Extracellular Enzymes Research Articles

Determination of soil phenanthrene degradation through a fungal-bacterial consortium.

Fungal-bacterial consortia enhance organic pollutant removal, but the underlying mechanisms are unclear. We used stable isotope probing (SIP) to explore the mechanism of bioaugmentation involved in polycyclic aromatic hydrocarbon (PAH) biodegradation in petroleum-contaminated soil by introducing the indigenous fungal strain Aspergillus sp. LJD-29 and the bacterial strain Pseudomonas XH-1. While each strain alone increased phenanthrene (PHE) degradation, the simultaneous addition of both strains showed no significant enhancement compared to treatment with XH-1 alone. Nonetheless, the assimilation effect of microorganisms on PHE was significantly enhanced. SIP revealed a role of XH-1 in PHE degradation, while the absence of LJD-29 in 13C-DNA indicated a supporting role. The correlations between fungal abundance, degradation efficiency, and soil extracellular enzyme activity indicated that LJD-29, while not directly involved in PHE assimilation, played a crucial role in the breakdown of PHE through extracellular enzymes, facilitating the assimilation of metabolites by bacteria. This observation was substantiated by the results of metabolite analysis. Furthermore, the combination of fungus and bacterium significantly influenced the diversity of PHE degraders. Taken together, this study highlighted the synergistic effects of fungi and bacteria in PAH degradation, revealed a new fungal-bacterial bioaugmentation mechanism and diversity of PAH-degrading microorganisms, and provided insights for in situ bioremediation of PAH-contaminated soil.IMPORTANCEThis study was performed to explore the mechanism of bioaugmentation by a fungal-bacterial consortium for phenanthrene (PHE) degradation in petroleum-contaminated soil. Using the indigenous fungal strain Aspergillus sp. LJD-29 and bacterial strain Pseudomonas XH-1, we performed stable isotope probing (SIP) to trace active PHE-degrading microorganisms. While inoculation of either organism alone significantly enhanced PHE degradation, the simultaneous addition of both strains revealed complex interactions. The efficiency plateaued, highlighting the nuanced microbial interactions. SIP identified XH-1 as the primary contributor to in situ PHE degradation, in contrast to the limited role of LJD-29. Correlations between fungal abundance, degradation efficiency, and extracellular enzyme activity underscored the pivotal role of LJD-29 in enzymatically facilitating PHE breakdown and enriching bacterial assimilation. Metabolite analysis validated this synergy, unveiling distinct biodegradation mechanisms. Furthermore, this fungal-bacterial alliance significantly impacted PHE-degrading microorganism diversity. These findings advance our understanding of fungal-bacterial bioaugmentation and microorganism diversity in polycyclic aromatic hydrocarbon (PAH) degradation as well as providing insights for theoretical guidance in the in situ bioremediation of PAH-contaminated soil.

Effects of local farming practices on soil organic carbon content, enzymatic activities, and microbial community structure in semi-arid soils of Morocco

Soil organic carbon (SOC) is essential in semi-arid agricultural land for enhancing soil health, particularly through the promotion of microbial activities. This study assessed the impact of different agronomic practices on soil properties, microbial communities, and SOC levels in semi-arid Moroccan wheat fields. Three treatments were investigated: eucalyptus (Eucalyptus spp.) companion planting (EU), and fallowing with harvest residue mulching (FA), with the latter involving both short (3 months; FAS) and long (15 months; FAL) fallow periods. The study revealed significant variation in soil characteristics and microbial communities between these agronomic management regimes. Notably, soils managed with FAL contained elevated SOC levels (1.2%) compared to other treatments (FAS and EU) which show lower SOC range (0.62–0.86%). Both labile C (water-soluble carbon) and recalcitrant C (humic substances) were increased by FAL. Additionally, soil microbial biomass and dehydrogenase activity were observed to be high in FAL-managed soils, along with increased levels of extracellular enzymes related to nutrient cycling (β-glucosidase, alkaline phosphatase, and urease). Phospholipid fatty acid (PLFA) analysis indicated positive correlation between carbon content in soils and microbial populations. In contrast, soils managed with EU had significantly lower SOC levels, possibly due to differences in carbon fractionation. FAL increased soil enzymatic activities and enriched the microbial community when compared to EU management. In conclusion, this study indicated the importance of fallowing and fallowing period for conservation of SOC, and potential to mitigate negative effects of biophysical constraints on agricultural productivity in semi-arid soils of Northwest Africa.

