Denitrificimonas halotolerans sp. nov., a novel species isolated from UASB sludge treating landfill leachate.

A Gram-stain-negative, facultative anaerobe, rod-shaped strain JX-1T was isolated from UASB sludge treating landfill leachate in Wuhan, China. The isolate is capable of growing under conditions of pH 6.0-11.0 (optimum, pH 7.0-8.0), temperature 4-42 â„ƒ (optimum, 20-30 â„ƒ), 0-8.0% (w/v) NaCl (optimum, 5.0%), and ammonia nitrogen concentration of 200-5000 mg/L (optimum, 500 mg/L) on LB plates. The microorganism can utilize malic acid, D-galactose, L-rhamnose, inosine, and L-glutamic acid as carbon sources, but does not reduce nitrates and nitrites. The major fatty acids are C18:1ω7c/C18:1ω6c, iso-C15:0, and anteiso-C15:0. The respiratory quinones are Q9 (91.92%) and Q8 (8.08%). Polar lipids include aminolipid, aminophospholipid, diphosphatidylglycerol, glycolipid, phosphatidylethanolamine, phosphatidylglycerol, and phospholipid. Compared with other strains, strain JX-1T and Denitrificimonas caeni HY-14T have the highest values in terms of 16S rRNA gene sequence similarity (96.79%), average nucleotide identity (ANI; 76.06%), and average amino acid identity (AAI; 78.89%). Its digital DNA-DNA hybridization (dDDH) result is 20.3%. The genome of strain JX-1T, with a size of 2.78 Mb and 46.12 mol% G + C content, lacks genes for denitrification and dissimilatory nitrate reduction to ammonium (DNRA), but contains genes for ectoine synthesis as a secondary metabolite. The results of this polyphasic study allow genotypic and phenotypic differentiation of the analysed strain from the closest related species and confirm that the strain represents a novel species within the genus Denitrificimonas, for which the name Denitrificimonas halotolerans sp. nov. is proposed with JX-1T (= MCCC 1K08958T = KCTC 8395T) as the type strain.

Parachitinimonas caeni gen. nov., sp. nov., a novel member of family Burkholderiaceae isolated from activated sludge collected in Shenzhen, PR China.

A Gram-stain-negative, strictly aerobic and filamentous bacterial strain, designated as DQS-5T, was isolated from the activated sludge of a municipal sewage treatment plant in Shenzhen, PR China. Optimal growth was observed at 28 °C and pH 7.5. Catalase and oxidase activities were detected. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain DQS-5T was most closely related to the genera Chitinimonas and Chitinivorax (91.0-93.4 % and 92.5 % 16S rRNA gene sequence similarity, respectively) and was close to the member of the family Burkholderiaceae. The complete genome sequence of strain DQS-5T contains 5 653 844 bp and 57.3 mol% G+C. The average nucleotide identity, digital DNA-DNA hybridization and average amino acid identity values between the genome of strain DQS-5T and those of its close relatives were 75.9-77.2, 19.0-20.3 and 57.2-61.8 %, respectively. Chemotaxonomic analysis of strain DQS-5T indicated that the sole respiratory quinone was ubiquinone-8, the predominant cellular fatty acids were C16 : 0 and summed feature 3 (C16 : 1 ω7c and/or C16 : 1 ω6c), and the major polar lipids consisted of phosphatidylethanolamine, phosphatidylglycerol, aminophospholipid and aminolipid. The phylogenetic, genotypic, phenotypic and chemotaxonomic data demonstrate that strain DQS-5T represents a novel species in a novel genus within the family Burkholderiaceae, for which the name Parachitinimonas caeni gen. nov., sp. nov., is proposed. Strain DQS-5T (=KCTC 92788T=CCTCC AB 2022320T) is the type and only strain of P. caeni.

Tessaracoccus caeni sp. nov., a novel member of Propionibacteriaceae isolated from activated sludge in Hefei, PR China.

A floc-forming bacterial strain, designated HF-7T, was isolated from the activated sludge of an industrial wastewater treatment plant in Hefei, PR China. Cells of this strain were Gram-stain-positive, catalase- and oxidase-negative, facultatively anaerobic, and rod-shaped. Growth occurred at 20-42 °C (optimum, 28 °C), at pH 5.5-10.5 (optimum, pH 7.5) and with 0-8.0 % (w/v) NaCl (optimum, 1 %). The major fatty acid was anteiso-C15 : 0. The polar lipid profile contained phosphatidylglycerol, diphosphatidylglycerol and phosphatidylinositol. The DNA G+C content was 67 mol% from whole genomic sequence analysis. Based on the results of 16S rRNA gene sequence analysis, this strain should be assigned to the genus Tessaracoccus and is closely related to Tessaracoccus arenae CAU 1319T (95.87 % similarity), Tessaracoccus lapidicaptus IPBSL-7T (95.19 %) and Tessaracoccus bendigoensis Ben 106T (94.63 %) but separated from them by large distances in different phylogenetic trees. Based on whole genome analysis, the orthologous average nucleotide identity and in silico DNA-DNA hybridization values against two of the closest relatives were 75.21-76.50 % and 14.2-24.4 %, respectively. The phylogenetic, genotypic, phenotypic and chemotaxonomic data demonstrated that strain HF-7T could be distinguished from its phylogenetically related species and represents a novel species within the genus Tessaracoccus, for which the name Tessaracoccus caeni sp. nov. is proposed. The type strain is HF-7T (=KCTC 49959T=CCTCC AB 2023019T).

