rDNA Sequence Analysis Research Articles

Novel insights into aerobic 17β-estradiol degradation by enriched microbial communities from mangrove sediments

Various persistent organic pollutants (POPs) including estrogens are often enriched in mangrove regions. This research investigated the estrogens pollution levels in six mangroves located in the Southern China. The estrogen levels were found to be in the range of 5.3–24.9 ng/g dry weight, suggesting that these mangroves had been seriously contaminated. The bacterial communities under estrogen stress were further enriched by supplementing 17β-estradiol (E2) as the sole carbon source. The enriched bacterial communities showed an excellent E2 degradation capacity > 95 %. These communities were able to transform E2 into estrone (E1), 4-hydroxy-estrone, and keto-estrone, etc. 16 S rDNA sequencing and metagenomics analysis revealed that bacterial taxa Oleiagrimonas, Pseudomonas, Terrimonas, and Nitratireductor etc. were the main contributors to estrogen degradation. Moreover, the genes involved in E2 degradation were enriched in the microbial communities, including the genes encoding 17β-hydroxysteroid dehydrogenase, estrone 4-hydroxylase, etc. Finally, the analyses of functional genes and binning genomes demonstrated that E2 was degraded by bacterial communities via dehydrogenation into E1 by 17β-hydroxysteroid dehydrogenase. E1 was then catabolically converted to 3aα-H-4α(3′-propanoate)− 7aβ-methylhexahydro-1,5-indanedione via 4,5-seco pathway. Alternatively, E1 could also be hydroxylated to keto-estrone, followed by B-ring cleavage. This study provides novel insights into the biodegradation of E2 by the bacterial communities in estrogen-contaminated mangroves.

Screening of rhizosphere nitrogen fixing, phosphorus and potassium solubilizing bacteria of Malus sieversii (Ldb.) Roem. and the effect on apple growth

Nitrogen, phosphorus and potassium inorganic fertilizers are commonly used when growing apples, but their excessive application has resulted in a decline in soil fertility and therefore an inability to maintain sustainable cultivation systems. It is possible to compensate for this with biofertilizers. Nitrogen-fixing, phosphorus solubilizing, and potassium solubilizing bacteria are biofertilizers with a broad range of possible uses. In this study, beneficial microorganisms were screened from the rhizosphere soil of the apple tree, Malus sieversii (Ldb.) Roem., which is rich in microbial diversity in natural environments. It was essential to investigate their effects on the growth of apple seedlings. Eight populations of organic phosphorus solubilizing bacteria (56), inorganic phosphorus solubilizing bacteria (13), nitrogen-fixing bacteria (22), and potassium solubilizing bacteria (24) were isolated from eight populations of Xinjiang wild apple rhizosphere in Ili by medium culture. Their morphological characteristics were recorded and their activity was determined. The most active strains were Pseudomonas migulae, Pseudomonas poae, and Pseudomonas extremaustralis, which was determined by physiological and biochemical properties and 16S rDNA sequence analysis. The L16 (45) orthogonal experiment, which used the three strains as testing materials, was created to assess the effects of the strains on apple physiological indicators, soil nutrients, leaf nutrients, and biomass, as well as to identify the ideal combination, concentration, timing, and application method. The results indicated that the peroxidase (POD) and catalase (CAT) activities of apple seedlings increased significantly under the 10 treatment, and the (SOD) activities of the 0 (control) and 1 (inorganic fertilizer only) treatments were significantly lower than the other treatments; soluble sugar, soluble protein and chlorophyll contents increased in all treatments compared to 0 and 1, while malondialdehyde and proline contents increased or decreased compared to 0 and 1; apple seedlings in treatment 10 had the highest soil N, P, and K content, leaf N, P, and K content and biomass were also all highest in treatment 10. In summary, the strains screened for the test can be used as biofertilizers and the optimum application was determined for treatment 10, meaning that the results also provide a theoretical basis for their application in artificially grown orchards.

Long-read sequencing reveals the shell microbiome of apparently healthy American lobsters Homarus americanus from Atlantic Canada.

