Paucibacter sediminis sp. nov., isolated from sediment in a freshwater pond.

A Gram-stain-negative, aerobic, oxidase-positive, weakly catalase-positive, motile by means of a single polar flagellum, rod-shaped bacterium designated as strain S2-9T was isolated from sediment sampled in Wiyang pond, Republic of Korea. Growth of this strain was observed at 10-40 °C (optimum, 35 °C) and pH 5.5-9.5 (optimum, pH 7.0-8.0) and in the presence of 0-0.5 % NaCl in Reasoner's 2A broth. The major fatty acids (>10 %) of strain S2-9T were C16 : 0 and summed feature 3 (comprising a mixture of C16 : 1 ω7c and/or C16 : 1 ω6c). Ubiquinone-8 was detected as the respiratory quinone. The major polar lipids were phosphatidylethanolamine and phosphatidylglycerol. Strain S2-9T showed the highest 16S rRNA gene sequence similarity to Paucibacter oligotrophus CHU3T (98.7 %), followed by 'Paucibacter aquatile' CR182 (98.4 %), all type strains of Pelomonas species (98.1-98.3 %), Mitsuaria chitosanitabida NBRC 102408T (97.9 %), Kinneretia asaccharophila KIN192T (97.8 %), Mitsuaria chitinivorans HWN-4T (97.4 %), and Paucibacter toxinivorans 2C20T (97.4 %). Phylogenetic trees based on the 16S rRNA gene and whole-genome sequences showed that strain S2-9T formed a tight phylogenetic lineage with Paucibacter species (CHU3T, CR182, and 2C20T). Average nucleotide identity and digital DNA-DNA hybridization values between strain S2-9T and Paucibacter strains were 76.6-79.3% and 19.5-21.5 %, respectively. The genomic DNA G+C content of strain S2-9T was 68.3 mol%. Notably, genes responsible for both sulphur oxidation and reduction and denitrification were found in the genome of strain S2-9T, suggesting that strain S2-9T is involved in the nitrogen and sulphur cycles in pond ecosystems. Based on the polyphasic taxonomic results, strain S2-9T represents a novel species of the genus Paucibacter, for which the name Paucibacter sediminis sp. nov. is proposed. The type strain is S2-9T (= KACC 22267T= JCM 34541T).

Fine-scale spatiotemporal variations in bacterial community diversity in agricultural pond water.

Microbial communities in surface waters are affected by environmental conditions and can influence changes in water quality. To explore the hypothesis that the microbiome in agricultural waters associates with spatiotemporal variations in overall water quality and, in turn, has implications for resource monitoring and management, we characterized the relationships between the microbiota and physicochemical properties in a model irrigation pond as a factor of sampling time (i.e., 9:00, 12:00, 15:00) and location within the pond (i.e., bank vs. interior sites and cross-sectional depths at 0, 1, and 2 m). The microbial communities, which were defined by 16S rRNA gene sequencing analysis, significantly varied based on all sampling factors (PERMANOVA P < 0.05 for each). While the relative abundances of dominant phyla (e.g., Proteobacteria and Bacteroidetes) were relatively stable throughout the pond, subtle yet significant increases in α-diversity were observed as the day progressed (ANOVA P < 0.001). Key water quality properties that also increased between the morning and afternoon (i.e., pH, dissolved oxygen, and temperature) positively associated with relative abundances of Cyanobacteria, though were inversely proportional to Verrucomicrobia. These properties, among additional parameters such as bioavailable nutrients (e.g., NH3, NO3, PO4), chlorophyll, phycocyanin, conductivity, and colored dissolved organic matter, exhibited significant relationships with relative abundances of various bacterial genera as well. Further investigation of the microbiota in underlying sediments revealed significant differences between the bank and interior sites of the pond (P < 0.05 for α- and ß-diversity). Overall, our findings emphasize the importance of accounting for time of day and water sampling location and depth when surveying the microbiomes of irrigation ponds and other small freshwater sources.

