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Bioreactors rich in Mn-oxidizing bacteria hold the potential for removing toxic Mn(II) and generating valuable bio-manganese oxides. Various environmental factors, such as pH, temperature, initial Mn(II) concentration, and dissolved oxygen, could affect microbial Mn(II) oxidation activity. However, limited information is available concerning the effect of residual Mn(II) concentrations on the optimization of bioreactor operations for efficient Mn(II) oxidation. This study aims to explore the influence of residual Mn(II) on the biological Mn(II) oxidation performance in the reactors, employing two identical reactors (R-1 and R-2) operating at varying Mn(II) loading rates. Interestingly, during Phase 2, despite both reactors receiving an equivalent Mn(II) loading rate, R-1 exhibited no residual Mn(II) and demonstrated a higher Mn(II) oxidation rate than R-2, which had a high residual Mn(II) concentration. The maximum Mn(II) oxidation rate in R-1 and R-2 was 0.91 and 0.61 kg m-3 d-1, respectively. The results unequivocally reveal that high concentrations of residual Mn(II) exert a substantial inhibitory effect on Mn(II) oxidation activity. This inhibition can be attributed to the diminished tolerance of specific Mn-oxidizing bacteria, such as Hyphomicrobium , to elevated Mn(II) levels. These findings underscore the critical importance of maintaining low residual Mn(II) concentrations within the reactors. This study carries practical implications for the effective operation of Mn(II) removal systems using bioreactors, highlighting the need to mitigate high residual Mn(II) concentrations for optimized performance. [Display omitted] • Mn-oxidizing bioreactors remove Mn(II) and generate valuable bio-manganese oxides. • Mn(II) oxidation rate of R-1 is higher than that of R-2 at comparable Mn(II) loading rates. • Elevated concentrations of residual Mn(II) significantly impede Mn(II) oxidation activity. • Hypomicrobium emerges as the predominant putative Mn-oxidizing bacteria. • High Mn(II) loading rates may induce stress on Mn-oxidizing bacteria. [ABSTRACT FROM AUTHOR]

• 6:2 FTCA biotransformation coupled with significant fluoride release (0.56∼1.83 f /molecule). • 5:3 FTCA was biotransformed with minimal fluoride release. • "Non-fluoride-releasing pathways" are dominant in 5:3 FTCA biotransformation. • Detection of less-fluorinated 5:3 FTUCA indicated defluorination at the γ carbon. • Hyphomicrobium and Dechloromonas were enriched and may contribute to aerobic FTCA biotransformation in activated sludge. Fluorotelomer carboxylic acids (FTCAs) represent an important group of per- and polyfluoroalkyl substances (PFAS) given their high toxicity, bioaccumulation potential, and frequent detection in landfill leachates and PFAS-impacted sites. In this study, we assessed the biodegradability of 6:2 FTCA and 5:3 FTCA by activated sludges from four municipal wastewater treatment plants (WWTPs) in the New York Metropolitan area. Coupling with 6:2 FTCA removal, significant fluoride release (0.56∼1.83 F-/molecule) was evident in sludge treatments during 7 days of incubation. Less-fluorinated transformation products (TPs) were formed, including 6:2 fluorotelomer unsaturated carboxylic acid (6:2 FTUCA), perfluorohexanoic acid (PFHxA), perfluoropentanoic acid (PFPeA), and perfluorobutanoic acid (PFBA). In contrast, little fluoride (0.01∼0.09 F-/molecule) was detected in 5:3 FTCA-dosed microcosms, though 25∼68% of initially dosed 5:3 FTCA was biologically removed. This implies the dominance of "non-fluoride-releasing pathways" that may contribute to the formation of CoA adducts or other conjugates over 5:3 FTCA biotransformation. The discovery of defluorinated 5:3 FTUCA revealed the possibility of microbial attacks of the C-F bond at the γ carbon to initiate the transformation. Microbial community analysis revealed the possible involvement of 9 genera, such as Hyphomicrobium and Dechloromonas, in aerobic FTCA biotransformation. This study unraveled that biotransformation pathways of 6:2 and 5:3 FTCAs can be divergent, resulting in biodefluorination at distinctive degrees. Further research is underscored to uncover the nontarget TPs and investigate the involved biotransformation and biodefluorination mechanisms and molecular basis. [Display omitted] [ABSTRACT FROM AUTHOR]

A soil bacterium, designated XQ2T, was isolated from Lang Mountain in Hunan province, P. R. China. The strain is Gram stain negative, facultative anaerobic, and the cells are motile and rod-shaped. The 16S rRNA gene sequence of strain XQ2T shared the highest similarities with Hyphomicrobium sulfonivorans S1T (97.1%), Pedomicrobium manganicum ACM 3038T (95.9%) and Hyphomicrobium aestuarii DSM 1564T (95.4%) and grouped with H. sulfonivorans S1T. The average nucleotide identity (ANI) values and the DNA–DNA hybridization (dDDH) values between strain XQ2T and H. sulfonivorans S1T were 86.6% and 55.4% respectively. Strain XQ2T had a genome size of 3.91 Mb and the average G+C content was 65.1%. The major fatty acids (> 5%) were C18:1ω6c, C18:1ω7c, C19:0 cyclo ω8c, C16:0 and C18:0. The major respiratory quinone was Q-9 (82.8%) and the minor one was Q-8 (17.2%). The polar lipids were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, phosphatidylcholine, unidentified phospholipid and two unidentified lipids. On the basis of phenotypic, chemotaxonomic and phylogenetic characteristics, strain XQ2T represents a novel species of the genus Hyphomicrobium, for which the name Hyphomicrobium album sp. nov. is proposed. The type strain is XQ2T (= KCTC 82378T = CCTCC AB 2020178T). The genus description is also emended. [ABSTRACT FROM AUTHOR]

