16S rRNA Gene Abundance Research Articles

Dicyandiamide Applications Mitigate the Destructive Effects of Graphene Oxide on Microbial Activity, Diversity, and Composition and Nitrous Oxide Emission in Agricultural Soil.

The widespread production and utilization of graphene oxide (GO) raise concerns about its environmental release and potential ecological impacts, particularly in agricultural soil. Effective nitrogen (N) management, especially through nitrification inhibitors like dicyandiamide (DCD), might mitigate the negative effects of GO exposure on soil microbes via N biostimulation. This study quantified changes in soil physicochemical properties, nitrous oxide (N2O) emissions, microbial activity, biomass, and community after treatments with GO and DCD. The GO exposure significantly reduced bacterial 16S rRNA gene abundance and the biomass of major bacterial phyla. It also stimulated pathways linked to human diseases. However, DCD application alleviated the negative effects of GO exposure on soil bacterial biomass. While DCD application significantly reduced soil N2O emission, the GO application tended to hinder the inhibiting performance of DCD. Our findings highlight the hazards of GO exposure to soil microbes and the potential mitigation strategy with soil N management.

Biogeographical distribution and community assembly of Myxococcota in mangrove sediments

BackgroundMyxococcota, characterized by their distinct social lifestyles, are widely distributed micro-predators in global sediments. They can feed on a wide range of bacterial, archaeal, and fungal prey. Myxococcota are capable of producing diverse secondary metabolites, playing key roles in microbial food webs, and regulating the microbial community structures in different ecosystems. However, Myxococcota are rarely pure cultured due to the challenging and stringent culturing conditions. Their natural distribution, niche differentiation, and predator–prey relationships in a specific habitat are poorly understood.ResultsIn this study, we conducted a comprehensive analysis of the 16S rRNA gene sequence data from public databases and our collection. We compared the abundance, diversity, and distribution patterns of Myxococcota in various habitats, with a specific focus on mangroves. We found that Myxococcota accounted for 1.45% of the total prokaryotes in global sediments based on the abundance of 16S rRNA genes. Myxococcota are abundant and diverse in mangrove sediments. They tend to be more generalistic in mangroves than in other habitats due to their wide niche breadth. Besides, the deterministic processes (variable selection) influenced the assembly of mangrove Myxococcota communities significantly more than stochastic processes. Further, we determined that environmental factors explained a greater amount of total community variation in mangrove Myxococcota than geographical variables (latitude and sediment depth). In the end, through the analysis of microbial co-occurrence networks, Myxococcota emerges as a key component and functions as a connector in the mangrove microbial community.ConclusionsOur study enhances comprehension of mangrove Myxococcota's biogeography, assembly patterns, driving factors, and co-occurrence relationships, as well as highlights their unique niche and ecological importance in mangrove sediments.

The Cellular Microbiome of Visceral Organs: An Inherent Inhabitant of Parenchymal Cells.

The cell is the basic unit of life. It is composed of organelles and various organic and inorganic biomolecules. Recent 16S ribosomal ribonucleic acid (16S rRNA) gene sequencing studies have revealed the presence of tissue bacteria in both tumor and normal tissues. Recently, we found that the liver microbiome resided in hepatocytes. Here, we further report on the cellular microbiome in the parenchymal cells of visceral organs as inherent inhabitants. We performed 16S rRNA gene sequencing on visceral organs of male adult Sprague Dawley (SD) rats, pregnant rats, newborn rats, and fetuses and placentas; then, we performed fluorescence in situ hybridization and immunofluorescence in visceral organs. Furthermore, we performed Western blotting on nuclear and cytoplasmic extractions of visceral organs of SD rats and cell lines HepG2, Huh-7, Hepa1-6, and HSC-T6. A high abundance of 16S rRNA gene was detected in the visceral organs of male adult, pregnant, newborn, and fetal rats as well as their placentas. The number of operational taxonomic units (OTUs) of visceral bacteria was higher than that of the feces and ileum bacteria. Bacterial 16S rRNA, lipopolysaccharide (LPS), and lipoteichoic acid (LTA) were found in the parenchymal cells of visceral organs, as well as in HepG2, Huh-7, HSC-T6, and Hepa1-6 cells. LPS consistently appeared in the nucleus of cells, while LTA was mainly found in the cytoplasm. In conclusion, the cellular microbiome is an intrinsic component of cells. Gram-negative bacteria are located in the nucleus, and Gram-positive bacteria are located in the cytoplasm. This differs from the gut microbiome and may be inherited.