Exploring the dynamics of algae-associated microbiome during the scale-up process of Tetraselmis sp. microalgae: A metagenomics approach.

Microalgae have become a key source of valuable compounds, promoting commercial scale applications. However, biological contamination is one of the most critical problems associated with large scale algal production, especially in open systems such as raceway ponds. The current research is the first to assess the effectiveness of open raceway ponds in maintaining a pure culture of Tetraselmis sp., starting from 20 L culture up to 10,000 L culture. Microbial profiling of each successive stage revealed lower abundance of eukaryotic organisms, whereas bacterial abundance increased notably resulting in a significant decrease in Tetraselmis sp. abundance. Furthermore, several bacteria with algae growth-promoting properties were found throughout the various culture stages including Balneola, Roseovarius, and Marinobacter. However, some algae-suppressive bacteria were evidenced at later stages such as Ulvibacter, Aestuariicoccus, and Defluviimonas. Overall, due to the increasing bacterial concentration, considerations limiting bacterial contamination need to be taken.

Microbiome analysis reveals Microcystis blooms endogenously seeded from benthos within wastewater maturation ponds.

Toxigenic Microcystis blooms periodically disrupt the stabilization ponds of wastewater treatment plants (WWTPs). Dense proliferations of Microcystis cells within the surface waters (SWs) impede the water treatment process by reducing the treatment efficacy of the latent WWTP microbiome. Further, water quality is reduced when conventional treatment leads to Microcystis cell lysis and the release of intracellular microcystins into the water column. Recurrent seasonal Microcystis blooms cause significant financial burdens for the water industry and predicting their source is vital for bloom management strategies. We investigated the source of recurrent toxigenic Microcystis blooms at Australia's largest lagoon-based municipal WWTP in both sediment core (SC) and SW samples between 2018 and 2020. Bacterial community composition of the SC and SW samples according to 16S rRNA gene amplicon sequencing showed that Microcystis sp. was dominant within SW samples throughout the period and reached peak relative abundances (32%) during the summer. The same Microcystis Amplicon sequence variants were present within the SC and SW samples indicating a potential migratory population that transitions between the sediment water and SWs during bloom formation events. To investigate the potential of the sediment to act as a repository of viable Microcystis cells for recurrent bloom formation, a novel in-vitro bloom model was established featuring sediments and sterilized SW collected from the WWTP. Microcystin-producing Microcystis blooms were established through passive resuspension after 12 weeks of incubation. These results demonstrate the capacity of Microcystis to transition between the sediments and SWs in WWTPs, acting as a perennial inoculum for recurrent blooms.IMPORTANCECyanobacterial blooms are prevalent to wastewater treatment facilities owing to the stable, eutrophic conditions. Cyanobacterial proliferations can disrupt operational procedures through the blocking of filtration apparatus or altering the wastewater treatment plant (WWTP) microbiome, reducing treatment efficiency. Conventional wastewater treatment often results in the lysis of cyanobacterial cells and the release of intracellular toxins which pose a health risk to end users. This research identifies a potential seeding source of recurrent toxigenic cyanobacterial blooms within wastewater treatment facilities. Our results demonstrate the capacity of Microcystis to transition between the sediments and surface waters (SWs) of wastewater treatment ponds enabling water utilities to develop adequate monitoring and management strategies. Further, we developed a novel model to demonstrate benthic recruitment of toxigenic Microcystis under laboratory conditions facilitating future research into the genetic mechanisms behind bloom development.

Sustainable treatment of primary and secondary effluent by algal-bacterial flocculent biomass in raceway ponds.

