Epiberberine: a potential rumen microbial urease inhibitor to reduce ammonia release screened by targeting UreG.

Rumen microbial urease inhibitors have been proposed for regulating nitrogen emission and improving nitrogen utilization efficiency in ruminant livestock industry. However, studies on plant-derived natural inhibitors of rumen microbial urease are limited. Urease accessory protein UreG, plays a crucial role in facilitating urease maturation, is a new target for design of urease inhibitor. The objective of this study was to select the potential effective inhibitor of rumen microbial urease from major protoberberine alkaloids in Rhizoma Coptidis by targeting UreG. Our results showed that berberine chloride and epiberberine exerted superior inhibition potential than other alkaloids based on GTPase activity study of UreG. Berberine chloride inhibition of UreG was mixed type, while inhibition kinetics type of epiberberine was uncompetitive. Furthermore, epiberberine was found to be more effective than berberine chloride in inhibiting the combination of nickel towards UreG and inducing changes in the second structure of UreG. Molecular modeling provided the rational structural basis for the higher inhibition potential of epiberberine, amino acid residues in G1 motif and G3 motif of UreG formed interactions with D ring of berberine chloride, while interacted with A ring and D ring of epiberberine. We further demonstrated the efficacy of epiberberine in the ruminal microbial fermentation with low ammonia release and urea degradation. In conclusion, our study clearly indicates that epiberberine is a promising candidate as a safe and effective inhibitor of rumen microbial urease and provides an optimal strategy and suitable feed additive for regulating nitrogen excretion in ruminants in the future. KEY POINTS: • Epiberberine is the most effective inhibitor of rumen urease from Rhizoma Coptidis. • Urease accessory protein UreG is an effective target for design of urease inhibitor. • Epiberberine may be used as natural feed additive to reducing NH3 release in ruminants.

Response of plants and soils to inundation duration and construction of the plant‒soil association mode in the hydro‒fluctuation belt of the reservoir wetland.

Hydro-Fluctuation Belt (HFB), a periodically exposed bank area formed by changes in water level fluctuations, is critical for damaging the reservoir wetland landscape and ecological balance. Thus, it is important to explore the mechanism of hydrological conditions on the plant-soil system of the HFB for protection of the reservoir wetland and landscape restoration. Here, we investigated the response of plant community characteristics and soil environment of the HFB of Tonghui River National Wetland Park (China), is a typical reservoir wetland, to the duration of inundation, as well as the correlation between the distribution of dominant plants and soil pH, nutrient contents, and enzyme activity by linear regression and canonical correlation analyses. The results show that as the duration of inundation decreases, the vegetation within the HFB is successional from annual or biennial herbs to perennial herbs and shrubs, with dominant plant species prominent and uneven distribution of species. Soil nutrient contents and enzyme activities of HFB decreased with increasing inundation duration. Dominant species of HFB plant community are related to soil environment, with water content, pH, urease, and available potassium being principle soil environmental factors affecting their distribution. When HFB was inundated for 0-30 days, soil pH was strongly acidic, with available potassium content above 150 mg kg-1 and higher urease activity, distributed with Arundo donax L., Polygonum perfoliatum L., Alternanthera philoxeroides (Mart.) Griseb., and Daucus carota L. communities. When inundated for 30-80 days, soil pH was acidic, with lower available potassium content (50-150 mg kg-1) and urease activity, distributed with Beckmannia syzigachne (Steud.) Fern.+ Polygonum lapathifolium L., Polygonum lapathifolium L., Medicago lupulina L. + Dysphania ambrosioides L. and Leptochloa panicea (Retz.) Ohwi communities. Using the constructed HFB plant-soil correlation model, changes in the wetland soil environment can be quickly judged by the succession of plant dominant species, which provides a simpler method for the monitoring of the soil environment in the reservoir wetland, and is of great significance for the scientific management and reasonable protection of the reservoir-type wetland ecosystem.

Feeding strategies for Sporosarcina pasteurii cultivation unlock more efficient production of ureolytic biomass for MICP.