The effect of the 13C abundance of soil microbial DNA on identifying labelled fractions after ultracentrifugation

DNA-based stable isotope probing (DNA-SIP) technology has been widely employed to trace microbes assimilating target substrates. However, the fractions with labelled universal genes are sometimes difficult to distinguish when detected by quantitative real-time PCR. In this experiment, three paddy soils (AQ, CZ, and NB) were amended with 0.1% glucose containing 13C at six levels, and DNA was then extracted after a 7-day incubation and subjected to isopycnic gradient centrifugation. The results showed that the amount of labelled DNA was notably related to the 13C-glucose percentage, while the separation spans of 18S rRNA and 16S rRNA genes between labelled and unlabelled treatments became notably clearer when the δ13C values of the total DNA were 90.9, 61.6, and 38.9‰ and 256.2, 104.5 and 126.1‰ in the AQ, CZ, and NB soils, respectively. Moreover, fractionated DNA was also labelled by determining the δ13C values while adding only 5 atom% 13C-glucose to the soil. The results suggest that the optimal labelling fractions were not always those fractions with the maximal gene abundance, and detecting the δ13C values of the total and fractionated DNA was beneficial in estimating the results of DNA-SIP.Key points• Appropriate 13C-DNA amount was needed for DNA-SIP.• Detecting the 13C ratio of fractionated DNA directly was an assistant method for identifying the labelled fractions.• Fractions with the maximal 18S or 16S rRNA gene abundance always were not labelled.

Regulation of nitrite-dependent anaerobic methane oxidation bacteria by available phosphorus and microbial communities in lake sediments of cold and arid regions

As global anthropogenic nitrogen inputs continue to rise, nitrite-dependent anaerobic methane oxidation (N-DAMO) plays an increasingly significant role in CH4 consumption in lake sediments. However, there is a dearth of knowledge regarding the effects of anthropogenic activities on N-DAMO bacteria in lakes in the cold and arid regions. Sediment samples were collected from five sampling areas in Lake Ulansuhai at varying depth ranges (0–20, 20–40, and 40–60 cm). The ecological characterization and niche differentiation of N-DAMO bacteria were investigated using bioinformatics and molecular biology techniques. Quantitative PCR confirmed the presence of N-DAMO bacteria in Lake Ulansuhai sediments, with 16S rRNA gene abundances ranging from 1.72 × 104 to 5.75 × 105 copies·g−1 dry sediment. The highest abundance was observed at the farmland drainage outlet with high available phosphorus (AP). Anthropogenic disturbances led to a significant increase in the abundance of N-DAMO bacteria, though their diversity remained unaffected. The heterogeneous community of N-DAMO bacteria was affected by interactions among various environmental characteristics, with AP and oxidation-reduction potential identified as the key drivers in this study. The Mantel test indicated that the N-DAMO bacterial abundance was more readily influenced by the presence of the denitrification genes (nirS and nirK). Network analysis revealed that the community structure of N-DAMO bacteria generated numerous links (especially positive links) with microbial taxa involved in carbon and nitrogen cycles, such as methanogens and nitrifying bacteria. In summary, N-DAMO bacteria exhibited sensitivity to both environmental and microbial factors under various human disturbances. This study provides valuable insights into the distribution patterns of N-DAMO bacteria and their roles in nitrogen and carbon cycling within lake ecosystems.