Background. Hyphomicrobium nitrativorans strain NL23 and Methylophaga nitratire- ducenticrescens strain JAM1 are the principal bacteria involved in the denitrifying activities of a methanol-fed, fluidized-bed marine denitrification system. Strain NL23 possesses the complete denitrification pathway, but cannot grow under marine conditions in pure cultures. Strain JAM1 is a marine bacterium that lacks genes encoding a dissimilatory nitrite (NO2 -) reductase and therefore cannot reduce NO2 -. Here, we report the characterization of some of their physiological traits that could influence their co-habitation. We also perform co-cultures to assess the potential synergy between the two strains under marine and denitrifying conditions. Methodology. Anoxic planktonic pure cultures of both strains were grown with different concentrations of nitrate (NO3 -). Anoxic planktonic co-cultures could only be cultured on low NaCl concentrations for strain NL23 to grow. Biofilm co-cultures were achieved in a 500-mL bioreactor, and operated under denitrifying conditions with increasing concentrations of NaCl. NO3 - and NO2 - concentrations and the protein content were measured to derive the denitrification rates. The concentrations of both strains in co-cultures were determined by quantitative PCR (qPCR). Ectoine concentration was measured by mass spectrometry in the biofilm co-culture. The biofilm was visualized by fluorescence in situ hybridization. Reverse-transcriptionqPCR and RNA-seq approaches were used to assess changes in the expression profiles of genes involved in the nitrogen pathways in the biofilm cultures. Results. Planktonic pure cultures of strain JAM1 had a readiness to reduce NO3 - with no lag phase for growth in contrast to pure cultures of strain NL23, which had a 2-3 days lag phase before NO3 - starts to be consumed and growth to occur. Compared to strain NL23, strain JAM1 has a higher mmax for growth and higher specific NO3 - reduction rates. Denitrification rates were twice higher in the planktonic co-cultures than those measured in strain NL23 pure cultures. The biofilm co-cultures showed sustained denitrifying activities and surface colonization by both strains under marine conditions. Increase in ectoine concentrations was observed in the biofilm co-culture with the increase of NaCl concentrations. Changes in the relative transcript levels were observed in the biofilm culture with genes encoding NapA and NapGH in strain NL23. The type of medium had a great impact on the expression of genes involved in the N-assimilation pathways in both strains. Conclusions. These results illustrate the capacity of both strains to act together in performing sustainable denitrifying activities under marine conditions. Although strain JAM1 did not contribute in better specific denitrifying activities in the biofilm co-cultures, its presence helped strain NL23 to acclimate to medium with NaCl concentrations >1.0%. [ABSTRACT FROM AUTHOR]

Background Hyphomicrobium nitrativorans strain NL23 and Methylophaga nitratireducenticrescens strain JAM1 are the principal bacteria involved in the denitrifying activities of a methanol-fed, fluidized-bed marine denitrification system. Strain NL23 possesses the complete denitrification pathway, but cannot grow under marine conditions in pure cultures. Strain JAM1 is a marine bacterium that lacks genes encoding a dissimilatory nitrite (NO2−) reductase and therefore cannot reduce NO2−. Here, we report the characterization of some of their physiological traits that could influence their co-habitation. We also perform co-cultures to assess the potential synergy between the two strains under marine and denitrifying conditions. Methodology Anoxic planktonic pure cultures of both strains were grown with different concentrations of nitrate (NO3−). Anoxic planktonic co-cultures could only be cultured on low NaCl concentrations for strain NL23 to grow. Biofilm co-cultures were achieved in a 500-mL bioreactor, and operated under denitrifying conditions with increasing concentrations of NaCl. NO3− and NO2− concentrations and the protein content were measured to derive the denitrification rates. The concentrations of both strains in co-cultures were determined by quantitative PCR (qPCR). Ectoine concentration was measured by mass spectrometry in the biofilm co-culture. The biofilm was visualized by fluorescence in situ hybridization. Reverse-transcription-qPCR and RNA-seq approaches were used to assess changes in the expression profiles of genes involved in the nitrogen pathways in the biofilm cultures. Results Planktonic pure cultures of strain JAM1 had a readiness to reduce NO3− with no lag phase for growth in contrast to pure cultures of strain NL23, which had a 2-3 days lag phase before NO3− starts to be consumed and growth to occur. Compared to strain NL23, strain JAM1 has a higher µmax for growth and higher specific NO3− reduction rates. Denitrification rates were twice higher in the planktonic co-cultures than those measured in strain NL23 pure cultures. The biofilm co-cultures showed sustained denitrifying activities and surface colonization by both strains under marine conditions. Increase in ectoine concentrations was observed in the biofilm co-culture with the increase of NaCl concentrations. Changes in the relative transcript levels were observed in the biofilm culture with genes encoding NapA and NapGH in strain NL23. The type of medium had a great impact on the expression of genes involved in the N-assimilation pathways in both strains. Conclusions These results illustrate the capacity of both strains to act together in performing sustainable denitrifying activities under marine conditions. Although strain JAM1 did not contribute in better specific denitrifying activities in the biofilm co-cultures, its presence helped strain NL23 to acclimate to medium with NaCl concentrations >1.0%.

In the present study, the continuous denitrification of water was carried out using Hyphomicrobium denitrificans coupled with methane in a packed bed reactor containing a mineral packing medium. The response surface methodology based on the Box–Behnken design was applied to examine the interaction effects of pH (6–8), temperature (20°C–30°C), and methane/air mixture flow rate (20–60 mL min−1) on the nitrate removal (initial concentration of 100 mg L−1) using the Hyphomicrobium coupled with the methane. Analysis of variance results indicated that the methane/air mixture flow rate had a maximum effect on the nitrate removal efficiency in comparison to the pH and temperature variations. The experimental design results indicated that the optimum values of pH 6.9, temperature 30°C, and flow rate 53 mL min−1 led to maximize the denitrification capacity of Hyphomicrobium in a packed bed reactor. The maximum experimental denitrification capacity was found to be 2.85 mg NO3 −N L−1 h−1 which was in good agreement with the predicted value by the Box–Behnken analysis (2.81 mg NO3 −N L−1 h−1). The effluent nitrate concentration reached less than 50 mg L−1 in the nitrate flow rates of 1.4 and 2.5 mL min−1. The obtained results demonstrated the high capability of Hyphomicrobium denitrificans coupled with methane for denitrification of water. [ABSTRACT FROM AUTHOR]

