Simultaneous degradation of triazophos, methamidophos and carbofuran pesticides in wastewater using an Enterobacter bacterial bioreactor and analysis of toxicity and biosafety.

Triazophos (TAP), methamidophos (MAP) and carbofuran (CF) pesticides are highly toxic, soluble and absorbable. Efficient co-degradation of multi-pesticides is rare reported. The objectives of this study were to investigate TAP, MAP and CF co-degradative ability of Enterobacter sp. Z1 and study the degradation mechanisms. Strain Z1 was shown to efficiently co-degrade TAP, MAP and CF when they were used as primary carbon sources. The degradation occurred over a wide range of temperatures, pH values and pesticide concentrations and followed first-order kinetics. Under the optimum conditions (37 °C, pH 7 and 100 mg/L of each pesticide), the degradation efficiencies were 100%, 100%, and 95.3% for TAP, MAP and CF, respectively. In addition, strain Z1 could simultaneously degrade TAP, MAP, CF and total nitrogen in wastewater in a batch bioreactor, with high removal efficiencies of 98.3%, 100%, 98.7% and 100%, respectively. Genomics, proteomics, qRT-PCR and gene overexpression analyses revealed that the degradation mechanisms involved the activities of multiple proteins, among which, organophosphorus hydrolase (Oph) and 3-hydroxyacyl-CoA dehydrogenase (PaaC) are primarily responsible for TAP and MAP degradation, while carbofuran hydrolase (Mcd) and amidohydrolase (RamA) primarily degrade CF. Among these enzymes, PaaC and RamA are newly identified pesticide-degrading enzymes. Toxicity assays of strain Z1 using reporter recombinase gene (recA) and zebrafish showed that there was no accumulation of toxic metabolites during the degradation process. Biosafety test using zebrafish showed that the strain was nontoxic toward zebrafish. Strain Z1 provides a good purification effect for pesticides-containing wastewater and novel microbial pesticide-degrading mechanisms were discovered.

NAD(P)H-dependent thioredoxin-disulfide reductase TrxR is essential for tellurite and selenite reduction and resistance in Bacillus sp. Y3.

Microbial reduction of selenite [Se(IV)] and tellurite [Te(IV)] to Se(0) and Te(0) can function as a detoxification mechanism and serve in energy conservation. In this study, Bacillus sp. Y3 was isolated and demonstrated to have an ability of simultaneous reduction of Se(IV) and Te(IV) during aerobic cultivation, with reduction efficiencies of 100% and 90%, respectively. Proteomics analysis revealed that the putative thioredoxin disulfide reductase (TrxR) and sulfate and energy metabolic pathway proteins were significantly upregulated after the addition of Se(IV) and Te(IV). qRT-PCR also showed an increased trxR transcription level in the presence of Se(IV) and Te(IV). Compared with a wild-type Escherichia coli strain, the TrxR-overexpressed E. coli strain showed higher Se(IV) and Te(IV) resistance levels and reduction efficiencies. Additionally, the TrxR showed in vitro Se(IV) and Te(IV) reduction activities when NADPH or NADH were present. When NADPH was used as the electron donor, the optimum conditions for enzyme activities were pH 8.0 and 37°C. The Km values of Te(IV) and Se(IV) were 16.31 and 2.91 mM, and the Vmax values of Te(IV) and Se(IV) were 12.23 and 11.20 µM min-1 mg-1, respectively. The discovery of the new reductive enzyme TrxR enriches the repertoire of the bacterial Se(IV) and Te(IV) resistance and reduction mechanisms. Bacillus sp. Y3 can efficiently reduce Se(IV) and Te(IV) simultaneously. Strain Y3 provides potential applications for selenite and tellurite bioremediation. The TrxR enzyme shows high catalytic activity for reducing Se(IV) and Te(IV). The discovery of TrxR enriches the bacterial Se(IV) and Te(IV) reduction mechanisms.

Sphingomonas montanisoli sp. nov., isolated from mountain soil.

A soil bacterium, designated ZX9611T, was isolated from Taihang Mountain in Henan province, PR China. The strain was Gram-stain-negative and strictly aerobic. The cells were motile, rod-shaped and formed light pink-colored colonies. The 16S rRNA gene sequence of ZX9611T shared the highest similarities with those of Sphingomonas crocodyli CCP-7T (97.0%), Sphingomonas jatrophae S5-249T (96.6%) and Sphingomonas starnbergensis 382T (95.9%). Phylogenetic analyses based on 16S rRNA gene sequences demonstrated that ZX9611T clustered with S. crocodyli CCP-7T, S. jatrophae S5-249T and S. starnbergensis 382T. The average nucleotide identity (ANI) values between ZX9611T and two type strains (S. crocodyli BCRC 81096T and S. jatrophae DSM 27345T) were 88.3 and 68.6% respectively. ZX9611T exhibited genome-sequence-based digital DNA-DNA hybridization (dDDH) values of 53.3 % and 15.3 %, compared with S. crocodyli BCRC 81096T and S. jatrophae DSM 27345T, respectively. ZX9611T had a genome size of 4.12 Mb and an average DNA G+C content of 64.8 %. ZX9611T had major fatty acids (>5 %) including summed feature 8 (C18 : 1 ω7c and/or C18 : 1 ω6c), C14 : 0 2-OH, C16 : 0 and summed feature 3 (C16 : 1 ω7c and/or C16 : 1 ω6c), and the major polyamine was sym-homospermidine. The only respiratory quinone was ubiquinone-10. The polar lipids were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, phosphatidylcholine and sphingoglycolipid. On the basis of phenotypic, chemotaxonomic and phylogenetic characteristics, strain ZX9611T represents a novel species of genus Sphingomonas, for which the name Sphingomonas montanisoli sp. nov. is proposed. The type strain is ZX9611T (=KCTC 72622T=CCTCC AB 2019350T).