Microbial detoxification of chlorpyrifos, profenofos, monocrotophos, and dimethoate by a multifaceted rhizospheric Bacillus cereus strain PM38 and its potential for the growth promotion in cotton.

Bacillus genera, especially among rhizobacteria, are known for their ability to promote plant growth and their effectiveness in alleviating several stress conditions. This study aimed to utilize indigenous Bacillus cereus PM38 to degrade four organophosphate pesticides (OPs) such as chlorpyrifos (CP), profenofos (PF), monocrotophos (MCP), and dimethoate (DMT) to mitigate the adverse effects of these pesticides on cotton crop growth. Strain PM38 exhibited distinct characteristics that set it apart from other Bacillus species. These include the production of extracellular enzymes, hydrogen cyanide, exopolysaccharides, Indol-3-acetic acid (166.8μg/mL), siderophores (47.3μg/mL), 1-aminocyclopropane-1-carboxylate deaminase activity (32.4μg/mL), and phosphorus solubilization (162.9μg/mL), all observed at higher concentrations. This strain has also shown tolerance to salinity (1200mM), drought (20% PEG-6000), and copper and cadmium (1200mg/L). The amplification of multi-stress-responsive genes, such as acdS, ituC, czcD, nifH, sfp, and pqqE, further confirmed the plant growth regulation and abiotic stress tolerance capability in strain PM38. Following the high-performance liquid chromatography (HPLC) analysis, the results showed striking compatibility with the first kinetic model. Strain PM38 efficiently degraded CP (98.4%), PF (99.7%), MCP (100%), and DMT (95.5%) at a concentration of 300ppm over 48h at 35°C under optimum pH conditions, showing high coefficients of determination (R2) of 0.974, 0.967, 0.992, and 0.972, respectively. The Fourier transform infrared spectroscopy (FTIR) analysis and the presence of opd, mpd, and opdA genes in the strain PM38 further supported the potential to degrade OPs. In addition, inoculating cotton seedlings with PM38 improved root length under stressful conditions. Inoculation of strain PM38 reduces stress by minimizing proline, thiobarbituric acid-reactive compounds, and electrolyte leakage. The strain PM38 has the potential to be a good multi-stress-tolerant option for a biological pest control agent capable of improving global food security and managing contaminated sites.

Enhanced denitrification by graphene oxide–modified cathode for the secondary effluent of wastewater treatment plants in three-dimensional biofilm electrode reactors

ABSTRACT In this study, a novel three-dimensional biofilm electrode reactor (3D-BER) with a graphene oxide (GO)–modified cathode was developed to enhance the denitrification performance of secondary effluent from wastewater treatment plants (SEWTPs). The effects of different hydraulic retention times (HRTs) and currents on the 3D-BER were explored. The results indicated that at the optimal HRT of 4 h and current of 350 mA/m2, the 3D-BER with GO-modified cathode had a higher denitrification rate (2.40 ± 0.1 mg TN/L/h) and less accumulation of intermediate products, especially with 3.34% total nitrogen (TN) molar conversion to N2O. The GO-modified cathode offered a large biocompatible specific surface area and enhanced the conductivity, which favored microbial growth and increased electron transfer efficiency and extracellular enzyme activities. Moreover, the activity of nitrite reductase increased more than that of nitrate reductase to accelerate nitrite reduction, thus facilitating the denitrification process. The proposed 3D-BER provided an effective solution to elevate tertiary denitrification in the SEWTP.

A systematic review on products derived from nematophagous fungi in the biological control of parasitic helminths of animals.

