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1.
Neuromolecular Med ; 26(1): 32, 2024 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-39090268

RESUMO

Parkinson's disease is a progressive neurodegenerative disorder marked by the death of dopaminergic neurons in the substantia nigra region of the brain. Aggregation of alpha-synuclein (α-synuclein) is a contributing factor to Parkinson's disease pathogenesis. The objective of this study is to investigate the neuroprotective effects of gut microbes on α-synuclein aggregation using both in silico and in vivo approaches. We focussed on the interaction between α-synuclein and metabolites released by gut bacteria that protect from PD. We employed three probiotic microbe strains against α-synuclein protein: Lactobacillus casei, Escherichia coli, and Bacillus subtilis, with their chosen PDB IDs being Dihydrofolate reductase (3DFR), methionine synthetase (6BM5), and tryptophanyl-tRNA synthetase (3PRH), respectively. Using HEX Dock 6.0 software, we examined the interactions between these proteins. Among the various metabolites, methionine synthetase produced by E. coli showed potential interactions with α-synuclein. To further evaluate the neuroprotective benefits of E. coli, an in vivo investigation was performed using a rotenone-induced Parkinsonian mouse model. The motor function of the animals was assessed through behavioural tests, and oxidative stress and neurotransmitter levels were also examined. The results demonstrated that, compared to the rotenone-induced PD mouse model, the rate of neurodegeneration was considerably reduced in mice treated with E. coli. Additionally, histopathological studies provided evidence of the neuroprotective effects of E. coli. In conclusion, this study lays the groundwork for future research, suggesting that gut bacteria may serve as potential therapeutic agents in the development of medications to treat Parkinson's disease. fig. 1.


Assuntos
Bacillus subtilis , Escherichia coli , Microbioma Gastrointestinal , Simulação de Acoplamento Molecular , Estresse Oxidativo , Probióticos , Rotenona , alfa-Sinucleína , Animais , Camundongos , Microbioma Gastrointestinal/fisiologia , Probióticos/uso terapêutico , Probióticos/farmacologia , alfa-Sinucleína/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Rotenona/toxicidade , Lacticaseibacillus casei/fisiologia , Metionina tRNA Ligase , Triptofano-tRNA Ligase/fisiologia , Masculino , Tetra-Hidrofolato Desidrogenase/metabolismo , Simulação por Computador , Transtornos Parkinsonianos/microbiologia , Humanos , Fármacos Neuroprotetores/uso terapêutico , Camundongos Endogâmicos C57BL , Modelos Animais de Doenças , Doença de Parkinson Secundária/induzido quimicamente , Neurônios Dopaminérgicos/efeitos dos fármacos , Doença de Parkinson/microbiologia
2.
Microbiology (Reading) ; 170(8)2024 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-39106481

RESUMO

The rhizosphere hosts complex and abundant microbiomes whose structure and composition are now well described by metagenomic studies. However, the dynamic mechanisms that enable micro-organisms to establish along a growing plant root are poorly characterized. Here, we studied how a motile bacterium utilizes the microhabitats created by soil pore space to establish in the proximity of plant roots. We have established a model system consisting of Bacillus subtilis and lettuce seedlings co-inoculated in transparent soil microcosms. We carried out live imaging experiments and developed image analysis pipelines to quantify the abundance of the bacterium as a function of time and position in the pore space. Results showed that the establishment of the bacterium in the rhizosphere follows a precise sequence of events where small islands of mobile bacteria were first seen forming near the root tip within the first 12-24 h of inoculation. Biofilm was then seen forming on the root epidermis at distances of about 700-1000 µm from the tip. Bacteria accumulated predominantly in confined pore spaces within 200 µm from the root or the surface of a particle. Using probabilistic models, we could map the complete sequence of events and propose a conceptual model of bacterial establishment in the pore space. This study therefore advances our understanding of the respective role of growth and mobility in the efficient colonization of bacteria in the rhizosphere.


Assuntos
Bacillus subtilis , Lactuca , Raízes de Plantas , Rizosfera , Microbiologia do Solo , Bacillus subtilis/crescimento & desenvolvimento , Bacillus subtilis/metabolismo , Bacillus subtilis/fisiologia , Raízes de Plantas/microbiologia , Lactuca/microbiologia , Biofilmes/crescimento & desenvolvimento , Plântula/microbiologia , Plântula/crescimento & desenvolvimento
3.
Biotechnol J ; 19(8): e2400280, 2024 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-39167550

RESUMO

BACKGROUND: D-Allulose is one of the most well-known rare sugars widely used in food, cosmetics, and pharmaceutical industries. The most popular method for D-allulose production is the conversion from D-fructose catalyzed by D-allulose 3-epimerase (DAEase). To address the general problem of low catalytic efficiency and poor thermostability of wild-type DAEase, D-allulose biosensor was adopted in this study to develop a convenient and efficient method for high-throughput screening of DAEase variants. RESULTS: The catalytic activity and thermostability of DAEase from Caballeronia insecticola were simultaneously improved by semi-rational molecular modification. Compared with the wild-type enzyme, DAEaseS37N/F157Y variant exhibited 14.7% improvement in the catalytic activity and the half-time value (t1/2) at 65°C increased from 1.60 to 27.56 h by 17.23-fold. To our delight, the conversion rate of D-allulose was 33.6% from 500-g L-1 D-fructose in 1 h by Bacillus subtilis WB800 whole cells expressing this DAEase variant. Furthermore, the practicability of cell immobilization was evaluated and more than 80% relative activity of the immobilized cells was maintained from the second to seventh cycle. CONCLUSION: All these results indicated that the DAEaseS37N/F157Y variant would be a potential candidate for the industrial production of D-allulose.


