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The aquaculture industry faces significant challenges due to bacterial infections caused by Edwardsiella tarda , Photobacterium damselae , and Vibrio harveyi . The extensive use of traditional antibiotics, has resulted in widespread antibiotic resistance. This study aimed to investigate the antibacterial potential of the brown seaweed Eisenia bicyclis , particularly its synergistic effects with antibiotics against these fish pathogenic bacteria. E. bicyclis were processed to obtain methanolic extracts and fractionated using different polar solvents. The antibacterial activities of these extracts and fractions were assessed through disc diffusion and minimum inhibitory concentration (MIC) assays. The study further evaluated the antibiotic susceptibility of the bacterial strains and the synergistic effects of the extracts combined with erythromycin and oxyteteracycline using the fractional inhibitory concentration index. Results showed that the ethyl acetate (EtOAc) fraction of E. bicyclis methanolic extract exhibited the highest antibacterial activity. The combination of the EtOAc fraction with erythromycin significantly enhanced its antibacterial efficacy against the tested strains. This synergistic effect was indicated by a notable reduction in MIC values, demonstrating the potential of E. bicyclis to enhance the effectiveness of traditional antibiotics. The findings suggest that E. bicyclis extracts, particularly the EtOAc fraction, could serve as a potent natural resource to counteract antibiotic resistance in aquaculture.

Understanding the efficacy of antimicrobials against pathogens from clinical samples is critical for their responsible use. The manuscript presents in vitro efficacy and antimicrobial resistance (AMR) genes in seven species of fish pathogens from the disease outbreaks of Indian aquaculture against oxytetracycline, florfenicol, oxolinic acid, and enrofloxacin. In vitro efficacy was evaluated by minimum inhibitory concentration and minimum bactericidal concentration. The gene-specific PCR screened AMR genes against quinolones (qnrA, qnrB, and qnrS) and tetracyclines (tetM, tetS, tetA, tetC, tetB, tetD, tetE, tetH, tetJ, tetG, and tetY). The results showed that Aeromonas veronii (45%) showed the maximum resistance phenotype, followed by Streptococcus agalactiae (40%), Photobacterium damselae (15%), Vibrio parahaemolyticus (10%), and Vibrio vulnificus (5%). There was no resistance among Vibrio harveyi and Vibrio alginolyticus against the tested antimicrobials. The positive association between tetA, tetB, tetC, tetM, or a combination of these genes to oxytetracycline resistance and qnrS to quinolone resistance indicated their potential in surveillance studies. The prevalence of resistance phenotypes (16.43%) and evaluated AMR genes (2.65%) against aquaculture antimicrobials was low. The resistance phenotype pattern abundance was 0.143. All the isolates showed susceptibility to florfenicol. The results help with the appropriate drug selection against each species in aquaculture practices., (© The Author(s) 2024. Published by Oxford University Press on behalf of Applied Microbiology International.)

Nitrite exposure has become a significant concern in the aquaculture industry, posing a severe threat to aquatic animals such as shrimp. While studies have reported the adverse effects of nitrite on shrimp growth, the part played by the gut microbiota in shrimp mortality resulting from nitrite exposure is poorly understood. Here, the effects of nitrite on shrimp gut bacterial community were investigated using 16S rRNA amplicon sequencing, bacterial isolation, genomic analysis, and infection experiments. Compared to the control_healthy group, changes in the bacterial composition of the nitrite_dead group were associated with reduced abundance of specific beneficial bacteria and increased abundance of certain pathogenic bacteria. Notably, members of the Photobacterium genus were found to be significantly enriched in the nitrite_dead group. Genomic analysis of a representative Photobacterium strain (LvS-8n3) revealed a variety of genes encoding bacterial toxins, including hemolysin, adhesin, and phospholipase. Furthermore, it was also found that LvS-8n3 exhibits strong pathogenicity, probably due to its high production of pathogenic factors and the ability to utilize nitrite for proliferation. Therefore, the proliferation of pathogenic Photobacterium species appears pivotal for driving shrimp mortality caused by nitrite exposure. These findings provide novel insights into the disease mechanism in shrimp under conditions of environmental change., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper, (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Luminous marine bacteria are traditionally used as a bioassay due to the convenience and high rate of registering the intensity of their physiological function - luminescence. This study aimed to develop the application of Photobacterium phosphoreum in traditional and novel fields - toxicity monitoring and biotechnology. We demonstrated (1) effects of selenite ions on bioluminescence, and (2) biotransformation of selenite to selenium(0) in the form of nanoparticles. The effects of selenite (SeO 3 2- ) on the intensity of bacterial bioluminescence were studied, and its dependencies on exposure time and concentration of Na 2 SeO 3 were analyzed. Bioluminescence activation and inhibition were revealed; dose-effect dependencies corresponded to the hormesis model. The toxicity of SeO 3 2- was characterized by an effective concentration of 10 -3 M. Effects of SeO 3 2- on reactive oxygen species (ROS) in bacterial suspensions were studied. High positive correlations were found between the bioluminescence intensity and ROS content, which indicates the decisive role of ROS and associated redox processes in the bioeffects of selenite ions. Scanning and transmission electron microscopy revealed the presence of nano-structures in the bacteria exposed to selenite. The energy dispersion spectrum detected a high content of selenium in the nanoparticles. The particle size distribution depended on Na 2 SeO 3 concentration; maxima of the distribution varied within 45-55 nm., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Nadezhda S. Kudryasheva reports financial support was provided by Russian Science Foundation and Krasnoyarsk Territory and Krasnoyarsk Regional Fund of Science. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Organophosphate esters (OPEs) have attracted extensive attention due to their toxic effects on human health and biological systems as plasticizers and flame retardants. This study focused on exploring a green approach to get toxicity mechanisms that used less solvents or organisms. The toxicity values of selected organophosphate esters including tri (2-chloroethyl) phosphate (TCEP), tri (1, 3-dichloro-2-propyl) phosphate (TDCP), tripropyl phosphate (TPrP), tri-n-butyl phosphate (TnBP), and tritolyl Phosphate (TCrP) to Photobacterium phosphoreum were determined. Their EC 50 range was between 7.27 × 10 -8 -5.12 × 10 -6  mol/L. Based on molecular dynamics simulation the data of binding affinity between OPEs and n-octanol / phospholipid bilayer were calculated respectively. Coupling with binding affinity energies and toxicity values, the mode of action (MOA) of OPEs including types of reactive or anesthetic mechanism could be fast deduced. The proposed hypothesis of n-octanol (C 8 H 18 O) as a virtual biofilm to replace phospholipid bilayer was verified. The alternative green approach could simplify toxicity assay and realize fast predicting MOA for different chemicals and model organisms., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Metal-organic framework (MOF) modified with iron oxide, Fe 3 O 4 -MOF, is a perspective drug delivery agent, enabling magnetic control and production of active hydroxyl radicals, •OH, via the Fenton reaction. This paper studies cytotoxic and radical activities of Fe-containing nanoparticles (NPs): Fe 3 O 4 -MOF and its components - bare Fe 3 O 4 and MOF (MIL-88B). Luminous marine bacteria Photobacteriumphosphoreum were used as a model cellular system to monitor bioeffects of the NPs. Neither the NPs of Fe 3 O 4 -MOF nor MOF showed cytotoxic effects in a wide range of concentrations (<10 mg/L); while Fe 3 O 4 was toxic at >3·10 -3  mg/L. The NPs of Fe 3 O 4 did not affect the bacterial bioluminescence enzymatic system; their toxic effect was attributed to cellular membrane processes. The integral content of reactive oxygen species (ROS) was determined using a chemiluminescence luminol assay. Bacteria mitigated excess of ROS in water suspensions of Fe 3 O 4 -MOF and MOF, maintaining bioluminescence intensity closer to the control; this resulted in low toxicity of these NPs. We estimated the activity of •OH radicals in the NPs samples with physical and chemical methods - spin capture technology (using electron paramagnetic resonance spectroscopy) and methylene blue degradation. Physico-chemical interpretation of cellular responses is provided in terms of iron content, iron ions release and •OH radical production., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Zearalenone (ZEN), a nonsteroidal estrogenic mycotoxin, causes enormous economic losses in the food and feed industries. Simple, rapid, low-cost, and quantitative analysis of ZEN is particularly urgent in the fields of food safety and animal husbandry. Using the bioluminescent bacterium Photobacterium phosphoreum T3, we propose a bioluminescence inhibition assay to evaluate ZEN levels quickly. The limit of detection (LOD), limit of quantification (LOQ), and quantitative working range of this bioluminescence inhibition assay were 0.1 µg/mL, 5 µg/mL, and 5-100 µg/mL, respectively. The concentration-response curve of the bioluminescence inhibition rate and ZEN concentration was plotted within the range 5 to 100 μg/mL, as follows: y = 0.0069x 2 - 0.0190x + 7.9907 (R 2 = 0.9943, y is luminescence inhibition rate, x is ZEN concentration). First, we used the bioluminescence inhibition assay to detect the remaining ZEN in samples treated with purified lactonohydrolase ZHD101. The bioluminescence inhibition assay results showed a strong correlation with the HPLC analysis. Furthermore, we successfully evaluated the overall toxicity of samples treated with purified peroxidase Prx and H 2 O 2 using the P. phosphoreum T3 bioluminescence inhibition assay. The results indicate that the degradation products of ZEN created by purified peroxidase Prx and H 2 O 2 showed little toxicity to P. phosphoreum T3. In this study, a simple, rapid, and low-cost assay method of zearalenone by bioluminescent P. phosphoreum T3 was developed. The bioluminescence inhibition assay could be used to estimate the efficiency of enzymatic degradation of ZEN.

