ABSTRACT
Bacterial DNA phosphorothioate (PT) modification provides a specific anchoring site for sulfur-binding proteins (SBDs). Besides, their recognition patterns include phosphate links and bases neighboring the PT-modified site, thereby bringing about genome sequence-dependent properties in PT-related epigenetics. Here, we analyze the contributions of the DNA backbone (phosphates and deoxyribose) and bases bound with two SBD proteins in Streptomyces pristinaespiralis and coelicolor (SBDSco and SBDSpr). The chalcogen-hydrophobic interactions remained constantly at the anchoring site while the adjacent bases formed conditional and distinctive non-covalent interactions. More importantly, SBD/PT-DNA interactions were not limited within the traditional "4-bp core" range from 5'-I to 3'-III but extended to upstream 5'-II and 5'-III bases and even 5''-I to 5''-III at the non-PT-modified complementary strand. From the epigenetic viewpoint, bases 3'-II, 5''-I, and 5''-III of SBDSpr and 3'-II, 5''-II, and 5''-III of SBDSco present remarkable differentiations in the molecular recognitions. From the protein viewpoint, H102 in SBDSpr and R191 in SBDSco contribute significantly while proline residues at the PT-bound site are strictly conserved for the PT-chalcogen bond. The mutual and make-up mutations are proposed to alter the SBD/PT-DNA recognition pattern, besides additional chiral phosphorothioate modifications on phosphates 5'-II, 5'-II, 3'-I, and 3'-II.
Subject(s)
Chalcogens , DNA , DNA/chemistry , DNA, Bacterial/chemistry , Bacterial Proteins/metabolism , Phosphates/chemistryABSTRACT
A bacterial strain was isolated from a brackish water sample of Tagus river, Alcochete, Portugal and was designated TO1_6T. It forms light pink colonies on M13 medium supplemented with N-acetylglucosamine. Cells are pear-shaped to spherical, form rosettes and divide by budding. Strain TO1_6T presents a mesophilic and neutrophilic profile, with optimum growth at 20 to 25 °C and pH 7.0 to 7.5, and vitamin supplementation is not required to promote its growth. The genome of the novel isolate is 7.77 Mbp in size and has a DNA G + C content of 56.3%. Based on its 16S rRNA gene sequence, this strain is affiliated with the phylum Planctomycetota. Further taxonomic characterization using additional phylogenetic markers, namely rpoB gene sequence (encoding the ß-subunit of the DNA-dependent RNA polymerase), as well as Percentage of conserved proteins, average nucleotide identity and average amino acid identity, suggest the affiliation of strain TO1_6T to the genus Stieleria, a recently described taxon in the family Pirellulaceae, order Pirellulales and class Planctomycetia. Based on the genotypic, phylogenetic and physiological characterization, we here describe a new species represented by the type strain TO1_6T (= CECT 30432T, = LMG 32465T), for which the name Stieleria tagensis sp. nov. is proposed.
Subject(s)
Fatty Acids , Rivers , Rivers/microbiology , Fatty Acids/analysis , Phospholipids/analysis , Planctomycetes , Sequence Analysis, DNA , Phylogeny , RNA, Ribosomal, 16S/genetics , Portugal , DNA, Bacterial/genetics , DNA, Bacterial/chemistry , Bacterial Typing TechniquesABSTRACT
The novel bacterial strain Marseille-P4005T was isolated from the stool sample of a healthy donor. It is a Gram-stain negative, non-motile, non-spore-forming rod. It grew optimally at 37 °C and at pH 7.0 on 5% sheep blood-enriched Columbia agar after preincubation in a blood-culture bottle supplemented with rumen and blood. This strain does not ferment monosaccharides (except D-tagatose), disaccharides, or polymeric carbohydrates. The major cellular fatty acids were hexadecenoic (24.6%), octadecanoic (22.8%), and tetradecanoic (20.1%) acids. Next-generation sequencing revealed a genome size of 3.2 Mbp with a 56.4 mol% G + C. Phylogenetic analysis based on the 16S rRNA gene highlighted Agathobaculum desmolans strain ATCC 43058T as the closest related strain. The OrthoANI, AAI, and digital DNA-DNA hybridization values were below the critical thresholds of 95%, 95-96%, and 70%, respectively, to define a novel bacterial species. Antibiotic resistance genes APH(3')-IIIa, erm(B), and tet(W) were detected with high identity percentages of 100%, 98.78%, and 97.18% for each gene, respectively. The APH(3')-IIIa gene confers resistance to amikacin, erm(B) gene confers resistance to erythromycin, lincomycin, and clindamycin, while tet(W) gene confers resistance to doxycycline and tetracycline. Based on KEGG BlastKOALA analyses, the annotation results showed that our strain could use glucose to produce L-lactate and pyruvate but not acetate or ethanol. Also, strain Marseille-P4005T was predicted to use phenylalanine to produce indole, a major intercellular signal molecule within the gut microbial ecosystem. Through having a gene coding for tryptophan synthase beta chain (trpB), strain Marseille-P4005T could produce L-tryptophan (an essential amino acid) from indole. Strain Marseille-P4005T showed its highest prevalence in the human gut (34.19%), followed by the reproductive system (17.98%), according to a query carried out on the Integrated Microbial NGS (IMNGS) platform. Based on phylogenetic, phenotypic, and genomic analyses, we classify strain Marseille-P4005T (= CSUR P4005 = CECT 9669), a novel species within the genus Agathobaculum, for which the name of Agathobaculum massiliense sp. nov. is proposed.
