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1.
Arch Microbiol ; 206(2): 63, 2024 Jan 13.
Artigo em Inglês | MEDLINE | ID: mdl-38217700

RESUMO

During the investigations of macrofungi resources in Zhejiang Province, China, an interesting wood rot fungus was collected. Based on morphological and molecular phylogenetic studies, it is described as a new species, Anthracophyllum sinense. A. sinense is characterized by its sessile, charcoal black and pleurotoid pileus, sparse lamellae occasionally branching, clavate basidia with long sterigmata [(3-)6-7(-8) µm], and non-heteromorphous cystidia. A. sinense establishes a separate lineage close to A. archeri and A. lateritium in the phylogenetic tree.


Assuntos
Agaricales , Basidiomycota , Filogenia , DNA Fúngico/genética , China
2.
Int J Mol Sci ; 25(11)2024 May 23.
Artigo em Inglês | MEDLINE | ID: mdl-38891868

RESUMO

Mycoviruses are usually transmitted horizontally via hyphal anastomosis and vertically through sporulation in natural settings. Oyster mushroom spherical virus (OMSV) is a mycovirus that infects Pleurotus ostreatus, with horizontal transmission via hyphal anastomosis. However, whether OMSV can be vertically transmitted is unclear. This study aimed to investigate the transmission characteristics of OMSV to progeny via basidiospores and horizontally to a new host. A total of 37 single-basidiospore offspring were obtained from OMSV-infected P. ostreatus and Pleurotus pulmonarius for Western blot detection of OMSV. The OMSV-carrying rate among monokaryotic isolates was 19% in P. ostreatus and 44% in P. pulmonarius. Then, OMSV-free and OMSV-infected monokaryotic isolates were selected for hybridization with harvested dikaryotic progeny strains. Western blot analyses of the offspring revealed that the OMSV transmission efficiency was 50% in P. ostreatus and 75% in P. pulmonarius, indicating vertical transmission via sexual basidiospores. Furthermore, we observed the horizontal transfer of OMSV from P. pulmonarius to Pleurotus floridanus. OMSV infection in P. floridanus resulted in significant inhibition of mycelial growth and yield loss. This study was novel in reporting the vertical transmission of OMSV through basidiospores, and its infection and pathogenicity in a new host P. floridanus.


Assuntos
Micovírus , Pleurotus , Esporos Fúngicos , Pleurotus/virologia , Esporos Fúngicos/crescimento & desenvolvimento , Micovírus/fisiologia
3.
Int J Mol Sci ; 24(13)2023 Jun 24.
Artigo em Inglês | MEDLINE | ID: mdl-37445762

RESUMO

Oyster mushroom spherical virus (OMSV) is a mycovirus with a positive-sense single-stranded RNA genome that infects the edible mushroom Pleurotus ostreatus. OMSV is horizontally transferred from an infected strain to a cured strain via mycelia. The infection results in significant inhibition of mycelial growth, malformation of fruiting bodies, and yield loss in oyster mushrooms. This study successfully transferred OMSV from P. ostreatus to Pleurotus pulmonarius. However, transmission was not successful in other Pleurotus species including P. citrinopileatus, P. eryngii, P. nebrodensis, and P. salmoneostramineus. The successful OMSV infection in P. pulmonarius was further verified with Western blot analysis using a newly prepared polyclonal antiserum against the OMSV coat protein. Furthermore, OMSV infection reduced the mycelial growth rate of P. pulmonarius. The OMSV-infected strain demonstrated abnormal performance including twisted mushrooms or irregular edge of the cap as well as reduced yield of fruiting bodies in P. pulmonarius, compared to the OMSV-free strain. This study is the first report on the infection and pathogenicity of OMSV to the new host P. pulmonarius. The data from this study therefore suggest that OMSV is a potential threat to P. pulmonarius.