Assuntos
Bacillus subtilis , Técnicas Biossensoriais , Estabilidade Enzimática , Frutose , Técnicas Biossensoriais/métodos , Frutose/metabolismo , Bacillus subtilis/enzimologia , Bacillus subtilis/genética , Carboidratos Epimerases/genética , Carboidratos Epimerases/metabolismo , Carboidratos Epimerases/química , Engenharia de Proteínas/métodos , Racemases e Epimerases/genética , Racemases e Epimerases/metabolismo , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Proteínas de Bactérias/química , Temperatura
4.
Int J Mol Sci ; 25(15)2024 Aug 05.
Artigo em Inglês | MEDLINE | ID: mdl-39126106

RESUMO

RNase Y is a key endoribonuclease that regulates global mRNA turnover and processing in Bacillus subtilis and likely many other bacteria. This enzyme is anchored to the cell membrane, creating a pseudo-compartmentalization that aligns with its role in initiating the decay of mRNAs primarily translated at the cell periphery. However, the reasons behind and the consequences of RNase Y's membrane attachment remain largely unknown. In our study, we examined a strain expressing wild-type levels of a cytoplasmic form of RNase Y from its chromosomal locus. This strain exhibits a slow-growth phenotype, similar to that of an RNase Y null mutant. Genome-wide data reveal a significant impact on the expression of hundreds of genes. While certain RNA substrates clearly depend on RNase Y's membrane attachment, others do not. We observed no correlation between mRNA stabilization in the mutant strains and the cellular location or function of the encoded proteins. Interestingly, the Y-complex, a specificity factor for RNase Y, also appears also recognize the cytoplasmic form of the enzyme, restoring wild-type levels of the corresponding transcripts. We propose that membrane attachment of RNase Y is crucial for its functional interaction with many coding and non-coding RNAs, limiting the cleavage of specific substrates, and potentially avoiding unfavorable competition with other ribonucleases like RNase J, which shares a similar evolutionarily conserved cleavage specificity.


Assuntos
Bacillus subtilis , Proteínas de Bactérias , Membrana Celular , Regulação Bacteriana da Expressão Gênica , Bacillus subtilis/genética , Bacillus subtilis/enzimologia , Bacillus subtilis/metabolismo , Membrana Celular/metabolismo , Proteínas de Bactérias/metabolismo , Proteínas de Bactérias/genética , Estabilidade de RNA , Endorribonucleases/metabolismo , Endorribonucleases/genética , RNA Mensageiro/genética , RNA Mensageiro/metabolismo
5.
Curr Microbiol ; 81(10): 305, 2024 Aug 12.
Artigo em Inglês | MEDLINE | ID: mdl-39133322

RESUMO

The bacterium Bacillus subtilis is a widely used study model and industrial workhorse organism that belongs to the group of gram-positive bacteria. In this study, we report the analysis of a newly sequenced complete genome of B. subtilis strain SRCM117797 along with a comparative genomics of a large collection of B. subtilis strain genomes. B. subtilis strain SRCM117797 has 4,255,638 bp long chromosome with 43.4% GC content and high coding sequence association with macromolecules, metabolism, and phage genes. Genomic diversity analysis of 232 B. subtilis strains resulted in the identification of eight clusters and three singletons. Of 147 B. subtilis strains included, 89.12% had strain-specific genes, of which 6.75% encoded strain-specific insertion sequence family transposases. Our analysis showed a potential role of strain-specific insertion sequence family transposases in intra-cellular accumulation of strain-specific genes. Furthermore, the chromosomal layout of the core genes was biased: overrepresented on the upper half (closer to the origin of replication) of the chromosome, which may explain the fast-growing characteristics of B. subtilis. Overall, the study provides a complete genome sequence of B. subtilis strain SRCM117797, show an extensive genomic diversity of B. subtilis strains and insights into strain diversification mechanism and non-random chromosomal layout of core genes.


Assuntos
Bacillus subtilis , Genoma Bacteriano , Bacillus subtilis/genética , Filogenia , Variação Genética , Composição de Bases , Genômica , Cromossomos Bacterianos/genética , Análise de Sequência de DNA
6.
Nat Commun ; 15(1): 6877, 2024 Aug 11.
Artigo em Inglês | MEDLINE | ID: mdl-39128925

RESUMO

The bactericidal activity of several antibiotics partially relies on the production of reactive oxygen species (ROS), which is generally linked to enhanced respiration and requires the Fenton reaction. Bacterial persister cells, an important cause of recurring infections, are tolerant to these antibiotics because they are in a dormant state. Here, we use Bacillus subtilis cells in stationary phase, as a model system of dormant cells, to show that pharmacological induction of membrane depolarization enhances the antibiotics' bactericidal activity and also leads to ROS production. However, in contrast to previous studies, this results primarily in production of superoxide radicals and does not require the Fenton reaction. Genetic analyzes indicate that Rieske factor QcrA, the iron-sulfur subunit of respiratory complex III, seems to be a primary source of superoxide radicals. Interestingly, the membrane distribution of QcrA changes upon membrane depolarization, suggesting a dissociation of complex III. Thus, our data reveal an alternative mechanism by which antibiotics can cause lethal ROS levels, and may partially explain why membrane-targeting antibiotics are effective in eliminating persisters.