Effects of profenofos on the growth, reproduction, behavior, and gene transcription of Daphnia magna.

Profenofos (PFF) is an organophosphorus pesticide frequently detected in surface waters, soil habitats, and even biota. Some studies have demonstrated the potential risks of PFF to aquatic organisms. However, most of these studies were focused on its acute rather than chronic impacts, and the subjects are usually large vertebrates. Here, we treated D. magna (< 24 h) with PFF at doses of 0, 0.07, 0.28, and 1.12 mg/L for 21 days to study its long-term toxic impacts. Exposure to PFF largely decreased the survival rate and inhibited the growth and reproduction of D. magna. Then, PCR arrays were used to evaluate the changes in the expression of 13 genes related to growth, reproduction, and swimming behavior. The results revealed that the expression of several genes was dramatically changed by exposure to each dose of PFF, which might be responsible for the observed toxic effects of PFF. In summary, our findings imply that long-term PFF exposure can be highly hazardous to the growth, development, and reproduction of D. magna.

Greater toxic potency of bisphenol AF than bisphenol A in growth, reproduction, and transcription of genes in Daphnia magna.

Limited studies were performed to evaluate the effects of bisphenol A (BPA) and bisphenol AF (BPAF) in low-trophic-level animals, such as Daphnia magna (D. magna). In this study, a 21-day standard reproductive toxicity test was carried out to assess the effects of the 2 bisphenols on development, reproduction, and transcription of genes in D. magna. The results demonstrated that only exposure to 5000 nM BPA significantly decreased the survival rate, while significant alternations were observed after exposure to 50, 500, or 5000 nM BPAF. Both the molting numbers of per female and body length of F0 generation were decreased after exposure to 500 nM BPAF, while no obvious effects were observed after treatment with lower concentrations of BPAF and BPA. Increased fecundity but decreased body length and swimming speed of F1 generation were observed after exposure to 5000 nM BPA or 500 nM BPAF. In addition, the results of the qRT-PCR showed the expressions of cyp314, ecra, ecrb, usp, vtg1, and vtg2 were significantly inhibited after exposure to 5000 nM BPA, while the expressions of cyp18a1 and vmo1 were obviously upregulated after exposure to 500 nM BPAF.

A comprehensive system for detection of behavioral change of D. magna exposed to various chemicals.

The purpose of the present study was to develop a sensitive and comprehensive method, based on D. magna swimming behavior, for toxicity assessment of environmental chemicals. Firstly, D. magna swimming in several chambers with different diameters were compared to determine the most suitable container, and then baseline behaviors during light/dark periods as well as reactions to light/dark switching and vibration stimulation were determined. Secondly, after exposure to sub-lethal concentrations of the selected 42 typical chemicals, which were classified into heavy metals, pesticides, fungicides and flame retardants, the alterations in the swimming parameters were evaluated. Our results indicated the 48-well plate was the most suitable chamber for behavioral monitoring of D. magna, and specific responsive patterns of D. magna neonates to light/dark switching and vibration stimulation were observed. The results of the behavioral assays of chemicals suggested that D. magna was the most sensitive to methylmercury-chloride and then to abamectin and chlorpyrifos. The three chemicals at several to dozens of ng/L significantly changed swimming behaviors of D. magna. Furthermore, the alteration in the behavioral parameters (average swimming speed, etc.) induced by the selected chemicals could be ascribed to various modes of actions, confirming the reliability and practicability of the monitoring method.

Toxicity of tris(2-chloroethyl) phosphate in Daphnia magna after lifetime exposure: Changes in growth, reproduction, survival and gene transcription.