Assuntos
Micovírus , Pleurotus , Vírus de RNA , Pleurotus/genética , Vírus de RNA/genética
4.
Curr Issues Mol Biol ; 44(11): 5778-5787, 2022 Nov 19.
Artigo em Inglês | MEDLINE | ID: mdl-36421676

RESUMO

Mycoviruses are viruses that specifically infect and replicate in fungi. Several mycoviruses have been previously reported in Pleurotus ostreatus, including the oyster mushroom spherical virus (OMSV), oyster mushroom isometric virus (OMIV), Pleurotus ostreatus spherical virus (POSV), and Pleurotus ostreatus virus 1 (PoV1). This study was designed to develop a multiplex RT-PCR for simultaneous detection and differentiation of the four P. ostreatus mycoviruses. Four pairs of primers were designed from conserved regions based on the reported sequences and the multiplex RT-PCR products were 672 bp for OMSV, 540 bp for OMIV, 310 bp for POSV, and 200 bp for PoV1. The optimal annealing temperature of the multiplex RT-PCR was 62 °C and the detection limits of the plasmids were 100 fg for OMSV and OMIV and 1 pg for POSV and PoV1. This technique was successfully applied for the detection of OMSV, OMIV, and POSV from different P. ostreatus strains and the plasmid containing the PoV1 sequence. This methodology can serve as a powerful diagnostic tool for the survey of the incidence and epidemiology of the four P. ostreatus mycoviruses, further contributing to the prevention and treatment of mycoviral diseases in P. ostreatus.

5.
Arch Microbiol ; 204(7): 381, 2022 Jun 10.
Artigo em Inglês | MEDLINE | ID: mdl-35687177

RESUMO

Based on morphological and phylogenetic analyses, an ectomycorrhizal fungus collected from Jiangxi, China, is described as a new species. R. atrofuscus is morphological characterized by a dark gray to black pileus, white to pale yellowish white hymenophore, a grayish brown to pale yellow context, which turning orange-yellow to rusty yellow when injured, a prominently and coarsely white to pale yellow to black reticulation on upper 3/4 or entire stipe, grayish yellow to pale yellow to yellowish brown basidiospores in KOH with measuring (7-)7.5-13(-13.5) × (3-)3.5-5(-5.5) µm. Phylogenetic analysis results showed that R. atrofuscus formed an independent lineage within Retiboletus. Descriptions and hand drawings of the new species and comparisons with similar species are presented.


Assuntos
Basidiomycota , Basidiomycota/genética , China , DNA Fúngico/genética , Filogenia , Análise de Sequência de DNA
6.
J Ind Microbiol Biotechnol ; 49(4)2022 Jul 30.
Artigo em Inglês | MEDLINE | ID: mdl-35595506

RESUMO

Validating the efficacy of sporicidal agents is a critical step in current good manufacturing practices for disinfection requirements. A limitation is that the poor quality of spores can lead to false positive sporicidal results. The aim of this study was to explore optimal sporulation and purification methods in Bacillus spores. Spores of 7 Bacillus strains were produced in 5 different sporulation media. After density centrifugation, spore yields were measured by phase-contrast microscopy and enumeration assays. Effects of purification methods including heat, sonication and lysozyme, and maturation on spore qualities were determined by sodium hypochlorite sporicidal assay. Difco sporulation media was identified as the preferred sporulation medium for 4 out of 7 tested Bacillus strains. Sporulation rates in B. cereus, B. sphaericus, and B. thuringiensis were higher at 30°C than the rates at 37°C at a difference of 5%, 65%, and 20%, respectively. Bacillus licheniformis favored Mn2+-amended 10% Columbia Broth at 37°C for sporulation with 40-72% higher sporulation rates than other media. The maximum sporulation rates of B. cereus and B. thuringiensis were observed on double-strength Schaeffer's-glucose broth. All studied purification methods improved the spore purity with strain variations. However, intense heat (80°C for 20 min) and lysozyme (100 µg/mL) treatment impaired the spore quality of specific Bacillus strains by sensitizing them against sodium hypochlorite. The length of the maturation period had an impact on the spore resistance, and the most optimal maturation periods ranged from 7 to 21 days in Bacillus strains. The results of this study will pave the way for further evaluation of the sporicidal activity of disinfectants.


