Classically, quorum sensing (QS) is defined as a form of bacterial communication that regulates gene expression in response to population density and other environmental conditions. While a variety of QS systems exist, N-acyl homoserine lactone-based quorum sensing (AHL-QS) is the best characterized in Gram-negative Pseudomonadota (formerly Proteobacteria), often in medically relevant taxa. However, it remains unclear what AHL-QS systems look like in underexplored environments, and how often or far the canonical model extends beyond the Pseudomonadota. Here, we investigated AHL production in environmental Bacteria from underexplored environments, including two newly described Pseudomonadota species, Brenneria uluponensis K61T, from a taro lo‘i in Hawai‘i, and Bradyrhizobium prioriisuperbiae BL16AT, from a lava cave on the Island of Hawai‘i, and two Gram-positive Actinomycetota (formerly Actinobacteria), Pseudonocardia alni GV4, from a culture of Gloeobacter violaceus, and Rhodococcus kroppenstedtii Y88A, from a dehydrated hypersaline mat on San Salvador Island, Bahamas. Custom hidden Markov models (HMMs) were used to identify putative luxI/R homologs, followed by analyses of genomic context, luxI/R-family protein phylogeny, and LuxR domain architecture. AHL production was assessed using liquid chromatography-multiple reaction monitoring-mass spectrometry (LC-MRM-MS), and targeted gene disruption was performed in R. kroppenstedtii Y88A. Putative luxI/R homologs were identified in all strains. In R. kroppenstedtii Y88A, disruption of the sole luxI homolog resulted in loss of all detectable AHLs, indicating that this gene (designated rhdI) is required for AHL production under the tested conditions. Across Actinomycetota genomes, luxI genes occurred as solitary elements rather than canonical luxI/R pairs and were associated with genes linked to metabolism and redox processes. Phylogenetic analyses revealed that Actinomycetota luxI/R-family proteins diverge from canonical systems, with LuxR-family proteins lacking canonical autoinducer-binding domains and instead comprising only helix-turn-helix regulatory architectures. Together, these findings expand the known diversity of AHL-QS systems into underexplored microbial lineages and environments and suggest broader ecological roles for AHL signaling. The canonical luxI/R model derived largely from Pseudomonadota may represent only one of several evolutionary architectures for AHL signaling, raising the possibility that AHL production in Actinomycetota operates through regulatory frameworks distinct from classical AHL-QS systems.

The potential opening of petroleum-related activities in the Arctic due to climate change has increased the risk of oil discharges and spills, which was already significant due to drilling and shipping. Oil spill response in this region is challenging due to harsh weather conditions and the current limitations of clean-up techniques. Therefore, a safe, rapid, and effective approach is needed for an efficient response to oil spills in Arctic, and other cold, regions. Quantitative oil removal from seawater using polyvinylpyrrolidone (PVP)-coated iron oxide nanoparticles (PVP-IONPs) under environmentally relevant conditions had been previously demonstrated. However, this method has only been tested at room temperature. This paper examined the efficacy of oil sorption by PVP-IONPs in low temperatures comparable to those of polar waters, in both synthetic and natural seawaters. Physico-chemical characteristics of the PVP-IONPs were consistent with previous batches. Oil removal results clearly indicated that removal of aliphatic and aromatic components was approximately 100% in synthetic marine waters with and without natural organic macromolecules (NOM), and there was no significant or systematic effect of temperature. Longer chain aliphatics were slightly less well removed (50–100% removal for C22-C26) as seen before, but unaffected by temperature. Interestingly, no change in removal effectiveness was observed even at −3 °C (vigorously mixed saline waters). The finding that nano-enabled removal was equally effective at all tested temperatures, suggests that this technology may be suitable for deployment in low temperature operational environments where the applicability of conventional methods is limited.