Suppression of the long non-coding RNA MALAT-1 impairs the growth and migration of human tongue squamous cell carcinoma SCC4 cells.

INTRODUCTION: Aberrant expression of long non-coding RNAs (lncRNAs) is associated with metastasis and poor prognosis in patients with various cancer types. However, few studies have assessed lncRNAs in oral squamous cell carcinoma (OSCC). This study aimed to investigate the expression and impact of lncRNAs in OSCC. MATERIAL AND METHODS: Real-time PCR analysis was used to examine the expression of four lncRNAs, MALAT-1, UCA1, BC200 and SRA, in 14 OSCC and adjacent normal tissue pairs. The impact of MALAT-1 suppression by siRNA on the proliferation, apoptosis, anchorage-independent growth and migration of the human tongue carcinoma cell line SSC4 was also determined. RESULTS: MALAT-1 levels were significantly higher in the OSCC tissue than in the normal tissues (p < 0.004); no significant differences in UCA1, BC200 or SRA RNA levels were observed. Knockdown of MALAT-1 by siRNA significantly suppressed proliferation of SSC4 cells (p < 0.004) and enhanced their apoptosis (p < 0.001). In addition, siRNA-mediated suppression of MALAT-1 inhibited SSC4 cell colony formation (p < 0.001) and migration (p < 0.004). CONCLUSIONS: Elevated expression of MALAT-1 in OSCC may play a role in tumorigenesis and/or metastasis. Further studies are necessary to identify the mechanism by which MALAT-1 influences SCC4 growth and migration and validate its increased expression in OSCC patients.

Cooperation between two strains of Enterobacter and Klebsiella in the simultaneous nitrogen removal and phosphate accumulation processes.

Two strains, Enterobacter sp. Z1 and Klebsiella sp. Z2, were exhibited great capacities for heterotrophic nitrification-aerobic denitrification (HNAD) and intracellular phosphate accumulation. Strikingly, the co-cultured strains enhanced the removal efficiency of total nitrogen and phosphate, with removal efficiencies of ammonia, nitrate, nitrite and soluble phosphate of 99.64%, 99.85%, 96.94% and 66.7% respectively. Furthermore, high removal efficiencies from wastewaters with high concentrations of ammonia (over 1000 mg/L) were achieved by inoculation with the co-strains, which left residual ammonia of less than 1 mg/L within 10 h. To elucidate the mechanism of HNAD in co-strains, quantitative PCR was carried out to examine the expression levels of hydroxylamine oxidase (Hao), nitrate reductase (NapA and NarG), nitrite reductase (NirS) and polyphosphate kinase (Ppk), and the results showed that the napA2, narG and ppk genes in the strains were significantly upregulated under the co-cultured conditions and provided an explanation for the nitrogen and phosphate removal.

Interaction between MALAT-1, CCR7 and correlated genes in oral squamous cell carcinoma.

OBJECTIVE: This study focuses on the feasible molecular mechanism of the interaction between MALAT-1, CCR7 and related genes in oral squamous cell carcinoma, to find new target molecules that can block the lymph node metastasis. METHODS: The expression of MALAT-1, miRNA-320s, SRSF1, YB-1 and CCR7 were detected in T3/T4-phase OSCC tissues of two groups with or without lymph node metastasis using real-time qPCR. CO-IP and western blot to test the interaction of RNAs (MALAT-1, miRNA-320s) with SRSF1 protein or YB-1 were evaluated by CO-IP, Western blot and real-time qPCR. The expression change of chemokine receptor CCR7 were investigated using CO-IP, Western blot and real-time qPCR after silencing miRNA-320d (one of the miRNA-320s family members) by transfection of miRNA mimics to explore related signaling pathway. RESULTS: The expression levels of MALAT-1 SRSF1 and CCR7 in OSCC tissues with were differentially higher compared with those of samples without lymph node metastasis as well as para-carcinoma tissues, exclusive of miRNA-320d. Moreover, it is confirmed that the target RNA (MALAT-1, miRNA-320s) and SRSF1 protein can combine with each other, based on the statistically significant difference compared with negative control group (P<0.05). In addition, the expression of CCR7 was higher than the negative control group after silencing miRNA-320d. CONCLUSION: SRSF1 is likely to mediate the interactive relationship between MALAT-1 and miRNA-320d. CCR7 expression can be distinctly increased by silencing miRNA-320d. The effect of long-chain non-coding RNA MALAT-1 on chemokine receptor CCR7 and possibly further influence on lymph node metastasis of oral squamous cell carcinoma are revealed in molecular level to offer help for prevention and treatment of OSCC in future.

