Three Microbial Musketeers of the Seas: Shewanella baltica, Aliivibrio fischeri and Vibrio harveyi, and Their Adaptation to Different Salinity Probed by a Proteomic Approach

Osmotic changes are common challenges for marine microorganisms. Bacteria have developed numerous ways of dealing with this stress, including reprogramming of global cellular processes. However, specific molecular adaptation mechanisms to osmotic stress have mainly been investigated in terrestrial model bacteria. In this work, we aimed to elucidate the basis of adjustment to prolonged salinity challenges at the proteome level in marine bacteria. The objects of our studies were three representatives of bacteria inhabiting various marine environments, Shewanella baltica, Vibrio harveyi and Aliivibrio fischeri. The proteomic studies were performed with bacteria cultivated in increased and decreased salinity, followed by proteolytic digestion of samples which were then subjected to liquid chromatography with tandem mass spectrometry analysis. We show that bacteria adjust at all levels of their biological processes, from DNA topology through gene expression regulation and proteasome assembly, to transport and cellular metabolism. The finding that many similar adaptation strategies were observed for both low- and high-salinity conditions is particularly striking. The results show that adaptation to salinity challenge involves the accumulation of DNA-binding proteins and increased polyamine uptake. We hypothesize that their function is to coat and protect the nucleoid to counteract adverse changes in DNA topology due to ionic shifts.

Environmental concerns about the massive use of disinfectants during COVID-19 pandemic: an overview on aquatic toxicity

Many public health measures to mitigate the spread of SARS-CoV-2 were adopted worldwide, and particularly to the environmental measure of regular cleaning and disinfection of surfaces, the increased use of disinfectant products raises environmental concerns. Quaternary ammonium compounds (QACs), povidone-iodine (PVP-I), chloroxylenol (PCMX) and chlorhexidine (CHX) are the active ingredients of most disinfectant products due to their effectiveness against various microbiological agents. Although presenting antimicrobial efficacy, these biocides have been associated with impacts on aquatic life. For instance, QACs can induce toxicity to Aliivibrio fischeri and fish (different species). Gill and liver damages are verified in Cyprinus carpio after exposure to PVP-I. CHX induces toxic effects on algae, crustaceans, and fish embryos. PCMX can induce genotoxicity to rainbow trout and malformations on zebrafish embryos, as well as it can reduce the reproduction rate of Caenorhabditis elegans. Thus, the potential to cause negative consequences on human and environmental health has resulted in activities from the U.S. and European agencies to favor the use of safer and greener disinfectant products during the COVID-19 pandemic. This review article summarizes the main findings on the impacts of disinfectants (the most used) on aquatic life. This information may help prioritize disinfectants with lower impacts on the aquatic environment for daily use, and especially for high-frequency use as verified in the COVID-19 pandemic. Furthermore, this review may help identify knowledge gaps on the aquatic hazard of disinfectants, which may drive future studies on this matter and, lastly, contribute to the development of sustainable products.

Mixture toxicity of six pharmaceuticals towards Aliivibrio fischeri, Daphnia magna, and Lemna minor

As the knowledge on the joint effects of pharmaceuticals towards different non-target organisms is still limited, the aim of our study was to evaluate the toxicity of mixtures of pharmaceuticals, as well as their baseline toxicity towards three selected organisms, namely the bioluminescent bacteria Aliivibrio fischeri, the crustacean Daphnia magna, and the duckweed Lemna minor. Different mixtures composed of three up to five pharmaceuticals having the same or different mechanisms of action in terms of their therapeutic activity (non-steroidal anti-inflammatory drugs, opioid analgesic, antibacterial and anti-epileptic drugs) were investigated. The observed EC50s were compared with those predicted using the concentration addition (CA) and independent action (IA) models. In general, the EC50 values for mixtures predicted with the CA model were lower than those obtained with the IA model, although, in some cases, test predictions of these two models were almost identical. Most of the experimentally determined EC50 values for the specific mixtures were slightly higher than those predicted with the CA model; hence, a less than additive effect was noted. Based on the obtained results, it might be concluded that the CA model assumes the worst-case scenario and gives overall closer predictions; therefore, it should be recommended also for modeling the mixture toxicity of pharmaceuticals with different modes of action.

Locomotor behavior of Neocaridina palmata: a study with leachates from UV-weathered microplastics

Weathering of plastics leads to the formation of increasingly smaller particles with the release of chemical compounds. The latter occurs with currently unknown environmental impacts. Leachate-induced effects of weathered microplastics (MPs) are therefore of increasing concern. To investigate the toxicity of the chemical mixtures from such plastics, we exposed the freshwater shrimp Neocaridina palmata to enriched leachates from unweathered and artificially weathered (UV-A/B light) MPs (≤1 mm) from recycled low-density polyethylene (LDPE-R) pellets and from a biodegradable, not fully bio-based starch blend (SB) foil. We analyzed the individual locomotor activity (moved distance and frozen events) on day 1, 3, 7 and 14 of exposure to five leachate concentrations equivalent to 0.40–15.6 g MPs L−1, representing the upper scale of MPs that have been found in the environment. The median moved distance did not change as a function of concentration, except for the unweathered SB treatment on day 14 that indicated hyperactivity with increasing concentrations. Significant impacts were solely detected for few concentrations and exposure days. Generally, no consistent trend was observed across the experiments. We further assessed the baseline toxicity of the samples in the Microtox assay and detected high bioluminescence inhibitions of the bacterium Aliivibrio fischeri. This study demonstrates that neither the recycled nor the biodegradable material are without impacts on test parameters and therefore cannot be seen as safe alternative for conventional plastics regarding the toxicity. However, the observed in vitro toxicity did not result in substantial effects on the behavior of shrimps. Overall, we assume that the two endpoints examined in the atyid shrimp N. palmata were not sensitive to chemicals leaching from plastics or that effects on the in vivo level affect other toxic endpoints which were not considered in this study.