The shell microbial community of lobsters-a key factor in the development of epizootic shell disease (ESD)-is still insufficiently researched in Atlantic Canada and many knowledge gaps remain. This study aimed to establish a baseline description and analysis of the shell microbiome of apparently healthy lobsters from four locations in the region. More than 180 lobster shell swab samples were collected from New Brunswick, Nova Scotia and Prince Edward Island (PEI). PacBio long-read 16S rDNA sequencing and bioinformatic analyses in QIIME2 identified the shell-associated bacteria. The shell microbiome of healthy lobsters consisted mainly of the bacterial classes Gammaproteobacteria, Saprospiria, Verrucomicrobiae, Alphaproteobacteria, Flavobacteriia, Acidimicrobiia and Planctomycetia. The microbial composition differed regionally and seasonally, with some classes showing decreased or increased relative abundances in the PEI samples as well as in the winter and spring samples in Nova Scotia. The core shell microbiome included potentially pathogenic as well as beneficial bacterial taxa, of which some were present only in certain regions. Bacterial taxa that have previously been associated with ESD were present on healthy lobsters in Atlantic Canada, but their frequency differed by location, sampling time, and moult stage. This study indicated that geographical and seasonal factors influenced the shell microbiome of apparently healthy lobsters more than host factors such as sex, size, and moult stage. Our results provide valuable reference microbial data from lobsters in a disease-free state.

Copper Tolerance of Novel Rhodotorula sp. Yeast Isolated from Gold Mining Ore in Gia Lai, Vietnam

In this study, twenty-five yeast strains were isolated from soil samples collected in the gold mining ore in Gia Lai, Vietnam. Among them, one isolate named GL1T could highly tolerate Cu2+ up to 10 mM, and the isolates could also grow in a wide range of pH (3–7), and temperature (10–40 °C). Dried biomass of GL1 was able to remove Cu2+ effectively up to 90.49% with a maximal biosorption capacity of 18.1 mg/g at pH 6, temperature 30 °C, and incubation time 60 min. Sequence analysis of rDNA indicated this strain was closely related to Rhodotorula mucilaginosa but with 1.53 and 3.46% nucleotide differences in the D1/D2 domain of the 28S rRNA gene and the ITS1-5.8S rRNA gene-ITS2 region sequence, respectively. Based on phylogenetic tree analysis and the biochemical characteristics, the strain appears to be a novel Rhodotorula species, and the name Rhodotorula aurum sp. nov. is proposed. This study provides us with more information about heavy metal-tolerant yeasts and it may produce a new tool for environmental control and metal recovery operations.

Integrative analysis of microbiota and metabolomics in chromium-exposed silkworm (Bombyx mori) midguts based on 16S rDNA sequencing and LC/MS metabolomics.

The gut microbiota, a complex ecosystem integral to host wellbeing, is modulated by environmental triggers, including exposure to heavy metals such as chromium. This study aims to comprehensively explore chromium-induced gut microbiota and metabolomic shifts in the quintessential lepidopteran model organism, the silkworm (Bombyx mori). The research deployed 16S rDNA sequence analysis and LC/MS metabolomics in its experimental design, encompassing a control group alongside low (12 g/kg) and high (24 g/kg) feeding chromium dosing regimens. Considerable heterogeneity in microbial diversity resulted between groups. Weissella emerged as potentially resilient to chromium stress, while elevated Propionibacterium was noted in the high chromium treatment group. Differential analysis tools LEfSe and random forest estimation identified key species like like Cupriavidus and unspecified Myxococcales, offering potential avenues for bioremediation. An examination of gut functionality revealed alterations in the KEGG pathways correlated with biosynthesis and degradation, suggesting an adaptive metabolic response to chromium-mediated stress. Further results indicated consequential fallout in the context of metabolomic alterations. These included an uptick in histidine and dihydropyrimidine levels under moderate-dose exposure and a surge of gentisic acid with high-dose chromium exposure. These are critical players in diverse biological processes ranging from energy metabolism and stress response to immune regulation and antioxidative mechanisms. Correlative analyses between bacterial abundance and metabolites mapped noteworthy relationships between marker bacterial species, such as Weissella and Pelomonas, and specific metabolites, emphasizing their roles in enzyme regulation, synaptic processes, and lipid metabolism. Probiotic bacteria showed robust correlations with metabolites implicated in stress response, lipid metabolism, and antioxidant processes. Our study reaffirms the intricate ties between gut microbiota and metabolite profiles and decodes some systemic adaptations under heavy-metal stress. It provides valuable insights into ecological and toxicological aspects of chromium exposure that can potentially influence silkworm resilience.