Pyrroloquinoline quinone (PQQ) is a natural antioxidant with diverse applications in food and pharmaceutical industries. A lot of effort has been devoted toward the discovery of PQQ high-producing microbial species and characterization of biosynthesis, but it is still challenging to achieve a high PQQ yield. In this study, a combined strategy of random mutagenesis and adaptive laboratory evolution (ALE) with fermentation optimization was applied to improve PQQ production in Hyphomicrobium denitrificans H4-45. A mutant strain AE-9 was obtained after nearly 400 generations of UV-LiCl mutagenesis, followed by an ALE process, which was conducted with a consecutive increase of oxidative stress generated by kanamycin, sodium sulfide, and potassium tellurite. In the flask culture condition, the PQQ production in mutant strain AE-9 had an 80.4% increase, and the cell density increased by 14.9% when compared with that of the initial strain H4-45. Moreover, batch and fed-batch fermentation processes were optimized to further improve PQQ production by pH control strategy, methanol and H 2 O 2 feed flow, and segmented fermentation process. Finally, the highest PQQ production and productivity of the mutant strain AE-9 reached 307 mg/L and 4.26 mg/L/h in a 3.7-L bioreactor, respectively. Whole genome sequencing analysis showed that genetic mutations in the ftfL gene and thiC gene might contribute to improving PQQ production by enhancing methanol consumption and cell growth in the AE-9 strain. Our study provided a systematic strategy to obtain a PQQ high-producing mutant strain and achieve high production of PQQ in fermentation. These practical methods could be applicable to improve the production of other antioxidant compounds with uncleared regulation mechanisms. KEY POINTS: • Improvement of PQQ production by UV-LiCl mutagenesis combined with adaptive laboratory evolution (ALE) and fermentation optimization. • A consecutive increase of oxidative stress could be used as the antagonistic factor for ALE to enhance PQQ production. • Mutations in the ftfL gene and thiC gene indicated that PQQ production might be increased by enhancing methanol consumption and cell growth., (© 2024. The Author(s).)

Background The biofilm of a methanol-fed, marine denitrification system is composed of a multi-species microbial community, among which Hyphomicrobium nitrativorans and Methylophaga nitratireducenticrescens are the principal bacteria involved in the denitrifying activities. To assess its resilience to environmental changes, the biofilm was cultivated in artificial seawater (ASW) under anoxic conditions and exposed to a range of specific environmental conditions. We previously reported the impact of these changes on the denitrifying activities and the co-occurrence of H. nitrativorans strain NL23 and M. nitratireducenticrescens in the biofilm cultures. Here, we report the impact of these changes on the dynamics of the overall microbial community of the denitrifying biofilm. Methods The original biofilm (OB) taken from the denitrification system was cultivated in ASW under anoxic conditions with a range of NaCl concentrations, and with four combinations of nitrate/methanol concentrations and temperatures. The OB was also cultivated in the commercial Instant Ocean seawater (IO). The bacterial diversity of the biofilm cultures and the OB was determined by 16S ribosomal RNA gene sequences. Culture approach was used to isolate other denitrifying bacteria from the biofilm cultures. The metatranscriptomes of selected biofilm cultures were derived, along with the transcriptomes of planktonic pure cultures of H. nitrativorans strain NL23 and M. nitratireducenticrescens strain GP59. Results High proportions of M. nitratireducenticrescens occurred in the biofilm cultures. H. nitrativorans strain NL23 was found in high proportion in the OB, but was absent in the biofilm cultures cultivated in the ASW medium at 2.75% NaCl. It was found however in low proportions in the biofilm cultures cultivated in the ASW medium at 0–1% NaCl and in the IO biofilm cultures. Denitrifying bacterial isolates affiliated to Marinobacter spp. and Paracoccus spp. were isolated. Up regulation of the denitrification genes of strains GP59 and NL23 occurred in the biofilm cultures compared to the planktonic pure cultures. Denitrifying bacteria affiliated to the Stappia spp. were metabolically active in the biofilm cultures. Conclusions These results illustrate the dynamics of the microbial community in the denitrifying biofilm cultures in adapting to different environmental conditions. The NaCl concentration is an important factor affecting the microbial community in the biofilm cultures. Up regulation of the denitrification genes of M. nitratireducenticrescens strain GP59 and H. nitrativorans strain NL23 in the biofilm cultures suggests different mechanisms of regulation of the denitrification pathway in the biofilm. Other denitrifying heterotrophic bacteria are present in low proportions, suggesting that the biofilm has the potential to adapt to heterotrophic, non-methylotrophic environments.

Background The biofilm of a methanol-fed denitrification system that treated a marine effluent is composed of multi-species microorganisms, among which Hyphomicrobium nitrativorans strain NL23 and Methylophaga nitratireducenticrescens strain JAM1 are the principal bacteria involved in the denitrifying activities. Here, we report the capacity of the denitrifying biofilm to sustain environmental changes, and the impact of these changes on the co-occurrence of H. nitrativorans and M. nitratireducenticrescens. Methods In a first set of assays, the original biofilm (OB) was cultivated in an artificial seawater (ASW) medium under anoxic conditions to colonize new carriers. The new formed biofilm was then subjected to short exposures (1–5 days) of a range of NaCl, methanol, nitrate (NO3−) and nitrite (NO2−) concentrations, and to different pHs and temperatures. In a second set of assays, the OB was cultivated in ASW medium for five weeks with (i) a range of NaCl concentrations, (ii) four combinations of NO3−/methanol concentrations and temperatures, (iii) NO2−, and (iv) under oxic conditions. Finally, the OB was cultivated for five weeks in the commercial Instant Ocean (IO) seawater. The growth of the biofilm and the dynamics of NO3− and NO2− were determined. The levels of M. nitratireducenticrescens and H. nitrativorans were measured by qPCR. Results In the first set of assays, the biofilm cultures had the capacity to sustain denitrifying activities in most of the tested conditions. Inhibition occurred when they were exposed to high pH (10) or to high methanol concentration (1.5%). In the second set of assays, the highest specific denitrification rates occurred with the biofilm cultures cultivated at 64.3 mM NO3− and 0.45% methanol, and at 30 °C. Poor biofilm development occurred with the biofilm cultures cultivated at 5% and 8% NaCl. In all biofilm cultures cultivated in ASW at 2.75% NaCl, H. nitrativorans strain NL23 decreased by three orders of magnitude in concentrations compared to that found in OB. This decrease coincided with the increase of the same magnitude of a subpopulation of M. nitratireducenticrescens (strain GP59 as representative). In the biofilm cultures cultivated at low NaCl concentrations (0% to 1.0%), persistence of H. nitrativorans strain NL23 was observed, with the gradual increase in concentrations of M. nitratireducenticrescens strain GP59. High levels of H. nitrativorans strain NL23 were found in the IO biofilm cultures. The concentrations of M. nitratireducenticrescens strain JAM1 were lower in most of the biofilms cultures than in OB. Conclusions These results demonstrate the plasticity of the marine methylotrophic denitrifying biofilm in adapting to different environmental changes. The NaCl concentration is a crucial factor in the dynamics of H. nitrativorans strain NL23, for which growth was impaired above 1% NaCl in the ASW-based biofilm cultures in favor of M. nitratireducenticrescens strain GP59.