The Hawaiian Islands are among the most geologically and volcanically active places on Earth. While the Hawaiian Archipelago is known for its animal and plant diversity, much less is known about microbial diversity in the area's diverse habitats. In this study, we focused on steam vent associated biofilms found on the most volcanically active island of Hawai'i, also known as the Big Island. From 46 samples from various biofilms and associated features around fumaroles emitting water steam, we generated amplicon and metagenomic sequences. This represents a total of 276 Gbp of raw sequencing data. From the shotgun metagenomic data, we constructed 363 non-redundant medium- to high-quality metagenome-assembled genomes (MAGs) that are at least 70% complete and with less than 5% contamination. Of these, ten MAGs belong in the domain Archaea, and 353 belong in the domain Bacteria. This dataset could provide valuable insights into microbial diversity and ecology around volcanic features in Hawai'i and elsewhere.

In this work, we explore the potency of poly(D-peptide) (D-PP) in combination with diverse classes of antibiotics to address multidrug-resistant (MDR) Gram-negative superbugs, including their mature biofilms, polymicrobial assemblies, and dormant subpopulations, which are among complex challenging targets. The polypeptide adjuvant displayed significant antibiotic potentiation, resulting in a moderate to rapid bactericidal activity against the Gram-negative pathogens in their planktonic stage. The adjuvant and its combination restored susceptibility in dormant bacterial subpopulations. In polymicrobial settings, these combinations were able to reduce both Gram-negative and Gram-positive bacterial species, unlike individual antibiotic exposure. Importantly, one of the lead combinations (D-PP-rifampicin) resulted in moderate biofilm eradication of P. aeruginosa. Transcriptomic analysis demonstrated the differentially expressed genes (DEGs) involved in membrane integrity, stress response, and transport systems. Together with our previous mechanistic findings, this further confirmed that bacterial membrane perturbation is caused by the adjuvant. Such perturbation facilitated antibiotic accumulation within microbial cells. Furthermore, polypeptide reduces the bacterial resistance development towards doxycycline and rifampicin, unlike polymyxin B. Taken together, these findings exhibit that membrane-active polypeptide adjuvants can enhance antibiotic efficacy across diverse bacterial states and environments, providing a promising strategy to combat persistent and multidrug-resistant infections.

Antimicrobial polypeptides (AMPs) are often engineered to adopt a secondary structure (e.g., α-helix, β-sheet, or turns), reinforcing the prevailing view that ordered conformations are preferred for enhanced activity. Here, we report a first-of-its-kind stereochemical series of cationic polypeptides with defined d/L ratios (100%L, 100%d, 80%d/20%L, 50%d/50%L, 20%d/80%L) via N-carboxyanhydride (NCA) polymerization and thiol-ene postfunctionalization. Circular dichroism spectroscopy confirms helix-like signatures for all compositions, except the racemic 50d/50L, which lacks a defined structure under membrane-mimicking conditions. Unexpectedly, the racemic polymer is strongly antimicrobial against Gram-positive pathogens (minimum inhibitory concentration, MIC = 4-16 μg/mL) and matches the proteolytic stability of the d-enantiomer analogues. As an adjuvant, it sensitizes multidrug-resistant Gram-negative bacteria, enhancing the efficacy of a wide variety of antibiotics by up to 128-fold across multiple classes and showing highly synergistic effects (fractional inhibitory concentration index, FICI≤0.5). Mechanistic assays reveal comparable outer-membrane perturbation, increased intracellular antibiotic accumulation, membrane depolarization, and reactive oxygen species (ROS) elevation across helical and nonhelical members, indicating that an ordered secondary structure is not required for either direct antimicrobial action or potentiation. By establishing enantiomeric-ratio control as a stereochemical design and identifying racemic composition as a practical, scalable alternative, this study provides guidance for developing robust, low-hemolytic AMP mimics and a universal adjuvant for antibiotics, particularly suited to low-resource settings where access to advanced therapeutics is limited.