Assuntos
Bacillus , Desinfetantes , Desinfetantes/farmacologia , Muramidase , Hipoclorito de Sódio/farmacologia , Esporos Bacterianos
7.
Can J Microbiol ; : 1-13, 2021 Sep 14.
Artigo em Inglês | MEDLINE | ID: mdl-34520677

RESUMO

Hyaluronic acid is a high-molecular-weight polysaccharide that is widely distributed in animal tissues. Bacterial hyaluronidases degrade hyaluronic acid as secreted enzymes and have been shown to contribute to infection. Staphylococcus aureus UAMS-1 is a clinical isolate that codes for two hyaluronidases (hysA1 and hysA2). Previous research has shown the presence of a full-length HysA1 protein from the S. aureus UAMS-1 strain with no evidence of enzymatic activity. In this study, the coding and upstream promoter regions of hysA1 from the S. aureus UAMS-1 strain were cloned, sequenced, and compared to the hysA1 gene from the S. aureus Sanger 252 strain. A single base change resulting in an E480G amino acid change was identified in the hysA1 gene from the S. aureus UAMS-1 strain when compared to the hysA1 gene from S. aureus Sanger 252. A plasmid copy of hysA1 from S. aureus Sanger 252 transduced into an S. aureus UAMS-1 hysA2 deletion mutant strain restored near wild-type levels of enzymatic activity. Homology modeling of the HysA1 hyaluronidase was performed with SWISS-MODEL using hyaluronidase from Streptococcus pneumoniae as the template, followed by a series of structural analyses using PyMOL, PLIP, PDBsum, and HOPE servers. This glutamic acid is highly conserved among hyaluronidases from Staphylococcus and other gram-positive bacteria. A series of structural analyses suggested that Glu-480 in HysA1 is critically responsible for maintaining the structural and functional ensemble of the catalytic and tunnel-forming residues, which are essential for enzyme activity. The missense mutation of Glu-480 to Gly introduces a loss of side chain hydrogen bond interactions with key residues Arg-360 and Arg-364, which are responsible for the tunnel topology, resulting in displacement of the substrate from an ideal position for catalysis through a localized conformational change of the active site. There is a high degree of relatedness among several gram-positive bacterial hyaluronidases; the loss of enzymatic activity of HysA1 in the S. aureus UAMS-1 strain is most likely caused by the mutation identified in our study. The role of hyaluronidase in staphylococcal infection and the redundancy of this gene are yet to be determined.

8.
Phytopathology ; 110(2): 336-344, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31524559

RESUMO

The endophytic bacterium Bacillus cereus BCM2 has shown great potential as a biocontrol organism against Meloidogyne incognita, which causes severe root-knot diseases in crops. In our previous study, the metabolite of BCM2 showed high nematicidal activity against the M. incognita second-stage juveniles. However, the mechanism employed by endophytic bacteria to infect and kill nematodes is still unclear. Here, we investigate both the endophytic bacterial extracellular proteins with nematicidal activity and their mechanism of killing nematodes. The first step was detecting the nematicidal activities of crude proteins. The results show that the nematode mortality rate reached 100% within 72 h, and the crude proteins damaged both the cuticle and eggshell, before finally destroying the targets. This suggests possible proteinaceous pathogeny in BCM2. Throughout the process, the fine-detail changes in the nematode cuticle and the intestinal structure were observed using scanning electron microscopy and transmission electron microscopy. These images show that BCM2 extracellular proteins did not damage the internal organization of the nematode but did severely damage its cuticle, which led to content leakage. From the crude proteins, chitosanase, alkaline serine protease, and neutral protease were purified and identified. The M. incognita-B. cereus BCM2 microenvironment simulation demonstrates that BCM2 adheres to the surface of nematodes and helps the metabolites that were produced by BCM2 to rapidly recognize and kill M. incognita. This relationship between plants, endophytic bacteria, and nematodes offers insight into the biological mechanisms that can be utilized for of nematode management.