Nematophagous fungi have been widely evaluated in the biological control of parasitic helminths in animals, both through their direct use and the use of their derived products. Fungal bioproducts can include extracellular enzymes, silver nanoparticles (AgNPs), as well as secondary metabolites. The aim of this study was to conduct a systematic review covering the evaluation of products derived from nematophagous fungi in the biological control of parasitic helminths in animals. In total, 33 studies met the inclusion criteria and were included in this review. The majority of the studies were conducted in Brazil (72.7%, 24/33), and bioproducts derived from the fungus Duddingtonia flagrans were the most commonly evaluated (36.3%, 12/33). The studies involved the production of extracellular enzymes (48.4%, 16/33), followed by crude enzymatic extract (27.2%, 9/33), secondary metabolites (15.1%, 5/33) and biosynthesis of AgNPs (9.1%, 3/33). The most researched extracellular enzymes were serine proteases (37.5%, 6/16), with efficacies ranging from 23.9 to 85%; proteases (31.2%, 5/16), with efficacies from 41.4 to 95.4%; proteases + chitinases (18.7%, 3/16), with efficacies from 20.5 to 43.4%; and chitinases (12.5%, 2/16), with efficacies ranging from 12 to 100%. In conclusion, extracellular enzymes are the most investigated derivatives of nematophagous fungi, with proteases being promising strategies in the biological control of animal helminths. Further studies under in vivo and field conditions are needed to explore the applicability of these bioproducts as tools for biological control.

Extracellular Enzyme Activity and Stoichiometry Across Vegetated and Non-Vegetated Coastal Ecosystems

The conversion of organic matter by extracellular enzymes can reveal important insights into carbon and nutrient cycling. The activity and stoichiometry of hydrolytic extracellular enzymes were investigated to assess the effects of vegetation cover and sediment characteristics on microbial-enzyme-mediated decomposition in coastal ecosystems. Extracellular enzyme activity (EEA) was quantified across transects extending from mangrove to tidal flat habitats in two New Zealand coastal ecosystems that differ in mud content (sandy: Hobson Bay, muddy: Snells Beach). We determined the activity of five key hydrolyzing enzymes: β-glucosidase (hydrolyzes cellulose to glucose); β-N-acetylglucosaminidase (catalyzes the terminal reaction in chitin degradation); alkaline phosphatase (releases soluble inorganic phosphate groups from organophosphates); β-D-cellobiohydrolase (hydrolyzes cellulose to generate cellobiose); and β-xylosidase (catalyzes hemicellulose). All enzymes involved in C acquisition and in N and P cycling had higher activity at the muddy site. No habitat differences in EEA were observed at the sandy site, whereas EEA was lower in the non-vegetated habitats for some enzymes at the muddy site. Models of microbial metabolic limitations highlighted that most habitats at both muddy and sandy sites were predominately C and P limited. The EEA in these coastal wetlands was generally lower than has been reported for other terrestrial, freshwater, and estuarine ecosystems, with values often one to two orders of magnitude lower than other wetland studies. These results can be used to advance our understanding of the biogeochemical processes underpinning the response of coastal ecosystems to land-derived nutrient and sediment inputs.

Community assembly of ectomycorrhizal fungal communities in pure and mixed Pinus massoniana forests

Ectomycorrhizal (EcM) fungi play an important role in nutrient cycling and community ecological dynamics and are widely acknowledged as important components of forest ecosystems. However, little information is available regarding EcM fungal community structure or the possible relationship between EcM fungi, soil properties, and forestry activities in Pinus massoniana forests. In this study, we evaluated soil properties, extracellular enzyme activities, and fungal diversity and community composition in root and soil samples from pure Pinus massoniana natural forests, pure P. massoniana plantations, and P. massoniana and Liquidambar gracilipes mixed forests. The mixed forest showed the highest EcM fungal diversity in both root and bulk soil samples. Community composition and co-occurrence network structures differed significantly between forest types. Variation in the EcM fungal community was significantly correlated with the activities of β-glucuronidase and β−1,4-N-acetylglucosaminidase, whereas non-EcM fungal community characteristics were significantly correlated with β-1,4-glucosidase and β-glucuronidase activities. Furthermore, stochastic processes predominantly drove the assembly of both EcM and non-EcM fungal communities, while deterministic processes exerted greater influence on soil fungal communities in mixed forests compared to pure forests. Our findings may inform a deeper understanding of how the assembly processes and environmental roles of subterranean fungal communities differ between mixed and pure plantations and may provide insights for how to promote forest sustainability in subtropical areas.