The escalating rise in antimicrobial resistance (AMR) coupled with a declining arsenal of new antibiotics is imposing serious threats to global public health. A pervasive aspect of many acquired AMR infections is that the pathogenic microorganisms exist as biofilms, which are equipped with superior survival strategies. In addition, persistent and recalcitrant infections are seeded with bacterial persister cells at infection sites. Together, conventional antibiotic therapeutics often fail in the complete treatment of infections associated with bacterial persisters and biofilms. Novel therapeutics have been attempted to tackle AMR, biofilms, and persister-associated complex infections. This review focuses on the progress in designing molecular biomaterials and therapeutics to address acquired and intrinsic AMR, and the fundamental microbiology behind biofilms and persisters. Starting with a brief introduction of AMR basics and approaches to tackling acquired AMR, the emphasis is placed on various biomaterial approaches to combating intrinsic AMR, including (1) semi-synthetic antibiotics; (2) macromolecular or polymeric biomaterials mimicking antimicrobial peptides; (3) adjuvant effects in synergy; (4) nano-therapeutics; (5) nitric oxide-releasing antimicrobials; (6) antimicrobial hydrogels; (7) antimicrobial coatings. Particularly, the structure-activity relationship is elucidated in each category of these biomaterials. Finally, illuminating perspectives are provided for the future design of molecular biomaterials to bypass AMR and cure chronic multi-drug resistant (MDR) infections.

The rise of antibiotic resistance, biofilm formation, and dormant bacterial populations poses serious global health threats. Synthetic antimicrobial peptide (AMP) mimics offer promising alternatives, though the impact of secondary structures in polymeric AMP mimics on antimicrobial efficacy is underexplored. This study investigates chirality-controlled α-peptide polymers (D-PP and DL-PP), synthesized via ring-opening polymerization of allylglycine N-carboxy anhydrides and post-polymerization modification through thiol-ene click chemistry. D-PP adopts a stable helical structure under biomimetic conditions, whereas DL-PP remains random. This helical structure enhanced D-PP's antibacterial and antibiotic potentiation activities, amplifying antibiotic efficacy by 2- to 256-fold across various classes-including tetracyclines, ansamycins, fusidanes, macrolides, cephalosporins, and monobactams-against multidrug-resistant Gram-negative pathogens, while maintaining low hemolytic activity and high protease stability. Mechanistic investigations revealed that D-PP exhibited greater membrane interaction. D-PP and antibiotic combinations eradicated dormant bacterial populations and disrupted biofilms with minimal antimicrobial resistance development. This study paves the way for the rational design of polypeptide-based antimicrobial agents, harnessing chirality and secondary structural features to enhance the efficacy of synthetic antimicrobial peptide mimics.

Climate-induced stressors, such as changes in temperature, salinity, and pH, contribute to the emergence of infectious diseases. These changes alter geographical constraint, resulting in increased Vibrio spread, exposure, and infection rates, thus facilitating greater Vibrio-human interactions. Multiple efforts have been developed to predict Vibrio exposure and raise awareness of health risks, but most models only use temperature and salinity as prediction factors. This study aimed to better understand the potential effects of temperature and pH on V. vulnificus and V. parahaemolyticus planktonic and biofilm growth. Vibrio strains were grown in triplicate at 25°, 30°, and 37°C in 96 well plates containing Modified Seawater Yeast Extract modified with CaCl2 at pH's ranging from 5 to 9.6. AMiGA software was used to model growth curves using Gaussian process regression. The effects of temperature and pH were evaluated using randomized complete block analysis of variance, and the growth rates of V. parahaemolyticus and V. vulnificus were modeled using the interpolation fit on the MatLab Curve Fitting Toolbox. Different optimal conditions involving temperature and pH were observed for planktonic and biofilm Vibrio growth within- and between-species. This study showed that temperature and pH factors significantly affect Vibrio planktonic growth rates and V. parahaemolyticus biofilm formation. Therefore, pH effects must be added to the Vibrio growth modeling efforts to better predict Vibrio risk in estuarine and coastal zones that can potentially experience the cooccurrence of Vibrio and harmful algal bloom outbreak events.