Two 5.5-L raceway open ponds were used to evaluate the removal of organic material and nutrients from wastewater. Algal-bacterial flocs were placed in the ponds to treat primary and secondary effluent. The organic loading rate ranged from 29 to 95 and 9 to 38 g sCOD m-3 d-1 for the reactor fed with primary and secondary effluent, respectively. The hydraulic retention time (HRT) gradually decreased in both reactors from 5.5 to 2.2 d during a period of 21 days, and after that, both reactors operated at an HRT of 1.1 d. A high biomass concentration of around 2.2 g L-1 was sustained using primary and secondary effluent after 130 days. The biomass, developed with both substrates was very active and completely removed organic material and nutrients in less than 12 h. The algal-bacteria biomass had excellent settling properties and could settle in less than 10 min.

Two machine learning approaches for predicting cyanobacteria abundance in aquaculture ponds.

Cyanobacteria blooms in aquaculture ponds harm the harvesting of aquatic animals and threaten human health. Therefore, it is crucial to identify key drivers and develop methods to predict cyanobacteria blooms in aquaculture water management. In this study, we analyzed monitoring data from 331 aquaculture ponds in central China and developed two machine learning models - the least absolute shrinkage and selection operator (LASSO) regression model and the random forest (RF) model - to predict cyanobacterial abundance by identifying the key drivers. Simulation results demonstrated that both machine learning models are feasible for predicting cyanobacterial abundance in aquaculture ponds. The LASSO model (R2 = 0.918, MSE = 0.354) outperformed the RF model (R2 = 0.798, MSE = 0.875) in predicting cyanobacteria abundance. Farmers with well-equipped aquaculture ponds that have abundant water monitoring data can use the nine environmental variables identified by the LASSO model as an operational solution to accurately predict cyanobacteria abundance. For crude ponds with limited monitoring data, the three environmental variables identified by the RF model provide a convenient solution for useful cyanobacteria prediction. Our findings revealed that chemical oxygen demand (COD) and total organic carbon (TOC) were the two most important predictors in both models, indicating that organic carbon concentration had a close relationship with cyanobacteria growth and should be considered a key metric in water monitoring and pond management of these aquaculture ponds. We suggest that monitoring of organic carbon coupled with phosphorus reduction in feed usage can be an effective management approach for cyanobacteria prevention and to maintain a healthy ecological state in aquaculture ponds.

Spatial Versus Nonspatial Variance in Fecal Indicator Bacteria Differs Within and Between Ponds.

Surface water environments are inherently heterogenous, and little is known about variation in microbial water quality between locations. This study sought to understand how microbial water quality differs within and between Virginia ponds. Grab samples were collected twice per week from 30 sampling sites across nine Virginia ponds (n = 600). Samples (100 mL) were enumerated for total coliform (TC) and Escherichia coli (EC) levels, and physicochemical, weather, and environmental data were collected. Bayesian models of coregionalization were used to quantify the variance in TC and EC levels attributable to spatial (e.g., site, pond) versus nonspatial (e.g., date, pH) sources. Mixed-effects Bayesian regressions and conditional inference trees were used to characterize relationships between data and TC or EC levels. Analyses were performed separately for each pond with ≥3 sampling sites (5 intrapond) while one interpond model was developed using data from all sampling sites and all ponds. More variance in TC levels were attributable to spatial opposed to nonspatial sources for the interpond model (variance ratio [VR] = 1.55) while intrapond models were pond dependent (VR: 0.65-18.89). For EC levels, more variance was attributable to spatial sources in the interpond model (VR = 1.62), compared to all intrapond models (VR < 1.0) suggesting that more variance is attributable to nonspatial factors within individual ponds and spatial factors when multiple ponds are considered. Within each pond, TC and EC levels were spatially independent for sites 56-87 m apart, indicating that different sites within the same pond represent different water quality for risk management. Rainfall was positively and pH negatively associated with TC and EC levels in both inter- and intrapond models. For all other factors, the direction and strength of associations varied. Factors driving microbial dynamics in ponds appear to be pond-specific and differ depending on the spatial scale considered.

Responses of bacterial and three sub-microeukaryote communities in the water of white shrimp Penaeus vannamei aquaculture ponds in two polyculture models.