Biodegradation of Pharmaceutical Pollutants Using Fungal Enzyme

Abstract: The potential of filamentous fungi and their enzymes are investigated for bioremediating pharmaceutical pollutants from wastewater thereby addressing the growing concern of emerging pollutants in water resources. Among the various microorganisms, the current abstract discusses the role of fungi in degrading antibiotics and persistent pharmaceutical compounds. These fungi allow the biodegradation via the formation of highly oxidative hydroxyl radicals. It summarizes the role of both intra (cytochrome P450) and extracellular (laccase) enzyme systems for the bioremediation of emerging clinical pollutants. It emphasizes the importance of both biosorption and biodegradation for effective pollutant removal by breaking down pharmaceuticals (e.g. antibiotics) and other hazardous compounds (e.g. endocrine-disrupting compounds, polycyclic aromatic hydrocarbons). Although the enzymes CYP450 are not directly engaged, their metabolic pathways execute mechanisms such as oxidation, the formation of reactive intermediates, and chemical modification, all of which result in the degradation of clinical pollutants. Amongst all enzyme systems in the case of laccase which oxidise a wide spectrum of substrates, converts them to free radicals. Additionally, the role of recombinant enzyme production and transgenic strains for pollutant degradation has also been elaborated. These considerations emphasise the need for ongoing research and biotechnological advancements to address the growing concern of pharmaceuticals and personal care product pollutants in water systems.

Diversity and Bioprospection of Gram-positive Bacteria Derived from a Mayan Sinkhole

Water-filled sinkholes known locally as cenotes, found on the Yucatán Peninsula, have remarkable biodiversity. The primary objective of this study was to explore the biotechnological potential of Gram-positive cultivable bacteria obtained from sediment samples collected at the coastal cenote Pol-Ac in Yucatán, Mexico. Specifically, the investigation aimed to assess production of hydrolytic enzymes and antimicrobial compounds. 16 S rRNA gene sequencing led to the identification of 49 Gram-positive bacterial isolates belonging to the phyla Bacillota (n = 29) and Actinomycetota (n = 20) divided into the common genera Bacillus and Streptomyces, as well as the genera Virgibacillus, Halobacillus, Metabacillus, Solibacillus, Neobacillus, Rossellomorea, Nocardiopsis and Corynebacterium. With growth at 55ºC, 21 of the 49 strains were classified as moderately thermotolerant. All strains were classified as halotolerant and 24 were dependent on marine water for growth. Screening for six extracellular hydrolytic enzymes revealed gelatinase, amylase, lipase, cellulase, protease and chitinase activities in 93.9%, 67.3%, 63.3%, 59.2%, 59.2% and 38.8%, of isolated strains, respectively. The genes for polyketide synthases type I, were detected in 24 of the strains. Of 18 strains that achieved > 25% inhibition of growth in the bacterial pathogen Staphylococcus aureus ATCC 6538, 4 also inhibited growth in Escherichia coli ATCC 35,218. Isolates Streptomyces sp. NCA_378 and Bacillus sp. NCA_374 demonstrated 50–75% growth inhibition against at least one of the two pathogens tested, along with significant enzymatic activity across all six extracellular enzymes. This is the first comprehensive report on the biotechnological potential of Gram-positive bacteria isolated from sediments in the cenotes of the Yucatán Peninsula.

Role of rhizospheric microbial enzymes in plant growth promotion, antagonism, and sustainable agriculture: A review

Plant growth-promoting rhizobacteria (PGPR) offer eco-friendly alternatives to chemical fertilizers, promoting sustainable agriculture by enhancing soil fertility, reducing pathogens, and aiding in stress resistance. In agriculture, they play a crucial role in plant growth promotion through the production of agroactive compounds and extracellular enzymes to promote plant health and protection against phytopathogens. In the rhizosphere, diverse microbial interactions, including those with bacteria and fungi, influence plant health by production of antimicrobial compounds. The antagonism displayed by rhizobacteria plays a crucial role in shaping microbial communities and has potential applications in developing a natural and environmentally friendly approach to pest control. The rhizospheric microbes showcase their ecological importance and potential for biotechnological applications in the context of plant-microbe interactions. The extracellular enzymes produced by rhizospheric microbes like amylases, chitinases, glucanases, cellulases, proteases, and ACC deaminase contribute to plant processes and stress response emphasizing their importance in sustainable agriculture. Moreover, this review highlights the new paradigm including artificial intelligence (AI) in sustainable horticulture and agriculture as a harmonious interaction between ecological networks for promoting soil health and microbial diversity that leads to a more robust and self-regulating agricultural system for protecting the environment in the future. Overall, this review emphasizes microbial interactions and the role of rhizospheric microbial extracellular enzymes which is crucial for developing eco-friendly approaches to enhance crop production and soil health.

