Long-term toxicity assessment of antibiotics against Vibrio fischeri: Test method optimization and mixture toxicity prediction.

The current luminescent bacteria test for acute toxicity with short contact time was invalid for antibiotics, and the non-uniformed contact times reported in the literature for long-term toxicity assessment led to incomparable results. Herein, a representative long-term toxicity assessment method was established which unified the contact time of antibiotics and Vibrio fischeri within the bioluminescence increasing period (i.e. 10-100% maximum luminescence) of control samples. The effects of excitation and detoxification of antibiotics such as ß-lactams were discovered. Half maximal inhibitory concentration (IC50) of toxic antibiotics (0.00069-0.061 mmol/L) obtained by this method was 2-3 orders of magnitude lower than acute test, quantifying the underestimated toxicity. As antibiotics exist in natural water as mixtures, an equivalent concentration addition (ECA) model was built to predict mixture toxicity based on physical mechanism rather than mathematical method, which showed great fitting results (R2 = 0.94). Furthermore, interaction among antibiotics was investigated. Antibiotics acting during bacterial breeding period had strong synergistic inhibition (IC50 relative deviation from 0.1 to 0.6) such as macrolides and quinolones. Some antibiotics produced increasing synergistic inhibition during concentration accumulation, such as macrolides. The discharge of antibiotics with severe long-term toxicity and strong synergistic inhibition effect should be seriously restricted.

A prototrophic suppressor of a Vibrio fischeri D-glutamate auxotroph reveals a member of the periplasmic broad-spectrum racemase family (BsrF).

Although bacterial peptidoglycan (PG) is highly conserved, some natural variations in PG biosynthesis and structure have evolved. Understanding the mechanisms and limits of such variation will inform our understanding of antibiotic resistance, innate immunity, and the evolution of bacteria. We have explored the constraints on PG evolution by blocking essential steps in PG biosynthesis in Vibrio fischeri and then selecting mutants with restored prototrophy. Here, we attempted to select prototrophic suppressors of a D-glutamate auxotrophic murI racD mutant. No suppressors were isolated on unsupplemented lysogeny broth salts (LBS), despite plating >1011 cells, nor were any suppressors generated through mutagenesis with ethyl methanesulfonate. A single suppressor was isolated on LBS supplemented with iso-D-gln, although the iso-D-gln subsequently appeared irrelevant. This suppressor has a genomic amplification formed by the creation of a novel junction that fuses proB to a gene encoding a putative broad-spectrum racemase of V. fischeri, bsrF. An engineered bsrF allele lacking the putative secretion signal (ΔSS-bsrF) also suppressed D-glu auxotrophy, resulting in PG that was indistinguishable from the wild type. The ΔSS-bsrF allele similarly suppressed the D-alanine auxotrophy of an alr mutant and restored prototrophy to a murI alr double mutant auxotrophic for both D-ala and D-glu. The ΔSS-bsrF allele increased resistance to D-cycloserine but had no effect on sensitivity to PG-targeting antibiotics penicillin, ampicillin, or vancomycin. Our work helps define constraints on PG evolution and reveals a periplasmic broad-spectrum racemase in V. fischeri that can be co-opted for PG biosynthesis, with concomitant D-cycloserine resistance. IMPORTANCE: D-Amino acids are used and produced by organisms across all domains of life, but often, their origins and roles are not well understood. In bacteria, D-ala and D-glu are structural components of the canonical peptidoglycan cell wall and are generated by dedicated racemases Alr and MurI, respectively. The more recent discovery of additional bacterial racemases is broadening our view and deepening our understanding of D-amino acid metabolism. Here, while exploring alternative PG biosynthetic pathways in Vibrio fischeri, we unexpectedly shed light on an unusual racemase, BsrF. Our results illustrate a novel mechanism for the evolution of antibiotic resistance and provide a new avenue for exploring the roles of non-canonical racemases and D-amino acids in bacteria.

Impact of modeled microgravity stress on innate immunity in a beneficial animal-microbe symbiosis.

