RESUMEN
Bacillus paranthracis, a Gram-positive conditional pathogen of Bacillus cereus group species, is capable of causing foodborne and waterborne illnesses, leading to intestinal diseases in humans characterized by diarrhoea and vomiting. However, documented cases of B. paranthracis infection outbreaks are rare in the world, and the genomic background of outbreak strains is seldom characterized. This study retrospectively analyzed strains obtained from an outbreak in schools, as well as from water systems in peri-urban areas, China, in 2020. In total, 28 B. cereus group isolates were retrieved, comprising 6 from stool samples and 22 from water samples. Epidemiological and phylogenetic investigations indicated that the B. paranthracis isolate from drinking water as the causative agent of the outbreak. The genomic comparison revealed a high degree of consistency among 8 outbreak-related strains in terms of antimicrobial resistance gene profiles, virulence gene profiles, genomic content, and multilocus sequence typing (MLST). The strains related to the outbreak show highly similar genomic ring diagrams and close phylogenetic relationships. Additionally, this study shed light on the pathogenic potential and complexity of B. cereus group through its diversity in virulence genes and mice infection model. The findings highlight the usefulness of B. paranthracis genomes in understanding genetic diversity within specific environments and in tracing the source of pathogens during outbreak situations, thereby enabling targeted infection control interventions.
Asunto(s)
Brotes de Enfermedades , Genoma Bacteriano , Filogenia , China/epidemiología , Animales , Humanos , Ratones , Virulencia , Estudios Retrospectivos , Infecciones por Bacterias Grampositivas/epidemiología , Infecciones por Bacterias Grampositivas/microbiología , Bacillus/genética , Bacillus/aislamiento & purificación , Bacillus/clasificación , Bacillus/patogenicidad , Tipificación de Secuencias Multilocus , Enfermedades Transmitidas por el Agua/epidemiología , Enfermedades Transmitidas por el Agua/microbiología , Masculino , Factores de Virulencia/genética , Bacillus cereus/genética , Bacillus cereus/aislamiento & purificación , Bacillus cereus/patogenicidad , Bacillus cereus/clasificación , Femenino , Genómica , Microbiología del AguaRESUMEN
The objective of this study was to analyse microbiological organisms in different locations and regions for physical activity in the city of João Pessoa, Brazil. Samples were collected on various objects used, such as: mattresses, drinking fountains, gloves, cell phones and others. The samples were collected in João Pessoa-PB, following the Standard Operating Procedure-SOP/ Microbiology of a specialized laboratory. The collection took place in the five macro-regions: North, South, East, West and Center. Foreach region samples were collected in one public place (square), a private one (gym) and one school (public or private), totaling fifteen collected sites and 450 samples. The following microorganisms were studied in all analyzed surfaces: Bacillus sp, Escherichia Coli, Klebsiella sppor Enterobacter sppand Coag. Neg. Staphylococcus.All regions had a high contamination level by some microorganism. The highest rates were found in the western, central and northern regions -96, 94 and 93% respectively. The Coag. Neg.Staphylococcus presented the highest and lowest incidence rates in the South and East regions, with 43.33 and 6.67%, respectively, as well as Klebsiella sppor Enterobacter spp, which presented high levels. It is concluded that there is a microorganisms' contamination in the most varied places and regions where physical activity practices are developed, with a predominance of Coag. Neg.Staphylococcusand Klebsiella sppor Enterobacter spp. These results lead to a warning about the hygiene importance in places for physical activity practice, especially in pandemic times (COVID-19), since almost all the evaluated surfaces were contaminated.
Asunto(s)
Higiene , Centros de Acondicionamiento/provisión & distribución , COVID-19/patología , Instituciones Académicas/provisión & distribución , Bacillus/patogenicidad , Ejercicio Físico/fisiología , Contaminación Biológica , Enterobacter/patogenicidad , Microbiología Ambiental , Escherichia/patogenicidad , Pandemias/estadística & datos numéricos , Klebsiella/patogenicidad , NoxasRESUMEN
This study elaborates inter-kingdom signaling mechanisms, presenting a sustainable and eco-friendly approach to combat biotic as well as abiotic stress in wheat. Fusarium graminearum is a devastating pathogen causing head and seedling blight in wheat, leading to huge yield and economic losses. Psychrophilic Bacillus atrophaeus strain TS1 was found as a potential biocontrol agent for suppression of F. graminearum under low temperature by carrying out extensive biochemical and molecular studies in comparison with a temperate biocontrol model strain Bacillus amyloliquefaciens FZB42 at 15 and 25 °C. TS1 was able to produce hydrolytic extracellular enzymes as well as antimicrobial lipopeptides, i.e., surfactin, bacillomycin, and fengycin, efficiently at low temperatures. The Bacillus strain-induced oxidative cellular damage, ultrastructural deformities, and novel genetic dysregulations in the fungal pathogen as the bacterial treatment at low temperature were able to downregulate the expression of newly predicted novel fungal genes potentially belonging to necrosis inducing protein families (fgHCE and fgNPP1). The wheat pot experiments conducted at 15 and 25 °C revealed the potential of TS1 to elicit sudden induction of plant defense, namely, H2O2 and callose enhanced activity of plant defense-related enzymes and induced over-expression of defense-related genes which accumulatively lead to the suppression of F. graminearum and decreased diseased leaf area.