Rhodomicrobium vannielii is a multicellular and differentiating member of the order Hyphomicrobiales in the class Alphaproteobacteria. Here, we report the complete genome of strain DSM166 obtained by PacBio SMRT sequencing. The results suggest that this strain is closely related to Rhodomicrobium lacus ., Competing Interests: The authors declare no conflict of interest.

Background. The biofilm of a methanol-fed denitrification system that treated a marine effluent is composed of multi-species microorganisms, among which Hyphomicro- bium nitrativorans strain NL23 and Methylophaga nitratireducenticrescens strain JAM1 are the principal bacteria involved in the denitrifying activities. Here, we report the capacity of the denitrifying biofilm to sustain environmental changes, and the impact of these changes on the co-occurrence of H. nitrativorans and M. nitratireducenticrescens. Methods. In a first set of assays, the original biofilm (OB) was cultivated in an artificial seawater (ASW) medium under anoxic conditions to colonize new carriers. The new formed biofilm was then subjected to short exposures (1-5 days) of a range of NaCl, methanol, nitrate (NO3-) and nitrite (NO2-) concentrations, and to different pHs and temperatures. In a second set of assays, the OB was cultivated in ASW medium for five weeks with (i) a range of NaCl concentrations, (ii) four combinations of NO3-/methanol concentrations and temperatures, (iii) NO2-, and (iv) under oxic conditions. Finally, the OB was cultivated for five weeks in the commercial Instant Ocean (IO) seawater. The growth of the biofilm and the dynamics of NO3- and NO2 - were determined. The levels of M. nitratireducenticrescens and H. nitrativorans were measured by qPCR. Results. In the first set of assays, the biofilm cultures had the capacity to sustain denitrifying activities in most of the tested conditions. Inhibition occurred when they were exposed to high pH (10) or to high methanol concentration (1.5%). In the second set of assays, the highest specific denitrification rates occurred with the biofilm cultures cultivated at 64.3 mM NO3 - and 0.45% methanol, and at 30 °C. Poor biofilm development occurred with the biofilm cultures cultivated at 5% and 8% NaCl. In all biofilm cultures cultivated in ASW at 2.75% NaCl, H. nitrativorans strain NL23 decreased by three orders of magnitude in concentrations compared to that found in OB. This decrease coincided with the increase of the same magnitude of a subpopulation of M. nitratireducenticrescens (strain GP59 as representative). In the biofilm cultures cultivated at low NaCl concentrations (0% to 1.0%), persistence of H. nitrativorans strain NL23 was observed, with the gradual increase in concentrations of M. nitratireducenticrescens strain GP59. High levels of H. nitrativorans strain NL23 were found in the IO biofilm cultures. The concentrations of M. nitratireducenticrescens strain JAM1 were lower in most of the biofilms cultures than in OB. Conclusions. These results demonstrate the plasticity of the marine methylotrophic denitrifying biofilm in adapting to different environmental changes. The NaCl concentration is a crucial factor in the dynamics of H. nitrativorans strain NL23, for which growth was impaired above 1% NaCl in the ASW-based biofilm cultures in favor of M. nitratireducenticrescens strain GP59. [ABSTRACT FROM AUTHOR]

Background. The biofilm of a methanol-fed, marine denitrification system is composed of a multi-species microbial community, among which Hyphomicrobium nitrativorans and Methylophaga nitratireducenticrescens are the principal bacteria involved in the denitrifying activities. To assess its resilience to environmental changes, the biofilm was cultivated in artificial seawater (ASW) under anoxic conditions and exposed to a range of specific environmental conditions. We previously reported the impact of these changes on the denitrifying activities and the co-occurrence of H. nitrativorans strain NL23 and M. nitratireducenticrescens in the biofilm cultures. Here, we report the impact of these changes on the dynamics of the overall microbial community of the denitrifying biofilm. Methods. The original biofilm (OB) taken from the denitrification system was cultivated in ASW under anoxic conditions with a range of NaCl concentrations, and with four combinations of nitrate/methanol concentrations and temperatures. The OB was also cultivated in the commercial Instant Ocean seawater (IO). The bacterial diversity of the biofilm cultures and the OB was determined by 16S ribosomal RNA gene sequences. Culture approach was used to isolate other denitrifying bacteria from the biofilm cultures. The metatranscriptomes of selected biofilm cultures were derived, along with the transcriptomes of planktonic pure cultures of H. nitrativorans strain NL23 and M. nitratireducenticrescens strain GP59. Results. High proportions of M. nitratireducenticrescens occurred in the biofilm cultures. H. nitrativorans strain NL23 was found in high proportion in the OB, but was absent in the biofilm cultures cultivated in the ASW medium at 2.75% NaCl. It was found however in low proportions in the biofilm cultures cultivated in the ASW medium at 0-1% NaCl and in the IO biofilm cultures. Denitrifying bacterial isolates affiliated to Marinobacter spp. and Paracoccus spp. were isolated. Up regulation of the denitrification genes of strains GP59 and NL23 occurred in the biofilm cultures compared to the planktonic pure cultures. Denitrifying bacteria affiliated to the Stappia spp. were metabolically active in the biofilm cultures. Conclusions. These results illustrate the dynamics of the microbial community in the denitrifying biofilm cultures in adapting to different environmental conditions. The NaCl concentration is an important factor affecting the microbial community in the biofilm cultures. Up regulation of the denitrification genes of M. nitratireducenticrescens strain GP59 and H. nitrativorans strain NL23 in the biofilm cultures suggests different mechanisms of regulation of the denitrification pathway in the biofilm. Other denitrifying heterotrophic bacteria are present in low proportions, suggesting that the biofilm has the potential to adapt to heterotrophic, non-methylotrophic environments. [ABSTRACT FROM AUTHOR]