The innate immune response is the first line of defense for all animals to not only detect invading microbes and toxins but also sense and interface with the environment. One such environment that can significantly affect innate immunity is spaceflight. In this study, we explored the impact of microgravity stress on key elements of the NFκB innate immune pathway. The symbiosis between the bobtail squid Euprymna scolopes and its beneficial symbiont Vibrio fischeri was used as a model system under a simulated microgravity environment. The expression of genes associated with the NFκB pathway was monitored over time as the symbiosis progressed. Results revealed that although the onset of the symbiosis was the major driver in the differential expression of NFκB signaling, the stress of simulated low-shear microgravity also caused a dysregulation of expression. Several genes were expressed at earlier time points suggesting that elements of the E. scolopes NFκB pathway are stress-inducible, whereas expression of other pathway components was delayed. The results provide new insights into the role of NFκB signaling in the squid-vibrio symbiosis, and how the stress of microgravity negatively impacts the host immune response. Together, these results provide a foundation to develop mitigation strategies to maintain host-microbe homeostasis during spaceflight.

Chlorination of antiviral drug ribavirin: Kinetics, nontargeted identification, and concomitant toxicity evolution.

Residual antiviral drugs in wastewater may increase the risk of generating transformation products (TPs) during wastewater treatment. Therefore, chlorination behavior and toxicity evolution are essential to understand the secondary ecological risk associated with their TPs. Herein, chlorination kinetics, transformation pathways, and secondary risks of ribavirin (RBV), one of the most commonly used broad-spectrum antivirals, were investigated. The pH-dependent second-order rate constants k increased from 0.18 M-1·s-1 (pH 5.8) to 1.53 M-1·s-1 (pH 8.0) due to neutral RBV and ClO- as dominant species. 12 TPs were identified using high-resolution mass spectrometry in a nontargeted approach, of which 6 TPs were reported for the first time, and their chlorination pathways were elucidated. The luminescence inhibition rate of Vibrio fischeri exposed to chlorinated RBV solution was positively correlated with initial free active chlorine, probably due to the accumulation of toxic TPs. Quantitative structure-activity relationship prediction identified 7 TPs with elevated toxicity, concentrating on developmental toxicity and bioconcentration factors, which explained the increased toxicity of chlorinated RBV. Overall, this study highlights the urgent need to minimize the discharge of toxic chlorinated TPs into aquatic environments and contributes to environmental risk control in future pandemics and regions with high consumption of antivirals.

Vibrio fischeri: a model for host-associated biofilm formation.

Multicellular communities of adherent bacteria known as biofilms are often detrimental in the context of a human host, making it important to study their formation and dispersal, especially in animal models. One such model is the symbiosis between the squid Euprymna scolopes and the bacterium Vibrio fischeri. Juvenile squid hatch aposymbiotically and selectively acquire their symbiont from natural seawater containing diverse environmental microbes. Successful pairing is facilitated by ciliary movements that direct bacteria to quiet zones on the surface of the squid's symbiotic light organ where V. fischeri forms a small aggregate or biofilm. Subsequently, the bacteria disperse from that aggregate to enter the organ, ultimately reaching and colonizing deep crypt spaces. Although transient, aggregate formation is critical for optimal colonization and is tightly controlled. In vitro studies have identified a variety of polysaccharides and proteins that comprise the extracellular matrix. Some of the most well-characterized matrix factors include the symbiosis polysaccharide (SYP), cellulose polysaccharide, and LapV adhesin. In this review, we discuss these components, their regulation, and other less understood V. fischeri biofilm contributors. We also highlight what is currently known about dispersal from these aggregates and host cues that may promote it. Finally, we briefly describe discoveries gleaned from the study of other V. fischeri isolates. By unraveling the complexities involved in V. fischeri's control over matrix components, we may begin to understand how the host environment triggers transient biofilm formation and dispersal to promote this unique symbiotic relationship.

Transcriptional pathways across colony biofilm models in the symbiont Vibrio fischeri.