Asunto(s)
Bacillus/genética , Fusarium/genética , Control Biológico de Vectores , Triticum/microbiología , Bacillus/crecimiento & desarrollo , Bacillus/patogenicidad , Resistencia a la Enfermedad/genética , Fusarium/patogenicidad , Glucanos/genética , Estrés Oxidativo/genética , Enfermedades de las Plantas/genética , Enfermedades de las Plantas/microbiología , Enfermedades de las Plantas/parasitología , Triticum/genética , Triticum/crecimiento & desarrolloRESUMEN
Fungal infection and synthesis of mycotoxins in coffee leads to significant economic losses. This study aimed to investigate the prevalence of toxigenic fungi, their metabolites, and the effect of traditional roasting and brewing on ochratoxin A (OTA) and aflatoxins (AFs) contents of naturally contaminated coffee samples. In addition, in vivo biocontrol assays were performed to explore the antagonistic activities of Bacillus simplex 350-3 (BS350-3) on the growth and mycotoxins synthesis of Aspergillus ochraceus and A. flavus. The relative density of A. niger, A. flavus, Penicillium verrucosum and A. carbonarius on green coffee bean was 60.82%, 7.21%, 3.09% and 1.03%, respectively. OTA contents were lowest in green coffee beans (2.15 µg/kg), followed by roasted (2.76 µg/kg) and soluble coffee (8.95 µg/kg). Likewise, AFs levels were highest in soluble coffee (90.58 µg/kg) followed by roasted (33.61 µg/kg) and green coffee (9.07 µg/kg). Roasting naturally contaminated coffee beans at three traditional methods; low, medium and high, followed by brewing resulted in reduction of 58.74% (3.50 µg/kg), 60.88% (3.72 µg/kg) and 64.70% (4.11 µg/kg) in OTA and 40.18% (34.65 µg/kg), 47.86% (41.17 µg/kg) and 62.38% (53.73 µg/kg) AFs contents, respectively. Significant inhibitions of AFs and OTA synthesis by A. flavus and A. carbonarius, respectively, on infected coffee beans were observed in presence of Bacillus simplex BS350-3 volatiles. Gas chromatography mass spectrochemistry (GC-MS/MS) analysis of head-space BS350-3 volatiles showed quinoline, benzenemethanamine and 1-Octadecene as bioactive antifungal molecules. These findings suggest that marketed coffee samples are generally contaminated with OTA and AFs, with a significant level of roasted and soluble coffee contaminated above EU permissible limits for OTA. Further, along with coffee roasting and brewing; microbial volatiles can be optimized to minimize the dietary exposure to mycotoxins.
Asunto(s)
Coffea/microbiología , Microbiota , Alquenos/metabolismo , Aspergillus/metabolismo , Aspergillus/patogenicidad , Bacillus/metabolismo , Bacillus/patogenicidad , Bencilaminas/metabolismo , Coffea/metabolismo , Micotoxinas/metabolismo , Penicillium/metabolismo , Penicillium/patogenicidad , Quinolinas/metabolismo , Semillas/microbiologíaRESUMEN
I report here the genome sequences of a Bacillus paranthracis strain isolated from book page surface. The presented data show a new study field for this species, which is frequently encountered in several environment, including soil, rhizosphere and notably human samples. I provide some insights about genomic content of Bacillus paranthracis, for example the presence of multiple antibiotic resistance genes, genes for polyhydroxybutyrate metabolism, 120 genes related to stress resistance and pathogenicity-related genes such as phospholipase C, metalloprotease and a cluster for non-hemolytic enterotoxin. In vitro tests showed that this isolate has motility, ability to produce biofilm, cytotoxic and enterotoxic ability, which makes this isolate a potential pathogen.