Assuntos
Bacillus cereus , Peptídeo Hidrolases , Tylenchoidea , Animais , Antinematódeos/metabolismo , Antinematódeos/farmacologia , Bacillus cereus/enzimologia , Peptídeo Hidrolases/metabolismo , Peptídeo Hidrolases/farmacologia , Doenças das Plantas/parasitologia , Tylenchoidea/efeitos dos fármacos
9.
Food Microbiol ; 82: 89-98, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31027824

RESUMO

Pseudomonas spp. have emerged as the main spoilage bacteria, with many strains easily forming biofilms on food-contact surfaces and causing cross-contamination. The efficacy of disinfectants against bacteria is usually tested with planktonic cells; however, the disinfection tolerance of biofilms, especially detached biofilms, remains unknown. Here, we investigated the tolerance responses of detached and adhered biofilms of Pseudomonas fluorescens to acidic electrolyzed water (AEW) by determining tolerance responses by plate counting, comparing them using a Weibull model, and verifying changes in bacterial morphology by scanning electron microscopy. The experimental data and the responses calculated using Weibull a (scale) and b (shape) parameters agreed well (R2 values: 0.974-0.999), and we found that AEW exhibited effective antimicrobial activity against P. fluorescens, with adhered biofilms were more resistant than detached biofilms and planktonic cells. Additionally, AEW increased the bacterial membrane permeability and decreased the membrane potential, intracellular ATP concentrations, and intracellular pH while also triggering the disruption of extracellular polymeric substances. These results demonstrated that the morphophysiological responses of detached and adhered biofilms differed significantly and provided information on disinfectant-resistance strategies potentially beneficial to the development of novel disinfection approaches.


Assuntos
Biofilmes/efeitos dos fármacos , Desinfetantes/farmacologia , Eletrólise , Pseudomonas fluorescens/efeitos dos fármacos , Água/farmacologia , Ácidos/química , Permeabilidade da Membrana Celular , Contagem de Colônia Microbiana , Desinfecção/métodos , Potenciais da Membrana/efeitos dos fármacos , Microscopia Eletrônica de Varredura , Pseudomonas fluorescens/fisiologia , Água/química
10.
Plant Dis ; 103(7): 1551-1558, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31059388

RESUMO

Root-knot nematodes (Meloidogyne spp.) cause serious crop losses worldwide. The colonization of tomato roots by endophytic bacteria Bacillus cereus BCM2 can greatly reduce Meloidogyne incognita damage, and tomato roots carrying BCM2 were repellent to M. incognita second-stage juveniles (J2). Here, the effects of BCM2 colonization on the composition of tomato root exudates was evaluated and potential mechanisms for BCM2-mediated M. incognita control explored using a linked twin-pot assay and GC-MS. On water agar plates, J2 preferentially avoided filter paper treated with tomato root exudates (organic phase only) from plants inoculated with BCM2, visiting these 67.1% less than controls. In a linked twin-pot assay, BCM2 treatment resulted in a 42.0% reduction in the number of nematodes in the soil, a 43.3% reduction in the number of galls and a 47.7% decrease in the density of M. incognita in root tissues. Analysis of root exudate composition revealed that BCM2 inoculation increased the number of components in exudates. Among these, 2,4-di-tert-butylphenol, 3,3-dimethyloctane, and n-tridecane secretions markedly increased. In repellency trials on water agar plates, J2 avoided 2,4-di-tert-butylphenol, n-tridecane, and 3,3-dimethyloctane at concentrations of 4 mmol/liter. In a linked twin-pot assay, inoculation with 2,4-di-tert-butylphenol or 3,3-dimethyloctane reduced the number of nematodes in the soil (by 54.9 and 70.6%, respectively), the number of galls (by 53.7 and 52.4%), and the number of M. incognita in root tissues (by 67.5 and 36.3%). BCM2 colonization in tomato roots affected the composition of root exudates, increasing the secretion of substances that appear to be repellent, thus decreasing M. incognita J2 infection of roots.