Polyculture operations in freshwater aquaculture ponds can disrupt microbial communities. High-throughput sequencing was used to assess the impact of polyculture operations on bacterial and three sub-microeukaryote communities (fungi, zooplankton, and eukaryotic phytoplankton) in Penaeus vannamei aquaculture ponds containing oriental river prawns and giant freshwater prawns, respectively. The results showed that the bacterial community was less sensitive than the microeukaryote communities to both the polyculture activity and environmental variations. The polyculture of giant freshwater prawns rather than oriental river prawns was the primary factor affecting the beta diversity of the three sub-microeukaryote communities. This may be due to the larger biomass of the polyculture varieties of giant freshwater prawns compared with oriental river prawns. The polyculture activity of giant freshwater prawns with a higher density and that of oriental river prawns with a lower density increased the stochasticity of the community assembly of the three sub-microeukaryote communities. It also affected the topological properties of the microbial communities, including greater correlations between ecosystem elements, and reducing the correlations among zooplanktons. The eukaryotic phytoplankton was the only microbial community that could also be explained by nutrient variation (mainly the total nitrogen). This highlights the potential role of the eukaryotic phytoplankton as a suitable indicator of the effects of nutrient input into ecosystems.

Strategy for the formation of microalgae-bacteria aggregates in high-rate algal ponds.

This work studied the formation of aggregates used for wastewater treatment in high-rate algal ponds (HRAP). For this, the establishment of microalgae-bacteria aggregates in these systems was evaluated, considering strategies for the inoculation and start-up. Two HRAP were operated in parallel, at first in batch mode and then in continuous flow. The wastewater treatment was efficient, with removal rates around 80% for COD and N-ammoniacal. Volatile suspended solids and chlorophyll for the culture grew continuously reached a concentration of 548 ± 11 mg L-1 and 7.8 mg L-1, respectively. Larger photogranules were observed when the system was placed in a continuous regime. The protein fraction of extracellular polymeric substances was identified as a determinant in photogranules formation. During the continuous regime, more than 50% of the biomass was higher than 0.2 mm, flocculation efficiency of 78 ± 6%, and the volumetric sludge index of 32 ± 5 mL g-1. The genetic sequencing showed the growth of cyanobacteria in the aggregate and the presence of microalgae from the chlorophytes and diatoms groups in the final biomass.

Exploring the mechanism of a novel recirculating aquaculture system based on water quality parameters and bacterial communities.

To mitigate the serious environmental problems caused by aquaculture wastewater discharge, the development of improved aquaculture systems with more self-purification capacity and less environmental impact has become essential. A novel recirculating aquaculture system (RAS) with ecological ponds was introduced. However, the mechanism of how the ecological ponds decompose the nutrients from the residual feed and excrement of fish is still unclear. Therefore, we designed a seven-week field experiment to explore the dynamic of water quality and the shift of bacterial communities during the initial stage of the RAS ecosystem to maintain the stability of the system. According to the result, the dissolved oxygen concentration maintained at 5.63 to 10.22 mg·L-1 in aquaculture water, and electrical conductivity increased by 100% to over 800 µs·cm-1. High-throughput sequencing showed that the abundance and diversity of the bacterial communities in sediment samples were significantly higher than in water samples, and the Proteobacteria, Bacteroidetes, Cyanobacteria, and Actinobacteria were dominant phyla in all samples. The relative abundance of ammonia-oxidizing archaea, ammonia-oxidizing bacteria, and nitrite-oxidizing bacteria was lower than 0.10, but rising. Redundancy analysis suggested that TOC, EC, NO3-, and NO2- were the most important factors shaping the bacterial communities in aquaculture water. Our studies assessed the RAS with ecological ponds for the first time based on water quality parameters and bacterial communities, which indicates that decomposition capacity is insufficient but improved at the initial stage of the operation, and potential risks like eutrophication require attention.

Apex Predators Enhance Environmental Adaptation but Reduce Community Stability of Bacterioplankton in Crustacean Aquaculture Ponds.