The bacterium Sporosarcina pasteurii is the most commonly used microorganism for Microbial Induced Calcite Precipitation (MICP) due to its high urease activity. To date, no proper fed-batch cultivation protocol for S. pasteurii has been published, even though this cultivation method has a high potential for reducing costs of producing microbial ureolytic biomass. This study focusses on fed-batch cultivation of S. pasteurii DSM33. The study distinguishes between limited fed-batch cultivation and extended batch cultivation. Simply feeding glucose to a S. pasteurii culture does not seem beneficial. However, it was exploited that S. pasteurii is auxotrophic for two vitamins and amino acids. Limited fed-batch cultivation was accomplished by feeding the necessary vitamins or amino acids to a culture lacking them. Feeding nicotinic acid to a nicotinic acid deprived culture resulted in a 24% increase of the specific urease activity compared to a fed culture without nicotinic acid limitation. Also, extended batch cultivation was explored. Feeding a mixture of glucose and yeast extract results in OD600 of ≈70 at the end of cultivation, which is the highest value published in literature so far. These results have the potential to make MICP applications economically viable.

In vitro anti-Helicobacter pylori activity and antivirulence activity of cetylpyridinium chloride.

The primary treatment method for eradicating Helicobacter pylori (H. pylori) infection involves the use of antibiotic-based therapies. Due to the growing antibiotic resistance of H. pylori, there has been a surge of interest in exploring alternative therapies. Cetylpyridinium chloride (CPC) is a water-soluble and nonvolatile quaternary ammonium compound with exceptional broad-spectrum antibacterial properties. To date, there is no documented or described specific antibacterial action of CPC against H. pylori. Therefore, this study aimed to explore the in vitro activity of CPC against H. pylori and its potential antibacterial mechanism. CPC exhibited significant in vitro activity against H. pylori, with MICs ranging from 0.16 to 0.62 µg/mL and MBCs ranging from 0.31 to 1.24 µg/mL. CPC could result in morphological and physiological modifications in H. pylori, leading to the suppression of virulence and adherence genes expression, including flaA, flaB, babB, alpA, alpB, ureE, and ureF, and inhibition of urease activity. CPC has demonstrated in vitro activity against H. pylori by inhibiting its growth, inducing damage to the bacterial structure, reducing virulence and adherence factors expression, and inhibiting urease activity.

Freeze-thaw aged polyethylene and polypropylene microplastics alter enzyme activity and microbial community composition in soil.

In cold regions, microplastics (MPs) in the soil undergo freeze-thaw (FT) aging process. Little is known about how FT aged MPs influence soil physico-chemical properties and microbial communities. Here, two environmentally relevant concentrations (50 and 500 mg/kg) of 50 and 500 µm polyethylene (PE) and polypropylene (PP) MPs treated soils were subjected to 45-day FT cycles (FTCs). Results showed that MPs experienced surface morphology, hydrophobicity and crystallinity alterations after FTCs. After 45-day FTCs, the soil urease (SUE) activity in control (MPs-free group that underwent FTCs) was 33.49 U/g. SUE activity in 50 µm PE group was reduced by 19.66 %, while increased by 21.16 % and 37.73 % in 500 µm PE and PP groups compared to control. The highest Shannon index was found in 50 µm PP-MPs group at 50 mg/kg, 2.26 % higher than control (7.09). Compared to control (average weighted degree=8.024), all aged MPs increased the complexity of network (0.19-1.43 %). Bacterial biomarkers of aged PP-MPs were associated with pollutant degradation. Aged PP-MPs affected genetic information, cellular processes, and disrupted the biosynthesis of metabolites. This study provides new insights into the potential hazards of MPs after FTCs on soil ecosystem in cold regions.

Genomic characterization of a novel ureolytic bacteria, Lysinibacillus capsici TSBLM, and its application to the remediation of acidic heavy metal-contaminated soil.