Assuntos
Bacillus cereus , Solanum lycopersicum , Tylenchoidea , Animais , Bacillus cereus/fisiologia , Cromatografia Gasosa-Espectrometria de Massas , Solanum lycopersicum/microbiologia , Solanum lycopersicum/parasitologia , Raízes de Plantas/microbiologia , Raízes de Plantas/parasitologia , Tylenchoidea/microbiologia
12.
Plant Dis ; 101(3): 448-455, 2017 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30677349

RESUMO

Root-knot nematodes (Meloidogyne spp.), which cause severe global agricultural losses, can establish a special niche in the root vascular cylinder of crops, making them difficult to control. Endophytic bacteria have great potential as biocontrol organisms against Meloidogyne incognita. Three endophytic bacteria were isolated from plant tissues and showed high nematicidal activity against M. incognita second-stage juveniles (J2) in vitro. The gyrB gene sequence amplification results indicated that the three isolates were Bacillus cereus BCM2, B. cereus SZ5, and B. altitudinis CCM7. The isolates colonized tomato roots rapidly and stably during the colonization dynamic experiment. Three pot experiments were designed to determine the potential of three endophytic bacterial isolates on control of root-knot nematodes. The results showed that the preinoculated B. cereus BCM2 experiment significantly reduced gall and egg mass indexes. The inhibition ratio of gall and egg mass was up to 81.2 and 75.6% on tomato roots and significantly enhanced shoot length and fresh weight. The other two experiments with inoculated endophytic bacteria and M. incognita at the same time or after morbidity had lower inhibition ratios compared with the preinoculated endophytic bacteria experiment. The confocal laser-scanning microscopy method was used to further study the possible mechanism of endophytic bacteria in the biocontrol process. The results showed the localization pattern of the endophytic bacteria B. cereus BCM2-(str')-pBCgfp-1 in tomato root tissues. Root tissue colonized by endophytic bacteria repelled M. incognita J2 infection compared with the untreated control in a repellence experiment. We isolated an endophytic B. cereus strain that stably colonized tomato and controlled M. incognita effectively. This strain has potential for plant growth promotion, successful ecological niche occupation, and M. incognita J2 repellent action induction. It plays an important role in endophytic bacteria against root-knot nematodes.

13.
J Adv Res ; 2024 Jun 07.
Artigo em Inglês | MEDLINE | ID: mdl-38852803

RESUMO

INTRODUCTION: Salmonella Enteritidis has brought great harm to public health, animal production and food safety worldwide. The biofilm formed by Salmonella Enteritidis plays a critical role in microbial cross-contamination. Small non-coding RNAs (sRNAs) have been demonstrated to be responsible for regulating the formation of biofilm. The sRNA SaaS has been identified previously, that promotes pathogenicity by regulating invasion and virulence factors. However, whether the SaaS is implicated in regulating biofilm formation in abiotic surfaces remains unclear. OBJECTIVES: This study aimed to clarify the effect of SaaS in Salmonella Enteritidis and explore the modulatory mechanism on the biofilm formation. METHODS: Motility characteristics and total biomass of biofilm of test strains were investigated by the phenotypes in three soft agar plates and crystal violet staining in polystyrene microplates. Studies of microscopic structure and extracellular polymeric substances (EPS) of biofilm on solid surfaces were carried out using confocal laser scanning microscope (CLSM) and Raman spectra. Transcriptomics and proteomics were applied to analyze the changes of gene expression and EPS component. The RNA-protein pull-down and promoter-reporter ß-galactosidase activity assays were employed to analyze RNA binding proteins and identify target mRNAs, respectively. RESULTS: SaaS inhibits biofilm formation by repressing the adhesion potential and the secretion of EPS components. Integration of transcriptomics and proteomics analysis revealed that SaaS strengthened the expression of the flagellar synthesis system and downregulated the expression of curli amyloid fibers. Furthermore, RNA-protein pull-down interactome datasets indicated that SaaS binds to Hfq (an RNA molecular chaperone protein, known as a host factor for phage Qbeta RNA replication) uniquely among 193 candidate proteins, and promoter-reporter ß-galactosidase activity assay confirmed target mRNAs including hilD, cheA, and csgA. CONCLUSION: SaaS inhibits the properties of bacterial mobility, perturbs the secretion of EPS, and contributes to the inhibition of biofilm formation by interacting with target mRNA (hilD, cheA, and csgA) through the Hfq-mediated pathway.