Aquaculture environments harbor complex bacterial communities that are critical for the growth and health of culture species. Apex predators are frequently added to aquaculture ponds to improve ecosystem stability. However, limited research has explored the effects of apex predators on the composition and function of bacterioplankton communities, as well as the underlying mechanisms of community assembly. Using 16S ribosomal RNA (rRNA) high-throughput sequencing, we investigated bacterioplankton communities of crustacean aquaculture ponds with and without apex predators (mandarin fish, Siniperca chuatsi) throughout the culture process. In addition to investigating differences in bacterioplankton communities, we also explored variations in environmental adaptation, functional redundancy, and community stability. Significant differences were observed in bacterioplankton composition among different cultural stages; there was an increase in Bacteriobota and fermentation-related bacteria, but a decrease in Firmicutes and pathogens in the middle stages of aquaculture. Apex predators increased the abundance of organic matter degradation bacteria and decreased pathogens. Bacterioplankton communities under apex predator disturbances had a wider environmental breadth, indicating broader environmental adaptation. Moreover, functional prediction and network analyses revealed that communities under apex predator disturbances were less functionally redundant and unstable. Based on the null model, stochastic processes drove community assembly during aquaculture, whereas apex predators elevated the contribution of deterministic processes. Greater changes in nitrate in culture ponds caused by apex predator disturbances were decisive in controlling the balance between stochasticity and determinism in community assembly. Our study provided insight into the mechanisms underlying bacterioplankton community assembly in aquaculture systems in response to apex predator disturbances.

Can water composition and weather factors predict fecal indicator bacteria removal in retention ponds in variable weather conditions?

Retention ponds provide benefits including flood control, groundwater recharge, and water quality improvement, but changes in weather conditions could limit the effectiveness in improving microbial water quality metrics. The concentration of fecal indicator bacteria (FIB), which is used as regulatory standards to assess microbial water quality in retention ponds, could vary widely based on many factors including local weather and influent water chemistry and composition. In this critical review, we analyzed 7421 data collected from 19 retention ponds across North America listed in the International Stormwater BMP Database to examine if variable FIB removal in the field conditions can be predicted based on changes in these weather and water composition factors. Our analysis confirms that FIB removal in retention ponds is sensitive to weather conditions or seasons, but temperature and precipitation data may not describe the variable FIB removal. These weather conditions affect suspended solid and nutrient concentrations, which in turn could affect FIB concentration in the ponds. Removal of total suspended solids and total P only explained 5% and 12% of FIB removal data, respectively, and TN removal had no correlation with FIB removal. These results indicate that regression-based modeling with a single parameter as input has limited use to predict FIB removal due to the interactive nature of their effects on FIB removal. In contrast, machine learning algorithms such as the random forest method were able to predict 65% of the data. The overall analysis indicates that the machine learning model could play a critical role in predicting microbial water quality of surface waters under complex conditions where the variation of both water composition and weather conditions could deem regression-based modeling less effective.

Bioremediation competence of Aspergillus flavus DDN on pond water contaminated by mining activities.

This research was performed to evaluate the possibilities of reducing the physicochemical properties of polluted pond water situated around the magnesite mine tailing through indigenous metal tolerant fungi. The physicochemical analysis results revealed that most of the physicochemical properties of pond water sample were crossing the permissible limits. From the muddy pond soil sample, Aspergillus flavus DDN was identified (through molecular characterization) as predominant metal tolerant fungal strain and it showed resistance to Cr(VI), Pb(II), Zn(II), Cd(II), and Mg(IV) up to 1000 µg mL-1 concentrations. This strain also effectively reduced (through biosorption) these metals in a short duration of the bioremediation process. In a lab-scale bioremediation study, the A. flavus DDN significantly reduced most of the physicochemical parameters crossing the permissible limit in polluted pond water in the presence of FM1 minimal media in 10 days of incubation. The dissolved oxygen level was significantly increased up to 74.91% from 5.86 ± 0.39 to 10.25 ± 0.95 in 10 days of treatment. The metal reduction and other physicochemical properties reduction were directly related to the biomass of A. flavus DDN. These findings suggest that A. flavus DDN can remove pollutants from magnesite mine tailing polluted pond water because elevated fungal biomass resulted in the highest percentage of pollutant reduction from the sample.