Members of the archaeal order Caldarchaeales (previously the phylum Aigarchaeota) are poorly sampled and are represented in public databases by relatively few genomes. Additional representative genomes will help resolve their placement among all known members of Archaea and provide insights into their roles in the environment. In this study, we analyzed 16S rRNA gene amplicons belonging to the Caldarchaeales that are available in public databases, which demonstrated that archaea of the order Caldarchaeales are diverse, widespread, and most abundant in geothermal habitats. We also constructed five metagenome-assembled genomes (MAGs) of Caldarchaeales from two geothermal features to investigate their metabolic potential and phylogenomic position in the domain Archaea. Two of the MAGs were assembled from microbial community DNA extracted from fumarolic lava rocks from Mauna Ulu, Hawai'i, and three were assembled from DNA obtained from hot spring sinters from the El Tatio geothermal field in Chile. MAGs from Hawai'i are high quality bins with completeness >95% and contamination <1%, and one likely belongs to a novel species in a new genus recently discovered at a submarine volcano off New Zealand. MAGs from Chile have lower completeness levels ranging from 27 to 70%. Gene content of the MAGs revealed that these members of Caldarchaeales are likely metabolically versatile and exhibit the potential for both chemoorganotrophic and chemolithotrophic lifestyles. The wide array of metabolic capabilities exhibited by these members of Caldarchaeales might help them thrive under diverse harsh environmental conditions. All the MAGs except one from Chile harbor putative prophage regions encoding several auxiliary metabolic genes (AMGs) that may confer a fitness advantage on their Caldarchaeales hosts by increasing their metabolic potential and make them better adapted to new environmental conditions. Phylogenomic analysis of the five MAGs and over 3,000 representative archaeal genomes showed the order Caldarchaeales forms a monophyletic group that is sister to the clade comprising the orders Geothermarchaeales (previously Candidatus Geothermarchaeota), Conexivisphaerales and Nitrososphaerales (formerly known as Thaumarchaeota), supporting the status of Caldarchaeales members as a clade distinct from the Thaumarchaeota.

Democratizing genomic data science, including bioinformatics, can diversify the STEM workforce and may, in turn, bring new perspectives into the space sciences. In this respect, the development of education and research programs that bridge genome science with "place" and world-views specific to a given region are valuable for Indigenous students and educators. Through a multi-institutional collaboration, we developed an ongoing education program and model that includes Illumina and Oxford Nanopore sequencing, free bioinformatic platforms, and teacher training workshops to address our research and education goals through a place-based science education lens. High school students and researchers cultivated, sequenced, assembled, and annotated the genomes of 13 bacteria from Mars analog sites with cultural relevance, 10 of which were novel species. Students, teachers, and community members assisted with the discovery of new, potentially chemolithotrophic bacteria relevant to astrobiology. This joint education-research program also led to the discovery of species from Mars analog sites capable of producing N-acyl homoserine lactones, which are quorum-sensing molecules used in bacterial communication. Whole genome sequencing was completed in high school classrooms, and connected students to funded space research, increased research output, and provided culturally relevant, place-based science education, with participants naming three novel species described here. Students at St. Andrew's School (Honolulu, Hawai'i) proposed the name Bradyrhizobium prioritasuperba for the type strain, BL16AT, of the new species (DSM 112479T = NCTC 14602T). The nonprofit organization Kauluakalana proposed the name Brenneria ulupoensis for the type strain, K61T, of the new species (DSM 116657T = LMG = 33184T), and Hawai'i Baptist Academy students proposed the name Paraflavitalea speifideiaquila for the type strain, BL16ET, of the new species (DSM 112478T = NCTC 14603T).

Marine bacteria often exist in biofilms as communities attached to surfaces, like plastic. Growing concerns exist regarding marine plastics acting as potential vectors of pathogenic Vibrio, especially in a changing climate. It has been generalized that Vibrio vulnificus and Vibrio parahaemolyticus often attach to plastic surfaces. Different strains of these Vibrios exist having different growth and biofilm-forming properties. This study evaluated how temperature and strain variability affect V. parahaemolyticus and V. vulnificus biofilm formation and characteristics on glass (GL), low-density polyethylene (LDPE), polypropylene (PP), and polystyrene (PS). All strains of both species attached to GL and all plastics at 25, 30, and 35°C. As a species, V. vulnificus produced more biofilm on PS (p ≤ 0.05) compared to GL, and biofilm biomass was enhanced at 25°C compared to 30° (p ≤ 0.01) and 35°C (p ≤ 0.01). However, all individual strains’ biofilm biomass and cell densities varied greatly at all temperatures tested. Comparisons of biofilm-forming strains for each species revealed a positive correlation (r = 0.58) between their dry biomass weight and OD570 values from crystal violet staining, and total dry biofilm biomass for both species was greater (p ≤ 0.01) on plastics compared to GL. It was also found that extracellular polymeric substance (EPS) chemical characteristics were similar on all plastics of both species, with extracellular proteins mainly contributing to the composition of EPS. All strains were hydrophobic at 25, 30, and 35°C, further illustrating both species’ affinity for potential attachment to plastics. Taken together, this study suggests that different strains of V. parahaemolyticus and V. vulnificus can rapidly form biofilms with high cell densities on different plastic types in vitro. However, the biofilm process is highly variable and is species-, strain-specific, and dependent on plastic type, especially under different temperatures.