Beneficial microbial symbionts that are horizontally acquired by their animal hosts undergo a lifestyle transition from free-living in the environment to associating with host tissues. In the model symbiosis between the Hawaiian bobtail squid and its microbial symbiont Vibrio fischeri, one mechanism used to make this transition during host colonization is the formation of biofilm-like aggregates in host mucosa. Previous work identified factors that are sufficient to induce V. fischeri biofilm formation, yet much remains unknown regarding the breadth of target genes induced by these factors. Here, we probed two widely used in vitro models of biofilm formation to identify novel regulatory pathways in the squid symbiont V. fischeri ES114. We discovered a shared set of 232 genes that demonstrated similar patterns in expression in both models. These genes comprise multiple exopolysaccharide loci that are upregulated and flagellar motility genes that are downregulated, with a consistent decrease in measured swimming motility. Furthermore, we identified genes regulated downstream of the key sensor kinase RscS that are induced independent of the response regulator SypG. Our data suggest that transcriptional regulator VpsR plays a strong role in expression of at least a subset of these genes. Overall, this study adds to our understanding of the genes involved in V. fischeri biofilm regulation while revealing new regulatory pathways branching from previously characterized signaling networks.IMPORTANCEThe V. fischeri-squid system provides an opportunity to study biofilm development both in the animal host and in culture-based biofilm models that capture key aspects of in vivo signaling. In this work, we report the results of the transcriptomic profiling of two V. fischeri biofilm models followed by phenotypic validation and examination of novel signaling pathway architecture. Remarkable consistency between the models provides a strong basis for future studies using either approach or both. A subset of the factors identified by the approaches were validated in the work, and the body of transcriptomic data provides a number of leads for future studies in culture and during animal colonization.

Sociomicrobiology: Coexistence of conflict and cooperation in the squid light organ.

The Hawaiian bobtail squid's Vibrio fischeri symbionts use quorum sensing for both bioluminescence and to modulate antagonism. New research finds quorum sensing unexpectedly represses V. fischeri's type 6 secretion system, highlighting intricate connections between cooperative and competitive microbial behaviors.

Comparative analysis of the capability of the extended biotic ligand model and machine learning approaches to predict arsenate toxicity.

Assessment of inorganic arsenate (As(V)) is critical for ensuring a sustainable environment because of its adverse effects on humans and ecosystems. This study is the first to attempt to predict As(V) toxicity to the bioluminescent bacterium Aliivibrio fischeri exposed to varying As(V) dosages and environmental factors (pH and phosphate concentration) using six machine learning (ML)-guided models. The predicted toxicity values were compared with those predicted using the extended biotic ligand model (BLM) we previously developed to evaluate the toxic effect of oxyanion (i.e., As(V)). The relationship between the variables (input features) and toxicity (output) was found to play an important role in the prediction accuracy of each ML-guided model. The results indicated that the extended BLM had the highest prediction accuracy, with a root mean square error (RMSE) of 12.997. However, with an RMSE of 14.361, the multilayer perceptron (MLP) model exhibited quasi-accurate prediction, despite having been trained with a relatively small dataset (n = 256). In view of simplicity, an MLP model is compatible with an extended BLM and does not require expert knowledge for the derivation of specific parameters, such as binding fraction and binding constant values. Furthermore, with the development and employment of reliable in-situ sensing techniques, monitoring data are expected to be augmented faster to provide sufficient training data for the improvement of prediction accuracy which may, thus, allow it to outperform the extended BLM after obtaining sufficient data.

Modelled microgravity impacts Vibrio fischeri population structure in a mutualistic association with an animal host.