Microbial diversity and composition in acidic sediments of freshwater finfish culture ponds fed with two types of feed: a metagenomic approach.

Microbial community profile associated with acidic pond sediments (APS) (pH = 3·0-4·5) of freshwater finfish aquaculture ponds (n = 8) was investigated. Sediment DNA extracted from the eight APS were subjected to high-throughput sequencing of V3 and V4 regions which yielded 7236 operational taxonomic units (OTUs) at a similarity of 97%. Overall results showed higher proportion of bacterial OTUs than archaeal OTUs in all the APS. Euryarchaeota (23%), Proteobacteria (19%), Chloroflexi (17%), Crenarchaeota (5·3%), Bacteroidetes (4·8%), Nitrospirae (3·2%), Nanoarchaeaeota (3%) which together constituted 75% of the microbial diversity. At the genus level, there was high preponderance of methanogens namely Methanolinea (5·4%), Methanosaeta (4·5%) and methanotrops, Bathyarchaeota (5%) in APS. Moreover, the abundant phyla in the APS were not drastically affected by the administration of chicken slaughter waste (R-group ponds) and commercial fish feed (C-group ponds), since 67% of the OTUs generated remained common in the APS of both the groups of ponds. There was a minimal difference of 24-26% of OTUs between C-group and R-group ponds, suggesting the existence of a core microbial community in these ponds driven by acidic pH over the years. This study concludes that microbial diversity in pond sediment was influenced to a lesser extent by the addition of chicken slaughter waste but was majorly driven by acidic nature of the pond.

Structural and functional comparisons of the environmental microbiota of pond and tank environments at different locations for the commercial aquaculture of American shad.

The environmental microbiota plays a significant role in the growth and development of aquatic life. In recent years, American shad has become an important economic aquaculture species in China. However, information on the correlation between the growth of American shad and the aquaculture environment is limited. Through 16S rDNA-based sequencing, the microbiota communities in ponds at different locations (Jiangyin and Yancheng in Jiangsu, China) were investigated. The results showed that the richness and diversity of the microbiota in the pond were greater than those in the tank at the same location. At the phylum level, Proteobacteria and Firmicutes were more abundant in the samples from Jiangyin than in those from Yancheng. In addition, the ratio of Firmicutes to Bacteroidetes was larger in the JYT samples than in the YCT samples, which implied that the American shad cultured in the Jiangyin environment tended to be fatter than those cultured in Yancheng. The body weight data also confirmed this finding. Moreover, the proportions of functional annotations in the samples from the Jiangyin and Yancheng environments were similar, but there were differences between the overall levels. Our results highlighted the correlations between the environmental microbiome and the growth tendency of American shad.

Anaerobes and methanogens dominate the microbial communities in water harvesting ponds used by Kenyan rural smallholder farmers.

Many rural smallholder farmers in Kenya use water-harvesting ponds, to collect rainwater, as sustainable sources of water for domestic and agricultural purposes. There is currently limited information regarding the microbial ecology in these ponds. Here, we used High Throughput Sequencing (HTS) to characterize the microorganisms present (including potential pathogens and indicator species) alongside ion chromatography to measure water chemistry (anion and cation concentration). Fluoride and magnesium concentration were the strongest predictor variables of the microbial community. Obligately or facultatively anaerobic bacterial genera (e.g. Spirochaeta and Opitutus) were abundant within the bacterial community, whilst Woesearchaeota and methanogens dominated the archaeal community. This suggests the water in the ponds is hypoxic or anoxic, and if used for irrigation, may potentially impact crop yield and viability. In addition, the opportunistic pathogen non-tuberculous mycobacteria (NTM), Mycobacterium fortuitum was found, comprising >1% of the bacterial community, suggesting a potential human health risk. Here we suggest low-cost changes to pond management, to improve or ameliorate pond anoxia and remove pathogens to benefit the livelihoods and welfare of these farms. This study also shows the applicability of HTS to broadly screen the microbial communities, assess water quality, and identify potentially pathogenic groups.