We report facially amphiphilic bile acid-based antimicrobials with a broad spectrum of activity against both bacterial and fungal pathogens and negligible detrimental effects on mammalian cells. Two lead compounds eliminated dormant subpopulations of various bacterial species, unlike conventional antibiotics. The lead compounds were also effective in eradicating biofilms of methicillin-resistant Staphylococcus aureus (MRSA), Pseudomonas aeruginosa, and Candida albicans. Additionally, these compounds substantially inhibited the formation of fungal biofilms (C. albicans). Mechanistic investigations revealed the membrane-active nature and endogenous reactive oxygen species (ROS) induction ability of these compounds. Finally, no detectable resistance was developed by the bacterial strains against this class of membrane-targeting antimicrobials.

Growing concerns exist regarding human ingestion of contaminated seafood that contains Vibrio biofilms on microplastics (MPs). One of the mechanisms enhancing biofilm related infections in humans is due to biofilm dispersion, a process that triggers release of bacteria from biofilms into the surrounding environment, such as the gastrointestinal tract of human hosts. Dispersal of cells from biofilms can occur in response to environmental conditions such as sudden changes in temperature, pH and nutrient conditions, as the bacteria leave the biofilm to find a more stable environment to colonize. This study evaluated how brief exposures to nutrient starvation, elevated temperature, different pH levels and simulated human media affect Vibrio parahaemolyticus and Vibrio vulnificus biofilm dispersal and processes on and from low-density polyethylene (LDPE), polypropylene (PP), and polystyrene (PS) MPs. Both species were able to adequately disperse from all types of plastics under most exposure conditions. V. parahaemolyticus was able to tolerate and survive the low pH that resembles the gastric environment compared to V. vulnificus. pH had a significantly (p ≤ 0.05) positive effect on overall V. parahaemolyticus biofilm biomass in microplates and cell colonization from PP and PS. pH also had a positive effect on V. vulnificus cell colonization from LDPE and PP. However, most biofilm biomass, biofilm cell and dispersal cell densities of both species greatly varied after exposure to elevated temperature, pH, and nutrient starvation. It was also found that certain exposures to simulated human media affected both V. parahaemolyticus and V. vulnificus biofilm biomass and biofilm cell densities on LDPE, PP and PS compared to exposure to traditional media of similar pH. Cyclic-di-GMP was higher in biofilm cells compared to dispersal cells, but exposure to more stressful conditions significantly increased signal concentrations in both biofilm and dispersal states. Taken together, this study suggests that human pathogenic strains of V. parahaemolyticus and V. vulnificus can rapidly disperse with high cell densities from different plastic types in vitro. However, the biofilm dispersal process is highly variable, species specific and dependent on plastic type, especially under different human body related environmental exposures.