In situ incubation of iron(II)-bearing minerals and Fe(0) reveals insights into metabolic flexibility of chemolithotrophic bacteria in a nitrate polluted karst aquifer

Groundwater nitrate pollution is a major reason for deteriorating water quality and threatens human and animal health. Yet, mitigating groundwater contamination naturally is often complicated since most aquifers are limited in bioavailable carbon. Since metabolically flexible microbes might have advantages for survival, this study presents a detailed description and first results on our modification of the BacTrap© method, aiming to determine the prevailing microbial community's potential to utilize chemolithotrophic pathways. Our microbial trapping devices (MTDs) were amended with four different iron sources and incubated in seven groundwater monitoring wells for ∼3 months to promote growth of nitrate-reducing Fe(II)-oxidizing bacteria (NRFeOxB) in a nitrate-contaminated karst aquifer. Phylogenetic analysis based on 16S rRNA gene sequences implies that the identity of the iron source influenced the microbial community's composition. In addition, high throughput amplicon sequencing revealed increased relative 16S rRNA gene abundances of OTUs affiliated to genera such as Thiobacillus, Rhodobacter, Pseudomonas, Albidiferax, and Sideroxydans. MTD-derived enrichments set up with Fe(II)/nitrate/acetate to isolate potential NRFeOxB, were dominated by e.g., Acidovorax spp., Paracoccus spp. and Propionivibrio spp. MTDs are a cost-effective approach for investigating microorganisms in groundwater and our data not only solidifies the MTD's capacity to provide insights into the metabolic flexibility of the aquifer's microbial community, but also substantiates its metabolic potential for anaerobic Fe(II) oxidation.

Vertical and temporal variations in activity, abundance, and composition of nitrite-driven anaerobic methanotrophs in a paddy field

Paddy soil is an important source of methane (CH4) emissions, and the nitrite-driven anaerobic oxidation of methane (AOM) mediated by Candidatus Methylomirabilis bacteria is a crucial process for CH4 mitigation. However, the vertical and temporal fluctuations in nitrite-driven AOM activity, as well as the abundance and composition of Methylomirabilis bacteria in paddy soil, remain poorly understood. We investigated the potential rate of nitrite-driven AOM and the composition of Methylomirabilis bacteria at varying depths (0–40 cm, at 10 cm intervals) within a paddy soil during different growth stages of rice, including tillering, jointing, flowering, and milky. The potential rate of nitrite-driven AOM ranged from 0.24 to 20.6 nmol CO2 g−1 (dry soil) d−1, and the abundance of Methylomirabilis bacterial 16S rRNA genes varied from 5.8 × 106 to 4.4 × 107 copies g−1 dry soil. The 10–20 cm soil depth showed higher potential activity and bacterial abundance compared to other depths, and the tillering and jointing stages exhibited greater activity and abundance. The composition of Methylomirabilis bacteria showed significant variation across different soil depths but remained stable throughout the growth stages of rice. Soil organic carbon content was positively correlated with potential nitrite-driven AOM rate, and nitrite content was negatively correlated with the abundance of Methylomirabilis bacteria. Soil organic carbon and ammonium content had a significant impact on the composition of Methylomirabilis bacteria. The results underscore the substantial influence of soil depth and the growth stage of rice on nitrite-driven AOM in paddy ecosystems.