In recent years, with the elimination of brominated flame retardants (BFRs), the product volume of tris(2-chloroethyl) phosphate (TCEP), as a main substitute of BFRs, was increasing and frequently detected in natural waters. However, the current toxicological studies on TCEP were mainly focused on the partial life stage assessment of model animals, and thus it might underestimate the impact of TCEP on environmental risks. Therefore, the whole-life-stage effects of TCEP on growth, reproduction, survival and gene transcription in Daphnia magna (D. magna) were studied in this study after exposure to environmentally relevant or greater concentrations (500 or 5000 ng/L). It was found that chronic exposure to TCEP at environmental relevant or greater concentrations promoted growth of D. magna and the expressions of genes involved in the pathways associated with growth were significantly up-regulated. TCEP did not affect reproduction of D. magna, but the expressions of some genes screened in reproduction stage were significantly changed. Furthermore, the expressions of genes involved in two heart disease-related pathways were down-regulated at the death stage of D. magna after TCEP exposure for 62 days, suggesting that TCEP delayed the death of D. magna by retarding their heart senility.

Effects of tris (2-chloroethyl) phosphate (TCEP) on growth, reproduction and gene transcription in the protozoan Tetrahymena thermophila.

As a typical organophosphorus flame retardant, tris (2-chloroethyl) phosphate (TCEP) has been widely detected in various environmental media. Toxicity of TCEP to vertebrates have been investigated, but potential effects on lower trophic level species were unknown to date. In this study, toxic effects and molecular mechanisms of toxic actions of TCEP on the aquatic protozoan Tetrahymena thermophila were evaluated by use of phenotypic observations, transcriptome sequencing analysis and real-time quantitative PCR detection. Exposure to 0.044, 0.411 or 4.26 mg/L TCEP for 5 days decreased the theoretical population, cell viability, number of cilia and cell size of Tetrahymena thermophila in a time- and dose-dependent manner. Meanwhile, RNA-Seq analysis indicated that exposure to 4.26 mg/L TCEP significantly changed expression of 2932 genes (up-regulation: 1228; down-regulation: 1704). Of these, expressions of 9, 10 and 17 genes that were enriched in soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) interaction in vesicular transport, proteasome and endocytosis pathway respectively were down-regulated. Data collected during this study suggested that exposure to high concentrations of TCEP might affect growth and reproduction of Tetrahymena thermophila through down-regulating transcriptional levels of genes encoding proteins associated with vesicle trafficking, proteasome and endocytosis.

Effects of triphenyl phosphate on growth, reproduction and transcription of genes of Daphnia magna.

The additive flame retardant triphenyl phosphate (TPHP) has been frequently detected in environments and biota. Evidences indicate that TPHP has potential risks to aquatic organisms. Seldom has been reported about its chronic effects to aquatic organism at low trophic levels, such as Cladocera. In the present study, <12 h old Daphnia magna (D. magna) were exposed to 0, 5, 50 or 500 µg/L TPHP for 21 days to investigate the chronic effects of TPHP on body length, fecundity and survival. Meanwhile, D. magna PCR arrays were used to evaluate the transcriptional responses of 155 genes involved in 40 pathways. Exposure to 500 µg/L TPHP for 21 days significantly decreased the body lengths of both F0 and F1 generation and inhibited the fecundity of F0 generation. Results of RT-qPCR showed that the expressions of 76 genes involved in 15 pathways were significantly altered after exposure to 500 µg/L TPHP for 21 days. The significantly altered pathways related to genetic information processing, cellular process and metabolism might be responsible for the observed effects of TPHP. Overall, our results showed that chronic exposure to TPHP caused developmental and reproductive toxicities to D. magna.

Whole-Life-Stage Characterization in the Basic Biology of Daphnia magna and Effects of TDCIPP on Growth, Reproduction, Survival, and Transcription of Genes.

Toxicity tests of chemicals have mainly focused on the partial life-cycle evaluation of model animals. Limited information is available for the evaluation of effects of chemicals from a whole-life-stage exposure perspective. The objective of this study was to perform a whole-life-stage characterization in the basic biology of Daphnia magna (D. magna) and evaluate the effects of a known organophosphate ester (OPE) contaminant, tris(1,3-dichloro-2-propyl) phosphate (TDCIPP), on growth, reproduction, survival, and transcription of genes. The whole-life-stage characterization in growth, reproduction, and survival of D. magna was conducted, and representative sampling time points for the three developmental stages were identified (day 6, day 32, and day 62). Transcriptomic profiles for these three stages were compared, and stage-specific PCR arrays of D. magna were developed. The whole-life-stage exposure to environmentally relevant or greater concentrations of TDCIPP significantly inhibited growth and reproduction of D. magna and decreased survival at the later stage of the exposure experiment (≥32 days). Such adverse effects were not observed in the early stage of the exposure (<32 days), suggesting that short-term toxicity tests, such as the standard 21-day test, might underestimate the environmental risk of TDCIPP. Furthermore, expressions of genes selected at day 6, day 32, and day 62 were significantly changed after TDCIPP exposure, and the changes in the expressions of partial genes were correlated to the inhibitory effects on growth, reproduction, and survival.