Study on the Effects of Different Light Supply Modes on the Development and Extracellular Enzyme Activity of Ganoderma lucidum

Study on the Effects of Different Light Supply Modes on the Development and Extracellular Enzyme Activity of Ganoderma lucidum

Integration of engineering microbial consortium and boosting extracellular amylase of B. amyloliquefaciens for intensifying bioconversion of food waste into lipopeptides

Lipopeptides possess significant antifungal activity and serve as eco-friendly biopesticides. However, their widespread application is limited by exorbitant production costs and low yields. Food waste emerges as a promising substrate for lipopeptide production. To efficiently harness the starch in food waste during bioconversion it into lipopeptides, the engineered Bacillus amyloliquefaciens HM618, constructed by knocking out multiple extracellular proteolytic enzyme genes (aprE, nprE, vpr, and epr) and overexpressing gene amyA with Phag promoter, capable of producing both high-level extracellular amylase and lipopeptides. It was found that the recombinant B. amyloliquefaciens HM3–5 in the pure culture containing starch exhibited notable amylase activity about 17046.37 U/mL, a total of 2530.72 mg/L lipopeptides, and 80.02 % degradation rate of starch. Considering the crucial roles of fatty acids and amino acids precursors in lipopeptide synthesis, these engineered B. amyloliquefaciens producing higher-level extracellular amylase, Yarrowia lipolytica producing fatty acids, and Corynebacterium glutamicum releasing proline and serine were integrated to construct an artificial consortium for efficiently bio-converting food waste into lipopeptides. After optimized inoculation times and ratios of these engineered strains, the levels of surfactin, fengycin, and iturin A in the four-strains artificial consortium during food waste bioconversion reached 8690.67, 635.83, and 74.19 mg/L, respectively. The total lipopeptides in the four-strain artificial consortium reached 9400.69 mg/L, 2.38-fold increase compared to that in the pure culture of strain HM3–5. Furthermore, the degradation efficiencies of starch and oil in food waste were 96.41 % and 98.35 %, respectively. This work provides a new strategy for efficiently bio-transforming food waste into lipopeptides.

Co-fermentation of acid treated coconut wastes using mixed Bacillus cultures for enhanced production of extracellular enzymes: Application in bioconversion of raw coconut fibers

Higher cost of production and functioning efficiency are the two most important aspects of cellulolytic enzymes, as they directly affect the bioconversion of cellulosic waste into fermentable sugars. Thus, the current study aims to investigate the production of cellulolytic enzymes from low-cost waste biomass for use in the sustainable hydrolysis of biomass to yield fermentable sugars. Since coconut waste is a major contributor to biodegradable waste, two different parts of coconut, including the hard shell of coconut (CS) and the fibers of the shell (FS), have been used for enzyme production as feedstock. Further, enzyme production has been carried out on co-fermentative and acid-pretreated coconut substrates (CS and FS) at different quantities using mixed bacterial cultures. Following this, a crude enzyme with a maximum 20 U/gds FPA, 62 U/gds BGL, and 167 U/gds EG has been produced by mixed Bacillus cultures under co-fermentation of 0.5 % acid-pretreated CS and FS substrates at a quantity of 5:5 (g:g) at 35 oC and pH 5.0. The crude enzyme showed its thermal stability for 16 h at 50 oC and released ∼34 g/L of sugars from the hydrolysis of untreated FS biomass in 24 h. This work may have significant applications in the area of biomass-based biorefining as well as waste management.

Investigating Sulforaphane's anti-virulence and anti-quorum sensing properties against Pseudomonas aeruginosa.