Effects of transitioning from conventional to organic farming on soil organic carbon and microbial community: a comparison of long-term non-inversion minimum tillage and conventional tillage

The combination of conservation tillage (non-inversion and no-till) with organic farming is rare due to weed problems. However, both practices have the potential to improve soil quality and increase soil organic C (SOC). This study investigated the changes in SOC, microbial biomass, and microbial composition during the transition from conventional to organic farming (from 2014 to 2020) in a long-term tillage trial established in 1999. Non-inversion minimum tillage to a depth of 10 cm (MT) resulted in SOC stratification, whilst conventional soil tillage with 25-cm-deep mouldboard ploughing (CT) maintained an even SOC distribution in the plough layer. After 12 years of contrasting tillage in 2011, the uppermost soil layer under MT had a 10% higher SOC content (1.6% w/w) than CT (1.45% w/w). This difference became even more pronounced after introducing organic farming in 2014. By the fall of 2020, the SOC content under MT increased to 1.94%, whilst it decreased slightly to 1.36% under CT, resulting in a 43% difference between the two systems. Conversion to organic farming increased microbial biomass under both tillage systems, whilst SOC remained unchanged in CT. Abundances of total bacterial and Crenarchaeal 16S rRNA and fungal ITS genes indicated shifts in the microbial community in response to tillage and depth. Fungal communities under MT were more responsive to organic farming than bacterial communities. The improved soil quality observed under MT supports its adoption in both organic and conventional systems, but potentially large yield losses due to increased weed cover discourage farmers from combining MT and organic farming.

Key taxa and mobilome-mediated responses co-reshape the soil antibiotic resistome under dazomet fumigation stress

Agrochemicals are emergingly being implicated in the widespread dissemination of antibiotic resistance genes (ARGs) in agroecosystems. However, minimal research exists on the disturbance of fumigant on soil ARGs. Focusing on a typical fumigant dazomet in a simulated soil microcosm, we characterized the dazomet-triggered timely response and longstanding dynamic of ARGs at one-fold and two-fold field recommended doses using metagenome and quantitative PCR. Dazomet treatments reduced 13.17%-69.98% of absolute abundance of 16S rRNA gene and targeted ARGs, but, awfully, boosted diversity and relative abundance of ARGs up to 1.33–1.60 and 1.62–1.90 folds, respectively. Approximately 77.28% of changes in relative abundance of ARGs could be explained by bacterial community and mobile genetic elements (MGEs). Mechanistically, primary hosts of ARGs shifted from Proteobacteria (control) to Firmicutes and Actinobacteria (treatments) accompanied with corresponding changes in their abundance by combining community analysis, host tracking analysis and antibiotic resistant bacteria assay. Meanwhile, dazomet exposure significantly increased the incidence of MGEs and stimulated the conjugation of antibiotic-resistant plasmid. In addition, absolute abundance of targeted ARGs gradually recovered in the post-fumigation stage. Collectively, our results elucidate the dazomet-triggered emergence and spread of soil ARGs and highlight the importance of navigating toward rational use of fumigant in agricultural fields.

Microbial gene abundance mirrors soil nitrogen mineralization intensity across an age gradient in Chinese-fir plantations

Stand development affects soil properties, nitrogen (N) dynamics, and soil microbial community composition, but the question remains whether differences in N mineralization rates are mirrored by the abundance of relevant functional genes. In this study, we used the 15N pool-dilution method to estimate N mineralization (i.e., ammonification and nitrification) rates across a Chinese fir (Cunninghamia lanceolata) chronosequence, with stands aged 7, 16, 29, 36, and >80 years. Gene copy numbers of bacteria (16S rRNA), fungi (ITS), ammonia-oxidizing archaea (AOA) and bacteria (AOB) (amoA), denitrifiers (nirS, nirK), N2 fixers (nifH) and organic N decomposers (chiA) were quantified by qPCR. Gross ammonification and nitrification rates increased linearly with stand age in the topsoil (0–5 cm depth) and were strongly positively correlated with the abundance of the bacterial 16S rRNA gene and AOA amoA, respectively. Higher net nitrification but lower NO3− immobilization rates in older stands (32 and > 80 years) drove higher N availability for vegetation than in young stands (7 years). Older stands also had higher rates of NH4+ consumption than younger stands due to the increased fungal ITS abundance and higher microbial biomass N (MBN), and AOA amoA was more abundant and active than AOB amoA due to the more acid conditions characteristic of mature forests. Redundancy analysis showed that functional gene abundance was strongly affected by soil properties such as pH, NH4+-N content, and MBN. We also found that microbial N storage potential was lower, and the NO3−-N leaching and gaseous N loss potential were higher in older stands than in younger stands. Collectively, stand developmental stage gave rise to the observed spatial gradient of gross ammonification and nitrification rates by altering the abundance of microbial functional genes, which affected plantation productivity via its modulation of the supply of bioavailable N.