Removal of fluorescent dissolved organic matter in biologically treated textile effluents by NDMP anion exchange process: efficiency and mechanism.

The efficiency and mechanism of anion exchange resin Nanda Magnetic Polymer (NDMP) for removal of fluorescent dissolved organic matter in biologically treated textile effluents were studied. The bench-scale experiments showed that as well as activated carbon, anion exchange resin could efficiently remove both aniline-like and humic-like fluorescent components, which can be up to 40 % of dissolved organic matter. The humic-like fluorescent component HS-Em460-Ex3 was more hydrophilic than HS-Em430-Ex2 and contained fewer alkyl chains but more acid groups. As a result, HS-Em460-Ex3 was eliminated more preferentially by NDMP anion exchange. However, compared with adsorption resins, the polarity of fluorescent components had a relatively small effect on the performance of anion exchange resin. The long-term pilot-scale experiments showed that the NDMP anion exchange process could remove approximately 30 % of the chemical oxygen demand and about 90 % of color from the biologically treated textile effluents. Once the issue of waste brine from resin desorption is solved, the NDMP anion exchange process could be a promising alternative for the advanced treatment of textile effluents.

Characterization of fluorescent-dissolved organic matter and identification of specific fluorophores in textile effluents.

This study focused on the characterization of fluorescent-dissolved organic matter and identification of specific fluorophores in textile effluents. Samples from different textile wastewater treatment plants were characterized by high-performance liquid chromatography and size exclusion chromatography as well as fluorescence excitation-emission matrix spectra. Despite the highly heterogeneous textile effluents, the fluorescent components and their physicochemical properties were found relatively invariable, which is beneficial for the combination of biological and physicochemical treatment processes. The humic-like substance with triple-excitation peaks (excitation (Ex) 250, 310, 365/emission (Em) 460 nm) presented as the specific fluorescence indicator in textile effluents. It was also the major contributor to UV absorbance at 254 nm and resulted in the brown color of biologically treated textile effluents. By spectral comparison, the specific fluorophore in textile effluents could be attributed to the intermediate structure of azo dyes 1-amino-2-naphthol, which was transferred into the special humic-like substances during biological treatment.

Characterization of dissolved organic matter in municipal wastewater using fluorescence PARAFAC analysis and chromatography multi-excitation/emission scan: a comparative study.

Dissolved organic matter (DOM) in municipal wastewater was mainly characterized using high-performance liquid chromatography (HPLC) and size exclusion chromatography (HPSEC) with multi-excitation/emission fluorescence scan. Meanwhile, fluorescence excitation-emission-matrix combined with parallel factor analysis (EEM-PARAFAC) was also applied. Compared with chromatography fluorescence fingerprints, the EEM-PARAFAC model could not reflect the variety of DOM species with similar fluorescence but different physicochemical properties. The chromatography results showed that the protein-like species were variable among different municipal wastewater treatment plants, some of which are in combination with humic-like species; while there were two major humic-like species fractionated by hydrophilicity and molecular weight (MW), which are also the major contributors to UV absorbance at 254 nm. It was also identified that the relatively hydrophilic humic fractions were slightly larger than the relatively hydrophobic humic fractions. In all the investigated wastewater treatment plants, the relatively hydrophilic and larger MW humic fraction mainly contributed to the fluorescence intensity of humic-like EEM-PARAFAC components. As well as facilitating interpretations of EEM-PARAFAC components, the HPLC/HPSEC fluorescence fingerprints also contributed to a better understanding of fluorescent DOM species in municipal wastewater.

HPLC/HPSEC-FLD with multi-excitation/emission scan for EEM interpretation and dissolved organic matter analysis.

The need to track and characterize dissolved organic matter (DOM) has made fluorescence excitation-emission matrix (EEM) spectroscopy extensively used. In this work, reverse phase high-performance liquid chromatography (RP-HPLC) and high-performance size exclusion chromatography (HPSEC) with fluorescence detector (FLD) were used for EEM interpretation and DOM analysis. Given that fluorescence detectors can scan with multi-excitation or multi-emission mode, HPLC-FLD with multi-excitation scan directly verified the prevalence of multi-peak fluorophores in EEM, which provides a corrective insight for the current fluorescence regional integration (FRI) methods; whereas HPLC-FLD with multi-emission scan provided more informative fluorescence fingerprints for identification of DOM species, which is a chromatographic surrogate for determining the proper number of PARAFAC components. Besides providing a deep insight for the current EEM interpretation, the HPLC/HPSEC-FLD results also directly related physiochemical properties to DOM species, including polarity and molecular weight (MW) distribution, which is helpful for further characterization their behavior in water and wastewater treatment process. A chromatography technique with multi-excitation and multi-emission fluorescence scan can be an informative method for EEM interpretation and DOM identification and characterization.