Paenibacillus polymyxa A26 and Its Surfactant-Deficient Mutant Degradation of Polycyclic Aromatic Hydrocarbons

We compared the ability of two bacterial strains, Paenibacillus polymyxa A26 and P. polymyxa A26Sfp, for biodegradation of naphthalene (NAP). The studies were performed under simulated laboratory conditions, in liquid medium and soil with different carbon sources, pH and salt contents. Changes in the luminescence inhibition of Aliivibrio fischeri, as an indicator of the baseline toxicity, were observed in degradation mixtures during 7 days of incubation. While both strains expressed the best growth and NAP degradation ability in the minimal salt medium containing sucrose and 5% NaCl at pH 7 and 8, the mutant strain remained effective even under extreme conditions. A26Sfp was found to be an efficient and potentially industrially important polycyclic aromatic hydrocarbon degradation strain. Its extracellular polysaccharide production is 30%, and glucan production is twice that of the wild type A 26. The surface tension reduction ability was ascertained as 25–30% increased emulsification ability.

The Influence of Herbicides to Marine Organisms Aliivibrio fischeri and Artemia salina

The aim of this work was to determine the toxic effect of the most used herbicides on marine organisms, the bacterium Aliivibrio fischeri, and the crustacean Artemia salina. The effect of these substances was evaluated using a luminescent bacterial test and an ecotoxicity test. The results showed that half maximal inhibitory concentration for A. fischeri is as follows: 15minIC50 (Roundup® Classic Pro) = 236 µg.l−1, 15minIC50 (Kaput® Premium) = 2475 µg.l−1, 15minIC50 (Banvel® 480 S) = 2637 µg.l−1, 15minIC50 (Lontrel 300) = 7596 µg.l−1, 15minIC50 (Finalsan®) = 64 µg.l−1, 15minIC50 (glyphosate) = 7934 µg.l−1, 15minIC50 (dicamba) = 15,937 µg.l−1, 15minIC50 (clopyralid) = 10,417 µg.l−1, 15minIC50 (nonanoic acid) = 16,040 µg.l−1. Median lethal concentrations for A. salina were determined as follows: LC50 (Roundup® Classic Pro) = 18 µg.l−1, LC50 (Kaput® Premium) = 19 µg.l−1, LC50 (Banvel® 480 S) = 2519 µg.l−1, LC50 (Lontrel 300) = 1796 µg.l−1, LC50 (Finalsan®) = 100 µg.l−1, LC50 (glyphosate) = 811 µg.l−1, LC50 (dicamba) = 3705 µg.l−1, LC50 (clopyralid) = 2800 µg.l−1, LC50 (nonanoic acid) = 7493 µg.l−1. These findings indicate the need to monitor the herbicides used for all environmental compartments.

LitR directly upregulates autoinducer synthesis and luminescence in Aliivibrio logei

LitR is a master-regulator of transcription in the ainS/R and luxS/PQ quorum sensing (QS) systems of bacteria from Vibrio and Aliivibrio genera. Here, we for the first time directly investigated the influence of LitR on gene expression in the luxI/R QS system of psychrophilic bacteria Aliivibrio logei. Investigated promoters were fused with Photorhabdus luminescens luxCDABE reporter genes cassette in a heterological system of Escherichia coli cells, litR A. logei was introduced into the cells under control of Plac promoter. LitR has been shown to upregulate genes of autoinducer synthase (luxI), luciferase and reductase (luxCDABE), and this effect doesn’t depend on presence of luxR gene. To a much lesser degree, LitR induces luxR1, but not the luxR2 — the main luxI/R regulator. Enhanced litR expression leads to an increase in a LuxI-autoinducer synthesis and a subsequent LuxR-mediated activation of the luxI/R QS system. Effect of LitR on luxI transcription depends on lux-box sequence in luxI promoter even in absence of luxR (lux-box is binding site of LuxR). The last finding indicates a direct interaction of LitR with the promoter in the lux-box region. Investigation of the effect of LitR A. logei on luxI/R QS systems of mesophilic Aliivibrio fischeri and psychrophilic Aliivibrio salmonicida showed direct luxR-independent upregulation of luxI and luxCDABE genes. To a lesser degree, it induces luxR A. fischeri and luxR1 A. salmonicida. Therefore, we assume that the main role of LitR in cross-interaction of these three QS systems is stimulating the expression of luxI.

A multi-bioassay integrated approach to assess antifouling potential of extracts from the Mediterranean sponge Ircinia oros

The phylum Porifera and their symbionts produce a wide variety of bioactive compounds, playing a central role in their ecology and evolution. In this study, four different extracts (obtained by non-polar and semi-polar extraction methodologies) of the Mediterranean sponge Ircinia oros were tested through a multi-bioassay integrated approach to assess their antifouling potential. Tests were performed using three common species, associated with three different endpoints: the marine bacterium Aliivibrio fischeri (inhibition of bioluminescence), the marine diatom Phaeodactylum tricornutum (inhibition of growth), and different development stages of the brackish water serpulid Ficopomatus enigmaticus (gametes: sperm motion, vitality inhibition and cellular damage; larvae: development; adults: AChE (acetylcholinesterase)-inhibitory activity). The effects of extracts were species specific and did not vary among different extraction methodologies. In particular, no significant reduction of bioluminescence of A. fischeri was observed for all tested samples. By contrast, extracts inhibited P. tricornutum growth and had toxic effects on different F. enigmaticus’ developmental stages. Our results suggest that the proposed test battery can be considered a suitable tool as bioactivity screening of marine natural products.