Microbial signatures in amniotic fluid at preterm birth and association with bronchopulmonary dysplasia

BackgroundMicrobiome dysbiosis can have long-lasting effects on our health and induce the development of various diseases. Bronchopulmonary dysplasia (BPD) is a multifactorial disease with pre- and postnatal origins including intra-amniotic infection as main risk factor. Recently, postnatal pathologic lung microbiota colonization was associated with BPD. The objectives of this prospective observational cohort study were to describe differences in bacterial signatures in the amniotic fluid (AF) of intact pregnancies without clinical signs or risk of preterm delivery and AF samples obtained during preterm deliveries and their variations between different BPD disease severity stages.MethodsAF samples were collected under sterile conditions during fetal intervention from intact pregnancies (n = 17) or immediately before preterm delivery < 32 weeks (n = 126). Metabarcoding based approaches were used for the molecular assessment of bacterial 16S rRNA genes to describe bacterial community structure.ResultsThe absolute amount of 16S rRNA genes was significantly increased in AF of preterm deliveries and detailed profiling revealed a reduced alpha diversity and a significant change in beta diversity with a reduced relative abundance of 16S rRNA genes indicative for Lactobacillus and Acetobacter while Fusobacterium, Pseudomonas, Ureaplasma and Staphylococcus 16S rRNA gene prevailed. Although classification of BPD by disease severity revealed equivalent absolute 16S rRNA gene abundance and alpha and beta diversity in no, mild and moderate/severe BPD groups, for some 16S rRNA genes differences were observed in AF samples. Bacterial signatures of infants with moderate/severe BPD showed predominance of 16S rRNA genes belonging to the Escherichia-Shigella cluster while Ureaplasma and Enterococcus species were enriched in AF samples of infants with mild BPD.ConclusionsOur study identified distinct and diverse intrauterine 16S rRNA gene patterns in preterm infants immediately before birth, differing from the 16S rRNA gene signature of intact pregnancies. The distinct 16S rRNA gene signatures at birth derive from bacteria with varying pathogenicity to the immature lung and are suited to identify preterm infants at risk. Our results emphasize the prenatal impact to the origins of BPD.

Influence of sedimentary deposition on the microbial assembly process in Arctic Holocene marine sediments.

The sea-level rise during the Holocene (11-0 ky BP) and its resulting sedimentation and biogeochemical processes may control microbial life in Arctic sediments. To gain further insight into this interaction, we investigated a sediment core (up to 10.7 m below the seafloor) from the Chuckchi Shelf of the western Arctic Ocean using metabarcoding-based sequencing and qPCR to characterize archaeal and bacterial 16S rRNA gene composition and abundance, respectively. We found that Arctic Holocene sediments harbor local microbial communities, reflecting geochemical and paleoclimate separations. The composition of bacterial communities was more diverse than that of archaeal communities, and specifically distinct at the boundary layer of the sulfate-methane transition zone. Enriched cyanobacterial sequences in the Arctic middle Holocene (8-7 ky BP) methanogenic sediments remarkably suggest past cyanobacterial blooms. Bacterial communities were phylogenetically influenced by interactions between dispersal limitation and environmental selection governing community assembly under past oceanographic changes. The relative influence of stochastic and deterministic processes on the bacterial assemblage was primarily determined by dispersal limitation. We have summarized our findings in a conceptual model that revealed how changes in paleoclimate phases cause shifts in ecological succession and the assembly process. In this ecological model, dispersal limitation is an important driving force for progressive succession for bacterial community assembly processes on a geological timescale in the western Arctic Ocean. This enabled a better understanding of the ecological processes that drive the assembly of communities in Holocene sedimentary habitats affected by sea-level rise, such as in the shallow western Arctic shelves.