Subject(s)
Lactobacillales , Tryptophan , Humans , Tryptophan/genetics , Phylogeny , RNA, Ribosomal, 16S/genetics , Ecosystem , Kanamycin Kinase/genetics , Base Composition , Genomics , Bacteria/genetics , Lactobacillales/genetics , Fatty Acids/chemistry , Indoles , DNA , DNA, Bacterial/genetics , DNA, Bacterial/chemistry , Sequence Analysis, DNA , Bacterial Typing TechniquesABSTRACT
DNA phosphorothioation (PT) exists in many pathogenic bacteria; however, the mechanism of PT-DNA resistance to the immune response is unclear. In this work, we meticulously investigated the peroxynitrite (PN) tolerance using PT-bioengineered E. coli strains. The in vivo experiment confirms that the S+ strain survives better than the S- strain under moderately oxidative stress. The LCMS, IC, and GCMS experiments demonstrated that phosphorothioate partially converted to phosphate, and the byproduct included sulfate and elemental sulfur. When O,O-diethyl thiophosphate ester (DETP) was used, the reaction rate k1 was determined to be 4.3 ± 0.5 M-1 s-1 in the first-order for both phosphorothioate and peroxynitrite at 35 °C and pH of 8.0. The IC50 values of phosphorothioate dinucleotides are dramatically increased by 400-700-fold compared to DETP. The SH/OH Yin-Yang mechanism rationalizes the in situ DNA self-defense against PN-mediated oxidative stress at the extra bioenergetic cost of DNA modification.
Subject(s)
DNA, Bacterial/metabolism , Oxidative Stress/drug effects , Peroxynitrous Acid/pharmacology , Phosphorothioate Oligonucleotides/metabolism , DNA, Bacterial/chemistry , DNA, Bacterial/genetics , Escherichia coli/drug effects , Escherichia coli/genetics , Escherichia coli/metabolism , Kinetics , Multigene Family , Phosphorothioate Oligonucleotides/chemistry , Phosphorothioate Oligonucleotides/geneticsABSTRACT
The bacteria inhabiting brackish lake environments in arid or semi-arid regions have not been thoroughly identified. In this study, the 454 pyrosequencing method was used to study the sedimentary bacterial community composition (BCC) and diversity in Lake Bosten, which is located in the arid regions of northwestern China. A total of 210,233 high-quality sequence reads and 8,427 operational taxonomic units (OTUs) were successfully obtained from 20 selected sediment samples. The samples were quantitatively dominated by members of Proteobacteria (34.1% ± 11.0%), Firmicutes (21.8% ± 21.9%) and Chloroflexi (13.8% ± 5.2%), which accounted for more than 69% of the bacterial sequences. The results showed that (i) Lake Bosten had significant spatial heterogeneity, and TOC(total organic carbon), TN(total nitrogen) and TP(total phosphorus) were the most important contributors to bacterial diversity; (ii) there was lower taxonomic richness in Lake Bosten, which is located in an arid region, than in reference lakes in eutrophic floodplains and marine systems; and (iii) there was a low percentage of dominant species in the BCC and a high percentage of unidentified bacteria. Our data help to better describe the diversity and distribution of bacterial communities in contaminated brackish lakes in arid regions and how microbes respond to environmental changes in these stable inland waters in arid or semi-arid regions.
Subject(s)
Chloroflexi/isolation & purification , Ecosystem , Firmicutes/isolation & purification , Lakes/microbiology , Proteobacteria/isolation & purification , Biodiversity , China , Chloroflexi/genetics , DNA, Bacterial/chemistry , DNA, Bacterial/metabolism , Firmicutes/genetics , Geologic Sediments/microbiology , Phosphorus/chemistry , Phosphorus/metabolism , Proteobacteria/genetics , Seasons , Sequence Analysis, DNA/methods , Water QualityABSTRACT
Although many literatures have reported that biomass materials had been used for water treatment, most of the biomass materials were directly used for adsorbing toxic or organic substances. In this research, modified pine bark and corn straw were used to prepare the high-efficiency and low-cost activated sludge immobilized materials. By treating phenol wastewater and ordinary organic wastewater, various factors influencing the treatment effect were investigated. The results showed that the immobilized biological carriers had good effects on the treatment of two kinds of wastewater. The removal efficiency of the phenol wastewater reached 100% in 24â¯h, and the removal efficiency of ordinary organic wastewater reached 95.5% in 96â¯h. The results of microbial community analysis showed that the abundance of the immobilized carrier and that of the original sludge was similar. But when treating different wastewater, the number and proportion of microorganisms were significantly different.