Human and environmental ecosystem beings are exposed to multicomponent compound mixtures but the toxicity nature of compound mixtures is not alike to the individual chemicals. This work introduces four models for the prediction of the negative logarithm of median effective concentration (pEC 50 ) of individual chemicals to marine bacteria Photobacterium Phosphoreum (P. Phosphoreum) and algal test species Selenastrum Capricornutum (S. Capricornutum) as well as their mixtures to P. Phosphoreum, and S. Capricornutum. These models provide the simplest approaches for the forecast of pEC 50 of some classes of organic compounds from their interpretable structural parameters. Due to the lack of adequate toxicity data for chemical mixtures, the largest available experimental data of individual chemicals (55 data) and their mixtures (99 data) are used to derive the new correlations. The models of individual chemicals are based on two simple structural parameters but chemical mixture models require further interaction terms. The new model's results are compared with the outputs of the best accessible quantitative structure-activity relationships (QSARs) models. Various statistical parameters are done on the new and comparative complex QSAR models, which confirm the higher reliability and simplicity of the new correlations., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

This study investigated the acute inflammatory response induced by subcutaneous injection of carrageenin (1%) or phosphate-buffered saline (control) in gilthead seabream (Sparus aurata). Skin mucus, serum, head kidney (HK) and liver were sampled at 1.5, 3 and 6 hr post-injection (p.i.) to determine the immune and antioxidant status of this fish species. The skin mucus of the carrageenin group showed increased superoxide dismutase and peroxidase activities, lysozyme abundance, bactericidal activity against Vibrio anguillarum and Photobacterium damselae, and total immunoglobulins compared with those of the control group. However, the carrageenin-injected fish sampled at 6 hr p.i. showed decreased protease activity in the skin mucus and peroxidase activity in the HK leucocytes compared with the control. Moreover, the carrageenin injection had no effects on the systemic immune system, but it reduced the liver catalase activities at both 3 and 6 hr in the carrageenin group relative to those in the control group. The expression levels of several proinflammatory and cell marker genes in the HK and liver were also determined. In the HK, the expression levels of interleukin-1β and prostaglandin D synthase 1 were upregulated at 1.5 and 3 hr, respectively, in the carrageenin group compared with those in the control group. Contrarily, the expression of the NADPH oxidase subunit phox40 (an acidophilic granulocyte marker) in the carrageenin group at 6 hr was downregulated compared with that in the control group. These results suggested that subcutaneous injection of κ/λ-carrageenin in gilthead seabream triggered an acute skin inflammation characterized by the rapid recruitment of acidophilic granulocytes and the release of humoral mediators into the skin mucus., (© 2021 John Wiley & Sons Ltd.)

Fresh meat is commonly packaged in modified atmosphere to decelerate spoilage processes. The applied gas mixture affects the growth of spoilage organisms and selectively shapes the spoilage community. In this study, we investigated the impact of O 2 and CO 2 on the growth of Photobacterium (P.) phosphoreum and P. carnosum strains in situ on chicken meat by packaging under different modified atmospheres (air, 70% O 2 /30% CO 2 , 70% N 2 /30% CO 2 , 100% N 2 ). Combination of 70% O 2 and 30% CO 2 resulted in significant growth reduction of the analyzed strains, suggesting inhibitory effects of both gases in combination. In contrast, 30% CO 2 alone had only a minor effect and photobacteria are supposed to have a growth advantage over other meat spoilers in this atmosphere. Additionally, single growth of the strains in the different atmospheres was compared when challenged with the presence of Pseudomonas (Ps.) fragi or Brochothrix (B.) thermosphacta as prominent co-contaminants in different ratios (10:1, 1:1, 1:10). Presence of co-contaminants resulted in increased cell numbers of P. carnosum TMW2.2149 but reduced or unchanged cell numbers of P. phosphoreum TMW2.2103 in most packaging atmospheres. The initial ratio of photobacteria and co-contaminants defined the relative abundance during storage but did not change the type of the interaction. Our results suggest either a commensalistic (P. carnosum) or competitive interaction (P. phosphoreum) of photobacteria and co-contaminants on modified atmosphere packaged chicken, respectively. Furthermore, in a mix comprising seven prominent spoilers, strains of both Photobacterium species prevailed as a constant part of the spoilage microbiome during 7 days of refrigerated storage on chicken meat packaged under O 2 /CO 2 atmosphere., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Aims: This study describes the effect of phage therapy on hatching of longfin yellowtail (Seriola rivoliana) eggs challenged with Photobacterium damselae subsp. damselae., Methods and Results: A lytic phage (vB&#95;Pd&#95;PDCC-1) against P. damselae subsp. damselae was isolated and characterized. The use of phage vB&#95;Pd&#95;PDCC-1 increased the hatching rate of eggs, and reduced presumptive Vibrio species to non-detectable numbers, even in non-disinfected eggs. High-throughput 16S rRNA gene sequencing analysis revealed that phage vB&#95;Pd&#95;PDCC-1 caused significant changes in the composition and structure of the associated microbiota, allowing that members (e.g. those belonging to the family Vibrionaceae) of the class Gammaproteobacteria to be displaced by members of the class Alphaproteobacteria., Conclusions: To the best of our knowledge, this represents the first study evaluating phage therapy to control potential negative effects of P. damselae subsp. damselae during hatching of longfin yellowtail eggs., Significance and Impact of the Study: The Seriola genus includes several important commercial fish species due to its rapid growth and easy adaptability to confinement conditions. However, bacterial infections (especially those caused by Vibrio and Photobacterium species) are among the main limiting factors for the intensification of marine fish aquaculture, particularly during early development stages. Therefore, the use of phages, which are natural killers of bacteria, represents a promising strategy to reduce the mortality of farmed organisms caused by pathogenic bacteria., (© 2020 The Society for Applied Microbiology.)

Mass mortality has occurred among cultured Nile tilapia, Oreochromis niloticus, on fish farms in Manzala, Dakahlia province, Egypt, in the summer season, 2019. Moribund fish were reported with deep ulcers, septicaemic lesions and sampled for bacterial isolation. In this study, most isolates were subjected to bacteriological examination, antibiotic sensitivity test, 16S rRNA gene sequencing and histopathological examination. Following isolate identification, intraperitoneal challenge of Nile tilapia with a bacterial suspension 2 × 10 6  CFU/ml was performed. Samples from liver, spleen and kidney were collected for histological and biochemical analysis. The results showed a high similarity (99%) to Photobacterium damselae strains using phylogenetic analysis of 16S rRNA. P. damselae exhibited resistance to amoxicillin and erythromycin, as well it was highly sensitive to chloramphenicol and doxycycline. Moreover, haemorrhage, oedema, hemosiderosis and melanomacrophage activation in the liver and head kidney of infected fish were detected by light and electron microscopy. Also, significant higher levels of CAT and SOD in the spleen and head kidney, as well as the serum levels of NO were observed in experimentally challenged O. niloticus, compared to the control fish. Our data identified P. damselae for the first time from infected Nile tilapia, describing its sensitivity to a variety of antibiotics, histopathological alterations and oxidative stress impact, and it could be useful indicators for understanding P. damselae pathogenesis, which might provide a preventive efficacy for P. damselae., (© 2020 John Wiley & Sons Ltd.)