Assuntos
Antibacterianos , Bacillus subtilis , Membrana Celular , Espécies Reativas de Oxigênio , Bacillus subtilis/efeitos dos fármacos , Bacillus subtilis/metabolismo , Bacillus subtilis/fisiologia , Espécies Reativas de Oxigênio/metabolismo , Antibacterianos/farmacologia , Membrana Celular/metabolismo , Membrana Celular/efeitos dos fármacos , Superóxidos/metabolismo , Potenciais da Membrana/efeitos dos fármacos , Complexo III da Cadeia de Transporte de Elétrons/metabolismo , Complexo III da Cadeia de Transporte de Elétrons/genética , Proteínas de Bactérias/metabolismo , Proteínas de Bactérias/genética
7.
Front Immunol ; 15: 1415009, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-39139572

RESUMO

The beneficial effects of feeding probiotic Bacillus subtilis DSM 32315 (BS) and Bacillus velezensis CECT 5940 (BV) to chickens in vivo are well-documented, with potential immune modulation as a key mechanism. In this study, we investigated the direct interactions of chicken peripheral blood mononuclear cells (PBMCs) with BS or BV in vitro through whole transcriptome profiling and cytokine array analysis. Transcriptome profiling revealed 20 significantly differentially expressed genes (DEGs) in response to both Bacillus treatments, with twelve DEGs identified in BS-treated PBMCs and eight in BV-treated PBMCs. Pathway analysis using the Kyoto Encyclopedia of Genes and Genomes (KEGG) indicated significant regulation of immune-related pathways by both BS and BV. Notably, BS treatment upregulated genes associated with immune cell surface markers (CD4, CD25, CD28), anti-inflammatory cytokine interleukin-10 (IL-10), and C-C motif chemokine ligand 5 (CCL5), while downregulating the gene encoding pro-inflammatory IL-16. BV treatment similarly affected genes associated with immune cell surface markers, IL-16, and CCL5, with no impact on the gene encoding IL-10. Both treatments induced higher expression of the gene encoding the avian ß-defensin 1 (AvBD1). The results of this in vitro study indicate an immunomodulatory effect of BS and BV in chicken PBMCs by regulating genes involved in anti-inflammatory, bacteriostatic, protective, and pro-inflammatory responses. Consequently, BS and BV may serve to augment the immune system's capacity to defend against infection by modulating immune responses and cytokine expression. Thus, the administration of these probiotics holds promise for reducing reliance on antimicrobials in farming practices.


Assuntos
Bacillus , Galinhas , Citocinas , Leucócitos Mononucleares , Probióticos , Animais , Galinhas/imunologia , Leucócitos Mononucleares/imunologia , Leucócitos Mononucleares/metabolismo , Citocinas/metabolismo , Perfilação da Expressão Gênica , Imunomodulação , Bacillus subtilis/imunologia , Células Cultivadas , Transcriptoma
8.
Sci Rep ; 14(1): 18870, 2024 08 14.
Artigo em Inglês | MEDLINE | ID: mdl-39143137

RESUMO

The characteristics of dopamine self-polymerization were used to cover the nano-titanium dioxide (TiO2) surface and produce nano-titanium dioxide-polydopamine (TiO2-PDA). The reducing nature of dopamine was then used to reduce silver nitrate to silver elemental particles on the modified nano-titanium dioxide: The resulting TiO2-PDA-Ag nanoparticles were used as antimicrobial agents. Finally, the antibacterial agent was mixed with silicone to obtain an antibacterial silicone composite material. The composition and structure of antibacterial agents were analyzed by scanning electron microscopy, transmission electron microscopy, X-ray photoelectron energy spectroscopy, and X-ray diffraction. Microscopy and the antibacterial properties of the silicone antibacterial composites were studied as well. The TiO2-PDA-Ag antimicrobial agent had good dispersion versus nano-TiO2. The three were strongly combined with obvious characteristic peaks. The antibacterial agents were evenly dispersed in silicone, and the silicone composite has excellent antibacterial properties. Bacillus subtilis (B. subtilis) adhesion was reduced from 246 × 104 cfu/cm2 to 2 × 104 cfu/cm2, and colibacillus (E. coli) reduced from 228 × 104 cfu/cm2 leading to bacteria-free adhesion.