Soil heavy metal contamination is an essential challenge in ecological and environmental management, especially for acidic soils. Microbially induced carbonate precipitation (MICP) is an effective and environmentally friendly remediation technology for heavy metal contaminated sites, and one of the key factors for its realization lies in the microorganisms. In this study, Lysinibacillus capsici TSBLM was isolated from heavy metal contaminated soil around a gold mine, and inferred to be a novel ureolytic bacteria after phylogenomic inference and genome characterization. The urease of L. capsici TSBLM was analyzed by genetic analysis and molecular docking, and further applied this bacteria to the remediation of Cu and Pb in solution and acidic soils to investigate its biomineralization mechanism and practical application. The results revealed L. capsici TSBLM possessed a comprehensive urease gene cluster ureABCEFGD, and the encoded urease docked with urea at the lowest binding energy site (ΔG = -3.43 kcal/mol) connected to three amino acids threonine, aspartic, and alanine. The urease of L. capsici TSBLM is synthesized intracellularly but mainly functions extracellularly. L. capsici TSBLM removes Cu/Pb from the solution by generating heavy metal carbonates or co-precipitating with CaCO3 vaterite. For acidic heavy metal-contaminated soil, the carbonate-bound states of Cu and Pb increased significantly from 7 % to 16 % and from 23 % to 35 % after 30 days by L. capsici TSBLM. Soil pH improved additionally. L. capsici TSBLM maintained the dominant status in the remediated soil after 30 days, demonstrating good environmental adaptability and curing persistence. The results provided new strain resources and practical application references for the remediation of acidic heavy metal contaminated soil based on MICP.

Deterioration phenomenon of Pb-contaminated aqueous solution remediation and enhancement mechanism of nano-hydroxyapatite-assisted biomineralization.

Modern metallurgical and smelting activities discharge the lead-containing wastewater, causing serious threats to human health. Bacteria and urease applied to microbial-induced carbonate precipitation (MICP) and enzyme-induced carbonate precipitation (EICP) are denatured under high Pb2+ concentration. The nano-hydroxyapatite (nHAP)-assisted biomineralization technology was applied in this study for Pb immobilization. Results showed that the extracellular polymers and cell membranes failed to secure the urease activity when subjected to 60 mM Pb2+. The immobilization efficiency dropped to below 50% under MICP, whereas it due to a lack of extracellular polymers and cell membranes dropped to below 30% under EICP. nHAP prevented the attachment of Pb2+ either through competing with bacteria and urease or promoting Ca2+/Pb2+ ion exchange. Furthermore, CO32- from ureolysis replaced the hydroxyl (-OH) in hydroxylpyromorphite to encourage the formation of carbonate-bearing hydroxylpyromorphite of higher stability (Pb10(PO4)6CO3). Moreover, nHAP application overcame an inability to provide nucleation sites by urease. As a result, the immobilization efficiency, when subjected to 60 mM Pb2+, elevated to above 80% under MICP-nHAP and to some 70% under EICP-nHAP. The findings highlight the potential of applying the nHAP-assisted biomineralization technology to Pb-containing water bodies remediation.

Microbial-induced carbonate precipitation effectively prevents Pb2+ migration through the soil profile: Lab experiment and model simulation.

Due to the inappropriate disposal of waste materials containing lead (Pb) and irrigation with sewage containing Pb, the migration of Pb2+ within the soil profile has been extensively investigated. The conventional Pb2+ block method is challenging to implement due to its complex operational procedures and high construction costs. To address this issue, this study introduces the microbial-induced carbonate precipitation (MICP) technique as a novel approach to impede the migration of Pb2+ in the soil profile. Soil acclimatization with urea resulted in an increased proportion of urease-producing microorganisms, including Bacillus, Paenibacillus, and Planococcaceae, along with heightened expression of urea-hydrolyzing genes (UreA, UreB, UreC, and UreG). This indicates that urea-acclimatized soil (Soil-MICP) possesses the potential to induce carbonate precipitation. Batch Pb2+ fixation experiments confirmed that the fixation efficiency of Soil-MICP on Pb2+ exceeded that of soil without MICP, attributed to the MICP process within the Soil-MICP group. Dynamic migration experiments revealed that the MICP reaction transformed exchangeable lead into carbonate-bound Pb, effectively impeding Pb2+ migration in the soil profile. Additionally, the migration rate of Pb2+ in Soil-MICP was influenced by varying urea amounts, pH levels, and pore flow rates, leading to a slowdown in migration. The Two-site sorption model aptly described the Pb2+ migration process in the Soil-MICP column. This study aims to elucidate the MICP biomineralization process, uncover the in-situ blocking mechanism of MICP on lead in soil, investigate the impact of Pb on key genes involved in urease metabolism, enhance the comprehension of the chemical morphology of lead mineralization products, and provide a theoretical foundation for MICP technology in preventing the migration of Pb2+ in soil profiles.

Influence of enzyme activity on domestic wastewater amelioration in a hybrid subsurface flow constructed wetland.