Aqueous and ethanolic extracts of drumstick, Moringa oleifera, leaves were evaluated in vitro to ascertain their principal active components and determine their immunostimulant, cytotoxic, antitumoral, bactericidal and antioxidant activities. Phytochemical screening of M. oleifera leaf extracts showed a greater abundance of phenolic and cyanogenic glycosides in aqueous than in ethanolic extracts, characterized by several flavonoids, condensed tannins and saponins. No significant effects on gilthead seabream (Sparus aurata) head-kidney leucocyte activities (phagocytic ability and capacity, respiratory burst and peroxidase) were detected after incubation for 24 h with different concentrations (0.001/1 mg mL -1 ) of either extract. In addition, the aqueous extract showed a marked cytotoxic effect on both SAF-1 (at doses above 0.01 mg mL -1 ) and PLHC-1 (at doses above 0.25 mg mL -1 ) cell lines. The ethanolic extract improved the viability of SAF-1 cells and decreased the viability of PLHC-1 cells when used at higher concentrations. Both the ethanolic and, particularly, the aqueous extracts showed significant bactericidal activity on pathogenic Vibrio anguillarum and Photobacterium damselae strains. The antiradical activity of M. oleifera, as determined by the ABTS assay, increased in a linear dose-response with increasing extract concentrations. The results as a whole for the cytotoxic, bactericidal and antioxidant activities of M. oleifera leaf extracts point to their possible use as additives in functional diets for farmed fish., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Kathon is among the most common non-oxidative biocides, containing 5-chloro-2-methyl-4-isothiazolin-3-one (CMIT) and methylisothiazolone (MIT) as the active ingredients. In our previous work, MIT was shown to be efficiently removed by ozonation. In this work, we found that ozonation didn't readily degrade CMIT. Rate constants [Formula: see text] and k ·OH,CMIT , determined to be 6.43 L mol -1  s -1 and 7.8 × 10 9 L mol -1  s -1 , indicated that hydroxyl radicals played a more important role than ozone molecule in the CMIT ozonation which was also proved by the significant inhibition (55.7 %) when adding t-butanol (TBA). Graphene oxide (GO) greatly enhanced the CMIT ozonation, and degradation efficiency raised from 15 % to 100 % after 10 min through the increased production of hydroxyl radical. Basic conditions benefited the CMIT degradation compared with acidic and neutral conditions by promoting ozone decomposition and hydroxyl radical generation, while high carbonate and humic acid concentrations had slight influence on the CMIT degradation. In spite of the complex water matrix, CMIT degradation by GO enhanced ozonation was applicable in reverse osmosis concentrate (ROC). Based on the identification of the inorganic and organic products, a possible CMIT degradation pathway was proposed. However, CMIT transformation products still showed toxicity to Photobacterium phosphoreum and Daphnia magna even after a longer ozonation time., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Assessment of eco-toxicant using bioluminescent bacterial assay is a widely used and globally accepted method. In this work, a new luminescent bacterium was isolated from squid (Loligo duvauceli) and identified as Photobacterium leiognathi strain AK-MIE using 16S rRNA, phylogeny analysis. The predicted optimum conditions by RSM were 2.76% (w/v) NaCl, 2.28% (w/v) peptone, 0.34% (w/v) yeast extract, and pH 6.83 with 541,211.80 RLU of luminescent production whereas the predicted optimum conditions by ANN were 2.21% (w/v) NaCl, 2.27% (w/v) peptone, 0.39% (w/v) yeast extract, and pH 6.94 which produced 541,986.20 RLU. The validation analysis of both RSM and ANN show 0.60% and 0.69% deviation from the predicted results indicating that both models provided good quality predictions with ANN showing a superior data fitting capability for non-linear regression analysis. Toxicity tests show strain AK-MIE was sensitive to mercury (concentration causing 50% inhibition or IC 50 of 0.00978 mgL -1 ), followed by cadmium (IC 50 of 0.5288 mgL -1 ), copper IC 50 of (0.8117 mgL -1 ), silver (IC 50 of 1.109 mgL -1 ), and lead (IC 50 of 10.71 mgL -1 ) which are more sensitive than previously isolated luminescent bacteria, suggesting that strain AK-MIE has the potential to be used in toxicity assessment of heavy metals in the environment. Based on the field trial results, several sediment samples from industrial areas in Bangi, Selangor managed to inhibit the bioluminescence of strain AK-MIE. Validation method carried out using ICP-MS proved the presence of several toxic heavy metal elements., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Armoracia rusticana (AR) was tested for antimicrobial and antioxidants power. The compound demonstrated to inhibit fish pathogens such as Vibrio anguillarum, V. harvey, V. alginolyticus, Aeromonas hydrophila, A. salmonicida, Photobacterium damselae subspecie piscicida, Tenacibaculum marinum and Pseudomonas anguilliseptica, . The total phenolic content and the reducing power resulted higher in the water extract of AR, respect to the hydroalcoolic. In vitro test demonstrated that AR significantly protect cells against death, induced by oxidative stress.

This paper reports the proportion-dependent toxicity of binary surfactant mixtures containing anionic sodium dodecyl sulfate (SDS) and nonionic fatty alcohol-polyoxyethlene ether (AEO) toward Photobacterium phosphoreum. The crucial role of toxicity interactions was elucidated by spectroscopic probing the refolding of the unfolded bovine serum albumin (BSA) induced by SDS and theoretical calculating the interaction parameter of mixed surfactants based on Rubingh's model from the critical micelle concentrations. The SDS/AEO mixtures can be divided into two groups based on the toxicity response to the proportion of AEO in the mixtures: Group I contained low mass proportions of AEO, that is, SDS:AEO = 4:1, 3:1; Group II featured high AEO proportions, that is, SDS:AEO = 3:2, 1:1, 2:3, 1:4. The toxicity of SDS/AEO mixtures decreased with the enhanced proportion of AEO in Group I and then fluctuated slightly when the AEO proportion increased to that of Group II. The mixture with the mass ratio of 1:1 showed a slightly higher toxicity than the others in Group II. Scanning electron microscopy (SEM) images illustrated that the addition of AEO hindered the action of SDS against the cell membrane. Fluorescence measurement indicated that AEO could extract SDS molecules embedded in the BSA matrix, except for those bound to the highly active sites of BSA, and refold stepwise the unfolded protein. The results were in excellent analogy to the proportion-dependent toxicity of SDS/AEO mixture, indicating the formation of mixed micelles playing a key role. The interaction parameter further revealed that antagonism led to the mixture with equal mass ratio (1:1) showing higher toxicity than other mass ratios in Group II. These results can be useful for compounding SDS/AEO mixtures in application efficiently and eco-friendly., Competing Interests: Declaration of Competing Interest We declare that we have no financial and personal relationships with other people or organizations that can inappropriately influence our work, there is no professional or other personal interest of any nature or kind in any product, service and/or company that could be construed as influencing the position presented in, or the review of, the manuscript entitled., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Advanced oxidation processes (AOPs) can degrade heavy metal complexes in wastewater to improve the removal efficiency of metals. However, the influences of AOP treatments on toxicity induced by metal complexes are not well understood. This study compared the toxicity induced by EDTA-copper (Cu) after UV/persulfate (PS) and UV/H 2 O 2 treatments on luminescent bacteria and human HepG2 cells. The results showed that EDTA-Cu complexes decreased Cu toxicity in luminescent bacteria but increased the cytotoxicity in HepG2 cells, indicating species-specific toxicity. The UV/PS and UV/H 2 O 2 treatments under most pH values and [oxidant]/[EDTA-Cu] conditions decreased the toxicity of EDTA-Cu in HepG2 cells but increased the toxicity in luminescent bacteria. When the ratio of [oxidant] to [EDTA-Cu] was 10, low toxicity in treated solutions was observed in both UV treatment processes. The alkaline precipitation treatment had a significant influence on toxicity reduction after UV/PS treatment; however, it had minimal influence on the UV/H 2 O 2 treatment system. The Cu and total organic carbon (TOC) removal efficiency cannot completely explain the results of toxicity assays. EDTA-Cu intermediates might play important roles in changing the toxicity of EDTA-Cu after both UV treatments. This study provides insights into evaluating the treatment efficiency of UV/PS and UV/H 2 O 2 on EDTA-Cu decomplexation., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Horizontal gene transfer (HGT) between bacteria with different habitats and nutritional requirements is important for the spread of antibiotic resistance genes (ARG). The objective of the present study was to clarify the effects of organic matter on HGT between nourished and starved bacteria. We demonstrated that conjugation ability is affected by the nutritional conditions of the cell and environment. A filter mating HGT experiment was performed using Photobacterium damselae ssp. damselae, strain 04Ya311, a marine-origin bacterium possessing the multidrug-resistance plasmid pAQU1, as the donor, and Escherichia coli as the recipient. The donor and recipient were both prepared as nutrient-rich cultured and starved cells. Filter mating was performed on agar plates with and without organic nutrients. The transcription of the plasmid-borne genes tet(M) and traI was quantitated under eutrophic and oligotrophic conditions. The donor P. damselae transferred the plasmid to E. coli at a transfer rate of 10 -4 under oligotrophic and eutrophic conditions. However, when the donor was starved, HGT was not detected under oligotrophic conditions. The addition of organic matter to starved cells restored conjugative HGT even after 6 d of starvation. The transcription of traI was not detected in starved cells, but was restored upon the addition of organic matter. The HGT rate appears to be affected by the transcription of plasmid-associated genes. The present results suggest that the HGT rate is low in starved donors under oligotrophic conditions, but is restored by the addition of organic matter.