Aflatoxins, a family of fungal secondary metabolites, are toxic and carcinogenic compounds that pose an enormous threat to global food safety and agricultural sustainability. Specifically agricultural products in African, Southeast Asian and hot and humid regions of American countries suffer most damage from aflatoxin producing molds due to the ideal climate conditions promoting their growth. Our recent studies suggest that Vibrio gazogenes (Vg), an estuarine bacterium non-pathogenic to plants and humans, can significantly inhibit aflatoxin biosynthesis in the producers. In this study, we investigated the mechanism underlying Vg-dependent aflatoxin inhibition using the prominent aflatoxin producer, Aspergillus flavus. We show that aflatoxin inhibition upon Vg treatment was associated with fungal uptake of Vg-prodigiosin, a red pigment, which was consistently visible inside fungal hyphae during treatment. The association of prodigiosin with aflatoxin inhibition was further evident as Serratia marcescens, another prodigiosin producer, significantly inhibited aflatoxin, while non-producers like Escherichia coli, Staphylococcus aureus, Vibrio harveyi, and Vibrio fischeri did not. Also, pure prodigiosin significantly inhibited aflatoxin biosynthesis. Endocytosis inhibitors, filipin and natamycin, reduced the Vg-prodigiosin uptake by the fungus leading to a significant increase in aflatoxin production, suggesting that uptake is endocytosis-dependent. The Vg treatment also reduced hyphal fusion (>98% inhibition) and branching, which are both endosome-dependent processes. Our results, therefore, collectively support our theory that Vg-associated aflatoxin inhibition is mediated by an endocytosis-dependent uptake of Vg-prodigiosin, which possibly leads to a disruption of normal endosomal functions.

Compared with single-network hydrogels, double-network hydrogels offer higher mechanical strength and toughness. Integrating useful functions into double-network hydrogels can expand the portfolios of the hydrogels. We report the preparation of double-network metallopolymer hydrogels with remarkable hydration, antifouling, and antimicrobial properties. These cationic hydrogels are composed of a first network of cationic cobaltocenium polyelectrolytes and a second network of polyacrylamide, all prepared via radical polymerization. Antibiotics were further installed into the hydrogels via ion-complexation with metal cations. These hydrogels exhibited significantly enhanced hydration, compared with polyacrylamide-based hydrogels, while featuring robust mechanical strength. Cationic metallopolymer hydrogels exhibited strong antifouling against oppositely charged proteins. These antibiotic-loaded hydrogels demonstrated a synergistic effect on the inhibition of bacterial growth and antifouling of bacteria, as a result of the unique ion complexation of cobaltocenium cations.

The Puquios of the Salar de Llamara in the Atacama Desert, northern Chile, is a system of small lakes that is characterized by evaporitic mineral deposition and that commonly hosts microbial communities. This region is known for its extreme aridity, solar irradiance, and temperature fluctuations. The Puquios are a highly diverse ecosystem with a variety of sedimentary bottom types. Our previous results identified electrical conductivity (EC) as a first-order environmental control on bottom types. In the present paper, we extend our analysis to examine the effects of additional environmental parameters on bottom types and to consider reasons for the importance of EC as a control of sedimentology. Our results identify microbially produced extracellular polymeric substances (EPS) as a major player in the determination of bottom types. The relative amounts and properties of EPS are determined by EC. EPS, in turn, determines the consistency of bottom types, exchange of bottom substrate with the overlying water column, and mineral precipitation within the substrate. Low-EC ponds in the Puquios system have flocculent to semi-cohesive bottom types, with low-viscosity EPS that allows for high-exchange with the surrounding waters and mineral precipitation of granular gypsum, carbonate, and Mg–Si clay in close association with microbes. Ponds with elevated EC have bottom types that are laminated and highly cohesive with high-viscosity EPS that restricts the exchange between sediments and the surrounding waters; mineral precipitation in these high-EC ponds includes granular to laminated gypsum, carbonate and Mg–Si, which also form in close association with microbes. Bottom types in ponds with EC above the threshold for thriving benthic microbial communities have insufficient EPS accumulations to affect mineral precipitation, and the dominant mineral is gypsum (selenite). The variations in EPS production throughout the Puquios, associated with heterogeneity in environmental conditions, make the Puquios region an ideal location for understanding the controls of sedimentary bottom types in evaporative extreme environments that may be similar to those that existed on early Earth and beyond.