Assuntos
Bacillus subtilis , Escherichia coli , Silicones , Prata , Titânio , Titânio/química , Titânio/farmacologia , Silicones/química , Prata/química , Prata/farmacologia , Escherichia coli/efeitos dos fármacos , Bacillus subtilis/efeitos dos fármacos , Nanopartículas Metálicas/química , Antibacterianos/farmacologia , Antibacterianos/química , Anti-Infecciosos/farmacologia , Anti-Infecciosos/química , Polímeros/química , Polímeros/farmacologia , Difração de Raios X , Testes de Sensibilidade Microbiana , Aderência Bacteriana/efeitos dos fármacos , Indóis
9.
ACS Chem Biol ; 19(8): 1794-1802, 2024 Aug 16.
Artigo em Inglês | MEDLINE | ID: mdl-39096241

RESUMO

Protein degradation is a tightly regulated biological process that maintains bacterial proteostasis. ClpPs are a highly conserved family of serine proteases that associate with the AAA + ATPase (an ATPase associated with diverse cellular activities) to degrade protein substrates. Identification and biochemical characterization of protein substrates for the AAA + ATPase-dependent ClpP degradation systems are considered essential for gaining an understanding of the molecular operation of the complex ClpP degradation machinery. Consequently, expanding the repertoire of protein substrates that can be degraded in vitro and within bacterial cells is necessary. Here, we report that AAA + ATPase-ClpP proteolytic complexes promote degradation of the secondary metabolite surfactin synthetases SrfAA, SrfAB, and SrfAC in Bacillus subtilis. On the basis of in vitro and in-cell studies coupled with activity-based protein profiling of nonribosomal peptide synthetases, we showed that SrfAC is targeted to the ClpC-ClpP proteolytic complex, whereas SrfAA is hydrolyzed not only by the ClpC-ClpP proteolytic complex but also by different ClpP proteolytic complexes. Furthermore, SrfAB does not appear to be a substrate for the ClpC-ClpP proteolytic complex, thereby implying that other ClpP proteolytic complexes are involved in the degradation of this surfactin synthetase. Natural product biosynthesis is regulated by the AAA + ATPase-ClpP degradation system, indicating that protein degradation plays a role in the regulatory stages of biosynthesis. However, few studies have examined the regulation of protein degradation levels. Furthermore, SrfAA, SrfAB, and SrfAC were identified as protein substrates for AAA + ATPase-ClpP degradation systems, thereby contributing to a better understanding of the complex ClpP degradation machinery.


Assuntos
Bacillus subtilis , Proteínas de Bactérias , Produtos Biológicos , Endopeptidase Clp , Proteólise , Endopeptidase Clp/metabolismo , Produtos Biológicos/metabolismo , Produtos Biológicos/química , Bacillus subtilis/enzimologia , Bacillus subtilis/metabolismo , Proteínas de Bactérias/metabolismo , Peptídeo Sintases/metabolismo , Adenosina Trifosfatases/metabolismo
10.
ACS Appl Mater Interfaces ; 16(32): 41800-41809, 2024 Aug 14.
Artigo em Inglês | MEDLINE | ID: mdl-39088721

RESUMO

In this study, we propose a self-limiting growth model forBacillus subtilisspores confined within porous polyacrylamide (PA) hydrogels. We observed thatB. subtilisspores germinate into vegetative cells within the hydrogel matrix, forming spherical colonies. These colonies expand until the mechanical stress they exert on their environment surpasses the yield stress of the hydrogel, leading to formation of a nonpermeable layer that halts nutrient diffusion and forces the bacteria to resporulate. These novel observations suggest a model to explain why bacterial growth in confined environments and material interfaces may be limited, providing insight for natural phenomena and biotechnological applications involving bacterial encapsulation.


Assuntos
Resinas Acrílicas , Hidrogéis , Estresse Mecânico , Resinas Acrílicas/química , Hidrogéis/química , Bacillus subtilis/crescimento & desenvolvimento , Porosidade , Modelos Biológicos
11.
PLoS Genet ; 20(8): e1011349, 2024 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-39088561

RESUMO

Cellular processes require precise and specific gene regulation, in which continuous mRNA degradation is a major element. The mRNA degradation mechanisms should be able to degrade a wide range of different RNA substrates with high efficiency, but should at the same time be limited, to avoid killing the cell by elimination of all cellular RNA. RNase Y is a major endoribonuclease found in most Firmicutes, including Bacillus subtilis and Staphylococcus aureus. However, the molecular interactions that direct RNase Y to cleave the correct RNA molecules at the correct position remain unknown. In this work we have identified transcripts that are homologs in S. aureus and B. subtilis, and are RNase Y targets in both bacteria. Two such transcript pairs were used as models to show a functional overlap between the S. aureus and the B. subtilis RNase Y, which highlighted the importance of the nucleotide sequence of the RNA molecule itself in the RNase Y targeting process. Cleavage efficiency is driven by the primary nucleotide sequence immediately downstream of the cleavage site and base-pairing in a secondary structure a few nucleotides downstream. Cleavage positioning is roughly localised by the downstream secondary structure and fine-tuned by the nucleotide immediately upstream of the cleavage. The identified elements were sufficient for RNase Y-dependent cleavage, since the sequence elements from one of the model transcripts were able to convert an exogenous non-target transcript into a target for RNase Y.