Organisms are frequently exposed to mixtures of heavy metals because of their persistence in the environment. The mixture toxicity of heavy metals should therefore be evaluated to perform a rational environmental risk assessment for organisms. In this study, we determined the inhibition toxicity of five heavy metals (Cu2+, Co2+, Zn2+, Fe3+ and Cr3+) and their binary mixtures to Photobacterium phosphoreum (P. phosphoreum). We obtained the following results: (1) the order of individual toxicity was Zn2+>Cu2+>Co2+>Cr3+>Fe3+, and (2) different combined effects (additive, synergistic and antagonistic) were observed in the binary mixtures of heavy metals, with toxicity unit (TU) values ranging from 0.15 to 3.50. To predict the mixture toxicity of heavy metals, we derived the ion characteristic parameters of heavy metal mixtures and explored the ion-characteristic-based quantitative structure-activity relationship (QSAR) model (R2 = 0.750, Q2 = 0.649). The developed QSAR model indicated that the mixture toxicity of heavy metals is related to the change in ionization potential ((ΔIP)mix), the first hydrolysis constant (log(KOH)mix) and the formation constant value ([Formula: see text])., Competing Interests: The authors have declared that no competing interests exist.

Surfactant mixtures have extensive industrial applications due to their ideal properties and low ecotoxicity. However, the ecotoxicity of surfactant mixtures with different proportions and their correlation with surface properties have remained poorly investigated. In this study, the ecotoxicity and surface activity of the composites of anionic surfactant sodium dodecylbenzene sulfonate (SDBS) and nonionic surfactant fatty alcohol-polyoxyethylene ether (AEO) in various mass ratios were assessed, and the correlation between ideal application properties and safe ecological perspective of the composites was explored. The ecotoxicity of individual SDBS, AEO, and SDBS/AEO mixtures was determined using the bioluminescence inhibition assay with Photobacterium phosphoreum, and the critical micelle concentrations (CMC) were measured by surface tension method and steady-state fluorescence spectroscopy. Sodium dodecylbenzene sulfonate (SDBS) showed a considerably higher toxicity than individual AEO and SDBS/AEO mixtures. Scanning electron microscope images illustrated the rupture of bacteria membrane induced by SDBS, and the addition of AEO alleviated the damage. According to the dose-response relationship on luminous bacteria, SDBS/AEO mixtures were divided into three groups (group I with a high proportion of SDBS, SDBS:AEO = 4:1 and 3:2; group II, SDBS:AEO = 1:1; group III with a high proportion of AEO, SDBS:AEO = 2:3 and 1:4). The sequence of toxicity of the SDBS/AEO mixtures was group II > group III > group I, demonstrating that the toxicity of the composites was related to the mixture proportion instead of the amount of AEO added. The CMC order of SDBS/AEO mixtures was group II > group I > group III, and it was proportion dependent. Furthermore, ΔC M was defined as the difference of the experimental (C M ) and ideal CMC (C M i deal ) of the mixed system, indicating the interaction between the two kinds of surfactants. The order of the ΔC M was group II > group III > group I, which was consistent with the sequence of the toxicity. Therefore, ΔC M can be a potential indicator for the hazardous assessment of surfactant mixtures involving high ionic strength., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Degradation of imipramine (IMI) in the VUV system (VUV 185  + UV 254 ) was firstly evaluated in this study. Both HO • oxidation and UV 254 direct photolysis accounted for IMI degradation. The quantum yields of UV 254 direct photolysis of deprotonated and protonated IMI were 1.31×10 -2 and 3.31×10 -3 , respectively, resulting in the higher degradation efficiency of IMI at basic condition. Increasing the initial IMI concentration lowered the degradation efficiency of IMI. While elevating reaction temperature significantly improved IMI degradation efficiency through the promotion of both the quantum yields of HO • and the UV 254 direct photolysis rate. The apparent activation energy was calculated to be about 26.6 kJ mol -1 . Negative-linear relationships between the k obs of IMI degradation and the concentrations of HCO 3 - /CO 3 2- , NOM and Cl - were obtained. The degradation pathways were proposed that cleavage of side chain and hydroxylation of iminodibenzyl and methyl groups were considered as the initial steps for IMI degradation in the VUV system. Although some high toxic intermediate products would be produced, they can be further transformed to other lower toxic products. The good degradation efficiency of IMI under realistic water matrices further suggests that the VUV system would be a good method to degrade IMI in aquatic environment., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

The tert-butylphenols (TBPs) are one group of alkylated phenolic compounds with wide applications in UV absorbers and antioxidants. They are becoming contaminants of emerging concern with residues frequently detected in natural surface water or drinking water. The direct sunlight may photolyze TBPs in waters and affect their aquatic toxicities; however, such data are very limited. In the present study, we investigate the photodegradation of 2,6-DTBP by direct sunlight in water and compare the aquatic toxicities of 2,6-DTBP with that of its product toward Photobacterium phosphoreum. 2,6-DTBP is photodegraded by 71.31 ± 2.64% under simulated sunlight following a pseudo-first-order kinetics with rate constant (k) of 0.061 h -1 . Density functional theory simulations at M06-2X/def2-SVP level reveal that the photodegradation occurred sequentially through oxidation, photo-isomerization and hydrogenation. The degradation product 2,5-DTBP is toxic to P. phosphoreum (EC 50 3.389 × 10 -5  mol/L) whereas 2,6-DTBP is not harmful (EC 50 3.917 × 10 -3  mol/L) as designated by the European Union Standard, indicating the enhanced toxicities driven by the direct sunlight photodegradation. We demonstrate the enhanced toxicities of 2,6-DTBP by natural sunlight, suggesting that negligence of photodegradation of TBPs-related contaminants will underestimate the comprehensive risk of these emerging contaminant in natural waters., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

The single toxicity (IC 50 ) of zinc (Zn) and 11 nitro-substituted benzenes to Photobacterium phosphoreum were determined, respectively. On basis of single toxicity, the joint toxicity of binary mixtures of Zn and 11 nitro-substituted benzenes at different Zn concentrations of 0.2 IC 50 , 0.5 IC 50 , and 0.8 IC 50 were measured. The joint toxicity was evaluated by toxic unit (TU) and additive index (AI) methods. The results indicated that the joint toxicity was not only depending on the Zn concentrations but also on the substituted groups of nitro-substituted benzenes. The quantitative structure-activity relation (QSAR) equations were developed and the results showed that the toxicity of nitro-substituted benzenes has different joint effect at the different Zn concentrations. At the Zn concentration of 0.2 IC 50 , the binary joint effects were mainly antagonism and the joint toxicity was negatively related to descriptors called VE2&#95;B(p) and TIC3. At the Zn concentration of 0.5 IC 50 and 0.8 IC 50 , the binary joint effects were mainly antagonism and simple addition, and the joint toxicity was related to the same descriptor Eig06&#95; AEA(dm). It indicated that the joint toxic actions were similar when combined at the medium and high concentrations of Zn.