Abstract Effects of sulfadiazine and ciprofloxacin on microorganisms in biofilm of drinking water distribution systems (DWDSs) were studied. The results verified that the increases of 16S rRNA for total bacteria and bacterial genus Hyphomicrobium were related to the promotion of antibiotic resistance genes (ARGs) and class 1 integrons (int1) in DWDSs with sulfadiazine and ciprofloxacin. Moreover, the bacteria showed higher enzymatic activities in DWDSs with sulfadiazine and ciprofloxacin, which resulted in more production of extracellular polymeric substances (EPS). The higher contents of EPS proteins and secondary structure β -sheet promoted bacterial aggregation and adsorption onto surface of pipelines to form biofilm. EPS can serve as a barrier for the microorganisms in biofilm. Therefore, the biofilm bacterial communities shifted and the 16S rRNA for total bacteria increased in DWDSs with antibiotics, which also drove the ARGs promotion. Furthermore, the two antibiotics exhibited stronger combined effects than that caused by sulfadiazine and ciprofloxacin alone. Graphical abstract Image 1 Highlights • Response of biofilm to sulfadiazine and ciprofloxacin in DWDSs was studied. • Bacterial community shift and total bacteria increase related to ARGs promotion. • The bacteria exhibited higher enzymatic activity and produced more EPS. • The higher contents of proteins and β -sheet in EPS promoted biofilm formation. • Sulfadiazine and ciprofloxacin exhibited stronger combined effects on biofilm. [ABSTRACT FROM AUTHOR]

Graphical abstract Highlights • Wastewater types showed obvious effects on bacterial communities and functions. • Among 12 potential pathogens, Mycobacterium showed highest relative abundance. • Efflux pumps were the most common resistance mechanisms for 69 predicted ARGs. • ARGs/MRGs in industrial and mixed wastewater differed from municipal wastewater. • ARGs were linked to MRGs, functional genera and potential pathogens in WWTPs. Abstract This study aimed to investigate the bacterial communities and antibiotic resistance genes (ARGs) in 16 wastewater treatment plants (WWTPs) treating municipal, industrial and mixed wastewater. Wastewater types showed obvious effects on bacterial communities and functions. Nitrosomonas , Nitrospira , Hyphomicrobium and Accumulibacter were the main functional genera. Mycobacterium was the dominant potential pathogens. A total of 69 ARGs were obtained, and the dominant ARGs subtypes were similar in different WWTPs. Efflux pumps were the most common resistance mechanisms. Copper and zinc resistance genes were the main metal resistance genes (MRGs). Wastewater types affected the distributions of ARGs and MRGs, and they were more similar in industrial and mixed wastewater. The co-occurrence of ARGs existed within or across ARG types, and they were also positively linked to MRGs, some functional and pathogenic genera or environmental factors. This study furthers the understanding of interactions between bacterial communities, ARGs and MRGs in different WWTPs. [ABSTRACT FROM AUTHOR]

Several bacteria are able to degrade the major industrial solvent dichloromethane (DCM) by using the conserved dehalogenase DcmA, the only system for DCM degradation characterised at the sequence level so far. Using differential proteomics, we rapidly identified key determinants of DCM degradation for Hyphomicrobium sp. MC8b, an unsequenced facultative methylotrophic DCM-degrading strain. For this, we designed a pan-proteomics database comprising the annotated genome sequences of 13 distinct Hyphomicrobium strains. Compared to growth with methanol, growth with DCM induces drastic changes in the proteome of strain MC8b. Dichloromethane dehalogenase DcmA was detected by differential pan-proteomics, but only with poor sequence coverage, suggesting atypical characteristics of the DCM dehalogenation system in this strain. More peptides were assigned to DcmA by error-tolerant search, warranting subsequent sequencing of the genome of strain MC8b, which revealed a highly divergent set of dcm genes in this strain. This suggests that the dcm enzymatic system is less strongly conserved than previously believed, and that substantial molecular evolution of dcm genes has occurred beyond their horizontal transfer in the bacterial domain. Our study showed the power of pan-proteomics for quick characterization of new strains belonging to branches of the Tree of Life that are densely genome-sequenced.

Different environmental conditions such as pH and dissolved elements of mine stream induce precipitation of different minerals and their associated microbial community may vary. Therefore, mine precipitates from various environmental conditions were collected and their associated microbiota were analyzed through metagenomic DNA sequencing. Various Fe and Mn minerals including ferrihydrite, schwertmannite, goethite, birnessite, and Mn-substituted δ-FeOOH (δ-(Fe 1-x , Mn x )OOH) were found in the different environmental conditions. The Fe and Mn minerals were enriched with toxic metal(loid)s including As, Cd, Ni and Zn, indicating they can act as scavengers of toxic metal(loid)s in mine streams. Under acidic conditions, Acidobacteria was dominant phylum and Gallionella (Fe oxidizing bacteria) was the predominant genus in these Fe rich environments. Manganese oxidizing bacteria, Hyphomicrobium , was found in birnessite forming environments. Leptolyngbya within Cyanobacteria was found in Fe and Mn oxidizing environments, and might contribute to Fe and Mn oxidation through the production of molecular oxygen. The potential interaction of microbial community with minerals in mine sites can be traced by analysis of microbial community in different Fe and Mn mineral forming environments. Iron and Mn minerals contribute to the removal of toxic metal(loid)s from mine water. Therefore, the understanding characteristics of mine precipitates and their associated microbes helps to develop strategies for the management of contaminated mine water. [ABSTRACT FROM AUTHOR]

The potential environmental risks of chromium oxide nanoparticles (Cr 2 O 3 NPs) have caused great concerns. However, their possible impacts on activated sludge process are very limited. In this study, we carried out long-term exposure experiments to evaluate the influence of Cr 2 O 3 NPs on wastewater nutrient removal, bacterial community and molecular ecological network (MEN) in the sequencing batch reactor (SBR). It was found that 1 mg/L Cr 2 O 3 NPs increased the effluent concentrations of NO 3 − -N and TP from 6.5 to 15.4 mg/L, and 0.6–2.7 mg/L, indicating the serious deterioration of denitrification and phosphorus removal. Cr 2 O 3 NPs significantly decreased the bacterial richness in terms of the number of different OTUs (626 OTUs in Cr 2 O 3 samples and 728 OTUs in controls). Detrended correspondence analysis (DCA) showed that the overall taxonomic structure of bacterial community was altered at Cr 2 O 3 NPs in activated sludge systems. Further analysis revealed that three genera related to denitrification ( Desulfovibrio, Pseudomonas and Hyphomicrobium ) and two genera related to phosphorus removal ( Accumulibacter and Rhodobacter ) decreased significantly, which was consistent with the observed influences of Cr 2 O 3 NPs on denitrification and phosphorus removal. MEN analysis showed that the overall architecture of the network under Cr 2 O 3 NPs was substantially alerted. β- Proteobacteria , playing an important role in nutrients removal, had less complex interactions in the presence of Cr 2 O 3 NPs, which may be associated with the deterioration of denitrification and phosphorus removal. This study provides insights into our understanding of shifts in the bacteria community and their MEN under Cr 2 O 3 NPs in activated sludge systems. [ABSTRACT FROM AUTHOR]