Assuntos
Bacillus subtilis , Regulação Bacteriana da Expressão Gênica , Clivagem do RNA , Estabilidade de RNA , RNA Bacteriano , Staphylococcus aureus , Staphylococcus aureus/genética , Staphylococcus aureus/enzimologia , Bacillus subtilis/genética , Bacillus subtilis/enzimologia , Bacillus subtilis/metabolismo , RNA Bacteriano/metabolismo , RNA Bacteriano/genética , Estabilidade de RNA/genética , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Proteínas de Bactérias/metabolismo , Proteínas de Bactérias/genética , Endorribonucleases/metabolismo , Endorribonucleases/genética , Conformação de Ácido Nucleico , Sequência de Bases
12.
World J Microbiol Biotechnol ; 40(9): 287, 2024 Aug 02.
Artigo em Inglês | MEDLINE | ID: mdl-39090427

RESUMO

Bacteriocins are antimicrobial peptides produced by bacteria to prevent the growth of pathogens. Combining bacteriocins with metal nanoparticles, like silver nanoparticles (AgNPs), has developed into a viable strategy to get over bacteriocin limitations. In this study, bacteriocin BacZY05 was extracted from Bacillus subtilis ZY05 and purified using various techniques. The resulting purified bacteriocin was then combined with silver nanoparticles to form bacteriocin silver nanoconjugates (BacZY05-AgNPs). The physicochemical properties of the BacZY05-AgNPs were characterized using various analytical techniques. The mean diameter of the synthesized AgNPs was approximately 20-60 nm with an oval or spherical shape. The antimicrobial activity of the BacZY05-AgNPs was evaluated against several indicator strains by their zone of inhibition (ZOI), using the agar well diffusion method. Compared to bacteriocin (ZOI- 13 to 20 mm) and AgNPs (ZOI- 10-22 mm) alone, the antibacterial activity data demonstrated a 1.3-1.5-fold increase in the activity of bacteriocin-nanoconjugates (ZOI- 22 to 26 mm). For Staphylococcus aureus MTCC3103 and Klebsiella pneumoniae MTCC109, BacZY05-capped AgNPs exhibited the lowest minimum inhibitory concentration (MIC), measuring 10.93 µg/mL. For Salmonella typhi NCIM2501, the MIC was 28.75 µg/mL. The highest MIC value was 57.5 µg/mL for Escherichia coli DH5α and Vibrio cholerae MTCC3909. With BacZY05-capped AgNPs, the lowest minimum bactericidal concentration (MBC) of 28.75 µg/mL was observed for Staphylococcus aureus MTCC31003. In the cases of Salmonella typhi NCIM2501 and Klebsiella pneumoniae MTCC109 concentration was 57.5 µg/mL. Vibrio cholerae MTCC3909 and Escherichia coli DH5α had the highest MBC values at 115 µg/mL.


Assuntos
Antibacterianos , Bacillus subtilis , Bacteriocinas , Klebsiella pneumoniae , Nanopartículas Metálicas , Testes de Sensibilidade Microbiana , Nanoconjugados , Prata , Staphylococcus aureus , Antibacterianos/farmacologia , Antibacterianos/química , Prata/farmacologia , Prata/química , Bacteriocinas/farmacologia , Bacteriocinas/química , Bacteriocinas/biossíntese , Nanopartículas Metálicas/química , Staphylococcus aureus/efeitos dos fármacos , Nanoconjugados/química , Bacillus subtilis/efeitos dos fármacos , Klebsiella pneumoniae/efeitos dos fármacos , Escherichia coli/efeitos dos fármacos
13.
BMC Vet Res ; 20(1): 351, 2024 Aug 07.
Artigo em Inglês | MEDLINE | ID: mdl-39113050

RESUMO

Probiotics are becoming increasingly popular as eco-friendly alternatives in aquaculture. However, there is limited research on their impacts on the reproductive efficiency of Red Tilapia (Oreochromis niloticus x O. mossambicus) broodstock. Therefore, this experiment aimed to explore the combined effects of selective probiotics Bacillus subtilis and B. licheniformis (BSL; 1:1) added to water on blood hematology, serum metabolites, gonadal histology, reproductive performance, and reproductive associated genes in Red Tilapia broodstock. Tilapia broodfish weighing 140-160 g were stocked in four treatment groups: control (T0), and the other three groups were added different levels of BSL to the water as follows: T1 (0.01 g/m3), T2 (0.02 g/m3), and T3 (0.03 g/m3), respectively. Results indicate that BSL administration significantly improved RBCs, hemoglobin, hematocrit, MCH, and MCHC, with the highest improvement seen in the T3 group (P < 0.05). BSL added to the fish water significantly enhanced serum protein fractions (total protein, albumin, and globulins), while AST, ALT, ALP, creatinine, uric acid, and glucose were significantly diminished in a dose-dependent way (P < 0.05). Adding 0.02-0.03 g/ m3 of BSL resulted in higher antioxidant status (superoxide dismutase and catalase) compared to other groups (P < 0.05). Testosterone levels were higher in T3 than in other groups (P < 0.05). All female hormones (LH, FSH, estradiol, and progesterone) were substantially augmented by the addition of BSL. Additionally, the BSL groups exhibited higher GSI, HSI, VSI (male only), egg diameter (mm), mean number of fry/fish, and mean fry weight (g) compared to the control group (P < 0.05). Expression of reproductive-associated genes (vasa, nanos1a, nanos2, dnd1, pum1, AMH, and vtg) were significantly up-regulated in the gonads of fish in the 0.03 g/m3 treatment. The histological gonadal structure exhibited that BSL improved gonad maturation in both genders of Tilapia fish. Overall, adding a mixture of B. subtilis and B. licheniformis (0.03 g/m3 water) can accelerate reproductive performance in Red Tilapia through up-regulation of reproductive genes and enhance the health profile.