This work investigated the biotoxicity and the residual dissolved organic matter (DOM) of the effluents from nine typical full-scale textile plants located at Pearl River Delta (PRD) in Guangdong province, China. The fluorescence regional integration (FRI) analysis showed that the tryptophan-like (II), soluble microbial product-like (IV) and fulvic acid-like substances (III) were the dominant compounds in the DOM. The acute toxicity test showed toxic effects still remained in most textile effluents, which might attribute to the undegraded dyes or aromatic compounds. Combining with the results from multiple methods, it indicated that the selected nine textile wastewater treatment plants (tWWTPs) all contained some residual micro organic pollutants in their effluents, and the residual benzene-derived products or aromatic amines were probably the toxicity-causing substances. Both ozonization and membrane filtration were capable of further decreasing the content of residual DOM, but by comprehensively considering the effects of removing DOM and biotoxicity, membrane filtration was better than ozonization., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Diclofenac was frequently found in various waters, indicating conventional wastewater treatment methods ineffective in its removal. In this study, LaFeO 3 (LFO) was synthesized and its catalytic activity of LFO as the activator of different oxidants such as persulfate (PS), hydrogen peroxide and peroxylmonosulfate (PMS) was evaluated in terms of DCF degradation. The influence of calcination temperature was examined on the catalytic activity of LFO. The effects of various parameters including pH levels, PMS concentration, LFO dose and initial DCF concentration were investigated on DCF degradation rate. The marginal effects of PMS concentration and LFO dose were compared. Langmuir-Hinshelwood (LH) model was used to quantitatively describe DCF degradation reaction in LFO/PMS system. The two constants, k (Limiting reaction rate at maximum coverage) and K (Equilibrium adsorption constant), were determined on the basis of LH model. The performance of LFO/PMS process was also estimated in the presence of various inorganic anions. The potential toxicity of LFO and PMS were evaluated using phytoplankton and the toxicity evolution during DCF degradation was also investigated using luminescent bacteria. This contribution provides a basic study regarding the potential application of heterogeneous PMS activation by perovskite LFO for both DCF removal and toxicity elimination., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

A high-efficiency pyrethroid-degrading bacterium, Photobacterium ganghwense strain 6046 (PGS6046), was first isolated from an offshore seawater environment. Metabolomics method was used to investigate the biotransformation pathway of PGS6046 to cyfluthrin wherein 156 metabolites were identified. The growth rates of the PGS6046 cultivated in nourishing media were much higher than those cultivated in seawater, regardless of the presence of cyfluthrin. Statistical analyses revealed that the metabolic profile of PGS6046 was associated with the culture medium, the presence of cyfluthrin, and culture time. The PGS6046 cultivated in a nourishing medium was characterized by higher levels of amino acids, a lower abundance of intermediates in the tricarboxylic acid cycle, and the presence of some fatty acids than those cultivated in seawater. The effects of cyfluthrin on PGS6046 metabolism varied based on the culture medium, whereas the cyanoalanine levels increased under both culture conditions. Culture time significantly affected the metabolism of amino acids and carbohydrates in PGS6046. The present study revealed the metabolic characteristics of PGS6046 under different culture conditions and will further facilitate the exploration of the fundamental questions regarding PGS6046 and its potential applications in environmental bioremediation.

Degradation of atrazine by catalytic ozonation in the presence of iron scraps (ZVI/O 3 ) was carried out. The key operational parameters (i.e., initial pH, ZVI dosage, and ozone dosage) were optimized by the batch experiments, respectively. This ZVI/O 3 system exhibited much higher degradation efficiency of atrazine than the single ozonation, ZVI, and traditional ZVI/O 2 systems. The result shows that the pseudo-first-order constant (0.0927 min -1 ) and TOC removal rate (86.6%) obtained by the ZVI/O 3 process were much higher than those of the three control experiments. In addition, X-ray diffraction (XRD) analysis indicates that slight of γ-FeOOH and Fe 2 O 3 were formed on the surface of iron scrap after ZVI/O 3 treatment. These corrosion products exhibit high catalytic ability for ozone decomposition, which could generate more hydroxyl radical (HO • ) to degrade atrazine. Six transformation intermediates were identified by liquid chromatography-mass spectrometry (LC-MS) analysis in ZVI/O 3 system, and the degradation pathway of atrazine was proposed. Toxicity tests based on the inhibition of the luminescence emitted by Photobacterium phosphoreum and Vibrio fischeri indicate the detoxification of atrazine by ZVI/O 3 system. Finally, reused experiments indicate the approving recyclability of iron scraps. Consequently, the ZVI/O 3 system could be as an effective and promising technology for pesticide wastewater treatment.

In this work, PM 10 samples previously subjected to thorough chemical speciation and receptor modelling, have been investigated for their bio-toxicity using an inhibition test based on bacterial luminescence modulation when in contact with airborne particulate samples. The variation of light emission intensity from a luminescent bacteria strain, the Photobacterium phosphoreum, is proposed as an efficient proxy for the quantification of bio-toxic effects induced by airborne particulate matter. PM 10 samples characterized by definite levels of pollutants from the pertaining air shed were found to induce a decrease in the bacterial bioluminescence intensity, expressed as percentage of Inhibition Ratio (IR%). This behaviour suggests the decay of this energy-consuming activity because of a toxic effect. Cluster analysis on chemical composition and IR% data provides evidence of a statistically significant association between the adverse effects on living cells and the range of specific chemical species in PM 10 ., (Copyright © 2018 Elsevier B.V. All rights reserved.)

A novel tool, the DAMTA analyzer (Device for Analytical Monitoring and Toxicity Assessment), designed for fully automated toxicity measurements based on luminescent bacteria as well as for concomitant determination of chemical parameters, was developed and field-tested. The instrument is a robotic water analyzer equipped with a luminometer and a spectrophotometer, integrated on a thermostated reaction plate which contains a movable carousel with 80 cuvettes. Acute toxicity is measured on-line using a wild type Photobacterium phosphoreum strain with measurable bioluminescence and unaltered sensitivity to toxicants lasting up to ten days. The EC50 values of reference compounds tested were consistent with A. fischeri and P. phosphoreum international standards and comparable to previously published data. Concurrently, a laboratory trial demonstrated the feasibility of use of the analyzer for the determination of nutrients and metals in parallel to the toxicity measurements. In a prolonged test, the system was installed only in toxicity mode at the premises of the World Fair "Expo Milano-2015″, a high security site to ensure the quality of the supplied drinking water. The monitoring program lasted for six months during which ca. 2400 toxicity tests were carried out; the results indicated a mean non-toxic outcome of -5.5 ± 6.2%. In order to warrant the system's robustness in detecting toxic substances, Zn was measured daily with highly reproducible inhibition results, 70.8 ± 13.6%. These results assure that this novel toxicity monitor can be used as an early warning system for protection of drinking water sources from emergencies involving low probability/high impact contamination events in source water or treated water., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Origanum vulgare is a well-known medicinal plant that has been used since ancient times as an additive in foods and cosmetic preparations. The possible application of O. vulgare extracts in fish was assessed by using gilthead seabream (Sparus aurata) as a marine fish model due to its importance in aquaculture. The in vitro effects of aqueous and ethanolic leaf extracts of O. vulgare were tested in order to observe any immunostimulant, cytotoxic, bactericidal or antioxidant properties. The results showed that medium or high concentration of aqueous extracts and low concentrations of ethanolic extract, increased head kidney leucocyte activities as well as the number of SAF-1 cells. However, moderate to high concentrations of ethanolic extracts decreased both leucocyte activities and the number of viable SAF-1 cells, suggesting some possible toxic effect towards them. Only the highest concentration of the aqueous extract and medium to high concentrations of the ethanolic extracts showed cytotoxic activity against the tumor PLHC-1 cell line. Bactericidal activity was only detected against Vibrio harveyi, V. anguillarum and Photobacterium damselae when using the highest concentration of aqueous extract and moderate to high concentrations of ethanolic extract. Finally, both plant extracts presented antioxidant activity particularly the aqueous extract. Overall, the results suggest that both extracts (when used at the appropriate concentration) have immunostimulant, cytotoxic, bactericidal and antioxidant properties, making O. vulgare an interesting candidate for incorporation as additive in functional diets for farmed fish., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