Identification of Diagnostic Biomarkers for Compensatory Liver Cirrhosis Based on Gut Microbiota and Urine Metabolomics Analyses

Liver cirrhosis is one of the most prevalent chronic liver disorders with high mortality. We aimed to explore changed gut microbiome and urine metabolome in compensatory liver cirrhosis (CLC) patients, thus providing novel diagnostic biomarkers for CLC. Forty fecal samples from healthy volunteers (control: 19) and CLC patients (patient: 21) were undertaken 16S rDNA sequencing. Chromatography-mass spectrometry was performed on 40 urine samples (20 controls and 20 patients). Microbiome and metabolome data were separately analyzed using corresponding bioinformatics approaches. The diagnostic model was constructed using the least absolute shrinkage and selection operator regression. The optimal diagnostic model was determined by five-fold cross-validation. Pearson correlation analysis was applied to clarify the relations among the diagnostic markers. 16S rDNA sequencing analyses showed changed overall alpha diversity and beta diversity in patient samples compared with those of controls. Similarly, we identified 841 changed metabolites. Pathway analysis revealed that the differential metabolites were mainly associated with pathways, such as tryptophan metabolism, purine metabolism, and steroid hormone biosynthesis. A 9-maker diagnostic model for CLC was determined, including 7 microorganisms and 2 metabolites. In this model, there were multiple correlations between microorganisms and metabolites. Subdoligranulum, Agathobacter, norank_f_Eubacterium_coprostanoligenes_group, Butyricicoccus, Lachnospiraceae_UCG_004, and L-2,3-Dihydrodipicolinate were elevated in CLC patients, whereas Blautia, Monoglobus, and 5-Acetamidovalerate were reduced. A novel diagnostic model for CLC was constructed and verified to be reliable, which provides new strategies for the diagnosis and treatment of CLC.

Physiological characteristics, rhizosphere soil properties, and root-related microbial communities of Trifolium repens L. in response to Pb toxicity

Trifolium repens L. (T. repens) is considered a potential phytoremediation species due to its large biomass and ability to accumulate and tolerate heavy metals. Lead (Pb) is an important heavy metal pollutant that can affect plant growth, photosynthesis, and enzyme activity. However, response mechanism of microorganisms in three root niches of metal tolerant plants to Pb is not completely understood. Therefore, in this study, a Pb poisoning model of T. repens was established with a Pb gradient (0, 1000 mg/kg, 2000 mg/kg, and 3000 mg/kg), and was used to evaluate growth and physiological responses, as well as enrichment and transport coefficients in T. repens, and explore the characteristics of rhizosphere soil and microbial composition of three root niches. We found that Pb stress caused oxidative injury, and inhibited photosynthesis in T. repens. 16S rDNA sequencing analysis showed that the richness of microbial communities in bulk soil was higher than that in rhizosphere soil both under Pb stress and Pb nonstress conditions. Moreover, Proteobacteria was dominant phylum in bulk and rhizosphere soils, and Proteobacteria and Cyanobacteria were dominant phylum in endophytic bacteria. For the first time, we systematically investigated the response of Pb from bulk soil to plant leaves. The results showed that microbial interaction existed between bulk and rhizosphere soil. Rhizosphere bacterium Haliangium was positively correlated with urease activity and soil nutrients. Endophytic bacterium Pseudomonas was positively correlated with plant biomass and played an important role in Pb tolerance of T. repens. In addition, endophytic bacteria formed complex correlation networks with growth and physiological indexes of both root and shoot, moreover the network in root was more complicated. Taken together, Pb stress dose-dependently inhibited the growth of plants. This study provided a theoretical basis for the further development of microbial cooperation with plant remediation of heavy metal contaminated soil.

Effects of Branched-Chain Fatty Acids Derived from Yak Ghee on Lipid Metabolism and the Gut Microbiota in Normal-Fat Diet-Fed Mice.