Following a request from the European Commission, the EFSA Panel on Dietetic Products, Nutrition and Allergies (NDA) was asked to deliver an opinion on pyrroloquinoline quinone disodium salt (PQQ), trade name BioPQQTM, as a novel food pursuant to Regulation (EC) No 258/97. PQQ is produced by fermentation using Hyphomicrobium denitrificans CK-275 and purification process. PQQ has a minimum purity of 99.0%. The information provided on the composition, specifications, batch-to-batch variability, stability and production process of PQQ is sufficient and does not raise safety concerns. The applicant intends to market PQQ for use in food supplements for healthy adults, except pregnant and lactating women, at a maximum proposed level of consumption of 20 mg/day (corresponding to 0.29 mg/kg bw per day for a 70-kg person). The proposed level of consumption is at least 250 times higher than the estimated background intake of PQQ occurring naturally in foods. Information on the absorption, distribution, metabolism and excretion of PQQ in animals and humans is limited. Considering the no-observed-adverse-effect-level (NOAEL) of 100 mg/kg bw per day from a 90-day repeated dose oral toxicity study with BioPQQ™, and the maximum proposed level of consumption, the Panel concludes that the margin of exposure (of 344) is sufficient. The Panel concludes that the novel food, pyrroloquinoline quinone disodium salt (BioPQQ™), is safe under the intended conditions of use as specified by the applicant. [ABSTRACT FROM AUTHOR]

The interaction of ciprofloxacin chlorination products (CIP-CPs) with bacteria in drinking water distribution systems (DWDSs) was investigated. The piperazine ring of CIP was destroyed by chlorination. Among of CIP-CPs, by the bacterial role, 7.63% of the derivative with two carboxylic groups went through decarboxylation to form desethylene ciprofloxacin, and then loss of C 2 H 5 N group generated aniline compound. Furthermore, 12.3% of the aniline compound, 7.60% of chlorinated aniline compound and 1.35% of defluorinated product were bio-mineralized. Therefore, the chlorine and bacteria played synergistic effects on transformation of CIP-CPs in DWDSs, contributing to the obvious decrease of genotoxicity in effluents. Correspondingly, the TEQ 4-NQO decreased from 667 μg/L to 9.41 μg/L. However, compared with DWDSs without CIP-CPs, the relative abundance of mexA and qnrS increased 1-fold in effluents and the relative abundance of qnrA and qnrB increased 3-fold in biofilms in DWDSs with CIP-CPs. mexA and qnrS positively correlated with Hyphomicrobium , Sphingomonas and Novosphingobium (p < 0.05), while qnrA and qnrB positively correlated with Shewanella and Helicobacter (p < 0.05), indicating the increase of antibiotic resistance genes (ARGs) came from the growth of these bacterial genera by transformation of CIP-CPs in DWDSs. These results suggested that biotransformation of antibiotics might increase ARGs risk in DWDSs. [ABSTRACT FROM AUTHOR]

In the Xinjiang region of Eurasia, sustained long-term and continuous cropping of cotton over a wide expanse of land is practiced, which requires application of high levels of pyrethroid and other classes of pesticides-resulting in high levels of pesticide residues in the soil. In this study, soil samples were collected from areas of long-term continuous cotton crops with the aim of obtaining microbial resources applicable for remediation of pyrethroid pesticide contamination suitable for the soil type and climate of that area. Soil samples were first used to culture microbial flora capable of degrading beta-cypermethrin using an enrichment culture method. Structural changes and ultimate microbial floral composition during enrichment were analyzed by high-throughput sequencing. Four strains capable of degrading beta-cypermethrin were isolated and preliminarily classified. Finally, comparative rates and speeds of degradation of beta-cypermethrin between relevant microbial flora and single strains were determined. After continuous subculture for 3 weeks, soil sample microbial flora formed a new type of microbial flora by rapid succession, which showed stable growth by utilizing beta-cypermethrin as the sole carbon source (GXzq). This microbial flora mainly consisted of Pseudomonas, Hyphomicrobium, Dokdonella, and Methyloversatilis. Analysis of the microbial flora also permitted separation of four additional strains; i.e., GXZQ4, GXZQ6, GXZQ7, and GXZQ13 that, respectively, belonged to Streptomyces, Enterobacter, Streptomyces, and Pseudomonas. Under culture conditions of 37 °C and 180 rpm, the degradation rate of beta-cypermethrin by GXzq was as high as 89.84% within 96 h, which exceeded that achieved by the single strains GXZQ4, GXZQ6, GXZQ7, and GXZQ13 and their derived microbial flora GXh. [ABSTRACT FROM AUTHOR]

Molecular monitoring of bacterial communities can explain and predict the stability of bioprocesses in varying physicochemical conditions. To study methanol-fed denitrification biofilters of municipal wastewater treatment plants, bacterial communities of two full-scale biofilters were compared through fingerprinting and sequencing of the 16S rRNA genes. Additionally, 16S rRNA gene fingerprinting was used for 10-week temporal monitoring of the bacterial community in one of the biofilters. Combining the data with previous study results, the family Methylophilaceae and genus Hyphomicrobium were determined as suitable target groups for monitoring. An increase in the relative abundance of Hyphomicrobium-related biomarkers occurred simultaneously with increases in water flow, NO load, and methanol addition, as well as a higher denitrification rate, although the dominating biomarkers linked to Methylophilaceae showed an opposite pattern. The results indicate that during increased loading, stability of the bioprocess is maintained by selection of more efficient denitrifier populations, and this progress can be analyzed using simple molecular fingerprinting. [ABSTRACT FROM AUTHOR]