This paper evaluates the oxidation of an antibacterial agent, chlorophene (4-chloro-2-(phenylmethyl)phenol, CP), by permanganate (Mn(VII)) in water. Second-order rate constant (k) for the reaction between Mn(VII) and CP was measured as (2.05 ± 0.05) × 10 1  M -1  s -1 at pH 7.0 for an initial CP concentration of 20.0 μM and Mn(VII) concentration of 60.0 μM. The value of k decreased with increasing pH in the pH range of 5.0-7.0, and then increased with an increase in solution pH from 7.0 to 10.0. The presence of MnO 2 and Fe 3+ in water generally enhanced the removal of CP, while the effect of humic acid was not obvious. Fourteen oxidation products of CP were identified by an electrospray time-of-flight mass spectrometer, and direct oxidation, ring-opening, and decarboxylation were mainly observed in the reaction process. The initial reaction sites of CP by Mn(VII) oxidation were rationalized by density functional theory calculations. Toxicity changes of the reaction solutions were assessed by the luminescent bacteria P. phosphoreum, and the intermediate products posed a relatively low ecological risk during the degradation process. The efficient removal of CP in secondary clarifier effluent and river water demonstrated the potential application of this Mn(VII) oxidation method in water treatment., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Microorganisms have been very useful in environmental monitoring due to their constant sensing of the surrounding environment, their easy maintenance and low cost. Some freeze-dried toxicity kits based on naturally bioluminescent bacteria are commercially available and commonly used to assess the toxicity of environmental samples such as Microtox (Aliivibrio fischeri) or ToxScreen (Photobacterium leiognathi), however, due to the marine origin of these bacteria, they could not be the most appropriate for fresh-waters monitoring. Cyanobacteria are one of the most representative microorganisms of aquatic environments, and are well suited for detecting contaminants in aqueous samples. This study presents the development and application of the first freeze-dried cyanobacterial bioassay for fresh-water contaminants detection. The effects of different cell growth phases, cryoprotectant solutions, freezing protocols, rehydration solutions and incubation conditions methods were evaluated and the best combination of these parameters for freeze-drying was selected. The study includes detailed characterization of sensitivity towards reference pollutants, as well as, comparison with the standard assays. Moreover, long-term viability and sensitivity were evaluated after 3 years of storage. Freeze-dried cyanobacteria showed, in general, higher sensitivity than the standard assays and viability of the cells remained after 3 years of storage. Finally, the validation of the bioassay using a wastewater sample was also evaluated. Freeze-drying of cyanobacteria in 96-well plates presents a simple, fast and multi-assay method for environmental monitoring., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Mitochondria-targeted antioxidants are known to alleviate mitochondrial oxidative damage that is associated with a variety of diseases. Here, we showed that SkQ1, a decyltriphenyl phosphonium cation conjugated to a quinone moiety, exhibited strong antibacterial activity towards Gram-positive Bacillus subtilis, Mycobacterium sp. and Staphylococcus aureus and Gram-negative Photobacterium phosphoreum and Rhodobacter sphaeroides in submicromolar and micromolar concentrations. SkQ1 exhibited less antibiotic activity towards Escherichia coli due to the presence of the highly effective multidrug resistance pump AcrAB-TolC. E. coli mutants lacking AcrAB-TolC showed similar SkQ1 sensitivity, as B. subtilis. Lowering of the bacterial membrane potential by SkQ1 might be involved in the mechanism of its bactericidal action. No significant cytotoxic effect on mammalian cells was observed at bacteriotoxic concentrations of SkQ1. Therefore, SkQ1 may be effective in protection of the infected mammals by killing invading bacteria.

Propolis has been used as a medicinal agent for centuries. The chemical composition of four propolis samples collected from four locations of the Sétif region, Algeria, using gas chromatography-mass spectrometry was determined. More than 20 compounds and from 30 to 35 compounds were identified in the aqueous and ethanolic extracts, respectively. Furthermore, the antimicrobial activity of the propolis extracts against two marine pathogenic bacteria was evaluated. Finally, the in vitro effects of propolis on gilthead seabream (Sparus aurata L.) leucocyte activities were measured. The bactericidal activity of ethanolic extracts was very high against Shewanella putrefaciens, average against Photobacterium damselae and very low against Vibrio harveyi. The lowest bactericidal activity was always that found for the aqueous extracts. When the viability of gilthead seabream head-kidney leucocytes was measured after 30 min' incubation with the different extracts, both the ethanolic and aqueous extracts of one of the propolis samples (from Babor) and the aqueous extract of another (from Ain-Abbassa) provoked a significant decrease in cell viability when used at concentrations of 100 and 200 μg ml -1 . Furthermore, significant inhibitory effects were recorded on leucocyte respiratory burst activity when isolated leucocytes where preincubated with the extracts. This effect was dose-dependent in all cases except when extracts from a third propolis sample (from Boutaleb) were used. Our findings suggest that some of Algerian propolis extracts have bactericidal activity against important bacterial pathogens in seabream and significantly modulate in vitro leucocyte activities, confirming their potential as a source of new natural biocides and/or immunomodulators in aquaculture practice., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Ozonation of a commonly used non-steroidal anti-inflammatory drug indomethacin (IM) was studied. Kinetic constants of IM with ozone and hydroxyl radicals were measured at an order of magnitude of 10 5 M -1 s -1 and 10 9 M -1 s -1 , respectively. IM was degraded within 7min under the lowest ozone dose, but TOC removal was only 50% even under the highest ozone dose used in the experiments. Ozone rather than hydroxyl radicals was found to be the main oxidant during reaction, with a contribution rate of 80% under pH 7. Six intermediates were identified by high resolution mass spectrometer. Nitrogen atom, CC double bond and benzene ring were found to be the main reaction sites. Electrophilic attack or Criegee cyclo-addition were proved to be the most probable pathways at the first step. The formation mechanism of one of the ozone products was first proposed during the experiment, then confirmed by the density functional theory (DFT) calculation. Acetic acid, formic acid and oxalic acid were detected as small molecule organic products. The toxicity change during ozonation was measured by luminescent bacterium with results showing that the toxicity can be reduced to zero when ozone dose was high enough., (Copyright © 2016 Elsevier B.V. All rights reserved.)

In China, a considerable part of industrial wastewater effluents are discharged into the municipal wastewater treatment plants (WWTPs) after pretreatment in their own wastewater treatment plants. Even though the industrial effluents meet the professional emission standards, many micro-pollutants still remained, and they could be resistant in the municipal WWTPs with conventional activated sludge process. Pigment wastewater was chosen in this study, and the acute toxicity reduction and identification of the pigment-contaminated wastewater treated by the conventional anaerobic-anoxic-oxic (A/A/O) process were evaluated. Results indicated that the raw pigment-contaminated wastewater was acutely toxic to Photobacterium phosphoreum (P. phosphoreum), Daphnia magna (D. magna) and Danio rerio (D. rerio). The acute toxicity was decreased in some degree after A/A/O treatment, but the final effluent still exhibited acute toxicity to D. magna and D. rerio with the toxic units (TU) of 1.1 and 2.0, respectively. Chemical analyses showed the presence of various refractory and toxic nitrogen-containing polycyclic and heterocyclic compounds in the pigment-contaminated wastewater. Toxicity identification by combining chemical analyses and correlation analysis showed that N-containing refractory organic toxicants were the main toxicity source for the pigment-contaminated wastewater, and several toxicants showed significant correlation with P. phosphoreum and D. magna. This study indicated that the A/A/O process was not efficient for pigment-contaminated wastewater treatment, and it was irradiative for technology improvement in the WWTPs receiving pretreated industrial wastewater effluents., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

We compared the TBT-resistant ability of resting cells prepared from isolates that formed colonies on nutrient agar plates containing 100 µM tributyltin (TBT) chloride, such as Photobacterium sp. TKY1, Halomonas sp. TKY2, and Photobacterium sp. NGY1, with those from taxonomically similar type strains. Photobacterium sp. TKY1 showed the highest ability among those three isolates. The number of surviving Photobacterium sp. TKY1 cells was hardly decreased after 1 h of exposure to 100 µM TBTCl, regardless of the number of resting cells in the range from 10 9.4 to 10 4.2 CFU mL -1 . In such an experimental condition, the maximum number of TBT molecules available to associate with a single cell was estimated to be approximately 6.0 x 10 11.8 . Resting cells prepared from type strains Photobacterium ganghwense JCM 12487 T and P. halotolerans LMG 22194 T , which have 16S rDNA sequences highly homologous with those of Photobacterium sp. TKY1, showed sensitivity to TBT, indicating that TBT-resistant marine bacterial species are not closely related in spite of their taxonomic similarity. We also estimated the impact of TBT-resistant bacterial species to indigenous microbial populations of TBT-polluted surface sediments. The number of surviving TBT-sensitive Vibrio natriegens ATCC 14048 T cells, 10 6.2±0.3 CFU mL -1 , was reduced to 10 4.4±0.4 CFU mL -1 when TBT-resistant Photobacterium sp. TKY1 cells, 10 9.1±0.2 CFU mL -1 , coexisted with 10 9.4±0.2 CFU mL -1 of V. natriegens ATCC 14048 T cells in the presence of 100 µM TBTCl. These results indicate that the toxicity of TBT to TBT-sensitive marine bacterial populations might be enhanced when a TBT-resistant marine bacterial species inhabits TBT-polluted surface sediments.