Wastewater treatment in a constructed wetland is achieved by the presence of plant species, the metabolism of microorganisms, and the enzyme activities. Three small-scale hybrid subsurface flow constructed wetlands (HSFCWs) planted with Arundo donax and one unplanted HSFCW were constructed near a water resource recovery facility at Guru Gobind Singh Indraprastha University. The purpose of the study was to determine the correlation between soil enzymatic activities and the removal of contaminants from domestic wastewater. Enzyme activity of phosphatase, protease, urease, and cellulase increased with an increase in temperature. A strong correlation between enzyme activities and TKN and surfactant removal was observed, whereas moderate correlation was observed with phosphate in planted HSFCW during the study. The correlation between COD removal and enzyme activities was low to moderate. In unplanted HSFCW, the correlation between enzyme activities and COD removal was negative, negligible to moderate to strong in the case of TKN, low to moderate in the case of phosphate, and negligible to low in the case of surfactants. The increased removal efficiency of the planted system compared with that of the unplanted system indicated a positive impact on enzyme activities with the growth of plants and their roots. PRACTITIONER POINTS: Protease, urease, and cellulase activities: Planted HSFCW exhibited higher protease, urease, and cellulase activities than unplanted, signifying enhanced breakdown. July displayed maximum enzyme activities, correlating with heightened biological breakdown in both systems. Fluctuations in enzyme activities reflected seasonal changes, influencing nutrient degradation rates. Planted HSFCW consistently showed higher enzymatic activities across protease, urease, and cellulase than unplanted.

Influence of nitrogen substitution at an equivalent total nitrogen level on bacterial and fungal communities, as well as enzyme activities of the ditch bottom soil in a rice-fish coculture system.

BACKGROUND: Rice-fish coculture system (RFS) operates through effectively utilizing water and land resources in a complementary form, but it requires more efficient utilization of fertilizer and feed without compromising rice yield. However, the knowledge of how to regulate the proportion of nitrogen (N) supplied from fertilizer and feed at an equivalent total N level to improve the benefits of RFS remains limited. Therefore, four treatments (S0: RFS with 0% N from fertilizer and 100% N from feed; S25: RFS with 25% N from fertilizer and 75% N from feed; S50: RFS with 50% N from fertilizer and 50% N from feed; S75: RFS with 75% N from fertilizer and 25% N from feed) were conducted to assess the variation of ditch bottom soil properties, microbial communities and enzyme activities, as well as to obtain the optimal ratio of N supplied from fish feed and fertilizer. RESULTS: The experiments showed that the contents of soil organic matter, total carbon and total N, and the activities of urease, N-acetyl-ß-D-glucosaminidase, protease, ß-1,4-glucosidase and catalase in the ditch bottom soil significantly reduced in S25 treatment, compared with the other three treatments. Ammonium N content decreased with increasing percentage of the basal fertilizer, whereas nitrate N content and pH value showed an adverse trend. However, the bacterial and fungal communities were unaffected by the ratio shifts between fertilizer-N and feed-N, but their dominant phyla were influenced by the ditch bottom soil N level. Moreover, the bacterial community composition was positively related to nitrate N, whereas fungal diversity was positively correlated with pH, ammonium N and nitrate N, and urease. We also found that the treatment of N input with 25% N from fertilizer and 75% N from feed can reduce N deposition in the ditch bottom soil in the rice-fish coculture system. CONCLUSION: Our findings indicate that under the equivalent total N input level, the relative higher ratio of N from fish feed increased (S0 treatment) or reduced (S25 treatment) the deposition of N in the ditch bottom soil, and improved fish production, but decreased rice yield; while the higher ratio of N from basal fertilizer increased the transportation of nutrients into the ditch bottom soil and rice yield, but reduced fish production. So when considering multi-balance and multiple benefits, we recommend that a selective substitution ratio within 50% ~ 75% from fish feed to substitute for the basal fertilizer under the equivalent total N input may achieve a good balance of rice and fish production improvement, and reduce nutrients wastage to the ditch bottom, as well as alleviate the potential of non-point source pollution. This study also provides an evidence for regulating and optimizing the ratio of N supplied from fertilizer and fish feed at an equivalent total N level through monitoring the nutrient accumulation in ditch bottom soil in the rice-fish coculture system. © 2024 Society of Chemical Industry.

