The present study has used soil samples from Nigeria, contaminated with Brass crude-oil, to determine its biodegradation through enhanced biostimulation with cow dung and periodic aeration. Over a period of twenty-eight days, the hydrocarbon-utilizing bacteria (HUB) and hydrocarbon-utilizing fungi (HUF) have been counted and identified. Results from biodegradation of the brass crude-oil over the aforementioned period show that amended crude-oil-spiked soil has had 54.82% degradation while for amendment and periodic turning this has been 55.90%, not significantly higher than the former at p≤0.05. Also degradation of spiked soil without cow dung amendment has been 16.13%. The identified HUB are Staphylococcus aureus, Pseudomonas aeruginosa, Bacillus subtilis, and Streptococcus thermophillus, with individual occurrence of 18.52% as well as Proteus vulgaris and Micrococcus luteus with 11.11% and 14.81% occurrence, respectively. Also, the occurrence rate of HUF like Aspergillus flavus, A. niger, Penicillium chrysogenum, Trichothecium roseum, and Penicillium citrinum have been 15.63% each;whilefor Alternaria alternata and Neurospora crazza it has been 6.25% and for Saccharomyces cerevisae and A. fumigatus, 9.38%and3.13%, respectively. The study concludes that amendment with cow dung and periodic turning of the soil enhance degradation of Brass crude-oil significantly. What is more, aeration by periodic turning slightly improves degradation only with cow dung treatment on Days 21 and 28.

Petroleum hydrocarbon contamination is a major global prevalent issue in the petroleum sector. This research focuses on evaluating biodegradation of three Gram-negative bacilli, isolated from cowpea planted soil, contaminated with kerosene. The Gram negative bacilli strains have been characterized and identified, using MicrobactTM ID24E systems for the identification of Enterobacteriaceae and common Miscellaneous Gram-Negative Bacilli (MGNB). The identified organisms include Aeromonas hydrophila, Vibrio parahaemolyticus, and Actinobacillus sp. with the biodegradation indices, monitored for the evaluation of their degrading abilities, being Optical density at 600 nm (OD600nm), pH, and emulsification stability. The chemical profile of single cultures and mixed cultures (consortia) on the jet fuel hydrocarbon has been determined by means of Gas Chromatography Mass Spectrometry (GC-MS), the results of which indicate that all the isolates have undergone above 70% reduction of the hydrocarbon substrates in terms of residual compounds. There has been 48 hydrocarbon compounds in the undegraded jet fuel which, following degradation process, decrease to 5, 13, 7, 10, 6, 9, and 10 compounds for Aeromonas hydrophila, Vibrio parahaemolyticus, Actinobacillus sp., Aeromonas hydrophila and Vibrio parahaemolyticus, Aeromonas hydrophila and Actinobacillus sp., Vibrio parahaemolyticus and Actinobacillus sp., Aeromonas hydrophila, Vibrio parahaemolyticus,and Actinobacillus sp., respectively. The degradation efficiency of the isolates have been relatively high and comparable to the control. Results from this study indicate that all the strains, especially the consortia, are potential candidates for remediating the problem of hydrocarbon contamination in the environment.

International audience | Microplastics (<5 mm) exhibit intrinsic features such as density, hydrophobic surface, or high surface/volume ratio, that are known to promote microbial colonization and biofilm formation in marine ecosystems. Yet, a relatively low number of studies have investigated the nature of microplastic associated bacterial communities in coastal ecosystems and the potential factors influencing their composition and structure. Here, we characterized microplastics collected in the Bay of Brest by manual sorting followed by Raman spectroscopy and studied their associated bacterial assemblages using 16S amplicon high-throughput sequencing. Our methodology allowed discriminating polymer type (polyethylene, polypropylene and polystyrene) within small size ranges (0.3-1 vs. 1-2 vs. 2-5 mm) of microplastics collected. Data showed high species richness and diversity on microplastics compared to surrounding seawater samples encompassing both free living and particle attached bacteria. Even though a high proportion of operational taxonomic units (OTU; 94 ± 4%) was shared among all plastic polymers, polystyrene fragments exhibited distinct bacterial assemblages as compared to polyethylene and polypropylene samples. No effect of microplastic size was revealed regardless of polymer type, site and date of collection. The Vibrio genus was commonly detected in the microplastic fraction and specific PCR were performed to determine the presence of potentially pathogenic Vibrio strains (namely V. aestuarianus and the V. splendidus polyphyletic group). V. splendidus related species harboring

Florfenicol is one of the most-used antimicrobial agents in global fish farming. Nevertheless, in most countries, its use is not conducted in accordance with good practices. The aim of this work was to evaluate the leaching of florfenicol from coated fish feed into the water. Analytical methods were developed and validated for the quantitation of florfenicol in medicated feed and water by UHPLC-MS/MS. Florfenicol residues in the water were quantified after 5- and 15-min exposures of the medicated feed in the water at 22 and 28 °C and at pH 4.5 and 8.0. The influence of pellet size and three coating agents (vegetable oil, carboxymethylcellulose, and low-methoxylated pectin) on the leaching of the drug was also assessed. Pellet size, coating agent, water temperature, and time of exposure significantly (p < 0.05) affected florfenicol leaching, while water pH did not interfere with the leaching. Coating with vegetable oil was the most efficient method to reduce florfenicol leaching, while coating with carboxymethylcellulose presented the highest leaching (approximately 60% after 15 min at 28 °C). Thus, the coating agent has a significant effect on the florfenicol leaching rate and, consequently, on the necessary dose of the drug to be administered. Moreover, it is worth mentioning that higher florfenicol leaching will pose a greater risk to environmental health, specifically in terms of the development of bacteria resistant to florfenicol. Additional studies are needed with other polymers and veterinary drugs used in medicated feed for fish farming.