Assuntos
Probióticos , Reprodução , Animais , Probióticos/administração & dosagem , Probióticos/farmacologia , Feminino , Masculino , Ciclídeos/fisiologia , Bacillus subtilis , Suplementos Nutricionais , Aquicultura/métodos , Tilápia/fisiologia , Ração Animal/análise , Água/química
14.
Molecules ; 29(15)2024 Jul 25.
Artigo em Inglês | MEDLINE | ID: mdl-39124897

RESUMO

The goal of this research was to analyse the synergistic effect between selected plant extracts with zinc oxide particles, and zinc stearate. The influence of ZnO on the antimicrobial effectiveness of the selected extracts was confirmed in previous research carried out by the authors. However, the impact of zinc stearate on extract activity has yet to be analysed. The aim was to cover PLA films with active coatings based on hydroxy-propyl-methyl-cellulose (HPMC), or/and ethyl cellulose (EC) containing plant extracts and ZnO which has a synergistic effect. An additional aim was to use a CO2 extract of raspberry seed (RSE) with zinc stearate as active additives within the coatings. An examination of the antimicrobial properties (against Staphylococcus aureus, Escherichia coli, Bacillus subtilis, Pseudomonas syringae and Φ6 bacteriophage) of the covered films, as well as an investigation of layer presence with regards to PLA morphology (SEM, ATR-FTIR analysis) was carried out. The research work that was performed indicated that black chokeberry extract (ChE) and zinc oxide particles were effective against S. aureus, P. syringae and B. subtilis strains. In addition, the ChE with zinc stearate (ZnSt) was active against all analysed strains. The HPMC with ChE and ZnO as additives had antimicrobial properties against S. aureus, P. syringae and E. coli strains. The ChE was found to inhibit the growth of all of the analysed bacterial strains. When considering the coatings based on EC with the CO2 extract of raspberry seed (RSE) and ZnO, it was noted that they were only active against Gram-negative bacteria. The results of the experiments confirmed that AC1 (EC with RSE with ZnO) and AC2 (EC with RSE with ZnSt) coatings were not active against a phi6 bacteriophage. The HPMC coating containing the AC3 layer (ChE and ZnO) eliminated Φ6 particles, confirming its antiviral properties. In addition, the presence of the active (AC1, AC2 and AC3) coatings was confirmed by SEM and FTIR analysis.


Assuntos
Extratos Vegetais , Rubus , Óxido de Zinco , Óxido de Zinco/química , Óxido de Zinco/farmacologia , Extratos Vegetais/farmacologia , Extratos Vegetais/química , Rubus/química , Testes de Sensibilidade Microbiana , Ácidos Esteáricos/química , Ácidos Esteáricos/farmacologia , Anti-Infecciosos/farmacologia , Anti-Infecciosos/química , Antibacterianos/farmacologia , Antibacterianos/química , Escherichia coli/efeitos dos fármacos , Escherichia coli/crescimento & desenvolvimento , Bacillus subtilis/efeitos dos fármacos , Staphylococcus aureus/efeitos dos fármacos
15.
Molecules ; 29(15)2024 Jul 28.
Artigo em Inglês | MEDLINE | ID: mdl-39124959

RESUMO

The objective of this study was to analyze the chemical composition and evaluate the biological capabilities of the essential oils (EOs) extracted from leaves and stems of wild Aeschynomene indica L. plants by the hydrodistillation method. By using GC-FID/MS, fifty-six and fifty-five compounds, representing 95.1 and 97.6% of the essential oils in the leaves and stems, respectively, were characterized. The predominant constituents of A. indica EOs were (E)-caryophyllene, linalool, viridiflorol, phytol, hexadecanoic acid, trans-verbenol, and α-guaiene. The antibacterial and synergistic activities of the EOs were assessed by microdilution and checkerboard assays. The results revealed a potent inhibition and bactericidal activity against Staphylococcus aureus and Bacillus subtilis with MICs of 0.312-0.625 mg/mL. When combined with traditional antibiotics, the essential oils of A. indica possessed excellent synergistic effects against all tested bacteria. Additionally, the EOs of A. indica leaves showed higher antioxidant activity (IC50 = 0.11 ± 0.01 µg/mL) compared to the stem oil (IC50 = 0.19 ± 0.01 µg/mL) using the ABTS radical scavenging assay. The in vitro cytotoxicity of EOs against human cancer cell lines HepG2, MCF-7, A-549, and HCT-116 was examined, and MTT assays showed that the EOs possessed a significant cytotoxic potential against MCF-7 breast cancer cells, with IC50 values of 10.04 ± 1.82 and 15.89 ± 1.66 µg/mL, and a moderate cytotoxic activity against other tested cells. In conclusion, the A. indica EOs could be considered a potential source of pharmacologically active compounds.