Oil sands tailings ponds in northern Alberta, Canada have been producing biogenic gases via microbial metabolism of hydrocarbons for decades. Persistent methanogenic activity in tailings ponds without any known replenishment of nutrients such as fixed nitrogen (N) persuaded us to investigate whether N 2 fixation or polyacrylamide (PAM; used as a tailings flocculant) could serve as N sources. Cultures comprising mature fine tailings (MFT) plus methanogenic medium supplemented with or deficient in fixed N were incubated under an N 2 headspace. Some cultures were further amended with citrate, which is used in oil sands processing, as a relevant carbon source, and/or with PAM. After an initial delay, N-deficient cultures with or without PAM produced methane (CH 4 ) at the same rate as N-containing cultures, indicating a mechanism of overcoming apparent N-deficiency. Acetylene reduction and 15 N 2 incorporation in all N-deficient cultures (with or without PAM) suggested active N 2 fixation concurrently with methanogenesis but inability to use PAM as a N source. 16S rRNA gene pyrosequencing revealed little difference between archaeal populations regardless of N content. However, bacterial sequences in N-deficient cultures showed enrichment of Hyphomicrobiaceae and Clostridium members that might contain N 2 -fixing species. The results are important in understanding long-term production of biogenic greenhouse gases in oil sands tailings. [ABSTRACT FROM AUTHOR]

In biological wastewater treatments, microbial populations of the so-called activated sludge work together in the abatement of pollutants. In this work, the metabolic behavior of the biomass of a lab-scale plant treating industrial pharmaceutical wastewater was investigated through a metaproteomic approach. The complete treatment process included a membrane biological reactor (MBR) coupled with an advanced oxidation process (AOP) for partial breakdown of non-biodegradable molecules. Proteins from biomass samples collected pre- and post-AOP application were investigated by two-dimensional gel electrophoresis (2DE), mass spectrometry (MS), and finally identified by database search. Results showed that most proteins remained constant between pre- and post-AOP. Methanol dehydrogenase (MDH) belonging to Hyphomicrobium zavarzinii appeared as the most constantly expressed protein in the studied consortium. Other identified proteins belonging to Hyphomicrobium spp. revealed a predominant methylotrophic metabolism, and H. zavarzinii appeared as a key actor in the studied microbial community. [ABSTRACT FROM AUTHOR]

This paper presents the results of DNA-based molecular analyses of the microbial community responsible for biological iron (Fe) and manganese (Mn) removal in slow sand filters (SSF). A lab-scale SSF was operated in 55-day sets under different operating conditions in order to evaluate long-term performance of the filter. The concentrations of Fe and Mn in synthetic feed water were increased from 1 mg/L to 2 mg/L at two different filtration rates (0.1 and 0.3 m/h). Daily samples were taken from influent and effluent for turbidity and Fe-Mn concentration measurements. 90-95% removal efficiencies were achieved with very low effluent concentrations. PCR-DGGE analyses were performed on samples, and Gallionella, Leptothrix, Crenothrix, and Hyphomicrobium were identified as the main microbial strains responsible for iron and manganese oxidation in SSF. Results also revealed that microbial activity was the main mechanism for Fe and Mn removal in the early stages of operation. [ABSTRACT FROM AUTHOR]

Methyl acrylate is a widely used raw chemical materials and it is toxic in humans. In order to treat the methyl acrylate waste gas, a 3-layer BTF packed with ceramic particles and immobilized with activated sludge was set up. The BTF exhibited excellent removal efficiency that no methyl acrylate could be detected when EBRT was larger than 266 s and inlet concentration was lower than 0.19 g/m 3 . The 1st layer performed the best at fixed inlet concentration of 0.42 g/m 3 . PCR combined with DGGE was performed to detect the differences in different layers of the BTF. Phylum Proteobacteria (e.g. α-, β-, γ-, δ-) was predominantly represented in the bacterial community, followed by Actinobacteria and Firmicutes . Desulfovibrio gigas , Variovorax paradoxus , Dokdonella koreensis , Pseudoxanthomonas suwonensis , Azorhizobium caulinodans , Hyphomicrobium denitrificans , Hyphomicrobium sp. and Comamonas testosteroni formed the bacteria community to treat methyl acrylate waste gas in the BTF. [ABSTRACT FROM AUTHOR]

Background and aims: High-throughput 454 pyrosequencing was applied to investigate differences in bacterial and fungal communities between replant and closely situated control non-replant (fallow) soils. Methods: The V1-V3 region of the bacterial 16S rRNA gene and the ITS1 region of fungi from the different soils were sequenced using 454 pyrosequencing (Titanium chemistry), and data were analysed using the MOTHUR pipeline. Results: The bacterial phyla Proteobacteria, Actinobacteria and Acidobacteria dominated in both fallow and replant apple orchard soils, and community composition at both phylum and genus level did not significantly differ according to NP-MANOVA. The fungal phyla Ascomycota, Zygomycota and Basidiomycota were dominant, and communities also did not differ in composition at either phylum or genus level. High positive Pearson correlations with plant growth in a plant growth assay performed with apple rootstocks plantlets were detected for the bacterial genera Gp16 and Solirubrobacter (r: >0.82) and fungal genera Scutellinia, Penicillium, Lecythophora and Paecilomyces (r: >0.65). Strong negative correlations with plant growth were detected for the bacterial genera Chitinophaga and Hyphomicrobium (r: <−0.78) and the fungal genera Acremonium, Fusarium and Cylindrocarpon (r: <−0.81). Conclusions: Study findings are in part consistent with those of previous research, but also highlight associations between apple plants and certain microbial genera. The functional role of these genera in affecting soil health and fertility should be further investigated. [ABSTRACT FROM AUTHOR]