14.
Adv Mater ; 36(13): e2311206, 2024 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-38104266

RESUMO

Metal halide scintillators serve as promising candidates for X-ray detection due to their high attenuation coefficients, high light yields, and low-cost solution-processable characteristics. However, the issues of humidity/thermal quenching and mechanical fragility, remain obstacles to the broad and diversified development of metal halide scintillators. Here, this work reports a lead-free, water-stable, stretchable, and self-healing (ethylenebis-triphenylphosphonium manganese (II) bromide (C38H34P2)MnBr4 organogel scintillator that meets X-ray imaging in complex scenarios. The robust organogel scintillator can be stretched with elongation up to 1300% while maintaining the scintillation properties. Activated by the dynamic hydrogen bonds and coordination bonds design, the organogel scintillator exhibits excellent self-healing properties at room temperature to alleviate the vignetting problem of the rigid scintillator films, the X-ray imaging resolution can reach 16.7 lp mm-1. The organogel scintillator can also realize flexible and self-healing X-ray imaging in water, providing a design path for portable devices in harsh conditions.

15.
MycoKeys ; 106: 117-132, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38948914

RESUMO

The rotting wood in freshwater is a unique eco-environment favoring various fungi. During our investigation of freshwater fungi on decaying wood, three hyphomycetes were collected from Jiangxi and Guangxi Provinces, China. Based on the morphological observations and phylogenetic analysis of a combined DNA data containing ITS, LSU, SSU and tef1-α sequences, two new Trichobotrys species, T.meilingensis and T.yunjushanensis, as well as a new record of T.effusa, were introduced. Additionally, a comprehensive description of the genus with both morphological and molecular data was first provided.

16.
Microbiology (Reading) ; 159(Pt 4): 782-791, 2013 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-23393148

RESUMO

In a previous study, two proteins identified as hyaluronidases were detected in spent media by MS and found to be in greater quantity in the sarA and sarA agr mutant strains when compared with the parent and agr mutant strains of Staphylococcus aureus UAMS-1. In the present study, spent media and total RNA were isolated from UAMS-1 and its regulatory mutants and analysed for hyaluronidase activity and steady-state hyaluronidase (hysA) RNA message levels. Hyaluronidase activity was observed throughout all time points examined regardless of the regulatory effects of sarA and agr but activity was always substantially higher in the sarA and sarA agr mutant strains than in the UAMS-1 parent and agr mutant strains. Northern analysis did not detect hysA message for either the UAMS-1 parent or the agr mutant strains at any time point examined, while steady-state hysA message levels were detected throughout growth for the sarA mutant strain, but only at exponential and early post-exponential growth for the sarA agr mutant strain. An in vitro biofilm plate assay, pre-coated with human plasma as a source of hyaluronic acid, demonstrated no significant increase in biofilm for a sarA mutant strain of S. aureus UAMS-1 defective in hyaluronidase activity when compared with the sarA mutant strain. These data indicate that, while hysA message levels and hyaluronidase activity are elevated in the sarA mutant strains of S. aureus UAMS-1, the increase in activity did not contribute to the biofilm-negative phenotype observed in the sarA mutant strain of S. aureus UAMS-1.