Hidden risks: Simulated leakage of domestic sewage to secondary water supply systems poses serious microbiological risks

There are continuous reports about the pollution of the secondary water supply systems (SWSSs), among which domestic sewage leakage is the most serious. In this study, a pilot experiment lasting 70 days was conducted to explore the changes in physicochemical water quality and the microbial profiles in SWSSs polluted by different doses of domestic sewage through qPCR and high-throughput sequencing methods. The results showed that when domestic sewage entered the simulated water storage tank, a large amount of organic matter brought by domestic sewage quickly consumed chlorine disinfectants. High pollution levels (pollution index ≥ 1/1000) were accompanied by significant increases in turbidity and ammonia nitrogen concentration (p < 0.05) and by abnormal changes in sensory properties. Although different microbial community structures were found only at high pollution levels, qPCR results showed that the abundance of the bacterial 16S rRNA gene and some pathogenic gene markers in the polluted tank increased with the pollution level, and the specific gene marker of pathogens could be detected even at imperceptible pollution levels. In particular, the high detection frequency and abundance of Escherichia coli and Enterococcus faecails in polluted tank water samples demonstrated that they can be used for early warning. Moreover, it seems that the microorganisms that came with the domestic sewage lost their cultivability soon after entering SWSSs but could recover their activities during stagnation. In addition, the biofilm biomass in the polluted tank with high pollution levels increased faster at the initial stage, while after a longer contact time, it tended to remain at the same level as the control tank. This study emphasized the high microbial risk introduced by sewage water leakage even at imperceptible levels and could provide scientific suggestions for early warning and prevention of pollution to SWSSs.

Potential benefits and risks of solar photovoltaic power plants on arid and semi-arid ecosystems: an assessment of soil microbial and plant communities.

Exponential increase in photovoltaic installations arouses concerns regarding the impacts of large-scale solar power plants on dryland ecosystems. While the effects of photovoltaic panels on soil moisture content and plant biomass in arid ecosystems have been recognized, little is known about their influence on soil microbial communities. Here, we employed a combination of quantitative PCR, high-throughput sequencing, and soil property analysis to investigate the responses of soil microbial communities to solar panel installation. We also report on the responses of plant communities within the same solar farm. Our findings showed that soil microbial communities responded differently to the shading and precipitation-alternation effects of the photovoltaic panels in an arid ecosystem. By redirecting rainwater to the lower side, photovoltaic panels stimulated vegetation biomass and soil total organic carbon content in the middle and in front of the panels, positively contributing to carbon storage. The shade provided by the panels promoted the co-occurrence of soil microbes but inhibited the abundance of 16S rRNA gene in the soil. Increase in precipitation reduced 18S rRNA gene abundance, whereas decrease in precipitation led to decline in plant aboveground biomass, soil prokaryotic community alpha diversity, and dehydrogenase activity under the panels. These findings highlight the crucial role of precipitation in maintaining plant and soil microbial diversities in dryland ecosystems and are essential for estimating the potential risks of large-scale solar power plants on local and global climate change in the long term.