The tetrodecamycins are a group of secondary metabolites that are characterized by the presence of a tetronate ring in their structure. Originally discovered for their antibiotic activity against Photobacterium damselae ssp. piscicida, the causative agent of pseudotuberculosis in fish, this family of molecules has also been shown to have potent antibiotic activity against methicillin-resistant Staphylococcus aureus. Due to their small size and highly cyclized nature, they represent an unusual member of the much larger group of bioactive molecules called the tetronates. Herein, we review what is known about the mechanism of action of these molecules and also present a hypothesis for their biosynthesis. A deeper understanding of the tetrodecamycins will provide a more holistic view of the tetronate-family, provide new chemical probes of bacterial biology, and may provide therapeutic lead molecules., (Copyright © 2016 The Authors. Published by Elsevier Ltd.. All rights reserved.)

A bioluminescent bacterium, Photobacterium phosphoreum T3 (PPT3), was used as a bio-indicator for the atmospheric fine particulate matter (PM2.5) to determine the eco-toxicity of PM2.5. The PM2.5 contains toxic chemicals, which reduce light output. The PM2.5 samples were collected in the period from March 2014 to January 2015 in Nanjing and analyzed for the chemical composition versus their eco-toxicity. The eco-toxicological responses of each toxicant were detected in PM2.5 samples with PPT3. The dose-response curves obtained were verified using the Weibull fitting function. According to the measured EC50 values (EC50, the concentration of a toxicant that inhibits 50% of the bioluminescence), the toxicity sequence was: B[a]P>hexa-PCB>tetra-PCB>tri-PCB>Pb(2+)>DEHP>Cu(2+)>DBP>BDE209>Zn(2+)>DMP>DEP, where B[a]P is benzo(a)pyrene, PCB is polychlorinated biphenyl, DEHP is diethylhexyl phthalate, DBP is dibutyl phthalate, BDE209 is decabromodiphenyl ether, DMP is dimethyl phthalate, and DEP is diethyl phthalate. All the PM2.5 samples analyzed proved to be weak toxic for PPT3. The toxicity of PM2.5 was assessed by the dose-addition of organic species and heavy metallic elements existing in PM2.5 with PPT3. The bioluminescence test showed that the metals and organics detected in PM2.5 promoted PM2.5 toxicity. The total detectable organics (denoted by ΣOrs) exhibited slightly higher toxicity than the total metals (denoted by ΣMs). In contrast, the sum of water-soluble ions (denoted by ΣIons) was beneficial to PPT3. The PM2.5 toxicity increased as the PM2.5 trapped more organics or metallic elements from the industrial or densely populated urban areas, where the PM2.5 had a high inhibition rate of bioluminescence for PPT3 in contrast to the residential PM2.5 samples, where the minimum inhibition rate was observed. The toxicity of PM2.5 samples varied with the mass concentrations, chemical constituents, and sampling locations. The chemicals in PM2.5, especially organic species and metallic elements, affected its eco-toxicity. These data provided good understanding of the atmospheric PM2.5 pollution in the large portion of the East China., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Photobacterium damselae is a Gram negative bacterium causes photobacteriosis, a worldwide septicemic disease in aquaculture including sea bass (Dicentrarchus labrax) and sea bream (Sparus aurata). The pathogenicity of bacterial subspecies and the disease pathological changes in natural and experimental infections have thus far yielded inconsistency of effective preventive measures. This study aimed to represent a comprehensive analysis of the potential pathogenic capacities of the two subspecies of P. damselae in cultured sea bass and sea bream in the Northwestern region of Egypt. Diseased 321 sea bass and 257 sea bream, in addition to 99 healthy sea bass fingerlings were sampled from three farms located along the Mediterranean Sea. P. damselae subspecies were isolated from diseased fish and characterized using bacteriological, molecular, and antimicrobial susceptibility methods. Healthy fish were challenged by a virulent P. damselae subsp. piscicida, monitored for disease signs and mortality, and the histopathological abnormalities and hematological disorders were carried out. Clinical signs and gross lesions in naturally infected sea bass and sea bream showed great similarities with absence of a subspecies-specific characteristic sign or lesion. The two subspecies were recovered through the entire year from individual fish sample, suggests a coexistence of two subspecies endemic infection. In diseased sea bass, 38.32% and 16.20% were positive for P. damselae subsp. piscicida and subsp. damselae, respectively. However in diseased sea bream, 44.47% and 26.46% were positive for P. damselae subsp. piscicida and subsp. damselae, respectively. High mortalities and devastating clinicopathologic abnormalities represented by sever clinical signs, hematological disorders and histological abnormalities strengthen the pathogenicity of P. damselae subspecies in the two fish species and therefore, a vaccination strategy against both subspecies should be taken into account., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Di-n-butyl phthalate (DBP) and di-2-ethylhexyl phthalate (DEHP) are two widely used phthalates, while Cu(II) is a common valence state of copper. They have been ubiquitously detected in the aquatic environment, but information on their joint toxicity to aquatic organisms is scarce. In this study, we evaluated the combined effects of copper and these two phthalates to Daphnia magna and Photobacterium phosphoreum by quantifying the acute toxicity expressed by the EC50 (the concentration causing 50 % of maximal effect) value. The toxicity order was DEHP + Cu(II) > DBP + Cu(II) > Cu(II) > DEHP > DBP for both test species. Antagonism effects were found in the joint toxicity of Cu(II) combined with DBP or DEHP using the toxic unit method. These findings have important implications in environmental risk assessment for phthalates in the aquatic environment in the presence of heavy metals.

The present study focused on the kinetics, transformation pathways and toxicity of several benzophenone-type ultraviolet filters (BPs) during the water chlorination disinfection process. The transformation kinetics of the studied three BPs was found to be second-order reaction, which was dependent on the concentration of BPs and chlorine. The second-order rate constants increased from 86.7 to 975 M(-1) s(-1) for oxybenzone, 49.6-261.7 M(-1) s(-1) for 4-hydroxybenzophenone and 51.7-540 M(-1) s(-1) for 2-hydroxy-4-methoxybenzophenone-5-sulfonic acid with the increasing pH value from 6 to 8 of the chlorination disinfection condition. Then the transformation products (TPs) of these BPs were identified by HPLC-QTof analysis. Several transformation pathways, including electrophilic substitution, methoxyl substitution, ketone groups oxidation, hydrolysis, decarboxylation and ring cleavage reaction, were speculated to participate in the chlorination transformation process. Finally, according to the toxicity experiment on luminescent bacteria, Photobacterium phosphoreum, enhanced toxicity was observed for almost all the TPs of the studied BPs except for 2,2'-dihydroxy-4,4'-dimethoxybenzophenone; it suggested the formation of TPs with more toxic than the parent compounds during the chlorination process. The present study provided a foundation to understand the transformation of BPs during chlorination disinfection process, and was of great significance to the drinking water safety., (Copyright © 2016 Elsevier B.V. All rights reserved.)

A hydrolysis acidification (HA)-anoxic-oxic (A/O) process was adopted to treat a petrochemical wastewater. The operation optimization was carried out firstly by a bench scale experimental reactor. Then a full scale petrochemical wastewater treatment plant (PCWWTP, 6500 m(3) h(-1)) was operated with the same parameters. The results showed that the BOD5/COD of the wastewater increased from 0.30 to 0.43 by HA. The effluent COD was 54.4 mg L(-1) for bench scale reactor and 60.9 mg L(-1) for PCWWTP when the influent COD was about 480 mg L(-1) on optimized conditions. The organics measured by gas chromatography-mass spectrometry (GC-MS) reduced obviously and the total concentration of the 5 organics (1,3-dioxolane, 2-pentanone, ethylbenzene, 2-chloromethyl-1,3-dioxolane and indene) detected in the effluent was only 0.24 mg L(-1). There was no obvious toxicity of the effluent. However, low acute toxicity of the effluent could be detected by the luminescent bacteria assay, indicating the advanced treatment is needed. The clone library profiling analysis showed that the dominant bacteria in the system were Acidobacteria, Proteobacteria and Bacteriodetes. HA-A/O process is suitable for the petrochemical wastewater treatment., (Copyright © 2016 Elsevier B.V. All rights reserved.)