Penicillium oxalicum SL2-enhanced nanoscale zero-valent iron effectively reduces Cr(VI) and shifts soil microbiota.

Most current researches focus solely on reducing soil chromium availability. It is difficult to reduce soil Cr(VI) concentration below 5.0 mg kg-1 using single remediation technology. This study introduced a sustainable soil Cr(VI) reduction and stabilization system, Penicillium oxalicum SL2-nanoscale zero-valent iron (nZVI), and investigated its effect on Cr(VI) reduction efficiency and microbial ecology. Results showed that P. oxalicum SL2-nZVI effectively reduced soil total Cr(VI) concentration from 187.1 to 3.4 mg kg-1 within 180 d, and remained relatively stable at 360 d. The growth curve of P. oxalicum SL2 and microbial community results indicated that γ-ray irradiation shortened the adaptation time of P. oxalicum SL2 and facilitated its colonization in soil. P. oxalicum SL2 colonization activated nZVI and its derivatives, and increased soil iron bioavailability. After restoration, the negative effect of Cr(VI) on soil microorganisms was markedly alleviated. Cr(VI), Fe(II), bioavailable Cr/Fe, Eh, EC and urease (SUE) were the key environmental factors of soil microbiota. Notably, Penicillium significantly stimulated the growth of urease-positive bacteria, Arthrobacter, Pseudarthrobacter, and Microvirga, synergistically reducing soil chromium availability. The combination of P. oxalicum SL2 and nZVI is expected to form a green, economical and long-lasting Cr(VI) reduction stabilization strategy.

Genome Sequence Analysis of Calcifying Bacteria Bacillus paranthracis CT5 and Its Biomineralization Efficacy to Improve the Strength and Durability Properties of Civil Structures.

Bacteria producing urea amidohydrolases (UA) and carbonic anhydrases (CA) are of great importance in civil engineering as these enzymes are responsible for microbially induced calcium carbonate precipitation (MICCP). In this investigation, genomic insights of Bacillus paranthracis CT5 and the expression of genes underlying in MICCP were studied. B. paranthracis produced a maximum level of UA (669.3 U/ml) and CA (125 U/ml) on 5th day of incubation and precipitated 197 mg/100 ml CaCO3 after 7 days of incubation. After 28 days of curing, compressive strength of bacterial admixed and bacterial cured (B-B) specimens was 13.7% higher compared to water-mixed and water-cured (W-W) specimens. A significant decrease in water absorption was observed in bacterial-cured specimens compared to water-cured specimens after 28 days of curing. For genome analysis, reads were assembled de novo producing 5,402,771 bp assembly with N50 of 273,050 bp. RAST annotation detected six amidohydrolase and three carbonic anhydrase genes. Among 5700 coding sequences found in genome, COG gene annotation grouped 4360 genes into COG categories with highest number of genes to transcription (435 genes), amino acid transport and metabolism (362 genes) along with cell wall/membrane/envelope biogenesis and ion transport and metabolism. KEGG functional classification predicted 223 pathways consisting of 1,960 genes and the highest number of genes belongs to two-component system (101 genes) and ABC transporter pathways (98 genes) enabling bacteria to sense and respond to environmental signals and actively transport various minerals and organic molecules, which facilitate the active transport of molecules required for MICCP.

Nickel sources affect soil biological properties but do not affect sorghum growth.

Nickel (Ni) is an essential element, but it can be phytotoxic in high concentration, which may be caused by high availability in soil solution. The objective of this study was to evaluate the effect of sources and doses of Ni applied to a dystrophic Red Latosol cultivated with sorghum on i) the availability of the metal in the soil; ii) the impact on biological and biochemical properties of the soil; iii) the absorption and distribution in sorghum plants; and iv) crop productivity. The experiment was carried out within a completely randomized design with two nickel sources [nickel(II) nitrate, Ni(NO3)2 and nickel(III) oxide, Ni2O3], three doses (35, 70, and 140 mg Ni kg-1 soil), plus controls without Ni, with 3 replications. The concentrations of Ni in the soil, soil microbial biomass (SMB), basal soil respiration (BSR), metabolic quotient (qCO2), fluorescein diacetate (FDA) hydrolysis, and urease activity were determined. The concentrations of Ni in the leaf diagnostic and in the plant (shoot, root, and grains) were also measured. In the soil, the concentrations of available Ni remained between 0.21 and 54.01 mg Ni kg-1. Ni2O3 contributed very little to the increase in available Ni. SMB and the FDA hydrolysis were not affected by the Ni source or Ni dose, but BSR and qCO2 had significant increase with Ni application rates, suggesting the soil microorganisms faced stress. Soil urease activity was affected by Ni dose but not by Ni source. The dose of Ni as Ni(NO3)2 decreased the metal concentration in the plant, while that of Ni2O3 increased it. Nickel source did not affect dry mass production of the plants, but grain yield was affected in a dose-dependent manner when Ni2O3 was the source of Ni.