Methylisothiazolinone (MIT) has been widely used to control bacterial growth in reverse osmosis (RO) systems. However, MIT's toxicity on microalgae should be determined because residual MIT is concentrated into RO concentrate (ROC) and might have a severe impact on microalgae-based ROC treatment. This study investigated the tolerance of Scenedesmus sp. LX1 to MIT and revealed the mechanism of algal growth inhibition and toxicity resistance. Scenedesmus sp. LX1 was inhibited by MIT with a half-maximal effective concentration at 72 h (72 h-EC50) of 1.00 mg/L, but the strain recovered from the inhibition when its growth was not completely inhibited. It was observed that this inhibition's effect on subsequent growth was weak, and the removal of MIT was the primary reason for the recovery. Properly increasing the initial algal density significantly shortened the adaptation time for accelerated recovery in a MIT-containing culture. Photosynthesis damage by MIT was one of the primary reasons for growth inhibition, but microalgal cell respiration and adenosine triphosphate (ATP) synthesis were not completely inhibited, and the algae were still alive even when growth was completely inhibited, which was notably different from observations made with bacteria and fungi. The algae synthesized more chlorophyll, antioxidant enzymes of superoxide dismutase (SOD) and catalase (CAT), and small molecules, such as reduced glutathione (GSH), to resist MIT poisoning. The microalgae-based process could treat the MIT-containing ROC, since MIT was added for only several hours a week in municipal wastewater reclamation RO processes, and the MIT average concentration was considerably lower than the maximum concentration that algae could tolerate.

The phenylurea herbicide, linuron (LIN), is used to control various types of weeds. Despite its efficient role in controlling weeds, it presents a persistent problem to the environment. In the current study, phytoremediation properties of transgenic CYP1A2 Arabidopsis thaliana plants to LIN were assessed. CYP1A2 gene was firstly cloned and expressed in bacteria before proceeding to plants. In presence of LIN, The growth of CYP1A2 expressing bacteria was superior compared to control bacteria transformed with the empty bacterial expression vector pET22b(+). No clear morphological changes were detected on CYP1A2 transgenic plants. However, significant resistance to LIN herbicide application either via spraying the foliar parts of the plant or via supplementation of the herbicide in the growth medium was observed for CYP1A2 transformants. Plant growth assays under LIN stress provide strong evidence for the enhanced capacity of transgenic lines to grow and to tolerate high concentrations of LIN compared to control plants. HPLC analyses showed that detoxification of LIN by bacterial extracts and/or transgenic plant leaves is improved as compared to the corresponding controls. Our data indicate that over expression of the human CYP1A2 gene increases the phytoremediation capacity and tolerance of Arabidopsis thaliana plants to the phenylurea herbicide linuron.

Nitrous oxide (N₂O) is a potent greenhouse gas and tends to accumulate as an intermediate in the process of bacteria denitrification. To achieve complete reduction of nitrogen oxide (NOₓ) in bacteria denitrification, the structural gene nosZ encoding nitrous oxide reductase (N₂OR) was cloned from Alcaligenes denitrificans strain TB (GenBank JQ044686). The recombinant plasmid containing the nosZ gene was built, and the expression of nosZ gene in Escherichia coli was determined. Results show that the nosZ gene consisting of 1917 nucleotides achieves heterologous expression successfully by codon optimization strategy under optimal conditions (pre-induction inoculum OD₆₀₀ of 0.67, final IPTG concentration of 0.5 mM, inducing time of 6 h, and inducing temperature of 28 °C). Determination result of gas chromatography confirms that N₂O degradation efficiency of recombinant E. coli is strengthened by at least 1.92 times compared with that of original strain TB when treated with N₂O as substrate. Moreover, N₂OR activity in recombinant strain is 2.09 times higher than that in wild strain TB, which validates the aforementioned result and implies that the recombinant E. coli BL21 (DE3)-pET28b-nosZ is a potential candidate to control N₂O accumulation and alleviate greenhouse effect. In addition, the N₂OR structure and the possible N₂O binding site in Alcaligenes sp. TB are predicted, which open an avenue for further research on the relationship between N₂OR activity and its structure.