Subject(s)
Microbiota , Waste Disposal, Fluid/methods , Bacteria , Biodegradation, Environmental , Biological Oxygen Demand Analysis , Biomass , Bioreactors , DNA, Bacterial/chemistry , Hydrogen-Ion Concentration , Particle Size , Phenols/chemistry , Pinus , Plant Bark , Sewage , Temperature , Wastewater , Zea maysABSTRACT
The present study aimed to isolate the high-efficiency petrol metabolizing thermophilic bacteria from petrol contaminated soil samples. Isolation was carried out through enrichment culture, serial dilution and pour plate methods using the petrol supplemented minimal salt media. The isolated bacteria were analyzed to document growth behavior, petrol removal efficiencies, antibiotic resistance profile, and biochemical characteristics. The 16S rRNA based phylogenetic analysis helped to reveal the identity of isolated bacterial species and construct the phylogenetic trees. Total nine bacteria were isolated, out of which three (IUBP2, IUBP3, IUBP5) were identified as Brevibacillus formosus, one (IUBP1) was found similar to Brevibacillus agri, four (IUBP7, IUBP8, IUBP13, and IUBP14) shared homology with Burkholderia lata, and one (IUBP15) with Burkholderia pyrrocinia. All the isolates were fast growing and exhibited considerable petrol degradation potential. The highest petrol removal efficiency (69.5% ± 13.44/6 days) was recorded for the strain IUBP15 at a petrol concentration of 0.1% (v/v). All bacteria studied (100%) were positive for esculinase and phosphatase. Many strains exhibited positive responses for arginine dehydrolase (22%), ß-naphthylamidase (11%), ß-D-glucosaminide (33%), mannitol (55%), sorbitol (66%) and inulin (88%) fermentation test. While all were sensitive to the antibiotics, some of them were found resistant against chloramphenicol and oxacillin. The remarkable biochemical characteristics and considerable petrol removal potential (40-70%) highlights utilization of the bacteria isolated for petrol bioremediation, mineralization of organophosphates, dairy and food industry, and also as biofertilizers and biocontrol agents.The present study aimed to isolate the high-efficiency petrol metabolizing thermophilic bacteria from petrol contaminated soil samples. Isolation was carried out through enrichment culture, serial dilution and pour plate methods using the petrol supplemented minimal salt media. The isolated bacteria were analyzed to document growth behavior, petrol removal efficiencies, antibiotic resistance profile, and biochemical characteristics. The 16S rRNA based phylogenetic analysis helped to reveal the identity of isolated bacterial species and construct the phylogenetic trees. Total nine bacteria were isolated, out of which three (IUBP2, IUBP3, IUBP5) were identified as Brevibacillus formosus, one (IUBP1) was found similar to Brevibacillus agri, four (IUBP7, IUBP8, IUBP13, and IUBP14) shared homology with Burkholderia lata, and one (IUBP15) with Burkholderia pyrrocinia. All the isolates were fast growing and exhibited considerable petrol degradation potential. The highest petrol removal efficiency (69.5% ± 13.44/6 days) was recorded for the strain IUBP15 at a petrol concentration of 0.1% (v/v). All bacteria studied (100%) were positive for esculinase and phosphatase. Many strains exhibited positive responses for arginine dehydrolase (22%), ß-naphthylamidase (11%), ß-D-glucosaminide (33%), mannitol (55%), sorbitol (66%) and inulin (88%) fermentation test. While all were sensitive to the antibiotics, some of them were found resistant against chloramphenicol and oxacillin. The remarkable biochemical characteristics and considerable petrol removal potential (4070%) highlights utilization of the bacteria isolated for petrol bioremediation, mineralization of organophosphates, dairy and food industry, and also as biofertilizers and biocontrol agents.