Promoting nitrogen conversion in aerobic biotransformation of swine slurry with the co-application of manganese sulfate and biochar.

This study aimed to explore the co-application of MnSO4 (Mn) and biochar (BC) in nitrogen conversion during the composting process. A 70-day aerobic composting was conducted using swine slurry, supplemented with different levels of Mn (0, 0.25%, and 0.5%) and 5% BC. The results demonstrated that the treatment with 0.5MnBC had the highest levels of NH4+-N (3.07 g kg-1), TKN (29.90 g kg-1), and NO3--N (1.94 g kg-1) among all treatments. Additionally, the 0.5MnBC treatment demonstrated higher urease, protease, nitrate reductase, and nitrite reductase activities than the other treatments, with the peak values of 18.12, 6.96, 3.57, and 15.14 mg g-1 d-1, respectively. The addition of Mn2+ increased the total organic nitrogen content by 29.59%-47.82%, the acid hydrolyzed ammonia nitrogen (AN) content by 13.84%-57.86% and the amino acid nitrogen (AAN) content by 55.38%-77.83%. The richness of Chloroflexi and Ascomycota was also enhanced by the simultaneous application of BC and Mn. Structural equation modeling analysis showed that Mn2+ can promote the conversion of Hydrolyzed Unknown Nitrogen (HUN) into AAN, and there is a positive association between urease and NH4+-N according to redundancy analysis. Firmicutes, Basidiomycota, and Mortierellomycota showed significant positive correlations with ASN, AN, and NH4+-N, indicating their crucial roles in nitrogen conversion. This study sheds light on promoting nitrogen conversion in swine slurry composting through the co-application of biochar and manganese sulfate.

Biomineralization of heavy metals based on urea transport and hydrolysis within a new bacterial isolate, B. intermedia TSBOI.

A newly isolated ureolytic bacteria, Brucella intermedia TSBOI, exhibited microbially induced calcite precipitation (MICP) which is a promising technique for the remediation of heavy metals in polluted environments. Brucella intermedia TSBOI achieved 90-100% removal of 1 mmol/L Cu2+/Pb2+/Zn2+ within 72 h. A distinctive feature lies in B. intermedia TSBOI's capacity for the transport and hydrolysis of urea, considered to be critical for its strong urease activity. This study explored the mechanisms of this capacity at the genetic, molecular and protein levels through complete genome sequencing, molecular docking and enzymatic reaction kinetics. The results revealed that, for urea hydrolysis, B. intermedia TSBOI exhibited a comprehensive urease gene cluster, with the key gene ureC demonstrating an absolute expression level approximating to 4 × 104 copies/RNA ng under optimal conditions. Results also confirmed the strong spontaneous, energy-independent binding ability of it's urease to urea, with the lowest Gibbs free energy binding site linking to the three amino acids, alanine, asparagine and serine. The urea transport gene yut presented and expressed, with the absolute expression enhanced in response to increasing urea concentrations. The significant positive correlation between ureC/yut expression levels and urease activity provided a theoretical basis for B. intermedia TSBOI's heavy metal bioremediation potential. ENVIRONMENTAL IMPLICATION: Heavy metals (Cu, Pb and Zn) were studied in this study. Heavy metals are hazardous due to their toxicity, persistence, and ability to bioaccumulate in living organisms. They can cause severe health issues, harm ecosystems, and contaminate air, water, and soil. A novel ureolytic bacteria, Brucella intermedia TSBOI, exhibited microbially induced carbonate precipitation capability was isolated which removed 90-100% of 1 mmol/L Cu2+/Pb2+/Zn2+ within 72 h. Its advantages in urea hydrolysis and transport facilitate the remediation of actual heavy metal contaminated environments.