Assuntos
Proteínas de Bactérias/genética , Biofilmes/crescimento & desenvolvimento , Regulação Bacteriana da Expressão Gênica , Hialuronoglucosaminidase/metabolismo , Mutação , Staphylococcus aureus/crescimento & desenvolvimento , Transativadores/genética , Proteínas de Bactérias/metabolismo , Humanos , Hialuronoglucosaminidase/genética , Staphylococcus aureus/efeitos dos fármacos , Staphylococcus aureus/enzimologia , Transativadores/metabolismo
17.
Food Chem ; 424: 136365, 2023 Oct 30.
Artigo em Inglês | MEDLINE | ID: mdl-37207606

RESUMO

Microbial spoilage of meat products is a significant problem in the food industry. Aeromonas salmonicida is a significant microorganism responsible for spoilage in chilled meat. Its effector protein, hemagglutinin protease (Hap), has been identified as an effective substance for degrading meat proteins. The ability of Hap to hydrolyze myofibrillar proteins (MPs) in vitro demonstrated that Hap has obvious proteolytic activity, which could alter MPs' tertiary structure, secondary structure, and sulfhydryl groups. Moreover, Hap could significantly degrade MPs, focusing primarily on myosin heavy chain (MHC) and actin. Active site analysis and molecular docking revealed that the active center of Hap was bound to MPs via hydrophobic interaction and hydrogen bonding. It may preferentially cleave peptide bonds between Gly44-Val45 in actin, and Ala825-Phe826 in MHC. These findings suggest that Hap may be involved in the spoilage mechanism of microorganisms and provide crucial insights into the mechanisms of meat spoilage induced by bacteria.


Assuntos
Aeromonas salmonicida , Aeromonas salmonicida/genética , Aeromonas salmonicida/metabolismo , Simulação de Acoplamento Molecular , Actinas/metabolismo , Carne/análise , Proteólise , Cadeias Pesadas de Miosina/metabolismo
18.
Gut Microbes ; 15(1): 2211184, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-37158502

RESUMO

Salmonella Enteritidis is a foodborne enteric pathogen that infects humans and animals, utilizing complex survival strategies. Bacterial small RNA (sRNA) plays an important role in these strategies. However, the virulence regulatory network of S. Enteritidis remains largely incomplete and knowledge of gut virulence mechanisms of sRNAs is limited. Here, we characterized the function of a previously identified Salmonella adhesive-associated sRNA (SaaS) in the intestinal pathogenesis of S. Enteritidis. We found that SaaS promoted bacterial colonization in both cecum and colon of a BALB/c mouse model; it was preferentially expressed in colon. Moreover, our results showed that SaaS enhanced damage to mucosal barrier by affecting expressions of antimicrobial products, decreasing the number of goblet cells, suppressing mucin gene expression, and eventually reducing thickness of mucus layer; it further breached below physical barrier by strengthening invasion into epithelial cells in Caco-2 cell model as well as decreasing tight junction expressions. High throughput 16S rRNA gene sequencing revealed that SaaS also altered gut homeostasis by depleting beneficial gut microbiota while increasing harmful ones. Furthermore, by employing ELISA and western blot analysis, we demonstrated that SaaS regulated intestinal inflammation through sequential activation P38-JNK-ERK MAPK signaling pathway, which enabled immune escape at primary infection stage but strengthened pathogenesis at later stage, respectively. These findings suggest that SaaS plays an essential role in the virulence of S. Enteritidis and reveals its biological role in intestinal pathogenesis.