Perturbations to host-microbe interactions, such as environmental stress, can alter and disrupt homeostasis. In this study, we examined the effects of a stressor, simulated microgravity, on beneficial bacteria behaviours when colonising their host. We studied the bacterium Vibrio fischeri, which establishes a mutualistic association in a symbiosis-specific organ within the bobtail squid, Euprymna scolopes. To elucidate how animal-microbe interactions are affected by the stress of microgravity, squid were inoculated with different bacterial strains exhibiting either a dominant- or sharing-colonisation behaviour in High Aspect Ratio Vessels, which simulate the low-shear environment of microgravity. The colonisation behaviours of the sharing and dominant strains under modelled microgravity conditions were determined by counting light-organ homogenate of squids as well as confocal microscopy to assess the partitioning of different strains within the light organ. The results indicated that although the colonisation behaviours of the strains did not change, the population levels of the sharing strains were at lower relative abundance in single-colonised animals exposed to modelled microgravity compared to unit gravity; in addition, there were shifts in the relative abundance of strains in co-colonised squids. Together these results suggest that the initiation of beneficial interactions between microbes and animals can be altered by environmental stress, such as simulated microgravity.

Application of the Aliivibrio fischeri bacterium bioassay for assessing single and mixture effects of antibiotics and copper.

The Aliivibrio fischeri bioassay was successfully applied in order to evaluate the acute effect of sulfamethoxazole (SMX), ciprofloxacin (CIP), chlortetracycline (CTC) and copper (Cu), alone or in binary, ternary, and overall mixture. The toxicity results are reported in terms of both effective concentrations, which inhibited 50% of the bacterium bioluminescence (EC50%), and in Toxic Units (TUs). The TUs were compared with predicted values obtained using the Concentration Addition model (CA). Finally, the toxicity of water extracts from a soil contaminated by the three antibiotics (7 mg Kg-1 each) in the presence/absence of copper (30 mg Kg-1) was also evaluated. Copper was the most toxic chemical (EC50: 0.78 mg L-1), followed by CTC (EC50: 3.64 mg L-1), CIP (96 mg L-1) and SMX (196 mg L-1). Comparing the TU and CA values of the mixtures, additive effects were generally found. However, a synergic action was recorded in the case of the CIP+Cu co-presence and antagonistic effects in the case of CTC+Cu and the ternary mixture (containing each antibiotic at 0.7 mg L-1), were identified. Soil water extracts did not show any toxicity, demonstrating the buffering ability of the soil to immobilize these chemicals.

NSO-heterocyclic PAHs - Controlled exposure study reveals high acute aquatic toxicity.

Environmental occurrence and hazardous nature of heterocyclic polyaromatic hydrocarbons (heterocyclic PAHs) has the potential to threaten the health of aquatic ecosystems. Here, we investigate the acute toxicity of heterocyclic PAHs (log KOW 3.7-6.9) to aquatic organisms: marine bacteria (Aliivibrio fischeri), freshwater green algae (Raphidocelis subcapitata), and water fleas (Daphnia magna) using passive dosing to maintain stable exposure. The membrane-water partition coefficient (KMW) of the heterocycles was measured to elucidate its relationship with toxicity. Our findings show that the tested heterocycles had little inhibitory effect on A. fischeri, while most compounds were highly toxic to R. subcapitata and D. magna. Toxicity generally increased with increasing KMW values, and nonpolar narcosis was identified as the most likely mode of toxic action of the heterocycles. Comparison of standard protocols with passive dosing emphasizes the importance of maintaining a constant concentration during toxicity testing, as very high losses occurred in standard tests and passive dosing experiments revealed higher toxicities. These results indicate a potentially high risk to aquatic life and call for more in-depth investigation of the (eco)toxic effects of NSO-PAHs.

Spring water quality monitoring using multiple bioindicators from multiple collection sites.