Metagenomic analysis of the intestinal microbiome reveals the potential mechanism involved in Bacillus amyloliquefaciens in treating schistosomiasis japonica in mice.

Schistosomiasis japonica is one of the neglected tropical diseases characterized by chronic hepatic, intestinal granulomatous inflammation and fibrosis, as well as dysbiosis of intestinal microbiome. Previously, the probiotic Bacillus amyloliquefaciens has been shown to alleviate the pathological injuries in mice infected with Schistosoma japonicum by improving the disturbance of the intestinal microbiota. However, the underlying mechanisms involved in this process remain unclear. In this study, metagenomics sequencing and functional analysis were employed to investigate the differential changes in taxonomic composition and functional genes of the intestinal microbiome in S. japonicum-infected mice treated with B. amyloliquefaciens. The results revealed that intervention with B. amyloliquefaciens altered the taxonomic composition of the intestinal microbiota at the species level in infected mice and significantly increased the abundance of beneficial bacteria. Moreover, the abundance of predicted genes in the intestinal microbiome was also significantly changed, and the abundance of xfp/xpk and genes translated to urease was significantly restored. Further analysis showed that Limosilactobacillus reuteri was positively correlated with several KEGG Orthology (KO) genes and metabolic reactions, which might play important roles in alleviating the pathological symptoms caused by S. japonicum infection, indicating that it has the potential to function as another effective therapeutic agent for schistosomiasis. These data suggested that treatment of murine schistosomiasis japonica by B. amyloliquefaciens might be induced by alterations in the taxonomic composition and functional gene of the intestinal microbiome in mice. We hope this study will provide adjuvant strategies and methods for the early prevention and treatment of schistosomiasis japonica. IMPORTANCE: Targeted interventions of probiotics on gut microbiome were used to explore the mechanism of alleviating schistosomiasis japonica. Through metagenomic analysis, there were significant changes in the composition of gut microbiota in mice infected with Schistosoma japonicum and significant increase in the abundance of beneficial bacteria after the intervention of Bacillus amyloliquefaciens. At the same time, the abundance of functional genes was found to change significantly. The abundance of genes related to urease metabolism and xfp/xpk related to D-erythrose 4-phosphate production was significantly restored, highlighting the importance of Limosilactobacillus reuteri in the recovery and abundance of predicted genes of the gut microbiome. These results indicated potential regulatory mechanism between the gene function of gut microbiome and host immune response. Our research lays the foundation for elucidating the regulatory mechanism of probiotic intervention in alleviating schistosomiasis japonica, and provides potential adjuvant treatment strategies for early prevention and treatment of schistosomiasis japonica.

Medium optimization and dust suppression performance analysis of microbial-based dust suppressant compound by response surface curve method.

At present, microbial dust suppressants based on microbial communities lack necessary systematic analysis of factors affecting dust suppression performance. Therefore, in this study, the response surface curve method was used to optimize the culture conditions for enrichment of urease-producing microorganisms from activated sludge. The results indicated that when urea = 9.67 g L-1, NH4Cl = 5.21 g L-1, and pH = 9.57, the maximum urease activity of urease-producing microbial community (UPMC) was 8.22 mM min-1. The UPMC under optimized culture conditions reached a mineralization rate of 98.8% on the 1st day of mineralization. Ureolysis is one of the biological mechanisms that trigger microbial mineralization with the consequent effect of dust suppression. The analysis of microbial community structure indicated that the urease-producing bacteria Sporosarcina sp. had the highest abundance at the genus level in the microbial-based dust suppressant compound. Jeotgalicoccus sp. plays an important role in improving and maintaining the stability of urease. In addition, the optimal UPMC had low pathogenicity, which is extremely attractive for the safe application of microbial dust suppressants.

Proteus mirabilis UreR coordinates cellular functions required for urease activity.