Assuntos
Microbioma Gastrointestinal , Humanos , Animais , Camundongos , Células CACO-2 , RNA Ribossômico 16S , Células Epiteliais/microbiologia , Salmonella enteritidis/genética
19.
Microbiol Spectr ; 11(1): e0293822, 2023 02 14.
Artigo em Inglês | MEDLINE | ID: mdl-36688642

RESUMO

Salmonella enterica serovar Enteritidis is a common foodborne pathogen that infects both humans and animals. The S. Enteritidis virulence regulation network remains largely incomplete, and knowledge regarding the specific virulence phenotype of small RNAs (sRNAs) is limited. Here, we investigated the role of a previously identified sRNA, Salmonella adhesive-associated sRNA (SaaS), in the virulence phenotype of S. Enteritidis by constructing mutant (ΔsaaS) and complemented (ΔsaaS/psaaS) strains. SaaS did not affect S. Enteritidis; it was activated in the simulated intestinal environment (SIE), regulating the expression of virulence target genes. We discovered that it directly binds ssaV mRNA. Caco-2 and RAW 264.7 cell assays revealed that SaaS promoted S. Enteritidis invasion and damage to epithelial cells while suppressing macrophage overgrowth and destruction. Furthermore, a BALB/c mouse model demonstrated that the deletion of SaaS significantly reduced mortality and attenuated the deterioration of pathophysiology, bacterial dissemination into systemic circulation, and systemic inflammation. Our findings indicate that SaaS is required for S. Enteritidis virulence and further highlight its biological role in bacterial pathogenesis. IMPORTANCE Salmonella is a zoonotic pathogen with high virulence worldwide, and sRNAs have recently been discovered to play important roles. We explored the biological characteristics of the sRNA SaaS and developed two cell infection models and a mouse infection model. SaaS is an SIE-responsive sRNA that regulates the expression of virulence-targeted genes. Additionally, it differentially mediates invasion and intracellular growth for survival and infection of the epithelium and macrophages. We further found that SaaS enhanced bacterial virulence by promoting lethality, colonization, and inflammatory response. These findings provide a better understanding of the critical role of sRNA in bacterial virulence.


Assuntos
Pequeno RNA não Traduzido , Salmonelose Animal , Humanos , Animais , Camundongos , Virulência/genética , Fatores de Virulência/genética , Células CACO-2 , Salmonelose Animal/microbiologia , Salmonella enteritidis/genética , Pequeno RNA não Traduzido/genética , Pequeno RNA não Traduzido/metabolismo , Proteínas de Bactérias/genética
20.
Ecol Evol ; 13(5): e10047, 2023 May.
Artigo em Inglês | MEDLINE | ID: mdl-37139404

RESUMO

Earthworms modulate carbon and nitrogen cycling in terrestrial ecosystems, but their effect may be compromised by the deposition of pollutants from industrial emissions. However, studies investigating how deposited compounds affect the role of earthworms in carbon cycling such as litter decomposition are lacking, although the interactions of earthworms and deposited compounds are important for understanding the impact of pollutants on ecosystems and the potential of earthworms in bioremediation. We performed a 365-day in situ litterbag decomposition experiment in a deciduous (Quercus variabilis) and coniferous (Pinus massoniana) forest in southeast China. We manipulated nitrogen (N), sodium (Na), and polycyclic aromatic hydrocarbons (PAHs) as model compounds during litter decomposition with and without earthworms (Eisenia fetida). After one year, N, Na, and PAH all slowed down litter mass loss, with the effects of Na being the strongest. By contrast, E. fetida generally increased litter mass loss, and the positive effects were uniformly maintained irrespective of the type of compounds added. However, the pathways to how earthworms increased litter mass loss varied among the compounds added and the two forests studied. As indicated by structural equation modeling, earthworms mitigated the negative effects of deposited compounds by directly increasing litter mass loss and indirectly increasing soil pH and microbial biomass. Overall, the results indicate that the acceleration of litter mass loss by earthworms is little affected by deposited compounds, and that earthworms have the potential to mitigate negative impacts of pollutants on litter decomposition and ecosystem processes.

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