Hymenobacter piscis sp. nov., isolated from a fish pond.

Bacterial strain NST-14T, isolated from a freshwater fish pond in Taiwan, was characterized using a polyphasic taxonomy approach. The strain was Gram-stain-negative, strictly aerobic, non-motile, rod-shaped and formed pink colonies. Optimal growth occurred at 30 °C, pH 7 and in the absence of NaCl. Phylogenetic analyses based on 16S rRNA gene sequences and coding sequences of 92 protein clusters indicated that strain NST-14T formed a phylogenetic lineage in the genus Hymenobacter. Analysis of 16S rRNA gene sequences showed that strain NST-14T had the highest similarity to Hymenobacter actinosclerus CCUG 39621T (97.7%), Hymenobacter amundsenii P5136T (97.3%) and Hymenobacter humicola P6312T (96.9%). Strain NST-14T showed 75.1-85.3 % average nucleotide identity, 73.7-89.8 % average amino acid identity and 14.5-26.0 % digital DNA-DNA hybridization with the type strains of other closely related Hymenobacter species. Strain NST-14T contained iso-C15 : 0, C16 : 1 ω5c and summed feature 3 (C16 : 1 ω7c and/or C16 : 1 ω6c) as the predominant fatty acids. The major hydroxyl fatty acids were iso-C17 : 0 3-OH and iso-C15 : 0 3-OH. The polar lipids were phosphatidylethanolamine, one unidentified glycolipid, four unidentified aminophospholipids, one unidentified aminolipid, two unidentified phospholipids and three unidentified lipids. The major polyamine was homospermidine. The major isoprenoid quinone was MK-7. The DNA G+C content of the genomic DNA was 62.4 mol%. Differential phenotypic properties, together with the phylogenetic inference, demonstrate that strain NST-14T should be classified as a novel species of the genus Hymenobacter, for which the name Hymenobacter piscis sp. nov. is proposed. The type strain is NST-14T (=BCRC 81249T=LMG 31686T).

Halorubrum salinarum sp. nov., an extremely halophilic archaeon isolated from a saturated brine pond of a saltern.

A novel extremely halophilic archaeon, strain RHB-CT, was isolated from a saturated brine pond of a solar saltern in Bolinao, Pangasinan, Philippines. Colonies were orange-red-pigmented, smooth, convex and round on a solid modified growth medium containing 25 % (w/v) of total salts. Cells of strain RHB-CT on the solid modified growth medium were ovoid-shaped (0.89-2.66 µm long), while the cells in a liquid modified growth medium were rod-shaped (1.53-5.65 µm long and 0.45-1.03 µm wide). The strain was Gram-stain-negative, motile and strictly aerobic. Strain RHB-CT grew with NaCl concentrations ranging from 10 to 30 % (w/v; optimum, 20-25 %), at pH 6.5-8.5 (optimum, pH 7.0-7.5) and at 20-55 °C (optimum, 40-45 °C). Furthermore, the strain grew even in the absence of Mg2+; however, when supplemented with Mg2+, growth was observed optimally at 0.2-0.4 M Mg2+. The 16S rRNA gene phylogeny inferred that the strain is a member of the genus Halorubrum and was related to Halorubrum xinjiangense CGMCC 1.3527T (99.0 %), Halorubrum sodomense DSM 3755T (98.8 %), Halorubrum coriense Ch2T (98.8 %), Halorubrum trapanicum NRC 34021T (98.4 %) and Halorubrum distributum JCM 9100T (98.1 %). The rpoB' gene sequences also showed that strain RHB-CT is related to Hrr. xinjiangense JCM 12388T (97.1 %), Hrr. distributum JCM 9100T (97.1 %), Hrr. coriense JCM 9275T (96.5 %), Hrr. californiense JCM 14715T (96.5 %), Hrr. trapanicum JCM 10477T (96.3%), Hrr. sodomense JCM 8880T (96.2%) and Hrr. tebenquichense DSM 14210T (95.6 %). The DNA G+C content of strain RHB-CT was 68.7 mol% (genome). Digital DNA-DNA hybridization (dDDH) and average nucleotide identity (ANI) values between strain RHB-CT and the closely related species of Halorubrum were below 40 and 90 %, respectively, which are far below the thresholds to delineate a new species. The polar lipids of strain RHB-CT were phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester, phosphatidylglycerol sulphate and sulfated mannosyl glycosyl diether. Based on dDDH and ANI values, and the significant morphological and physiological differences from known taxa, it is hereby suggested that strain RHB-CT represents a novel species of the genus Halorubrum, for which the name Halorubrum salinarum sp. nov. is proposed. The type strain is RHB-CT (=KCTC 4274T=CMS 2103T).