Branched-chain fatty acids (BCFAs) are natural components with a variety of biological activities. However, the regulation of lipid metabolism by BCFAs is unknown. It was dedicated to examining the impacts of BCFAs inferred from yak ghee on the expression of qualities related to lipid metabolism, natural pathways, and intestinal microbiota in mice. The treatment group (purified BCFAs from yak ghee) exhibited a decrease in cholesterol levels; a decrease in HMGCR levels; downregulation of FADS1, FADS2, ACC-α, FAS, GAPT4, GPAM, ACSL1, THRSP, A-FABP, and PPARα gene expression; and upregulation of SCD1, ACSS1, FABP1, CPT1, and DGAT-1 gene expression. Gut microbiota 16S rDNA sequencing analysis showed that the treatment group improved the gut microbiota by increasing the relative abundances and increasing the short-chain fatty acid levels produced by the genera Akkermansia, Clostridium, Lachnospiraceae, Lactobacillus, Anaerotaenia, and Prevotella. After adding BCFAs to cultured breast cancer cells, pathways that were downregulated were found to be related to fatty acid degradation and fatty acid metabolism, while 20 other pathways were upregulated. Our results suggest that BCFAs reduce body fat in mice by modulating intestinal flora and lipid metabolism and modulating fatty acid metabolism in breast cancer cells.

Characterization of Cellulose-Degrading Bacteria Isolated from Silkworm Excrement and Optimization of Its Cellulase Production.

An abundance of refractory cellulose is the key limiting factor restricting the resource utilization efficiency of silkworm (Bombyx mori) excrement via composting. Screening for cellulose-degrading bacteria is likely to provide high-quality strains for the safe and rapid decomposition of silkworm excrement. In this study, bacteria capable of degrading cellulose with a high efficiency were isolated from silkworm excrement and the conditions for cellulase production were optimized. The strains were preliminarily screened via sodium carboxymethyl cellulose culture and staining with Congo red, rescreened via a filter paper enzyme activity test, and identified via morphological observation, physiological and biochemical tests, and phylogenetic analysis of the 16S rDNA sequence. Enzyme activity assay was performed using the 3,5-dinitrosalicylic acid method. DC-11, a highly cellulolytic strain, was identified as Bacillus subtilis. The optimum temperature and pH of this strain were 55 °C and 6, respectively, and the filter paper enzyme activity (FPase), endoglucanase activity (CMCase), and exoglucanase activity (CXase) reached 15.40 U/mL, 11.91 U/mL, and 20.61 U/mL. In addition, the cellulose degradation rate of the treatment group treated with DC-11 was 39.57% in the bioaugmentation test, which was significantly higher than that of the control group without DC-11 (10.01%). Strain DC-11 was shown to be an acid-resistant and heat-resistant cellulose-degrading strain, with high cellulase activity. This strain can exert a bioaugmentation effect on cellulose degradation and has the potential for use in preparing microbial inocula that can be applied for the safe and rapid composting of silkworm excrement.

The Effects of Mammary Gland ATIII Overexpression on the General Health of Dairy Goats and Their Anti-Inflammatory Response to LPS Stimulation.

Antithrombin III is an important anticoagulant factor with anti-inflammatory properties. However, few studies have explored its anti-inflammatory actions in ATIII overexpressed transgenic animals. In this study, the dairy goats with mammary overexpression of ATIII were used to investigate their general health, milk quality and particularly their response to inflammatory challenge. The results showed that transgenic goats have a normal phenotype regarding their physiological and biochemical parameters, including whole blood cells, serum protein levels, total cholesterol, urea nitrogen, uric acid, and total bilirubin, compared to the WT. In addition, the quality of milk also improved in transgenic animals compared to the WT, as indicated by the increased milk fat and dry matter content and the reduced somatic cell numbers. Under the stimulation of an LPS injection, the transgenic goats had elevated contents of IGA, IGM and superoxide dismutase SOD, and had reduced proinflammatory cytokine release, including IL-6, TNF-α and IFN-β. A 16S rDNA sequencing analysis also showed that the transgenic animals had a similar compositions of gut microbiota to the WT goats under the stimulation of LPS injections. Mammary gland ATIII overexpression in dairy goats is a safe process, and it did not jeopardize the general health of the transgenic animals; moreover, the compositions of their gut microbiota also improved with the milk quality. The LPS stimulation study suggests that the increased ATIII expression may directly or indirectly suppress the inflammatory response to increase the resistance of transgenic animals to pathogen invasion. This will be explored in future studies.