Subject(s)
Bacteria/isolation & purification , Bacteria/metabolism , Petroleum/metabolism , Soil Microbiology , Soil Pollutants/metabolism , Anti-Bacterial Agents/pharmacology , Bacteria/classification , Bacteria/genetics , Cluster Analysis , DNA, Bacterial/chemistry , DNA, Bacterial/genetics , DNA, Ribosomal/chemistry , DNA, Ribosomal/genetics , Drug Resistance, Bacterial , Enzymes/analysis , Microbial Sensitivity Tests , Petroleum Pollution , Phylogeny , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNAABSTRACT
The increased concentrate amounts in cow diets may initiate changes in both particle-associated (PaM) and epimural microbiota (EpM) with the potential for promoting the establishment of pathogens. Clay minerals have shown promising potentials in binding harmful microorganisms and metabolites due to their high adsorption capacity. This study evaluated the effects of a clay-mineral based product (CM) on PaM, EpM, fermentation parameters, and epithelial gene expression in cows fed a high-concentrate diet. Eight rumen-cannulated non-lactating Holstein cows received a concentrate mix supplemented with CM or not (CON) in a change-over design with an initial 100% roughage diet phase (RD, 1 week), followed by intermittent 65%-high-concentrate-diet phases (HC1, HC2; 1 and 2 week duration, respectively), interrupted by 1 week roughage only. Rumen samples for short-chain fatty acids, ammonia, and lactate quantification, as well as PaM, and epithelial biopsies for EpM examination and epithelial gene expression were collected via the cannula once during each feeding phase. Phylogenetic distance analysis of Illumina MiSeq sequencing of the 16S rRNA gene region V345 showed a clear clustering of RD microbiota compared to HC in PaM, showing the impact of the high-concentrate diet on the bacterial community. In the EpM this effect was less pronounced, due to higher variability in RD. In the PaM, a decrease (Pâ¯<â¯0.01) of community diversity occurred with the onset of HC feeding, while in the EpM there was an increase in diversity (Pâ¯<â¯0.05). In the PaM, CM increased the relative abundance of genus Butyrivibrio (Pâ¯<â¯0.01), a commensal bacterium of the rumen, which was, with 6.4%, the second most abundant genus. There, the CM supplementation decreased the genera Lactobacillus, Fusobacterium, and Treponema (Pâ¯=â¯0.05), which are potentially either lactate producing or opportunistic pathogens. In the EpM, CM decreased the relative abundance of Succiniclasticum genus (Pâ¯<â¯0.01), a possible endotoxin producer, and increased bacteria that are associated with a normobiotic rumen, such as Campylobacter (Pâ¯=â¯0.06). Barrier function genes were upregulated in HC2 and nutrient transport genes downregulated in HC1 (Pâ¯<â¯0.05); however, there was little effect on pro-inflammatory genes at the epithelium. The CM showed a significant decreasing effect on the cellular metabolism genes HMGCS1 (Pâ¯=â¯0.04). Our results suggest that CM supplementation can increase the relative abundance of commensal microbiota and decrease bacteria that could negatively impact the rumen milieu and health during high-concentrate feeding.
Subject(s)
Clay/chemistry , Diet/methods , Gene Expression Regulation/drug effects , Microbiota/drug effects , Minerals/administration & dosage , Rumen/microbiology , Ammonia/analysis , Animals , Cattle , Cluster Analysis , DNA, Bacterial/chemistry , DNA, Bacterial/genetics , DNA, Ribosomal/chemistry , DNA, Ribosomal/genetics , Fatty Acids, Volatile/analysis , Fermentation/drug effects , Lactates/analysis , Phylogeny , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNAABSTRACT
In order to better understand the composition and driving factors of the bacterial community in Mollisols, we selected 9 representative facility agricultural lands in Mollisol area of China for sampling, and described it on a larger spatial scale. Soil bacterial community structure in these 9 regions (determined by high-throughput sequencing analysis) showed significant differences at the genus level. The correlation between bacterial community composition and soil properties, contaminants and geographical latitude showed that the diversity of bacterial community was still strongly correlated with pH and SOM under the influence of phthalates (Pâ¯<â¯0.05). Principal component Analysis (PCA) showed that soil properties (i.e. pH, organic matter, stacking density, the content of nitrogen, potassium, phosphorus) and PAEs level rather than geographic latitude were main drivers of differences in bacterial community structure. These factors account for 73.04% of the total variation of the bacterial community. Among them, PAEs act as a typical pollutant is the main factor driving the composition of bacterial community in facility agriculture Mollisols. This shows that PAEs is a potential pollution risk factor, which has important guiding significance for the sustainable and healthy development of agriculture in Mollisol area.
Subject(s)
Environmental Pollution/analysis , Phthalic Acids/analysis , Soil Microbiology , Soil Pollutants/analysis , Agriculture , Bacteria/classification , Bacteria/drug effects , Bacteria/genetics , China , DNA, Bacterial/chemistry , Microbiota , Nitrogen/analysis , Phosphorus/analysis , Phthalic Acids/toxicity , Principal Component Analysis , Sequence Analysis, DNA , Soil/chemistry , Soil Pollutants/toxicityABSTRACT
A total of 150 rhizobacteria and endorhizobacteria previously isolated from three different horticultural crops; strawberry, apple and apricot were screened for antagonistic activitiy against Clavibacter michiganensis ssp. michiganensis. Among them strain S1, exhibiting significantly higher antagonistic and plant growth promoting ability was characterized as Bacillus amyloliquefaciens based on morphological, biochemical and partial gene sequence analysis of 16S rRNA. B. amyloliquefaciens strain S1 showed maximum growth inhibition of C. michiganensis (12â¯mm). Moreover, B. amyloliquefaciens strain S1 exhibit significant phosphorus solubilization (94.16 %SEl) and indole acetic acid (27⯵gâ¯ml-1) production under in vitro conditions. Antagonistic activity of Bacillus amyloliquefaciens strain S1 was compared with other four strains KU2S1, R2S(1), RG1(3) and AG1(7) against bacterial canker of tomato under net house conditions. Minimum bacterial canker disease incidence (30.0%) was recorded in B. amyloliquefaciens S1 followed by RG1(3) after 30 days of inoculation. The bio-control efficacy was higher in B. amyloliquefaciens S1 treated plants, followed by RG1(3).