The aim of this study was to examine the water quality of the Extrema River spring in a Brazilian Cerrado area. Three collection sites (P1 - P3) were sampled in the dry and rainy seasons, which are close to industries from different sectors. In the physicochemical analysis, a decrease in dissolved oxygen levels (<5 mg/L) and pH (< 6) at P3 was detected. An increase in heterotrophic bacteria count was recorded at all sites (> 500 colonies/ml). In ecotoxicological analyses, P2 and P3 exhibited toxicity using Vibrio fischeri (> 20%). In evaluating toxicity, the reduction in seed germination was significant utilizing Lactuca sativa at all locations and with Allium cepa only at P2; rootlet length was decreased at P3 on L. sativa and at all sites with A. cepa. In contrast, loss of membrane integrity and mitochondrial function of meristems was adversely affected at all locations using both L. sativa and A. cepa assays. Principal components analysis (PCA) approach indicated that seasonality apparently did not markedly interfere with the obtained data, but it is important to include more collection locations to be evaluated with multiple bioindicators in the spring region. Our data indicate the urgent need for more rigorous programs to monitor the discharge of effluents into water springs.

A Benchmark Study of Graph Models for Molecular Acute Toxicity Prediction.

With the wide usage of organic compounds, the assessment of their acute toxicity has drawn great attention to reduce animal testing and human labor. The development of graph models provides new opportunities for acute toxicity prediction. In this study, five graph models (message-passing neural network, graph convolution network, graph attention network, path-augmented graph transformer network, and Attentive FP) were applied on four toxicity tasks (fish, Daphnia magna, Tetrahymena pyriformis, and Vibrio fischeri). With the lowest prediction error, Attentive FP was reported to have the best performance in all four tasks. Moreover, the attention weights of the Attentive FP model helped to construct atomic heatmaps and provide good explainability.

Chemical and ecotoxicological indicators for evaluating the treatment of coal mining wastes.

This paper consists of the evaluation in regards to the ecotoxicological effectiveness of a treatment applied to a coal mining waste. The treatment consisted of separating the particles based on gravimetric concentration in spirals, generating three fractions: heavy, intermediate, and light, with high, moderate, and low pyrite content, respectively. The intermediate fraction represents the larger disposal volume of the waste on soils. To evaluate the effectiveness of the treatment, metal determination and bioassays Eisenia andrei, Folsomia candida, Lactuca sativa, Daphnia similis, and Raphidocelis subcapitata were applied to the intermediary fraction. To evaluate the toxicity to aquatic organisms, elutriates were generated from the unprocessed waste and the intermediate fraction. The intermediate fraction showed a decrease of metal concentrations compared to the untreated waste. Metal concentrations in the intermediate fraction were below the Brazilian thresholds for soil quality. Avoidance bioassay with E. andrei and germination tests of L. sativa showed no significant effects. The bioassay with F. candida indicated a significant reduction in reproduction at the highest doses used (24% and 50%). Bioassays with D. similis and R. subcapitata revealed a reduction in toxicity of the intermediate fraction compared to the untreated waste. However, the toxicity levels of the intermediate fraction to aquatic organisms still require attention, especially in regards to pH that played a crucial role in the toxicity. Finally, the results suggest that the treatment performed on the coal waste was efficient, even though significant toxicity have still been detected in the treated waste and additional steps are still required for adequate final disposal.

Quantification of the capacity of vibrio fischeri to establish symbiosis with Euprymna scolopes.

Most animals establish long-term symbiotic associations with bacteria that are critical for normal host physiology. The symbiosis that forms between the Hawaiian squid Euprymna scolopes and the bioluminescent bacterium Vibrio fischeri serves as an important model system for investigating the molecular mechanisms that promote animal-bacterial symbioses. E. scolopes hatch from their eggs uncolonized, which has led to the development of squid-colonization assays that are based on introducing culture-grown V. fischeri cells to freshly hatched juvenile squid. Recent studies have revealed that strains often exhibit large differences in how they establish symbiosis. Therefore, we sought to develop a simplified and reproducible protocol that permits researchers to determine appropriate inoculum levels and provides a platform to standardize the assay across different laboratories. In our protocol, we adapt a method commonly used for evaluating the infectivity of pathogens to quantify the symbiotic capacity of V. fischeri strains. The resulting metric, the symbiotic dose-50 (SD50), estimates the inoculum level that is necessary for a specific V. fischeri strain to establish a light-emitting symbiosis. Relative to other protocols, our method requires 2-5-fold fewer animals. Furthermore, the power analysis presented here suggests that the protocol can detect up to a 3-fold change in the SD50 between different strains.