A hallmark of Proteus mirabilis infection of the urinary tract is the formation of stones. The ability to induce urinary stone formation requires urease, a nickel metalloenzyme that hydrolyzes urea. This reaction produces ammonia as a byproduct, which can serve as a nitrogen source and weak base that raises the local pH. The resulting alkalinity induces the precipitation of ions to form stones. Transcriptional regulator UreR activates expression of urease genes in a urea-dependent manner. Thus, urease genes are highly expressed in the urinary tract where urea is abundant. Production of mature urease also requires the import of nickel into the cytoplasm and its incorporation into the urease apoenzyme. Urease accessory proteins primarily acquire nickel from one of two nickel transporters and facilitate incorporation of nickel to form mature urease. In this study, we performed a comprehensive RNA-seq to define the P. mirabilis urea-induced transcriptome as well as the UreR regulon. We identified UreR as the first defined regulator of nickel transport in P. mirabilis. We also offer evidence for the direct regulation of the Ynt nickel transporter by UreR. Using bioinformatics, we identified UreR-regulated urease loci in 15 Morganellaceae family species across three genera. Additionally, we located two mobilized UreR-regulated urease loci that also encode the ynt transporter, implying that UreR regulation of nickel transport is a conserved regulatory relationship. Our study demonstrates that UreR specifically regulates genes required to produce mature urease, an essential virulence factor for P. mirabilis uropathogenesis. IMPORTANCE: Catheter-associated urinary tract infections (CAUTIs) account for over 40% of acute nosocomial infections in the USA and generate $340 million in healthcare costs annually. A major causative agent of CAUTIs is Proteus mirabilis, an understudied Gram-negative pathogen noted for its ability to form urinary stones via the activity of urease. Urease mutants cannot induce stones and are attenuated in a murine UTI model, indicating this enzyme is essential to P. mirabilis pathogenesis. Transcriptional regulation of urease genes by UreR is well established; here, we expand the UreR regulon to include regulation of nickel import, a function required to produce mature urease. Furthermore, we reflect on the role of urea catalysis in P. mirabilis metabolism and provide evidence for its importance.

Synergizing structure and function: Cinnamoyl hydroxamic acids as potent urease inhibitors.

The current investigation encompasses the structural planning, synthesis, and evaluation of the urease inhibitory activity of a series of molecular hybrids of hydroxamic acids and Michael acceptors, delineated from the structure of cinnamic acids. The synthesized compounds exhibited potent urease inhibitory effects, with IC50 values ranging from 3.8 to 12.8 µM. Kinetic experiments unveiled that the majority of the synthesized hybrids display characteristics of mixed inhibitors. Generally, derivatives containing electron-withdrawing groups on the aromatic ring demonstrate heightened activity, indicating that the increased electrophilicity of the beta carbon in the Michael Acceptor moiety positively influences the antiureolytic properties of this compounds class. Biophysical and theoretical investigations further corroborated the findings obtained from kinetic assays. These studies suggest that the hydroxamic acid core interacts with the urease active site, while the Michael acceptor moiety binds to one or more allosteric sites adjacent to the active site.

Differences in the activities of six soil enzymes in response to cadmium contamination of paddy soils in high geological background areas.

East Yunnan province in southwest China is a region with elevated natural abundance (high geological background levels) of Cd due to high metal (loid) contents in the soils. Enzyme activities are useful indicators of metal (loid) toxicity in contaminated soils and whether Cd inhibits enzyme activities in paddy soils in high geological background areas is of considerable public concern. A pot experiment combined with field investigation was conducted to assess the effects of Cd on six soil enzymes that are essential to the cycling of C, N, and P in soils. Inhibitory effects of Cd fractions on enzyme activities were assessed using ecological dose-response models. The impact of soil properties on the inhibition of sensitive soil enzymes by Cd were assessed using linear and structural equation models. Cadmium was enriched in the paddy soils with 72.2 % of soil samples from high geological background areas exceeding the Chinese threshold values (GB 15618-2018) of Cd. Enzyme responses to Cd contamination varied markedly with a negative response by catalase but a positive response by invertase. Urease, ß-glucosidase, and alkaline phosphatase activities were stimulated at low Cd concentrations and inhibited at high concentrations. The average inhibition ratios of ß-glucosidase, urease, and catalase in high Cd levels were 19.9, 38.9, and 51.9%, respectively. Ecological dose-response models indicate that catalase and urease were the most Cd-sensitive of the enzymes studied and were suitable indicators of soil quality in high geological background areas. Structural equation modeling (SEM) indicates that soil properties influenced sensitive enzymes through various pathways, indicating that soil properties were factors determining Cd inhibition of enzyme activities. This suggests that Cd concentrations and soil physicochemical properties under a range of environmental conditions should be considered in addressing soil Cd pollution.