Subject(s)
Actinobacteria/growth & development , Antibiosis , Bacillus amyloliquefaciens/growth & development , Plant Diseases/microbiology , Plant Diseases/prevention & control , Solanum lycopersicum/microbiology , Bacillus amyloliquefaciens/classification , Bacillus amyloliquefaciens/genetics , Bacillus amyloliquefaciens/isolation & purification , Cluster Analysis , DNA, Bacterial/chemistry , DNA, Bacterial/genetics , DNA, Ribosomal/chemistry , DNA, Ribosomal/genetics , Indoleacetic Acids/metabolism , Phosphorus/metabolism , Phylogeny , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNAABSTRACT
This study aimed at testing the effects of three different formulations of feed supplements based on three different combinations of plant derived alkaloids, prebiotics, tannins, vitamins and minerals on rumen fermentation and the microbiome in vitro. A Rusitec experiment was conducted in 2 identical runs using a complete randomized design with 3 replicates per treatment resulting in total of 6 treatment combinations (nâ¯=â¯6). Each run lasted 12â¯d with sampling occurring in the last 5â¯d. Diets were a standard dairy ration (60:40; concentrate:forage) supplemented with one of 3 different plant-based combinations (PI, PII, and PIII) at a level of 100â¯mg/l and a non-supplemented control (basal diet, control). Microbial DNA samples were taken on the last day of each run and the 16S rRNA target gene sequenced using Illumina MiSeq technology. The supplementations had no effect on the pH, methane and carbon dioxide production. However, both total SCFA (Pâ¯=â¯0.08) and molar concentrations of acetate (Pâ¯=â¯0.06) tended to be increased in the treatment groups in comparison to control, with PII having the highest overall values (102.7â¯mmol/L and 43.3â¯mmol/L, respectively). Alpha diversity indices Shannon, Simpson and Chao1 showed no effect of supplementations or combinations. The addition of PII increased the relative abundance of Bacteroidetes compared to all other treatments (Pâ¯=â¯0.05). Supplementation with plant-based combinations reduced the relative abundance of Pyramidobacter from the family Dethiosulfovibrionaceae in comparison with the control diet (Pâ¯=â¯0.05). Evaluation of predicted gene function through PICRUSt analysis showed variation in predicted cellular function and metabolism between bacterial communities supplemented with plant-based combinations compared to the control diet. This shows that the addition of plant-based combinations can have the potential to modulate the metabolic function of rumen microbes, and likely the production of small-sized rumen metabolites, without disrupting the rumen microbial community structure and diversity.
Subject(s)
Animal Feed , Bacteria/classification , Bacteria/metabolism , Fermentation , Rumen/microbiology , Animals , Cluster Analysis , DNA, Bacterial/chemistry , DNA, Bacterial/genetics , DNA, Ribosomal/chemistry , DNA, Ribosomal/genetics , Metabolome , Metagenome , Models, Theoretical , Phylogeny , RNA, Ribosomal, 16S/genetics , Rumen/chemistry , Sequence Analysis, DNAABSTRACT
In the present study, we investigated the potential of Bifidobacterium spp., isolated from ruminal fluid samples from buffaloes (Bubalus bubalis) for conjugated linoleic acid (CLA) production. A total of 294 isolates were obtained from 86 ruminal fluid samples using Bifidus Selective Medium (BSM) medium, and based on phospoketolase assay, 24 isolates were presumptively confirmed to be Bifidobacterium species. Further, the isolates were confirmed morphologically, biochemically and by PCR assays for genus specific (16s rDNA) and transaldolase genes. All 24 strains were positive for conversion of linoleic acid (LA) to CLA by spectrophotometric screening. Gas chromatographic analysis showed that the strains produced cis9, trans11 and tran10, cis12 CLA isomers in LA-supplemented deMan-Rogosa-Sharpe (MRS) broth. The strains were identified as B. thermophilum (nâ¯=â¯21) and B. pseudolongum (nâ¯=â¯3) based on 16 rDNA sequence analysis. The study shows that Bifidobacterium spp., present in the rumens of buffaloes produce CLA from LA and the strains may have the potential to be used as probiotics to enhance the nutraceutical value of ruminant food products.