Hyaluronic acid production by Klebsiellapneumoniae strain H15 (OP354286) under different fermentation conditions

BackgroundHyaluronic acid (HA) has gained significant attention due to its unique physical, chemical, and biological properties, making it widely used in various industries. This study aimed to screen bacterial isolates for HA production, characterize favorable fermentation conditions, and evaluate the inhibitory effect of bacterial HA on cancer cell lines.ResultsA total of 108 bacterial isolates from diverse sources were screened for HA production using HPLC, turbidimetric, and carbazole determination methods. Among the HA-producing isolates, Klebsiella pneumoniae H15 isolated from an animal feces sample, was superior in HA production. The strain was characterized based on its morphological, cultural, and biochemical characteristics. Molecular identification using 16S rDNA sequencing and phylogenetic analysis confirmed its identity. Fermentation conditions, including pH, temperature, time, and agitation rate, were optimized to maximize HA production. The basal medium, comprising sucrose (7.0%) as carbon source and combined yeast extract with peptone (1.25% each) as nitrogen substrate, favored the highest HA production at pH 8.0, for 30 h, at 30 °C, under shaking at 180 rpm. The average maximized HA concentration reached 1.5 g L−1. Furthermore, bacterial HA exhibited a significant inhibitory effect on three cancer cell lines (MCF-7, HepG-2 and HCT), with the lowest concentration ranging from 0.98–3.91 µg mL−1.ConclusionsK. pneumoniae H15, isolated from animal feces demonstrated promising potential for HA production. The most favorable fermentation conditions led to a high HA production. The inhibitory effect of bacterial HA on cancer cell lines highlights its potential therapeutic applications. These findings contribute to a broader understanding and utilization of HA in various industries and therapeutic applications.

First report of Melampsora ferrinii (pucciniales) on Babylon willow (Salix babylonica) in Iran

As an agroforestry resource willow species (Salix spp.) becoming increasingly important in the protection of riverbanks, recovery of floodable lands, soil bioremediation, biomass production, and wicker furniture manufacture, Babylon willow (Salix babylonica) is widely planted all over the world and is found throughout Iran as part of the rural landscape and is mostly planted for ornamental or bioremediation purposes. Rust fungi in the genus Melampsora usually cause disease on Salix species. Identification of Melampsora species is often complicated because there are few differences in spore morphology and little publicly available comparative sequence data. Willow trees in Iran have been reported to be infected by 15 Melampsora species; however, most of these records are based on morphological characterization. During surveillance activities carried out in June 2007, a rust species was detected on isolated Babylon willow trees in an area in Northern Iran. Both the rust fungus and the host plant were identified on the basis of morphological features and DNA sequencing. Based on a combination of morphology, large subunit (LSU) rDNA sequencing and phylogenetic analyses, our rust specimen was identified as Melampsora ferrinii, which is newly reported from Iran on Salix babylonica. Melampsora ferrinii was originally described from the Americas and this is the first time this rust has been observed on its telial host, Babylon willow, in Asia. The documentation of the rust fungus was performed using both scanning electron microscopy (SEM) and light microscopy. Additionally, an identification key for Melampsora species reported on Salix spp. in Iran is included.

Rapid biosynthesis of biosurfactants by Bacillus tequilensisSL9 isolated from oily sludge: Characterization, optimization, and potential applications

AbstractThe demand of biosurfactants is increasing. It is of great significance to obtain nonpathogenic and high‐yielding microorganisms for the industrial‐scale production of biosurfactants. A biosurfactant‐producing strain SL9 was isolated from oily sludge. It was identified as Bacillus tequilensis based on colonial morphology and 16S rDNA sequence analysis. The biosurfactants produced by strain SL9 was extracted by modified acid precipitation method. A mixture of C12, C13, C14 and C15 surfactin homologues was detected based on HPLC–MS analysis. The optimum production of biosurfactants occurs at pH 7.0 and in the presence of 1.0 g/L NaCl. The optimal carbon source and nitrogen source were 40.0 g/L sucrose and 4.0 g/L NaNO3, respectively. The maximum yield of biosurfactants from strain SL9 was up to 1332.68 mg/L. Interestingly, B. tequilensis SL9 can rapidly synthesize biosurfactants with a yield of 739.36 mg/L in 12 h. The extracted biosurfactants reduced the surface tension of water from 72.1 to 27.5 mN/m with a CMC of 33 mg/L. The biosurfactants can also emulsify paraffin with a EI24 value of 57.58% and keep this good emulsifying activity after 96 h. The produced biosurfactants showed antimicrobial activities against Staphylococcus aureus, Cladosporium sp. and Alternaria sp. with inhibitory rate of 75.67%, 59.07% and 22.91%, respectively. The isolated strain B. tequilensis SL9 can be a nonpathogenic producer for rapid biosynthesis of biosurfactants with a relative high yield. Strain SL9 and its biosurfactant products are promising for application in the oil exploration, bioremediation, agricultural biocontrol, and pharmaceutical industries.