Hyphomicrobium spp. are commonly identified as major players in denitrification systems supplied with methanol as a carbon source. However, denitrifying Hyphomicrobium species are poorly characterized, and very few studies have provided information on the genetic and physiological aspects of denitrification in pure cultures of these bacteria. This is a comparative study of three denitrifying Hyphomicrobium species, H. denitrificans ATCC 51888, H. zavarzinii ZV622, and a newly described species, H. nitrativorans NL23, which was isolated from a denitrification system treating seawater. Whole-genome sequence analyses revealed that although they share numerous orthologous genes, these three species differ greatly in their nitrate reductases, with gene clusters encoding a periplasmic nitrate reductase (Nap) in H. nitrativorans, a membrane-bound nitrate reductase (Nar) in H. denitrificans, and one Nap and two Nar enzymes in H. zavarzinii. Concurrently with these differences observed at the genetic level, important differences in the denitrification capacities of these Hyphomicrobium species were determined. H. nitrativorans grew and denitrified at higher nitrate and NaCl concentrations than did the two other species, without significant nitrite accumulation. Significant increases in the relative gene expression levels of the nitrate (napA) and nitrite (nirK) reductase genes were also noted for H. nitrativorans at higher nitrate and NaCl concentrations. Oxygen was also found to be a strong regulator of denitrification gene expression in both H. nitrativorans and H. zavarzinii, although individual genes responded differently in these two species. Taken together, the results presented in this study highlight the potential of H. nitrativorans as an efficient and adaptable bacterium that is able to perform complete denitrification under various conditions. [ABSTRACT FROM AUTHOR]

This study deals with the potential of biological processes combining a biotrickler and a biofilter to treat a mixture of sulphur-reduced compounds including dimethyl sulphide (DMS), dimethyl disulphide (DMDS) and hydrogen sulphide (HS). As a reference, duplicated biofilters were implemented, and operating conditions were similar for all bioprocesses. The first step of this work was to determine the efficiency removal level achieved for each compound of the mixture and in a second step, to assess the longitudinal distribution of biodegradation activities and evaluate the total bacteria, Hyphomicrobium sp. and Thiobacillus thioparus densities along the bed height. A complete removal of hydrogen sulphide is reached at the start of the experiment within the first stage (biotrickler) of the coupling. This study highlighted that the coupling of a biotrickling filter and a biofilter is an interesting way to improve both removal efficiency levels (15-20 % more) and kinetics of recalcitrant sulphur compounds such as DMS and DMDS. The total cell densities remained similar (around 1 × 10 16S recombinant DNA (rDNA) copies g dry packing material) for duplicated biofilters and the biofilter below the biotrickling filter. The relative abundances of Hyphomicrobium sp. and T. thioparus have been estimated to an average of 10 ± 7.0 and 0.23 ± 0.07 %, respectively, for all biofilters. Further investigation should allow achieving complete removal of DMS by starting the organic sulphur compound degradation within the first stage and surveying microbial community structure colonizing this complex system. [ABSTRACT FROM AUTHOR]

Highlights: [•] A higher DMSO oxidation at a higher Hyphomicrobium spp. abundace. [•] Hyphomicrobium is important in complete DMSO oxidation to sulfate compared with Thiobacillus. [•] Thiobacillus spp. was more abundant during poor nitrification. [•] Hyphomicrobium spp. was more abundant during good nitrification. [•] Population dynamics is closely linking to both DMSO/DMS degradation and nitrification. [ABSTRACT FROM AUTHOR]

Background: The agrichemical 4-aminopyridine is used as a bird repellent in crop fields and has an epileptogenic action in a variety of animals, including man and mouse. 4-Aminopyridine is biodegraded in the environment through an unknown mechanism. Results: A 4-aminopyridine-degrading enrichment culture utilized 4-aminopyridine as a carbon, nitrogen, and energy source, generating 4-amino-3-hydroxypyridine, 3,4-dihydroxypyridine, and formate as intermediates. 4-Amino-3-hydroxypyridine could not be further metabolized and probably accumulated as a dead-end product in the culture. Biodegradability tests and partial sequence analysis of the enrichment culture indicated that 4-aminopyridine was mainly degraded via 3,4-dihydroxypyridine and that the metabolite is probably cleaved by 3-hydroxy-4-pyridone dioxygenase. Seven culturable predominant bacterial strains (strains 4AP-A to 4AP-G) were isolated on nutrient agar plates. Changes in the bacterial populations of 4-aminopyridine, 3,4-dihydroxypyridine, or formate/ammonium chloride enrichment cultures were monitored by denaturing gradient gel electrophoresis (DGGE) profiling of PCR-amplified 16S rRNA gene fragments. Sequence analysis of the 16S rRNA gene fragments derived from predominant DGGE bands indicated that Pseudomonas nitroreducens 4AP-A and Enterobacter sp. 4AP-G were predominant in the three tested enrichment cultures and that the unculturable strains Hyphomicrobium sp. 4AP-Y and Elizabethkingia sp. 4AP-Z were predominant in 4-aminopyridine and formate/ammonium chloride enrichment cultures and in the 3,4-dihydroxypyridine enrichment culture, respectively. Among the culturable strains, strain 4AP-A could utilize 3,4-dihydroxypyridine as a growth substrate. Although we could not isolate strain 4AP-Y on several media, PCR-DGGE analysis and microscopy indicated that the unique bi-polar filamentous bacterial cells gradually became more dominant with increasing 4-aminopyridine concentration in the medium. Conclusions: Hyphomicrobium sp. 4AP-Y, P. nitroreducens 4AP-A, and Elizabethkingia sp. 4AP-Z probably play important roles in 4-aminopyridine degradation in crop fields. In the enrichment culture, 3,4-dihydroxypyridine and its metabolites including formate might be shared as growth substrates and maintain the enrichment culture, including these indispensable strains [ABSTRACT FROM AUTHOR]

Sugarcane bagasse was used as a carrier material of microorganisms in two different biofilters used to remove dimethyl sulfide (DMS) from a gas stream. The first biofilter was inoculated withHyphomicrobiumVS and the second withThiobacillus thioparusTk-m. During the operation of the biofilters the empty bed residence time (EBRT) was varied from 90 to 180 seconds and the inlet concentration of DMS from 12 to 50 ppmv. The inlet load was varied in the range of 0.62 to 5.2 (g DMS/m3 h). The maximum elimination capacity (EC) of the biofilter inoculated withHyphomicrobiumVS was 5 g DMS/m3 h; however, for the biofilter inoculated withT. thioparusTk-m the maximum EC was 3.9 g DMS/m3 h. ForT. thioparusTK-m the maximum removal efficiency (RE) obtained was 85.1±5.2% at 12 ppmv DMS inlet concentration, inlet load of 0.62 g DMS/m3 h and 180 s EBRT. The highest removal efficiency forHyphomicrobiumVS was 97.6±4.8% at 12 ppmv DMS inlet concentration, load of 0.62 g DMS/m3h and 180 s EBRT. [ABSTRACT FROM PUBLISHER]