Inhibition of reductive transformation of arsenic (As) in flooded paddy soils is of fundamental importance for mitigating As transfer into food chain. Anaerobic arsenite (As(III)) oxidizers maintain As in less mobile fraction under nitrate-reducing conditions. In this study, we explored the dynamic profile of As speciation in porewater and As distribution among the pools of differential bioavailability in soil solid phase with and without nitrate treatment. In parallel, the abundance and diversity of As(III) oxidase gene (aioA) in flooded paddy soil with nitrate amendment was examined by quantitative PCR and aioA gene clone library. Furthermore, the impact of nitrate on As accumulation and speciation in rice seedlings was unraveled. With nitrate addition (25 mmol NO₃⁻ kg⁻¹ soil), porewater As(III) was maintained at a consistently negligible concentration in the flooded paddy soil and the reductive dissolution of As-bearing Fe oxides/hydroxides was significantly restrained. Specifically, nitrate amendment kept 81% of total soil As in the nonlabile fraction with arsenate (As(V)) dominating after 30 days of flooding, compared to only 61% in the unamended control. Nitrate treatment induced 4-fold higher abundance of aioA gene, which belonged to domains of bacteria and archaea under the classes α-Proteobacteria (6%), ß-Proteobacteria (90%), ɣ-Proteobacteria (2%), and Thermoprotei (2%). By nitrate addition, As accumulation in rice seedlings was decreased by 85% with simultaneously elevated As(V) ratio in rice plant relative to control after 22 days of growth under flooded conditions. These results highlight that nitrate application can serve an efficient method to inhibit reductive dissolution of As in flooded paddy soils, and hence diminish As uptake by rice under anaerobic growing conditions.

In respect to direct and indirect water reuse, the microbiological quality of treated wastewater is highly important. Conventional wastewater treatment plants are normally not equipped with advanced technologies for the elimination of bacteria. Molecular biology analyses were combined with live-dead discrimination analysis of wastewater population using Propidium monoazide (PMA) to study population shifts during ozonation (1 g ozone/g DOC) at a municipal wastewater treatment plant. Escherichia coli, enterococci, and Pseudomonas aeruginosa were quantified by polymerase chain reaction (qPCR) and the whole wastewater population was analyzed by metagenomic sequencing. The PMA-qPCR experiments showed that the abundances of P. aeruginosa didn't change by ozone treatment, whereas a reduction was observed for E. coli and enterococci. Results comparing conventional cultivation experiments with PMA-qPCR underlined the presence of viable but not culturable cells (VBNC) and their regrowth potential after ozone treatment. Illumina HiSeq sequencing results with and without PMA treatment demonstrated high population similarities in water samples originating from ozone inflow sampling sides. Upon using PMA treatment after ozonation, population shifts became visible and also underlined the importance of PMA treatment for the evaluation of elimination and selection processes during ozonation at WWTPs. Amongst a number of 14 most abundant genera identified in the inflow samples, 9 genera were found to be reduced, whereas 4 genera increased in relative abundance and 1 genus almost remained constant. The strongest increase in relative abundance after ozonation was detected for Oscillatoria spp., Microcoleus spp. and Nitrospira spp. Beside this, a continuous release of Pseudomonas spp. (including P. aeruginosa) to the downstream receiving body was confirmed. Regrowth experiments demonstrated a high prevalence of P. aeruginosa as part of the surviving bacterial population. Summing up, molecular biology analyses in combination with live-dead discrimination are comprehensive methods to evaluate the elimination processes targeting specific species and/or whole microbial populations.

The tidal flats near Sinduri beach in Taean, Korea, have been severely contaminated by heavy crude oils due to the Korea's worst oil spill accident, say the Hebei Spirit Oil Spill, in 2007. Crude oil compounds, including polycyclic aromatic hydrocarbons (PAHs), pose significant environmental damages due to their wide distribution, persistence, high toxicity, mutagenicity, and carcinogenicity. Microbial community of Sinduri beach sediments samples was analyzed by metagenomic data with 16S rRNA gene amplicons. Three phyla (Proteobacteria, Firmicutes, and Bacteroidetes) accounted for approximately ≥93.0% of the total phyla based on metagenomic analysis. Proteobacteria was the dominant phylum in Sinduri beach sediments. Cultivable bacteria were isolated from PAH-enriched cultures, and bacterial diversity was investigated through performing culture characterization followed by molecular biology methods. Sixty-seven isolates were obtained, comprising representatives of Actinobacteria, Firmicutes, α- and γ-Proteobacteria, and Bacteroidetes. PAH catabolism genes, such as naphthalene dioxygenase (NDO) and aromatic ring hydroxylating dioxygenase (ARHDO), were used as genetic markers to assess biodegradation of PAHs in the cultivable bacteria. The ability to degrade PAHs was demonstrated by monitoring the removal of PAHs using a gas chromatography mass spectrometer. Overall, various PAH-degrading bacteria were widely present in Sinduri beach sediments and generally reflected the restored microbial community. Among them, Cobetia marina, Rhodococcus soli, and Pseudoalteromonas agarivorans were found to be significant in degradation of PAHs. This large collection of PAH-degrading strains represents a valuable resource for studies investigating mechanisms of PAH degradation and bioremediation in oil contaminated coastal environment, elsewhere.