High-level production of poly-γ-glutamic acid by a newly isolated Bacillus sp. YJY-8 and potential use in increasing the production of tomato

Strain YJY-8, a new γ-polyglutamic acid producer, was separated from fermented soybean paste samples. The strain was identified as a genus of Bacillus by morphological and 16S rDNA sequence analysis and was named Bacillus sp. YJY-8. The optimal medium composition and cultural conditions were studied using a single-factor experiment and a response surface experiment. The optimized medium consisted of monosodium glutamate 70 g/L, glucose 54.3 g/L, glycerol 31.8 g/L, ammonium sulfate 11.1 g/L, yeast extract 3.2 g/L, tryptone 1.5 g/L, L-glutamic acid 6.8 g/L, MgSO4 7H2O 0.5 g/L, FeCl3 6H2O 0.02 g/L, KH2PO4 0.9 g/L, CaCl2 0.03 g/L, MnSO4 H2O 0.3 g/L, ammonium molybdate 0.02 g/L, pH 7.0. The optimal cultivation conditions were 35 °C and pH 7.0. Under the optimized conditions, after 48 hr of cultivation, the highest shaking flask fermentation level of γ-PGA reached 65.2 ± 0.36 g/L. In addition, through fed-batch fermentation in 30 L fermenters, the fermentation level of γ-PGA reached its highest level at 88.42 g/L and productivity was 1.23 g/(L hr) after 72 hr. Then, the effect of γ-PGA on tomato yield was investigated. At the seedling stage, the plant height and stem diameter of γ-PGA treated plants increased by 5.69 and 15.735% after spraying γ-PGA for 19 days. During the flowering and fruiting period, the stem diameter of the γ-PGA treatment group increased by 6.74%, with a maximum increase of 11.65%. The number of fruit branches increased by 0.56–16.29% and the number of fruit sets increased by 1.01–28.47%. At the fruit maturation stage, the yield of tomatoes increased by 10.51, 14.27, and 5.83%.

The effects of astragaloside IV on gut microbiota and serum metabolism in a mice model of intracerebral hemorrhage

BackgroundAstragaloside IV (AS-IV) is the main active component of “Astragalus membranaceus (Fisch.) Bunge, a synonym of Astragalus propinquus Schischkin (Fabaceae)”, which demonstrated to be useful for the treatment of intracerebral hemorrhage (ICH). However, due to the low bioavailability and barrier permeability of AS-IV, the gut microbiota may be an important key regulator for AS-IV to work. ObjectiveTo explore the influences of gut microbiota on the effects of AS-IV on ICH. MethodsMice were randomly divided into five groups: sham, ICH, and AS-IV-treated groups (25 mg/kg, 50 mg/kg, and 100 mg/kg). Behavioral tests, brain histopathology, and immunohistochemistry analysis were used to evaluate the degree of brain injury. Western blot was employed to verify peri‑hematoma inflammation. The plasma lipopolysaccharide (LPS) leakage, the fluorescein isothiocyanate-dextran permeability, the colonic histopathology, and immunohistochemistry were detected to evaluate the barrier function of intestinal mucosal. Moreover, 16S rDNA sequencing and metabolomic analysis was applied to screen differential bacteria and metabolites, respectively. The correlation analysis was adopted to determine the potential relationship between differential bacteria and critical metabolites or neurological deficits. ResultsAS-IV alleviated neurological deficits, neuronal injury and apoptosis, and blood-brain barrier disruption. This compound reduced tumor necrosis factor (TNF)-α expression, increased arginase (Arg)-1 and interleukin (IL)-33 levels around the hematoma. Next, 16S rRNA sequencing indicated that AS-IV altered the gut microbiota, and inhibited the production of conditional pathogenic bacteria. Metabolomic analysis demonstrated that AS-IV regulated the serum metabolic profiles, especially the aminoacid metabolism and peroxisome proliferator-activated receptor (PPAR) signaling pathway. Additionally, AS-IV mitigated intestinal barrier damage and LPS leakage. ConclusionThis study provides a new perspective on the use of AS-IV for the treatment of ICH. Among them, gut microbiota and its metabolites may be the key regulator of AS-IV in treating ICH.