The toxicity of metal oxide nanoparticles (NPs) has aroused great concern over the past few years. However, there still remains the question whether the toxicity of the metal oxide NPs originates from the released ions or the NPs themselves. In this study, the metal ion release of CuO, Fe2O3, ZnO, Co3O4, Cr2O3, and NiO NPs in aqueous media was investigated, and their contributions to the metal oxide NPs' inhibition on the bioluminescence of Photobacterium phosphoreum were studied. It was found that the ions release of the metal oxide NPs in aqueous media was complex, depending on both the dissolution and adsorption processes of the metal oxide NPs. The relationships between the metal oxide NPs' antibacterial effects and their released metal ions could be divided into three categories: (1) the ZnO NPs' antibacterial effect was due solely to the released Zn(2+); (2) the CuO NPs' antibacterial effect originated from both the released Cu(2+),and the CuO particles; and (3) the antibacterial effects of Fe2O3, Co3O4, Cr2O3, and NiO NPs were caused by the NPs themselves. Our findings suggest that the ions release and their contributions to the NPs' toxicity should be considered in the toxicity evaluations of the metal oxide NPs., (Copyright © 2016 Elsevier B.V. All rights reserved.)

The occurrence of benzophenone-4 (BP-4) in water environments may pose a serious public health hazard due to its potential endocrine disrupting effects. In this work, the intermediates, probable degradation pathways and toxicity changes during ozonation of BP-4 in aqueous solution were systematically investigated. Results revealed that alkaline conditions favored the oxidation of BP-4. However, inorganic anions (Cl(-), NO3(-), SO4(2-)), cations (K(+), Ca(2+), Mg(2+)) and humic acid had no remarkable effect on BP-4 removal within the tested concentrations. Ozonation was also effective for the fast removal of BP-4 in real waters. The TOC suggested a low mineralization rate, even after the complete BP-4 removal. Meanwhile, the treated mixtures exhibited an obvious inhibition to the bioluminescent bacteria Photobacterium phosphoreum, indicating the formation of transformation products with higher toxicities. Furthermore, fourteen products were identified by means of liquid chromatography-mass spectrometry. Notably, seven of them have not been reported previously. The quenching test indicated that the degradation processes probably were dominated by OH. Next, possible degradation pathways were proposed and further justified by theoretical calculations of frontier electron densities. This investigation will contribute to the systematic elucidation of the ozonation process of UV filters in aquatic environments., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Acyclovir and penciclovir, 2 antiviral drugs, are increasingly detected in aquatic environments. The present study explores the natural photochemical transformation mechanisms and fate of these drugs, examining direct and indirect photochemical transformation under simulated sunlight irradiation. The 2 antiviral drugs are photostable under certain conditions but significantly degrade in the presence of chromophoric dissolved organic matter (DOM). The degradation rate associated with the drugs' indirect photochemical transformation scaled with chromophoric DOM concentration. Quenchers and sensitizers were used to identify indirect photochemical transformation mechanism. Results suggested that both pharmaceuticals could be transformed by reacting with (1)O2, (•)OH, and excited chromophoric DOM. The (1)O2 played an important role in indirect photochemical transformation. Furthermore, the reaction kinetics between their substructural molecules, guanine, isocytosine, and imidazole, with different reactive oxygen species were evaluated to determine which substrate functionalities were most susceptible to singlet oxygenation. Imidazole was identified as the reaction site for (1)O2, and preliminary (1)O2 oxidation mechanisms were further evaluated based on liquid chromatographic-tandem mass spectrometric results. Finally, aquatic ecotoxicity assessment of phototransformed solutions revealed that the degradation of acyclovir and penciclovir may not ultimately diminish environmental risk because of either formation of more toxic intermediates than parent pharmaceuticals or some synergistic effects existing between the intermediates., (© 2015 SETAC.)

Chloride ion is known to affect on degradation kinetics in different ways during HO· and SO4(·-)-based advanced oxidation processes (AOPs). However, its effect on absorbable organic halogen (AOX) evolution and acute toxicity of treated water remains unknown, despite the importance of the two parameters in evaluating the applicability of AOPs. In the present study, Co/peroxymonosulfate (Co/PMS) and UV/hydrogen peroxide (UV/H2O2) treatment of 2,4,6-trichlorophenol was compared in terms of AOX formation, chlorinated byproducts and acute toxicity. Both Co/PMS and UV/H2O2 systems were more reactive under acidic conditions, resulting in elevated AOX levels when compared with those at neutral pH. The presence of high levels of chloride led to an accumulation and increase of AOX in the Co/PMS system. The toxicity of chlorinated byproducts was evaluated using Photobacterium phosphoreum, and the results revealed a sharp increase in acute toxicity of Co/PMS reaction solutions on addition of chloride ion. However, addition of Cl(-) had no apparent impact on AOX and toxicity of UV/H2O2 reaction solutions. These findings may have significant technical implications for selecting feasible technologies to treat high salinity wastewater., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Proliferation of microbial population on fresh poultry meat over time elicits spoilage when reaching unacceptable levels, during which process slime production, microorganism colony formation, negative organoleptic impact and meat structure change are observed. Spoilage organisms in raw meat, especially Gram-negative bacteria can be difficult to combat due to their cell wall composition. In this study, the natural antimicrobial agents ε-poly-L-lysine (ε-PL) and isoeugenol were tested individually and in combinations for their activities against a selection of Gram-negative strains in vitro. All combinations resulted in additive interactions between ε-PL and isoeugenol towards the bacteria tested. The killing efficiency of different ratios of the two antimicrobial agents was further evaluated in vitro against Pseudomonas putida. Subsequently, the most efficient ratio was applied to a raw turkey meat model system which was incubated for 96 h at spoilage temperature. Half of the samples were challenged with P. putida, and the bacterial load and microbial community composition was followed over time. CFU counts revealed that the antimicrobial blend was able to lower the amount of viable Pseudomonas spp. by one log compared to untreated samples of challenged turkey meat, while the single compounds had no effect on the population. However, the compounds had no effect on Pseudomonas spp. CFU in unchallenged meat. Next-generation sequencing offered culture-independent insight into population diversity and changes in microbial composition of the meat during spoilage and in response to antimicrobial treatment. Spoilage of unchallenged turkey meat resulted in decreasing species diversity over time, regardless of whether the samples received antimicrobial treatment. The microbiota composition of untreated unchallenged meat progressed from a Pseudomonas spp. to a Pseudomonas spp., Photobacterium spp., and Brochothrix thermosphacta dominated food matrix on the expense of low abundance species. We observed a similar shift among the dominant species in meat treated with ε-PL or the antimicrobial blend, but the samples differed markedly in the composition of less abundant species. In contrast, the overall species diversity was constant during incubation of turkey meat challenged with P. putida although the microbiota composition did change over time. Untreated or ε-PL treated samples progressed from a Pseudomonas spp. to a Pseudomonas spp. and Enterobacteriaceae dominated food matrix, while treatment with the antimicrobial blend resulted in increased relative abundance of Hafnia spp., Enterococcaceae, and Photobacterium spp. We conclude that the blend delayed the onset of spoilage of challenged meat, and that all antimicrobial treatments of unchallenged or challenged meat affect the progression of the microbial community composition. Our study confirms that the antimicrobial effects observed in vitro can be extrapolated to a food matrix such as turkey meat. However, it also underlines the consequence of species-to-species variation in susceptibility to antimicrobials, namely that the microbial community change while the CFU remains the same. Addition of antimicrobials may thus prevent the growth of some microorganisms, allowing others to proliferate in their place., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Anti-cancer drugs are discussed as high risk substances in regard to human health and considered as problematic for the environment. They are of potential environmental relevance due to their poor biodegradability and toxicological properties. Methotrexate (MTX) is an antimetabolite that was introduced in the pharmaceutical market in the 40's and still today is one of the most consumed cytotoxic compounds around the world. In the present study MTX was only partially biodegraded in the closed bottle test (CBT). Therefore, it was submitted to three different advanced oxidation processes (AOPs): UV/H2O2, UV/Fe(2+)/H2O2 and UV/TiO2. The irradiation was carried out with a Hg medium-pressure lamp during 256min whereas the analytical monitoring was done through LC-UV-MS/MS and DOC analysis. MTX was easily removed in all the irradiation experiments, while the highest mineralization values and rates were achieved by the UV/Fe(2+)/H2O2 treatment. The lowest resulted from the UV/H2O2 reactions. The UV/H2O2 treatment resulted in little biodegradable transformation products (TPs). However, the same treatment resulted in a reduction of the toxicity of MTX by forming less toxic TPs. Analysis by LC-UV-MS/MS revealed the existence of nine TPs formed during the photo-catalytic treatments. The pH of the solutions decreased from 6.4 (t 0min) to 5.15 in the UV/H2O2 and from 6.4 (t 0min) to 5.9 in the UV/TiO2 at the end of the experiments. The initial pH of the UV/Fe(2+)/H2O2 experiments was adjusted to 5 and after the addition of H2O2 the pH decreased to around 3 and remained in this range until the end of the treatments., (Copyright © 2015 Elsevier Ltd. All rights reserved.)