceted instrument in cancer therapy. They provide a promising avenue for cancer treatment, but at the same time, they also create significant obstacles and complications that contribute to cancer growth and development. This review article explores the role of bacteria and viruses in cancer treatment, examining their potential benefits and drawbacks. By amalgamating established knowledge and perspectives, this review offers an in-depth examination of the present research landscape within this domain and identifies avenues for future investigation.