Antifungal Trait and Plant Growth Promotion Potential of Bacillus Spp. from Rhizosphere Soils of Black Aromatic Rice, ‘Chakhao’

Manipuri black aromatic rice, ‘Chakhao’, is a nutrient rich unique local rice cultivar of Manipur, India. Rhizosphere bacteria may have potential as antifungal agent and seedling growth promoter. Hence, the current investigation was aimed at screening, for the above traits, using bacteria, Bacillus spp., isolated from the soil of Rhizosphere of six Chakhao rice cultivars. Altogether, 323 bacterial isolates were obtained from the rhizospheric soils of 6 different cultivars of Chakhao, namely Chakhao Amubi (CA), Chakhao Poireiton (CP), Chakhao Sempak (CS), Chakhao Angoubi (CAng), Chakhao Angangbi (CR) and Chakhao Wairi (CW). All the rhizobacterial isolates were screened for antifungal activity against 5 rice fungal pathogens viz. Rhizoctonia solani (RS), Fusarium oxysporum (FO), Curvularia oryzae (CO), Pyricularia oryzae (PO) and Aspergillus niger (AN). The isolates were also subjected to PGP (plant growth promotion) assays such as ammonia, indole acetic acid (IAA), siderophore and 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase production, and phosphate solubilization. Of 323 isolates obtained, 64 were found to exhibit antifungal activity while 69 showed PGP traits. Twenty-five (25) isolates were found to be positive for both antifungal and PGP traits. Of 25 bioactive isolates, 4 (CR12, CW11, CA2 and CP2) potent isolates were shortlisted for further studies. The shortlisted potent isolates were subjected to quantitative estimation of PGP activities like phosphate solubilization, IAA and siderophore production. The strains could produce significant amount of IAA and siderophore and solubilize phosphate. Molecular characterization by 16S rDNA sequence analyses revealed the identity of the isolates: CR12: Bacillus subtilis (Accn. No.OM866257), CW11: Bacillus paralicheniformis (Accn. No.OM868047), CA2: Bacillus sp. CCMB1014 (Accn. No.OM868070) and CP2: Bacillus licheniformis (Accn. No.OM892495). Seedling vigor assays of the shortlisted potent strains were carried out on Chakhao Amubi rice. The isolates exhibited higher seedling vigor indices (CR12: 668.00, CA2: 618.66, CP2: 510.92 and CW11: 478.91) over the control (164.48). These rhizospheric strains have the potential to be developed as bioinoculants or biostimulants for enhancing Chakhao rice growth.

Analysis of the Potassium-Solubilizing Priestia megaterium Strain NK851 and Its Potassium Feldspar-Binding Proteins.

Potassium-solubilizing bacteria are an important microbial group that play a critical role in releasing mineral potassium from potassium-containing minerals, e.g., potassium feldspar. Their application may reduce eutrophication caused by overused potassium fertilizers and facilitate plants to utilize environmental potassium. In this study, a high-efficiency potassium-solubilizing bacterium, named NK851, was isolated from the Astragalus sinicus rhizosphere soil. This bacterium can grow in the medium with potassium feldspar as the sole potassium source, releasing 157 mg/L and 222 mg/L potassium after 3 days and 5 days of incubation, respectively. 16S rDNA sequencing and cluster analysis showed that this strain belongs to Priestia megaterium. Genome sequencing further revealed that this strain has a genome length of 5,305,142 bp, encoding 5473 genes. Among them, abundant genes are related to potassium decomposition and utilization, e.g., the genes involved in adherence to mineral potassium, potassium release, and intracellular trafficking. Moreover, the strong potassium-releasing capacity of NK851 is not attributed to the acidic pH but is attributed to the extracellular potassium feldspar-binding proteins, such as the elongation factor TU and the enolase that contains potassium feldspar-binding cavities. This study provides new information for exploration of the bacterium-mediated potassium solubilization mechanisms.