High variability in organic carbon sources and microbial activities in the hadopelagic waters

AbstractHadal sediments are recognized as organic carbon depocenters with intensified microbial activity compared to adjacent abyssal sites due to focusing of relatively labile organic materials. However, the sources and turnover of hadopelagic organic carbon and its linkages to microbial activities have not been studied. We present the first synergic research on particulate organic carbon, dark carbon fixation, and size‐fractionated microbial community respiration proxy over the Atacama Trench. The results demonstrate that all parameters attenuate rapidly from surface to mesopelagic water (~ 1000 m). Progressing deeper, values remain relatively stable throughout bathypelagic (~ 4000 m) and abyssopelagic (~ 6000 m) waters. However, in the hadopelagic zone (&gt; 6000 m), highly variable values indicate dynamic organic carbon sources and microbial activities in the deepest trench. On average, 71% of the microbial community respiration proxy is attributable to particle‐associated communities, indicating importance of particles for microbial metabolism. No apparent relationship was observed between the microbial community respiration proxy and microbial 16S rRNA gene abundance below the epipelagic depth, indicating variable supply and quality of organic carbon likely constrained heterotrophic activities rather than microbial abundances in the deep ocean. The depth‐integrated dark carbon fixation (&gt; 1000 m) accounts for 11.5% ± 7.6% of the surface net primary production, of which 2.9% ± 0.4% is from hadopelagic depth. Dark carbon fixation is thus an important in situ organic carbon source for hadal life. This study suggests that high variability in organic carbon sources and microbial activities in the hadopelagic trench cannot be simply extrapolated from findings in the shallower dark ocean (e.g., 1000–6000 m).

Effects of Cropping Systems on Soil Physicochemical Properties and Abundances and Spatial Distributions of Nitrogen-Cycling Bacteria

Soil nitrogen (N) is a common limiting factor where soil N-cycling is a key component of agroecosystems. Soil N transformation processes are largely mediated by microbes, and understanding bacteria involvement in soil N-cycling in agricultural systems has both agronomic and environmental importance. This 2 yr field-scale study examined the abundances and spatial distributions of the total bacterial community (16S rRNA), bacteria involved in nitrification (amoA) and denitrification (narG, nirK, and nosZ), and soil physicochemical properties of winter wheat (Triticum aestivum L.)–soybean (Glycine max L.) double-crop with 2–3 weeks of spring grazing (WGS) and without grazing (WS) and tall fescue (Festuca arundinacea (L.) Schreb.) pasture (TF) managed to near-natural conditions with similar grazing. The TF soil had a significantly higher abundance of 16S rRNA, amoA, narG, nirK, and nosZ genes than the WS and WGS soils, which had similar levels between themselves. Soil organic matter (OM) and soil pH had stronger effects on the N-cycling bacteria gene abundance. All bacterial gene concentrations and soil pH showed nonrandom distribution patterns with a 141–186 m range autocorrelation. These results indicate that biological N transformation processes are more important in natural agricultural systems and the abundance of N-cycling bacteria can be manipulated by field-scale management strategies.

A state-of-the-art review and guidelines for enhancing nitrate removal in bio-electrochemical systems (BES)

Excess unmanaged nitrogen pollutes the environment. A sustainable wastewater treatment system must achieve better pollutant removal efficiency at a lower cost, and the feasibility of integrating biological nitrogen removal into bio-electrochemical systems (BES) has been reported as a tool in green technology. Nitrogen in the form of nitrate (NO3) is a common pollutant in both surface and ground waters, and a high level of NO3 makes water unsuitable for drinking water. This analysis and review of BES for treating NO3 polluted water investigate BES's components and operational factors and their importance on the NO3 removal efficiency to design more powerful but economic systems. The NO3 removal efficiencies were analyzed by the influence of electrode materials, working mode, number of chambers, type of inoculum, capacity, and microbial community structure. Overall, the electrode materials, significantly influence the NO3 removal rate. The operational parameters, such as working mode, the number of chambers, inoculum type and the systems' capacity, were deemed important and have significantly influenced the NO3 removal efficiencies when analyzed by the random forest classification algorithm. Proteobacteria and Firmicute were the prominent phyla observed in BES treating NO3 polluted water. Besides the denitrification (abundance of narG, nirS, nirK, nosZI, and nosZII genes) process in BES, there is evidence of electrochemical support for anaerobic ammonium oxidation (ANAMMOX) (abundance of hzsB or ANAMMOX-specific 16S rRNA genes) and dissimilatory NO3 reduction to ammonium (DNRA) (abundance of nrfA genes) processes. Our analysis suggest that BES, as a continuous two-chamber system with cathode and anode materials as granular carbon and carbon paper, respectively, with denitrifying microbes as inoculum type, would contribute to optimum NO3 removal efficiencies.