P. aeruginosa, a significant bacterium, can cause severe illness and resistance to antibiotics. Quorum sensing (QS) systems regulate virulence factors production. Targeting QS could reduce bacteria pathogenicity and prevent antibiotic resistance. Cruciferous vegetables contain sulforaphane, known for its anti-inflammatory, antioxidant, anticancer, and antimicrobial properties. We aimed to examine the inhibitory influences of sulforaphane, at a sub-inhibitory concentration (¼ minimum inhibitory concentration, MIC), on virulence and QS in P. aeruginosa. The sulforaphane's anti-virulence actions at sub-inhibitory concentrations were explored in vitro and in vivo. A sub-MIC concentration of sulforaphane was combined with anti-pseudomonal drugs, and the results of this combination were assessed. The virtual affinity of sulforaphane for the receptors of QS was studied, and its effect on the expression of QS genes was quantified. Sulforaphane significantly decreased the biofilm formation, motility, ability to withstand oxidative stress, and the synthesis of virulence extracellular enzymes such as proteases, hemolysins, and elastase, as well as other virulence factors like pyocyanin. In addition, sulforaphane lessened the severity of P. aeruginosa infection in mice. Sulforaphane reduced the antipseudomonal antibiotics' MICs when used together, resulting in synergistic effects. The observed anti-virulence impacts were attributed to the ability of sulforaphane to inhibit QS via suppressing the QS genes' expression. Sulforaphane shows promise as a potent anti-virulence and anti-QS agent that can be used alongside conventional antimicrobials to manage severe infections effectively. Furthermore, this study paves the way for further investigation of sulforaphane and similar structures as pharmacophores for anti-QS candidates.

Piceatannol: a renaissance in antibacterial innovation unveiling synergistic potency and virulence disruption against serious pathogens.

In the relentless battle against multi-drug resistant Gram-negative bacteria, piceatannol emerges as a beacon of hope, showcasing unparalleled antibacterial efficacy and a unique ability to disrupt virulence factors. Our study illuminates the multifaceted prowess of piceatannol against prominent pathogens-Proteus mirabilis, Pseudomonas aeruginosa, Acinetobacter baumannii, and Klebsiella pneumoniae. Notably, piceatannol demonstrated a remarkable ability to inhibit biofilm formation, reduce bacterial mobility, and diminish extracellular enzyme synthesis.Mechanistic insights into piceatannol's activity unraveled its impact on membrane potential, proton motive force, and ATP production. Furthermore, our study delved into piceatannol's anti-quorum sensing (QS) activity, showcasing its potential to downregulate QS-encoding genes and affirming its affinity to critical QS receptors through molecular docking. Crucially, piceatannol exhibited a low propensity for resistance development, positioning it as a promising candidate for combating antibiotic-resistant strains. Its mild effect on red blood cells (RBCs) suggests safety even at higher concentrations, reinforcing its potential translational value. In an in vivo setting, piceatannol demonstrated protective capabilities, significantly reducing pathogenesis in mice infected with P. aeruginosa and P. mirabilis. This comprehensive analysis positions piceatannol as a renaissance in antibacterial innovation, offering a versatile and effective strategy to confront the evolving challenges posed by resilient Gram-negative pathogens.

О СУБСТРАТНОЙ СПЕЦИФИЧНОСТИ И НЕКОТОРЫХ СВОЙСТВАХ ВНЕКЛЕТОЧНОЙ ОКСИДАЗЫ ИЗ БАЗИДИОМИЦЕТА NEONOTHOPANUS NAMBI

An extracellular enzyme with oxidase activity was isolated from the mycelium of the higher fungus Neonothopanus nambi by mild treatment of the biomass with β-glucosidase. A substrate specificity and some properties of the isolated extracellular oxidase were studied in the present work. Experiments revealed that the extracellular oxidase exhibited activity with most phenolic compounds chosen as model substrates. It is important to note that the enzyme exhibited a catalytic function in the reactions without the addition of exogenous hydrogen peroxide and other mediators. The highest catalytic activity of the enzyme was observed with veratryl alcohol and dihydric phenols, hydroquinone and guaiacol. The enzyme showed lower activity with aromatic azo compounds (ABTS, DAB, o-dianisidine). In reactions with dihydric phenol resorcinol and monophenol, the enzyme efficiency was extremely low. The kinetic parameters of the enzymatic reactions with actively oxidized substrates were determined. The addition of a divalent metal ion chelator (EDTA) did not affect the activity of the enzyme, while the addition of the SH reagent (DTT) increased the catalytic efficiency of the studied oxidase. The totality of the data obtained indicates that the extracellular oxidase of the N. nambi fungus catalyzes the oxidation of a wide range of aromatic compounds under slightly acidic and neutral conditions without the addition of additional mediators (in particular, hydrogen peroxide). This creates the prerequisites for studying the applicability of the enzyme in biomedical analytics.