Lava caves, tubes, and fumaroles in Hawai‘i present a range of volcanic, oligotrophic environments from different lava flows and host unexpectedly high levels of bacterial diversity. These features provide an opportunity to study the ecological drivers that structure bacterial community diversity and assemblies in volcanic ecosystems and compare the older, more stable environments of lava tubes, to the more variable and extreme conditions of younger, geothermally active caves and fumaroles. Using 16S rRNA amplicon-based sequencing methods, we investigated the phylogenetic distinctness and diversity and identified microbial interactions and consortia through co-occurrence networks in 70 samples from lava tubes, geothermal lava caves, and fumaroles on the island of Hawai‘i. Our data illustrate that lava caves and geothermal sites harbor unique microbial communities, with very little overlap between caves or sites. We also found that older lava tubes (500–800 yrs old) hosted greater phylogenetic diversity (Faith's PD) than sites that were either geothermally active or younger (<400 yrs old). Geothermally active sites had a greater number of interactions and complexity than lava tubes. Average phylogenetic distinctness, a measure of the phylogenetic relatedness of a community, was higher than would be expected if communities were structured at random. This suggests that bacterial communities of Hawaiian volcanic environments are phylogenetically over-dispersed and that competitive exclusion is the main driver in structuring these communities. This was supported by network analyses that found that taxa (Class level) co-occurred with more distantly related organisms than close relatives, particularly in geothermal sites. Network “hubs” (taxa of potentially higher ecological importance) were not the most abundant taxa in either geothermal sites or lava tubes and were identified as unknown families or genera of the phyla, Chloroflexi and Acidobacteria. These results highlight the need for further study on the ecological role of microbes in caves through targeted culturing methods, metagenomics, and long-read sequence technologies.

Facial amphiphilicity is an extraordinary chemical structure feature of a variety of antimicrobial peptides and polymers. Vast efforts have been dedicated to small molecular, macromolecular and dendrimer-like systems to mimic this highly preferred structure or conformation, including local facial amphiphilicity and global amphiphilicity. This work conceptualizes Facial Amphiphilicity Index (FAI) as a numerical value to quantitatively characterize the measure of chemical compositions and structural features in dictating antimicrobial efficacy. FAI is a ratio of numbers of charges to rings, representing both compositions of hydrophilicity and hydrophobicity. Cationic derivatives of multicyclic compounds were evaluated as model systems for testing antimicrobial selectivity against Gram-negative and Gram-positive bacteria. Both monocyclic and bicyclic compounds are non-antimicrobial regardless of FAIs. Antimicrobial efficacy was observed with systems having larger cross-sectional areas including tricyclic abietic acid and tetracyclic bile acid. While low and high FAIs respectively lead to higher and lower antimicrobial efficacy, in consideration of cytotoxicity, the sweet spot is typically suited with intermediate FAIs for each specific system. This can be well explained by the synergistic hydrophobic-hydrophobic and electrostatic interactions with bacterial cell membranes and the difference between bacterial and mammalian cell membranes. The adoption of FAI would pave a new avenue toward the design of next-generation antimicrobial macromolecules and peptides.

Bacterial extracellular polymeric substances (EPS) have been recently found to contribute most for metal removal in nanoenhanced bioremediation. However, the mechanism by which NPs affect EPS-metal interactions is not fully known. Here, Halomonas sp. was employed to explore the role of EPS after in vivo exposure to Cd/Pb and polyvinylpyrrolidone (PVP) coated iron oxide nanoparticles (IONPs, 20 mg L-1) for 72 h. Cd-IONPs produced the highest concentrations of EPS proteins (136.3 mg L-1), while Cd induced the most production of polysaccharides (241.0 mg L-1). IONPs increased protein/polysaccharides ratio from 0.2 (Cd) to 1.2 (Cd-IONPs). The increased protein favors the formation of protein coronas on IONPs surface, which would promote Cd adsorption during NP-metal-EPS interaction. FTIR analysis indicated that the coexistence of Cd and IONPs interacted with proteins more strongly than with polysaccharides. Glycosyl monomer analyses suggested mannose and glucose as target sugars for EPS complexation with metals, and IONPs reduced metal-induced changes in monosaccharide profiles. Protein secondary structures changed in all treatments, but we could not distinguish stresses induced by metals from those by IONPs. These findings provide greater understanding of the role of EPS in NP-metal-EPS interaction, providing a better underpinning knowledge for the application of NP-enhanced bioremediation.