Subject(s)
Bifidobacterium/isolation & purification , Bifidobacterium/metabolism , Buffaloes , Linoleic Acids, Conjugated/metabolism , Rumen/microbiology , Animals , Bacteriological Techniques , Bifidobacterium/classification , Bifidobacterium/genetics , Body Fluids/microbiology , Cluster Analysis , DNA, Bacterial/chemistry , DNA, Bacterial/genetics , DNA, Ribosomal/chemistry , DNA, Ribosomal/genetics , Phylogeny , Polymerase Chain Reaction , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA , Transaldolase/geneticsABSTRACT
Plant-microbe interactions such as rhizobacteria legumes are interesting in organic farming that has undergone significant expansion in the world. The organic agriculture is as an environment-friendly technique and a sustainable alternative to intensive agricultural system. Three types of soil were chosen, organic (ORG), conventional (CON), and fallow land (NA) to isolate soil bacteria-nodulating Medicago sativa, in order to develop microbial inoculants for use in agricultural sustainable system. Soil analysis revealed significant higher amounts of total nitrogen, organic carbon, total phosphorus, and matter detected in ORG. As for heavy metals, ORG showed high Cu content due to the authorized chemical use in organic farming. A sample of 130 bacteria was isolated from Medicago sativa nodule, genetically characterized by PCR/RFLP of ribosomal 16S RNAs, and a great dominance of Sinorhizobium meliloti (88.4%, 73.8%, and 55.5%) is obtained among NA-, CON-, and ORG-managed soils, respectively. The ORG showed the high bacterial diversity with 13.3% of non-identified strains. The resistance against five pesticides (Prosper, Cuivox, Fungastop, Nimbecidine, and Maneb) revealed a maximum of inhibitory concentration about 10 mg l-1 of Prosper, 12 mg l-1 of Cuivox, 6 ml l-1 of Fungastop, 7.5 ml l-1of Nimbecidine, and 25 ml l-1 of Maneb. The analysis of the symbiotic properties and plant growth-promoting potential revealed two efficient strains significantly increased alfalfa dry weight through producing siderophores, phosphorus, and indole acetic acid (13.6 mg ml-1 and 19.9 mg ml-1 respectively). Hence, we identify two tolerant and efficient strains, Achromobacter spanium and Serratia plymuthica, isolated from Medicago sativa nodule with valuable potential able to phytostabilize pesticide-contaminated soils.
Subject(s)
Bacteria/classification , Bacteria/isolation & purification , Medicago sativa/microbiology , Root Nodules, Plant/microbiology , Anti-Bacterial Agents/metabolism , Bacteria/genetics , Bacteria/metabolism , Carbon/analysis , Cluster Analysis , DNA, Bacterial/chemistry , DNA, Bacterial/genetics , DNA, Ribosomal/chemistry , DNA, Ribosomal/genetics , Microbial Sensitivity Tests , Nitrogen/analysis , Pesticides/metabolism , Phosphorus/analysis , Phylogeny , Polymerase Chain Reaction , Polymorphism, Restriction Fragment Length , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA , Soil/chemistry , SymbiosisABSTRACT
Chicken feather waste is generally insufficiently utilized despite its high content of protein, constituting an environmental issue. Biodegradation of the waste with enabling microbes provides an advantageous option among the available solutions. In this study, an efficient whole feather-degrading strain was strategically isolated from a soil sample taken from a local tea plantation that has little or nothing to do with feathers. The strain was identified as Bacillus thuringiensis (designated as FDB-10) according to the cloned complete 16S rRNA sequence. The FDB-10 could efficiently degrade briefly heat-treated whole feather (102 °C, 5 min; up to 90% of a maximum concentration of 30 g/L) in a salt medium supplemented with 0.1 g/L yeast extract within 24 h (37 °C, 150 rpm). Addition of carbon sources (glycerol, glucose, starch, Tween 20, Tween 80, 1.25 g/L as glycerol) to the fermentation medium could improve the degradation. However, significant inhibition could be observed when the added carbon source reached the amount usually adopted in the investigation of carbon source preference (1%). Nitrogen source (NH4Cl, (NH4)2SO4, peptone) adversely influenced the performance of the strain. When the molar concentrations of NH4+ were equal for the two salt, the inhibitory effect on degradation of whole feathers was similar. Entirely different from other reported feather-degrading strains showing a preference to melanin-free feather substrates, the strain isolated in this study could degrade melanin-containing feather equally efficiently, and higher protease activity could be detected in the digest mix. As a plus, the strain could degrade feathers in rice wash produced in daily cooking, indicating its potential use in the simultaneous treatment of rice cooker wastewater produced by a rice processing plant. All these results imply that the FDB-10 is a strain with great potential in the biodegradation of feather waste.
Subject(s)
Bacillus thuringiensis/isolation & purification , Bacillus thuringiensis/metabolism , Feathers/metabolism , Soil Microbiology , Animals , Bacillus thuringiensis/classification , Bacillus thuringiensis/genetics , Biotransformation , Chickens , Cluster Analysis , Culture Media , DNA, Bacterial/chemistry , DNA, Bacterial/genetics , DNA, Ribosomal/chemistry , DNA, Ribosomal/genetics , Fermentation , Industrial Waste , Keratins/metabolism , Phylogeny , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA , Tea/growth & development , TemperatureABSTRACT
The objective of the present study was to isolate Actinobacteria, preferably Streptomyces spp. from the rhizosphere soils of three ethno-medicinal plants collected in Serbia (Papaver rhoeas, Matricaria chamomilla, and Urtica dioica) and to screen their antifungal activity against Candida spp. Overall, 103 sporulating isolates were collected from rhizosphere soil samples and determined as Streptomyces spp. Two different media and two extraction procedures were used to facilitate identification of antifungals. Overall, 412 crude cell extracts were tested against Candida albicans using disk diffusion assays, with 42% (43/103) of the strains showing the ability to produce antifungal agents. Also, extracts inhibited growth of important human pathogens: Candida krusei, Candida parapsilosis, and Candida glabrata. Based on the established degree and range of antifungal activity, nine isolates, confirmed as streptomycetes by 16S rRNA sequencing, were selected for further testing. Their ability to inhibit Candida growth in liquid culture, to inhibit biofilm formation, and to disperse pre-formed biofilms was assessed with active concentrations from 8 to 250 µg/mL. High-performance liquid chromatographic profiles of extracts derived from selected strains were recorded, revealing moderate metabolic diversity. Our results proved that rhizosphere soil of ethno-medicinal plants is a prolific source of streptomycetes, producers of potentially new antifungal compounds.