Comparison of planktonic bacterial communities indoor and outdoor of aquaculture greenhouses.

AIMS: Greenhouses are widely used in agriculture systems to shield crops from unfavourable weather to achieve a year-round food supply. In recent years, aquaculture ponds have been placed in greenhouses in many regions. The impacts of the greenhouses on planktonic bacterial communities should be uncovered. METHODS AND RESULTS: In this study, two polyolefin film greenhouses accommodating aquaculture ponds were established and planktonic bacterial communities were compared from samples taken in aquaculture ponds inside and outside the greenhouses, using Illumina 16S rRNA sequencing. CONCLUSIONS: The results showed there were significant variations in bacterial community structure between indoor and outdoor samples. Obvious differences were also found between two greenhouses, whereas the differences in indoor samples were weaker than outdoor samples. Significantly higher temperature (in summer), pH and permanganate index were found in the outdoor pond samples. Results of redundancy analysis showed that Proteobacteria and Bacteroidota were positively related to the dissolved oxygen, total nitrogen and total phosphorus, and Actinobacteriota were positively related to pH, temperature and permanganate index, whereas Cyanobacteria were positively related to the salinity, conductivity, total dissolved solids and ammonia nitrogen. SIGNIFICANCE AND IMPACT OF THE STUDY: The results of this study revealed that greenhouses significantly influenced planktonic bacterial communities in aquaculture ponds. This study is expected to provide a scientific basis for aquaculture in greenhouses.

The dynamic of the potential pathogenic bacteria, antibiotic-resistant bacteria, and antibiotic resistance genes in the water at different growth stages of grass carp pond.

Pond aquaculture has become the most important and broadest breeding model in China, and an extremely important source of aquatic products, but the potential hazard factors of potential pathogenic bacteria (PPB), antibiotic resistance bacteria (ARB), and antibiotic resistance genes (ARGs) in aquaculture environment are largely invisible. In the present study, the bacterial communities in the larvae, juvenile, rearing, and harvesting culture stages of great grass carp (Ctenopharyngodon idellus) ponds were investigated and the structure of microbial flora analysis showed that the larvae culture stage has the highest abundance and the most dominant phyla were Proteobacteria (27.8%). A total of 123 significant Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway annotations and the relative abundance of nine bacterial phenotypes implied that the larvae culture stage had the most abundance of pathogenic potential and mobile elements. The correlation analyses of environmental factors showed that temperature, stocking density, pH, and transparency showed the significant impacts on both the distribution of microbiome and the PPB. More importantly, a total of 40 ARB were identified, and 16 ARGs have the detection rates of 100%, which revealed that they are widely distributed and highly enriched in the aquaculture production. Notably, this is the first robust report to analyze and understand the PPB, ARB, and ARGs characteristics and dynamic changes in the pond aquaculture.