Identification of Potential Biofertilizer and Bioremediator Bacteria from Upland Soil Based on 16s rDNA Sequence Analysis

The long-term presence of synthetic pesticides on agricultural land can lead to a decline in soil fertility. Synthetic pesticides inhibit the activity of essential enzymes in the soil and suppress beneficial microbial populations for plants. One potential approach to mitigate the extent of contamination caused by synthetic pesticides involves the utilization of indigenous pesticide-resistant bacteria. Several upland soil bacteria from Banyumas Regency, Central Java Province, Indonesia, were successfully isolated from a previous study. The isolated bacteria have the potential to be developed as pesticide bio-remediators and biofertilizers. The bacterial isolates are expected to have characters that support plant growth through their ability to provide dissolved phosphate. However, the potential bacterial isolates need to be identified by molecular approaches. This study was conducted to identify bacterial isolates of GT2, SR1, SW1, and PA1 by 16S rDNA sequencing analysis. The results showed that isolate GT2 was placed within a group of reference strains of Bacillus proteolyticus, isolate SR1 was placed within a group of B. paramycoides, isolate SW1 was set within a group of B. albus, and isolate PA1 was placed within a group of Acidovorax delafieldii. The genetic distance of isolate GT2 and B. B. proteolyticus, isolate SR1 and B. paramycoides, isolate SW1 and B. albus were 0.0000 each, and isolate PA1 and A. delafieldii were 0.0061.

Rapid arsenite oxidation by Paenarthrobacter nicotinovorans strain SSBW5: unravelling the role of GlpF, aioAB and aioE genes.

A novel arsenite resistant bacterial strain SSBW5 was isolated from the battery waste site of Corlim, Goa, India. This strain interestingly exhibited rapid arsenite oxidation with an accumulation of 5mM arsenate within 24h and a minimum inhibitory concentration (MIC) of 18mM. The strain SSBW5 was identified as Paenarthrobacter nicotinovorans using 16S rDNA sequence analysis. Fourier-transformed infrared (FTIR) spectroscopy of arsenite-exposed cells revealed the interaction of arsenite with several important functional groups present on the cell surface, possibly involved in the resistance mechanism. Interestingly, the whole genome sequence analysis also clearly elucidated the presence of genes, such as GlpF, aioAB and aioE encoding transporter, arsenite oxidase and oxidoreductase enzyme, respectively, conferring their role in arsenite resistance. Furthermore, this strain also revealed the presence of several other genes conferring resistance to various metals, drugs, antibiotics and disinfectants. Further suggesting the probable direct or indirect involvement of these genes in the detoxification of arsenite thereby increasing its tolerance limit. In addition, clumping of bacterial cells was observed through microscopic analysis which could also be a strategy to reduce arsenite toxicity thus indicating the existence of multiple resistance mechanisms in strain SSBW5. In the present communication, we are reporting for the first time the potential of P. nicotinovorans strain SSBW5 to be used in the bioremediation of arsenite via arsenite oxidation along with other toxic metals and metalloids.

BzATP reverses ferroptosis-induced gut microbiota disorders in collagen-induced arthritis mice

Recent studies suggested that altered gut microbiota may be related to the pathogenesis of rheumatoid arthritis (RA), albeit the exact mechanisms are unknown. In this study, we aimed to discover the particular mechanism of RA treatment by microbiota by investigating the effects of ferroptosis on gut microbiota and its metabolites in collagen-induced arthritis (CIA) mice. Mice were divided into five groups: control, CIA, erastin, BzATP, and BzATP + erastin group. We performed 16S rDNA sequencing and metabolomics analysis on mouse feces and found that erastin and BzATP altered the microbiota and metabolites. The findings demonstrated that the microbiota was significantly disturbed at the phylum (Proteobacteria, Firmicutes, and Bacteroidota) and genus level (Lachnospiraceae_NK4A136, Lactobacillus, and Bifidobacterium) in the CIA group, and erastin exacerbated this disturbance. Unexpectedly, BzATP treatment could repair the disruptive effects of erastin. Additionally, there were significant variations in metabolites between each group. Erastin worsened metabolite abnormalities in CIA mice, while BzATP mitigated them, consistent with the microbiota results. These findings provide novel perspectives and insights into the therapy of RA.