Subject(s)
Antifungal Agents/metabolism , Candida/drug effects , Plants, Medicinal/microbiology , Rhizosphere , Soil Microbiology , Streptomyces/isolation & purification , Streptomyces/metabolism , Candida/growth & development , Cluster Analysis , DNA, Bacterial/chemistry , DNA, Bacterial/genetics , DNA, Ribosomal/chemistry , DNA, Ribosomal/genetics , Phylogeny , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA , Serbia , Streptomyces/classification , Streptomyces/geneticsABSTRACT
A Gram-stain-negative, strictly aerobic, non-motile, ivory colored and rod-shaped bacterium (designated Gsoil 652T) isolated from ginseng cultivating soil, was characterized using a polyphasic approach to clarify its taxonomic position. Strain Gsoil 652T was observed to grow optimally at 30 °C and at pH 7.0 on R2A agar medium. Phylogenetic analysis, based on 16S rRNA gene sequences similarities, indicated that Gsoil 652T belongs to the genus Caballeronia of the family Burkholderiaceae and was most closely related to Caballeronia choica LMG 22940T (98.9%), Caballeronia udeis LMG 27134T (98.9%), Caballeronia sordidicola LMG 22029T (98.2%) and Caballeronia humi LMG 22934T (98.1%). The DNA G+C content was 62.1 mol% and Q-8 was the major isoprenoid quinone. The main polar lipids were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, unidentified aminophospholipid, and unidentified phospholipid. The predominant fatty acids were C16:0, C17:0 cyclo and C19:0 cyclo ω8c. The DNA-DNA relatedness value between strain Gsoil 652T and closely related type strains of Caballeronia species were less than 36.0%. Moreover, strain Gsoil 652T could be distinguished phenotypically from the recognized species of the genus Caballeronia. The novel isolate, therefore, represents a novel species, for which the name Caballeronia ginsengisoli sp. nov. is proposed, with the type strain Gsoil 652T (= KACC 19441T = LMG 30326T).
Subject(s)
Burkholderiaceae/classification , Soil Microbiology , Base Composition , Burkholderiaceae/chemistry , Burkholderiaceae/genetics , Burkholderiaceae/isolation & purification , DNA, Bacterial/chemistry , Fatty Acids/analysis , Panax , Phospholipids/analysis , Phylogeny , RNA, Ribosomal, 16S/genetics , SoilABSTRACT
Sediment microbial communities from plain river networks exert different effects on pollutant transformation and migration in lake basins. In this study, we examined millions of Illumina reads (16S rRNA gene amplicons) to compare lake, lake wetland, and estuary bacterial communities through a technically consistent approach. Results showed that bacterial communities in the sampled lake sediments had the highest alpha-diversity (Group B), than in sampled lake wetland sediments and estuary sediments. Proteobacteria was the most abundant (more than 30%) phyla in all the sediments. The lake sediments had more Nitrospirae (1.63%-11.75%) and Acidobacteria (3.46%-10.21%) than the lake wetland and estuary sediments, and estuary sediments had a greater abundance of the phylum Firmicutes (mean of 22.30%). Statistical analysis (LEfSe) revealed that lake wetland sediments contained greater abundances of the class Anaerolineaceae, orders Xanthomonadales, Pseudomonadales, and genera Flavobacterium, Acinetobacter. The lake sediments had a distinct community of diverse primary producers, such as phylum Acidobacteria, order Ignavibacteriales, and families Nitrospiraceae, Hydrogenophilaceae. Total phosphorus and organic matter were the main factors influencing the bacterial communities in sediments from several parts of the lake wetland and river estuary (p < .05). The novel insights into basin pollution control in plain river networks may be obtained from microbial distribution in sediments from different basin regions.