Soil erosion‐induced decline in aggregate stability and soil organic carbon reduces aggregate‐associated microbial diversity and multifunctionality of agricultural slope in the Mollisol region

AbstractSoil erosion has become a serious ecological problem hindering the agricultural and economic development in Northeast China. Soil aggregates are basic units that regulate soil fertility and microbial activity. However, there have been few reports on the role of soil erosion in regulating the microbial diversity, multiple ecosystem services, and functions (multifunctionality) of soil aggregates. In this study, we investigated the effects of soil erosion on microbial diversity and the activities of various extracellular enzymes associated with soil aggregates by using Mollisol soil in Northeastern China at different positions of a slope, and how soil erosion affects soil multifunctionality indexes obtained by normalized calculation of soil enzyme activities. The results showed that soil erosion could reduce the physicochemical properties, soil enzyme activity, and microbial diversity of the aggregates, resulting in accumulation of aggregate nutrients and increase in microbial diversity and enzyme activity in the sedimentation area. Compared with smaller soil aggregates, larger soil aggregates had higher nutrient content, enzyme activity, and microbial diversity. However, with the intensification of soil erosion, the differences in microbial diversity and soil enzyme activities among different sizes of aggregates were reduced. Soil erosion also significantly reduced soil multifunctionality (p < 0.001). The multifunctionality of 2000–250 μm, 250–53 μm, and <53 μm aggregates respectively declined from 0.52, 0.20, and 0.08 (no erosion sites) to −0.17, −0.66, and −0.77 (heavy erosion sites). Additionally, microbial diversity was found to have a linear positive correlation with soil multifunctionality (R2 = 0.23, p < 0.01). Structural equation modeling and random forest analysis revealed that soil organic carbon and aggregate water stability are main predictors of soil multifunctionality. Furthermore, soil physicochemical properties had a greater influence on soil multifunctionality than microbial diversity. Collectively, our results demonstrate that soil erosion negatively affects the structure, resource effectiveness, and microbial activity of soil aggregates, thereby reducing to differential soil multifunctionality.

Short rotation willow to restore degraded marginal land and enhance climate resiliency within the Prairie Pothole Region: A potential nature-based solution

Short rotation willow (SRW) is a land management strategy involving the cultivation of rapidly growing, biomass-rich herbaceous-woody plants. This practice holds promise for renewable energy production, water quality preservation, carbon sequestration, greenhouse gas (GHG) mitigation, enhancement of soil extracellular enzyme activities (EEAs), and promotion of overall soil health. The rapid growth of SRW demands substantial water and nutrient resources, posing concerns when cultivated in marginal riparian lands within the Prairie Pothole Region (PPR), potentially leading to alterations in groundwater table (GWT) depth fluctuations, elevated soil salinity levels, and disruptions to biogeochemical cycles. Hence, this study comprehensively evaluated the effects of establishing SRW as a degraded marginal riparian land use practice in the PPR and attempted to answer several vital questions in the field and microcosm scale on soil hydrology, salinity, nutrients, soil organic carbon (SOC), GHG emissions, and EEAs involved in biogeochemical cycling. In a field experiment, the effects of SRW were evaluated by measuring the depth to GWT, groundwater and soil electrical conductivity (EC), macronutrients (N, P, K, and S), and SOC content in different fractions and chemical compositions during the first rotation (3-year cycle) compared with adjacent annual crop and pasture in two semi-arid PPR sites. In a microcosm experiment, GHG (CO2, CH4, and N2O) emissions and EEAs [β-glucosidase (BG), N-acetyl glucosaminidase (NAG), and alkaline phosphatase (AP)] were measured in intact soil cores treated with declining water tables and different groundwater salinity levels. No consistent land use impacts on GWT or soil EC were observed between sites. Land use in site B significantly impacted GWT depth, implying site-specific factors, such as topography and soil characteristics, may be dominant over land use effects. Under SRW, the levels of macronutrients in the soil varied but did not significantly reduce the overall nutrient content of the soil. Total SOC was highest in pasture; light fraction organic carbon and particulate organic carbon followed a similar land use pattern, i.e., pasture > SRW = annual crop. Land uses affected GHG emissions significantly in the order of pasture > annual crop = SRW. GHG emission varied with salinity and GWT but there was no interaction with land use practices. Soil EEAs were significantly impacted by different land uses, i.e., pasture > annual crop = SRW, suggesting that the effects resulted from associated SOC. Our microcosm experiment suggests that the SRW land use practice holds promise as a sustainable Nature-Based Solution for enhancing climate resiliency in PPR. It exhibits a lower global warming potential compared to annual crop and pasture. Therefore, widespread implementation of the SRW land use practice in degraded marginal land could help mitigate the effects of climate change in the region.