Different isoforms of parabens into marine environment: Biological effects on the bacterium Aliivibrio fischeri and the marine mussel Mytilus galloprovincialis.

Different isoforms of alkyl esters of p-Hydroxybenzoic acid, also known as parabens, are of great concern due to their widespread presence into the aquatic environment, their high concentrations in wastewater discharges, as well as their ability to induce adverse effects on aquatic organisms. Considering the imperative need for assessing their fate and risk to aquatic environment, the present study investigated the biological effects of two isoforms of parabens, methyl- (MeP) and propyl- (PrP), on the bacterium Aliivibrio fischeri (using the Bioluminescence Inhibition/Microtox® bioassay) and the mussel Mytilus galloprovincialis (in terms of mussel mortality, cellular, oxidative and genotoxic stress indices). The assessment of MeP and PrP behavior into aquatic media (artificial sea water/ASW and 2 % NaCl), primarily performed by UHPLC-UV-MS analysis, showed only a slight hydrolysis of PrP to 4-Hydrobenzoic acid (4-HBA). Furthermore, exposure of both species to different concentrations of each paraben revealed differences among their toxic potential, as well as their ability to cause cellular, oxidative and genotoxic effects on hemocytes of challenged mussels. Interestingly, the Microtox® bioassay showed that PrP mediated toxicity in A. fischeri were more pronounced than MeP, as revealed by the estimated toxic endpoints (in terms of concentration that promote 50 % of bioluminescence inhibition, EC50). Similarly, in challenged mussels, a significant disturbance of mussel hemocytes' lysosomal membrane integrity, as well as enhanced levels of superoxides, nitric oxides, lipid peroxidation byproducts, and micronuclei formation were observed. These findings are of great interest, since MeP and PrP differential toxic potential, as well their ability to induce pre-pathological alterations in marine species, like mussels, give new evidence for their risk to aquatic biota.

Photolysis and photo-enhanced toxicity of three novel designed neonicotinoids: Impact of novel modifying groups.

Three novel neonicotinoids (cycloxaprid, flupyradifurone and sulfoxaflor) were designed to reduce the biotoxicity for non-target organisms. These neonicotinoids were photolyzed under light radiation, but it was unclear for the photo-enhanced toxicity and influences of the novel modifying group of the three neonicotinoids. The photolysis and photo-enhanced toxicity experiments were performed for the three neonicotinoids, coupled with quantum chemistry calculation, the mechanisms of photolysis, photo-enhanced toxicity and the influences of novel modifying groups were analyzed. The results showed the photolysis pathways were enriched as compared with previous neonicotinoids due to the composition of modifying groups, singlet oxygen and hydroxyl participated the photolysis of cycloxaprid and flupyradifurone. All tested neonicotinoids exhibited photo-enhanced toxicity to Vibrio fischeri. Due to the difference of photolysis mechanism and toxicity to V. fischeri, the photo-enhanced toxicity curves showed diverse variation when histidine, tert-butanol or dissolved organic matters was in presence of the test solutions. The impact of novel modifying groups over photolysis and photo-enhanced toxicity were analyzed based on the comparison with previous neonicotinoids, theoretically predicted UV-Vis spectra and photo-physical/chemical property descriptors. The data showed the composition of novel modifying group increased the light absorption and photo-chemical activities for the three neonicotinoids.

Large Dataset-Based Regression Model of Chemical Toxicity to Vibrio fischeri.

For the first time, a global regression quantitative structure-toxicity/activity relationship (QSTR/QSAR) model was developed for the toxicity of a large data set including 1236 chemicals towards Vibrio fischeri, by using random forest (RF) regression algorithm. The optimal RF model with RF parameters of mtry = 3, ntree = 150 and nodesize = 5 was based on 13 molecular descriptors. It can achieve accurate prediction for the toxicity of 99.1% of 1236 chemicals, and yield coefficients of determination R2 of 0.893 for 930 log(Mw/IBC50) in the training set, 0.723 for 306 log(Mw/IBC50) in the test se, and 0.865 for 1236 toxicity log(Mw/IBC50) in the total set. The optimal RF global model proposed in this work is comparable to other published local QSTR models on small datasets of the toxicity to Vibrio fischeri.