Subject(s)
Bacteria/classification , Bacteria/genetics , Geologic Sediments/microbiology , Lakes/microbiology , Microbiota , Rivers/microbiology , Cluster Analysis , DNA, Bacterial/chemistry , DNA, Bacterial/genetics , DNA, Ribosomal/chemistry , DNA, Ribosomal/genetics , Estuaries , High-Throughput Nucleotide Sequencing , Organic Chemicals/analysis , Phosphorus/analysis , Phylogeny , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA , Water/chemistry , WetlandsABSTRACT
Astragalus membranaceus is an herbaceous perennial plant, growing to about 2 feet tall, with sprawling stems and alternate leaves about 12-24 leaflets. In total, 24 cross bred (Duroc × Landrace × Yorkshire) piglets weaned at 4 weeks with an average body weight of 10.84 ± 1.86 kg, were divided into four groups and randomly assigned to dietary treatments containing different AMSLF levels (0.00%, 2.50%, 5.00%, and 7.50%). The piglets in the control group (0.00% AMSLF) were fed basal diet and other treatment groups were fed basal diet in addition to 2.50%, 5.00%, and 7.50% pulverized AMSLF. The results indicated that supplementation with AMSLF significantly (p < 0.05) decreased diarrheal incidence in piglets. There was significant difference between treatment in terms of ADFI, ADG and FCR. Both 5.00% and 7.50% treatments significantly increased growth performance. The digestibility of gross energy and dry matter increased (p > 0.05) with increasing AMSLF level. The level of blood IL-2 and TNF-α were significantly affected by AMSLF supplementation with 7.50% AMSLF group having higher (p < 0.05) IL-2 and TNF-α levels than the other treatment groups. The 16SrDNA sequencing results from the four treatments showed that the potentially active bacterial microbial population and diversity in pig cecum were dominated by the phyla Bacteriodetes and Firmicutes regardless of the AMSLF supplementation. The Shannon diversity, PD whole tree diversity indices and Chao analyses exhibited significant variability in species richness across the treatments. The principal coordinates analysis (PCoA) showed significant (p < 0.1) differences between bacterial communities in all treatment groups. Results from the current study suggested that AMSLF supplementation increased composition of bacterial microbiota in pig gut. In conclusion, dietary supplements with AMSLF could potentially be used to prevent diarrheal incidence and improved pig production.
Subject(s)
Animals, Newborn/growth & development , Animals, Newborn/immunology , Astragalus propinquus/chemistry , Bacteria/classification , Diarrhea/veterinary , Dietary Fiber/administration & dosage , Fatty Acids, Volatile/analysis , Animals , Bacteria/genetics , Biota , Cecum/microbiology , Cluster Analysis , Cytokines/blood , DNA, Bacterial/chemistry , DNA, Bacterial/genetics , DNA, Ribosomal/chemistry , DNA, Ribosomal/genetics , Diarrhea/prevention & control , Incidence , Phylogeny , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA , SwineABSTRACT
Nutritional iron deficiency (ID) causes not only anemia but also malfunction of the entire human organism. Recently, a role of the gut microbiota has been hypothesized, but limited data are available especially in infants. Here, we performed a pilot study to explore the gut microbiota in 10 patients with iron deficiency anemia (IDA) and 10 healthy controls aged 6-34 months. Fresh stool samples were collected from diapers, and the fecal microbiota was profiled by next-generation sequencing of the V3-V4 hypervariable region of the 16S rRNA gene. Except for diet diversity, the breastfeeding status at the enrollment, the exclusive breastfeeding duration, and the introduction of complementary foods did not differ between groups. Distinct microbial signatures were found in IDA patients, with increased relative abundance of Enterobacteriaceae (mean relative abundance, patients vs. controls, 4.4% vs. 3.0%) and Veillonellaceae (13.7% vs. 3.6%), and reduced abundance of Coriobacteriaceae (3.5% vs. 8.8%) compared to healthy controls. A decreased Bifidobacteriaceae/Enterobacteriaceae ratio was observed in IDA patients. Notwithstanding the low sample size, our data highlight microbiota dysbalance in IDA worth for further investigations, aimed at unraveling the ID impact on the microbiome trajectory in early life, and the possible long-term consequences.
Subject(s)
Anemia, Iron-Deficiency/complications , Dysbiosis , Gastrointestinal Microbiome , Microbiota , Bacteria/classification , Bacteria/genetics , Child , Child, Preschool , Cluster Analysis , DNA, Bacterial/chemistry , DNA, Bacterial/genetics , DNA, Ribosomal/chemistry , DNA, Ribosomal/genetics , Feces/microbiology , Female , Humans , Infant , Male , Metagenomics , Phylogeny , Pilot Projects , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNAABSTRACT
Barriers to microbial migrations can lead adaptive radiations and increased endemism. We propose that extreme unbalanced nutrient stoichiometry of essential nutrients can be a barrier to microbial immigration over geological timescales. At the oasis in the Cuatro Ciénegas Basin in Mexico, nutrient stoichiometric proportions are skewed given the low phosphorus availability in the ecosystem. We show that this endangered oasis can be a model for a lost world. The ancient niche of extreme unbalanced nutrient stoichiometry favoured survival of ancestral microorganisms. This extreme nutrient imbalance persisted due to environmental stability and low extinction rates, generating a diverse and unique bacterial community. Several endemic clades of Bacillus invaded the Cuatro Cienegas region in two geological times, the late Precambrian and the Jurassic. Other lineages of Bacillus, Clostridium and Bacteroidetes migrated into the basin in isolated events. Cuatro Ciénegas Basin conservation is vital to the understanding of early evolutionary and ecological processes.