Correlation between Bacterial Cell Density and Abundance of Antibiotic Resistance on Milking Machine Surfaces Assessed by Cultivation and Direct qPCR Methods

The relative abundance of antibiotic-resistant bacteria and antibiotic-resistance genes was surveyed for different parts of a milking machine. A cultivation approach based on swab samples showed a highly diverse microbiota, harboring resistances against cloxacillin, ampicillin, penicillin, and tetracycline. This approach demonstrated a substantial cloxacillin resistance of numerous taxa within milking machine microbiota coming along with regular use of cloxacillin for dry-off therapy of dairy cows. For the less abundant tetracycline-resistant bacteria we found a positive correlation between microbial cell density and relative abundance of tetracycline-resistant microorganisms (R2 = 0.73). This indicated an accelerated dispersion of resistant cells for sampling locations with high cell density. However, the direct quantification of the tetM gene from the swap samples by qPCR showed the reverse relation to bacterial density if normalized against the abundance of 16S rRNA genes (R2 = 0.88). The abundance of 16S rRNA genes was analyzed by qPCR combined with a propidium monoazide treatment, which eliminates 16S rRNA gene signals in negative controls.

Alizarin enhancement of the abundance of ARGs and impacts on the microbial community in water.

Alizarin, a dyestuff from herbs, showed effective inhibition effects on pathogenic bacteria, and thus has been frequently used in the world as the main alternative to antibiotics in the treatment of inflammations and pathogen infections. However, it was unclear whether alizarin played key a role in antibiotic-induced antibiotic-resistant gene (ARG) alterations and impacted microbial community shifts in aquatic environments. In this study, the effects of alizarin or co-exposure of alizarin with antibiotics on the fate of ARGs, class 1 integron-integrase gene (intI1), and microbial populations in lake water were investigated, and the potential hosts for ARGs were analyzed. The results showed that the absolute abundance of 16s rRNA gene, ARGs (tetA, tetC, and qnrS), and intI1 were increased during the treatment of alizarin. The combination of alizarin and antibiotics was superior to alizarin in its ability to promote population growth of bacteria and induce ARGs. Additionally, alizarin more significantly altered the community composition of microorganisms in water, which resulted in differences in bacterial communities and functions.

Impact of nitrogen and phosphorus on the growth and microcystin‐related gene expression of Microcystis aeruginosa PCC 7806

AbstractHarmful algal blooms (HABs) have incurred numerous health problems through drinking water or exposure. A better understanding of the growth kinetics and toxin production under different environmental conditions will help develop tools for early‐warning indicators for predicting HABs. Lab‐scale experiments were performed with Microcystis aeruginosa PCC 7806 under varying N:P ratios and temperatures. We observed different growth behaviors for cultures growing at the three temperatures (20, 25, and 30°C). M. aeruginosa exhibited higher growth rates at higher N:P ratios during the incubation at 25 and 30°C. Microcystis 16S rRNA gene abundance correlated well with the cell growth parameters. Based on RT‐qPCR data, the transcripts of mcyA, mcyD, and mcyE were occasionally detected with N:P of 76.86 showing highest detections of all gene transcripts. Intracellular microcystin (MC) production corresponded well with the presence of gene transcripts, while no extracellular MC was detected. These findings would provide valuable insights and beneficial guidance to further explore and predict the impacts of cyanobacterial blooms in freshwater systems.