Natronospira bacteriovora sp. nov., and Natronospira elongata sp. nov., extremely salt-tolerant predatory proteolytic bacteria from soda lakes and proposal to classify the genus Natronospira into Natronospiraceae fam. nov., and Natronospirales ord. nov., within the class Gammaproteobacteria

The genus Natronospira is represented by a single species of extremely salt-tolerant aerobic alkaliphilic proteolytic bacterium, isolated from hypersaline soda lakes. When cells of Gram-positive cocci were used as a substrate instead of proteins at extremely haloalkaline conditions, two new members of this genus were enriched and isolated in pure culture from the same sites. Strains AB-CW1 and AB-CW4 are obligate aerobic heterotrophic proteolytic bacteria able to feed on both live and dead cells of staphylococci and a range of proteins and peptides. Similar to the type species, N. proteinivora, the isolates are extremely salt-tolerant obligate alkaliphiles. However, N. proteinivora was unable to use bacterial cells as a substrate. Electron microscopy showed direct contact between the prey and predator cells. Functional analysis of the AB-CW1 and AB-CW4 genomes identified two sets of genes coding for extracellular enzymes potentially involved in the predation and proteolysis, respectively. The first set includes several copies of lysozyme-like GH23 peptidoglycan-lyase and murein-specific M23 [Zn]-di-peptidase enabling the cell wall degradation. The second set features multiple copies of secreted serine and metallopeptidases apparently allowing for the strong proteolytic phenotype. Phylogenomic analysis placed the isolates into the genus Natronospira as two novel species members, and furthermore indicated that this genus forms a deep-branching lineage of a new family (Natronospiraceae) and order (Natronospirales) within the class Gammaproteobacteria. On the basis of distinct phenotypic and genomic properties, strain AB-CW1T (JCM 335396 = UQM 41579) is proposed to be classified as Natronospira elongata sp. nov., and AB-CW4T (JCM 335397 = UQM 41580) as Natronospira bacteriovora sp. nov.

Development and evaluation of the inactivated vaccine against Aeromonas schubertii in hybrid snakehead

Aeromonas schubertii is a global pathogen of fish that causes internal nodular diseases and death in severe cases. In the present study, we isolated nine pathogenic strains from diseased hybrid snakehead (Channa argus♂ × Channa maculata♀) which were formally identified as A. schubertii by molecular identification, including 16S rRNA and gyrB gene sequencing. Subsequently, we determined the extracellular enzyme activity of six isolates and compared pathogenicity to investigate potential relationships with geographic characteristics. Two highly pathogenic strains, AS-14 and AS-16, were finally identified as inactivated vaccine candidates. Subsequently, we evaluated different concentrations of the antigen for each vaccine by performing experiments in hybrid snakehead; fish were inoculated with inactivated vaccine via the intraperitoneal route and a booster injection was administered 14 days later. Relative percent survival (RPS) for fish immunized with high doses of the two vaccines (1 × 108 CFU/fish) were 54.4% and 57.4%, respectively; these values were higher than other groups of fish receiving different doses (1 × 106 and 1 × 107 CFU/fish). The bacterial load of tissues harvested from the fish immunized with high doses of the vaccine groups were significantly lower than those of fish immunized with low doses. Titers of specific antibodies against A. schubertii in fish immunized with a high dose of AS-16 vaccine were maintained at high levels when tested six weeks after the booster injection. Furthermore, immune-related genes in the spleen, including MHC I, MHC II, CD4, CD8α and IgM, all exhibited high levels of expression, thus suggesting that hybrid snakehead underwent an adaptive immune response post-vaccination. Collectively, our findings enrich our understanding of the specific causative agents underlying nodular diseases in hybrid snakehead and provide scientific reference guidelines for the development of vaccines against A. schubertii. The development of such vaccines will be conducive to the sustainable development of hybrid snakehead in commercial aquaculture.