Assuntos
Antibacterianos , Antioxidantes , Testes de Sensibilidade Microbiana , Óleos Voláteis , Folhas de Planta , Caules de Planta , Óleos Voláteis/farmacologia , Óleos Voláteis/química , Antioxidantes/farmacologia , Antioxidantes/química , Folhas de Planta/química , Antibacterianos/farmacologia , Antibacterianos/química , Humanos , Caules de Planta/química , Bacillus subtilis/efeitos dos fármacos , Linhagem Celular Tumoral , Staphylococcus aureus/efeitos dos fármacos , Extratos Vegetais/farmacologia , Extratos Vegetais/química
16.
Int J Mol Sci ; 25(15)2024 Jul 23.
Artigo em Inglês | MEDLINE | ID: mdl-39125604

RESUMO

The growing activity in the textile industry has been demanding the search for new and innovative technologies to meet consumers' needs regarding more sustainable and ecological processes, with functionality receiving more attention. Bee products are known for their wide spectra of properties, including antioxidant and antibacterial activities. Propolis and honey are the most popular and used since ancient times for the most diverse applications due to their health benefits. With the increasing need for safer and more sustainable practices, the use of natural products for the functional finishing process can be a suitable alternative due to their safety and eco-friendly nature. For that, a biosolution, composed of a mixture of propolis and honey in water, was used to perform the functional finishing of cotton knits, both in the presence and in the absence of potassium alum as a chemical mordant. The fastness strength was also evaluated after three washing cycles. The antioxidant potential of the biosolution, assessed with the in vitro ABTS scavenging assay, provided textiles with the capacity to reduce more than 90% of the ABTS radical, regardless of the mordant presence and even after three washing cycles. Furthermore, biofunctional textiles decreased the growth of Bacillus subtilis, Propionibacterium acnes, Escherichia coli, and, particularly, Staphylococcus aureus cultures after 24 h of incubation with an increase in antibacterial activity when potassium alum was used. These findings show that bee products are promising and effective alternatives to be used in the textile industry to confer antioxidant and antibacterial properties to cotton textiles, thereby enhancing human health.


Assuntos
Antibacterianos , Antioxidantes , Mel , Própole , Própole/química , Própole/farmacologia , Mel/análise , Antioxidantes/farmacologia , Antioxidantes/química , Antibacterianos/farmacologia , Antibacterianos/química , Têxteis , Fibra de Algodão/análise , Staphylococcus aureus/efeitos dos fármacos , Staphylococcus aureus/crescimento & desenvolvimento , Testes de Sensibilidade Microbiana , Escherichia coli/efeitos dos fármacos , Escherichia coli/crescimento & desenvolvimento , Compostos de Alúmen/química , Bacillus subtilis/efeitos dos fármacos , Bacillus subtilis/crescimento & desenvolvimento
17.
Nat Commun ; 15(1): 7188, 2024 Aug 21.
Artigo em Inglês | MEDLINE | ID: mdl-39169056

RESUMO

The transcriptional control of sporulation in Bacillus subtilis is reasonably well understood, but its translational control is underexplored. Here, we use RNA-seq, ribosome profiling and fluorescence microscopy to study the translational dynamics of B. subtilis sporulation. We identify two events of translation silencing and describe spatiotemporal changes in subcellular localization of ribosomes during sporulation. We investigate the potential regulatory role of ribosomes during sporulation using a strain lacking zinc-independent paralogs of three zinc-dependent ribosomal proteins (L31, L33 and S14). The mutant strain exhibits delayed sporulation, reduced germination efficiency, dysregulated translation of metabolic and sporulation-related genes, and disruptions in translation silencing, particularly in late sporulation.


Assuntos
Bacillus subtilis , Proteínas de Bactérias , Regulação Bacteriana da Expressão Gênica , Biossíntese de Proteínas , Proteínas Ribossômicas , Ribossomos , Esporos Bacterianos , Bacillus subtilis/genética , Bacillus subtilis/metabolismo , Bacillus subtilis/fisiologia , Esporos Bacterianos/metabolismo , Esporos Bacterianos/genética , Esporos Bacterianos/crescimento & desenvolvimento , Ribossomos/metabolismo , Proteínas de Bactérias/metabolismo , Proteínas de Bactérias/genética , Proteínas Ribossômicas/metabolismo , Proteínas Ribossômicas/genética , Mutação , Microscopia de Fluorescência
18.
PLoS Biol ; 22(8): e3002744, 2024 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-39137235

RESUMO

Bacterial interactions are vital for adapting to changing environments, with quorum sensing (QS) systems playing a central role in coordinating behaviors through small signaling molecules. The RRNPPA family is the prevalent QS systems in Bacillota and mediating communication through secreted oligopeptides, which are processed into active pheromones by extracellular proteases. Notably, in several cases the propeptides show the presence of multiple putative pheromones within their sequences, which has been proposed as a mechanism to diversify peptide-receptor specificity and potentially facilitate new functions. However, neither the processes governing the maturation of propeptides containing multiple pheromones, nor their functional significance has been evaluated. Here, using 2 Rap systems from bacteriophages infecting Bacillus subtilis that exhibit different types of pheromone duplication in their propeptides, we investigate the maturation process and the molecular and functional activities of the produced pheromones. Our results reveal that distinct maturation processes generate multiple mature pheromones, which bind to receptors with varying affinities but produce identical structural and biological responses. These findings add additional layers in the complexity of QS communication and regulation, opening new possibilities for microbial social behaviors, highlighting the intricate nature of bacterial interactions and adaptation.