Genetic Analysis Reveals a Requirement for the Hybrid Sensor Kinase RscS in para-Aminobenzoic Acid/Calcium-Induced Biofilm Formation by Vibrio fischeri.

The marine bacterium Vibrio fischeri initiates symbiotic colonization of its squid host, Euprymna scolopes, by forming and dispersing from a biofilm dependent on the symbiosis polysaccharide locus (syp). Historically, genetic manipulation of V. fischeri was needed to visualize syp-dependent biofilm formation in vitro, but recently, we discovered that the combination of two small molecules, para-aminobenzoic acid (pABA) and calcium, was sufficient to induce wild-type strain ES114 to form biofilms. Here, we determined that these syp-dependent biofilms were reliant on the positive syp regulator RscS, since the loss of this sensor kinase abrogated biofilm formation and syp transcription. These results were of particular note because loss of RscS, a key colonization factor, exerts little to no effect on biofilm formation under other genetic and medium conditions. The biofilm defect could be complemented by wild-type RscS and by an RscS chimera that contains the N-terminal domains of RscS fused to the C-terminal HPT domain of SypF, the downstream sensor kinase. It could not be complemented by derivatives that lacked the periplasmic sensory domain or contained a mutation in the conserved site of phosphorylation, H412, suggesting that these cues promote signaling through RscS. Lastly, pABA and/or calcium was able to induce biofilm formation when rscS was introduced into a heterologous system. Taken together, these data suggest that RscS is responsible for recognizing pABA and calcium, or downstream consequences of those cues, to induce biofilm formation. This study thus provides insight into signals and regulators that promote biofilm formation by V. fischeri. IMPORTANCE Bacterial biofilms are common in a variety of environments. Infectious biofilms formed in the human body are notoriously hard to treat due to a biofilm's intrinsic resistance to antibiotics. Bacteria must integrate signals from the environment to build and sustain a biofilm and often use sensor kinases that sense an external signal, which triggers a signaling cascade to elicit a response. However, identifying the signals that kinases sense remains a challenging area of investigation. Here, we determine that a hybrid sensor kinase, RscS, is crucial for Vibrio fischeri to recognize para-aminobenzoic acid and calcium as cues to induce biofilm formation. This study thus advances our understanding of the signal transduction pathways leading to biofilm formation.

Impact of global change on environmental hazards of different clays: A case study on Aliivibrio fischeri.

The effects of global change in marine ecosystems are expected to lower pH from the current 8.1-7.5-7.0, which will have significant impacts on marine species. The purpose of this study is to investigate whether the ecotoxicity of ten different natural clays change significantly in response to the acidification process and what factors are associated with the observed changes. In this study, the ecotoxicological response of a bacterium (Aliivibrio fischeri) was tested under current (pH= 8.1) and acidified (pH 7.5 and 7.0) conditions. The ecotoxicity detected in the solid phase test (SPT protocol) and in the contact water was affected by the pH, which increased the ecotoxicity from 2/10 clays (pH 8.10) to 7/10 clays (pH 7.00), also shifting the detected effects from low to high toxicity values. The analyses performed on the natural clays studied show that pH can affect the release of metals, metalloids and rare earths from the clays into the contact water phase, affecting the toxicity observed. This phenomenon depends on the type of clay and is closely related to its mineralogical composition. As consequence, in a globally changing scenario, ecotoxicity, even of natural materials such as clay, cannot be considered stable, but must be accurately revaluated depending on the mineralogical and chemical composition of the clay. Moreover, the mineralogical composition of clays showed different efficiency in absorbing bacteria on the surface of clay particles. It was found that live bacterial cells were absorbed on the clay surface in numbers that were dependent on both clay types and pH levels.