Assuntos
Bacillus subtilis , Feromônios , Proteólise , Percepção de Quorum , Feromônios/metabolismo , Bacillus subtilis/metabolismo , Bacillus subtilis/genética , Proteínas de Bactérias/metabolismo , Proteínas de Bactérias/genética , Bacteriófagos/metabolismo , Bacteriófagos/genética , Sequência de Aminoácidos
19.
Int J Biol Macromol ; 277(Pt 2): 134306, 2024 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-39094860

RESUMO

Fungal infections pose severe and potentially lethal threats to plant, animal, and human health. Ergosterol has served as the primary target for developing antifungal medications. However, many antifungal drugs remain highly toxic to humans due to similarity in cell membrane composition between fungal and animal cells. Iturin A, lipopeptide produced by Bacillus subtilis, efficiently inhibit various fungi, but demonstrated safety in oral administration, indicating the existence of targets different from ergosterol. To pinpoint the exact antifungal target of iturin A, we used homologous recombination to knock out and overexpress erg3, a key gene in ergosterol synthesis. Saccharomyces cerevisiae and Aspergillus carbonarius were transformed using the LiAc/SS-DNNPEG and Agrobacterium-mediated transformation (AMT), respectively. Surprisingly, increasing ergosterol content did not augment antifungal activity. Furthermore, iturin A's antifungal activity against S. cerevisiae was reduced while it pre-incubation with voltage-gated potassium (Kv) channel inhibitor, indicating that Kv activation was responsible for cell death. Iturin A was found to activate the Kv protein, stimulating K+ efflux from cell. In vitro tests confirmed interaction between iturin A and Kv protein. This study highlights Kv as one of the precise targets of iturin A in its antifungal activity, offering a novel target for the development of antifungal medications.


Assuntos
Antifúngicos , Bacillus subtilis , Peptídeos Cíclicos , Saccharomyces cerevisiae , Antifúngicos/farmacologia , Antifúngicos/química , Peptídeos Cíclicos/farmacologia , Peptídeos Cíclicos/química , Bacillus subtilis/efeitos dos fármacos , Saccharomyces cerevisiae/metabolismo , Saccharomyces cerevisiae/efeitos dos fármacos , Saccharomyces cerevisiae/genética , Lipopeptídeos/farmacologia , Canais de Potássio/metabolismo , Canais de Potássio/genética , Ergosterol , Aspergillus/efeitos dos fármacos , Aspergillus/metabolismo , Potássio/metabolismo , Testes de Sensibilidade Microbiana
20.
Dev Comp Immunol ; 160: 105241, 2024 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-39121939

RESUMO

Probiotics play an essential role in the largemouth bass (Micropterus salmoides) aquaculture sector. They aid the fish in sickness prevention, intestinal structure improvement, food absorption, and immune system strengthening. In this experiment, Bacillus subtilis (BS, 107 CFU/g) and Lactobacillus reuteri (LR, 107 CFU/g) were added to the feed and then fed to M. salmoides for 35 days. The effects of two probiotics on the growth, immunity, and metabolism of M. salmoides organisms were studied. The results revealed that the BS group significantly increased the growth rate and specific growth rate of M. salmoides, while both the BS and LR groups significantly increase the length of villi M. salmoides intestines. The BS group significantly increased the levels of AKP, T-AOC, and CAT in the blood of M. salmoides, as well as AKP levels in the intestine. Furthermore, the BS group significantly increased the expression of intestinal genes Nrf2, SOD1, GPX, and CAT, while significantly decreasing the expression of the keap1 gene. M. salmoides gut microbial analysis showed that the abundance of Planctomycetota was significantly different in both control and experimental groups. Analyzed at the genus level, the abundance of Citrobacter, Paracoccus, Luedemannella, Sphingomonas, Streptomyces and Xanthomonas in the both control and experimental groups were significantly different. The BS group's differentially expressed genes were predominantly enriched in oxidative phosphorylation pathways in the intestine, indicating that they had a good influence on intestinal metabolism and inflammation suppression. In contrast, differentially expressed genes in the LR group were primarily enriched in the insulin signaling and linoleic acid metabolism pathways, indicating improved intestine metabolic performance. In conclusion, B. subtilis and L. reuteri improve the growth and health of M. salmoides, indicating tremendous potential for enhancing intestinal metabolism and providing significant application value.


Assuntos
Ração Animal , Bacillus subtilis , Bass , Suplementos Nutricionais , Microbioma Gastrointestinal , Limosilactobacillus reuteri , Probióticos , Animais , Probióticos/administração & dosagem , Bass/imunologia , Bass/crescimento & desenvolvimento , Bass/microbiologia , Limosilactobacillus reuteri/imunologia , Limosilactobacillus reuteri/fisiologia , Microbioma Gastrointestinal/imunologia , Intestinos/imunologia , Intestinos/microbiologia , Aquicultura , Proteínas de Peixes/metabolismo , Proteínas de Peixes/genética
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