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Bacillus/genética , Bacillus/patogenicidad , Libros , Farmacorresistencia Bacteriana/genética , Genoma Bacteriano , Antibacterianos/farmacología , Bacillus/aislamiento & purificación , Brasil , VirulenciaRESUMEN
Bacillus cytotoxicus belongs to the Bacillus cereus group that also comprises the foodborne pathogen Bacillus cereus sensu stricto, Bacillus anthracis causing anthrax, as well as the biopesticide Bacillus thuringiensis. The first B. cytotoxicus was isolated in the context of a severe food poisoning outbreak leading to fatal cases of diarrheal disease. Subsequent characterization of the outbreak strain led to the conclusion that this Bacillus strain was highly cytotoxic and eventually resulted in the description of a novel species, whose name reflects the observed toxicity: B. cytotoxicus. However, only a few isolates of this species have been characterized with regard to their cytotoxic potential and the role of B. cytotoxicus as a causative agent of food poisoning remains largely unclear. Hence, the aim of this study was to gain further insights into the toxicity of B. cytotoxicus. To this end, 19 isolates were obtained from mashed potato powders and characterized by toxin gene profiling and Vero cell cytotoxicity assays. All isolates harbored the cytK1 (cytotoxin K1) gene and species-specific variants of the nhe (non-hemolytic enterotoxin) gene. The isolates exhibited low or no toxicity towards Vero cells. Thus, this study indicates that the cytotoxic potential of B. cytotoxicus may be potentially lower than initially assumed.
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Bacillus/metabolismo , Enterotoxinas/metabolismo , Perfilación de la Expresión Génica , Riñón/microbiología , Solanum tuberosum/microbiología , Transcriptoma , Animales , Bacillus/genética , Bacillus/patogenicidad , Supervivencia Celular , Chlorocebus aethiops , Enterotoxinas/genética , Manipulación de Alimentos , Microbiología de Alimentos , Regulación Bacteriana de la Expresión Génica , Riñón/patología , Células VeroRESUMEN
An enzyme hyaluronidase (hyase) producing halotolerant bacterium was isolated from dental caries and identified as Brevibacterium halotolerans DC1. Higher growth and hyase production were observed in nutrient broth fortified with hyaluronic acid at pH 7.0, temperature 37 °C, 120 rpm upon 48 h of incubation. Hyase was purified using salt precipitation, DEAE cellulose ion exchange, and Sephadex G-100 gel filtration chromatography. The enzyme was purified to 13-fold with 67.19% recovery of activity and 26.37 U/mg of specific activity. SDS-PAGE and zymography revealed it to be near to homogeneity showing a relative molecular weight of about 43 kDa that was confirmed by MALDI-TOF MS. This hyase was very active and stable at pH 7.0 and temperature 40 °C. The presence of metal ions Ca2+ and Mg2+ increased its activity while Zn2+ and Cu2+ severely inhibited it. Being stable at 2 M NaCl, hyase exhibited its halotolerant nature. This enzyme showed wide substrate specificity where hyaluronic acid (HA) was the best substrate. The kinetic studies revealed that Km and Vmax were 91.3 µg/mL and 306.2 µg/mL/min respectively. This is the first report of hyaluronidase from a halotolerant Brevibacterium spp. which can find applications under high salinity.
Asunto(s)
Bacillus/enzimología , Proteínas Bacterianas/química , Hialuronoglucosaminidasa/química , Tolerancia a la Sal , Bacillus/aislamiento & purificación , Bacillus/patogenicidad , Proteínas Bacterianas/metabolismo , Cobre/química , Caries Dental/microbiología , Estabilidad de Enzimas , Humanos , Hialuronoglucosaminidasa/metabolismo , Cloruro de Sodio/química , Zinc/químicaRESUMEN
We aimed to profile the metabolism of soybean roots that were infected with soybean cyst nematodes and treated with Bacillus simplex to identify metabolic differences that may explain nematode resistance. Compared with control soybean roots, B. simplex-treated soybean roots contained lower levels of glucose, fructose, sucrose, and trehalose, which reduced the nematodes' food source. Furthermore, treatment with B. simplex led to higher levels of melibiose, gluconic acid, lactic acid, phytosphingosine, and noradrenaline in soybean roots, which promoted nematocidal activity. The levels of oxoproline, maltose, and galactose were lowered after B. simplex treatment, which improved disease resistance. Collectively, this study provides insight into the metabolic alterations induced by B. simplex treatment, which affects the interactions with soybean cyst nematodes.
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Bacillus/patogenicidad , Resistencia a la Enfermedad , Glycine max/parasitología , Metaboloma , Nematodos/patogenicidad , Animales , Metabolismo de los Hidratos de Carbono , Enfermedades de las Plantas/microbiología , Enfermedades de las Plantas/parasitología , Raíces de Plantas/metabolismo , Raíces de Plantas/microbiología , Raíces de Plantas/parasitología , Glycine max/metabolismo , Glycine max/microbiologíaRESUMEN
BACKGROUND: Nonspecific vaginitis, also known as Bacterial vaginosis, unlike genital candidiasis and trichomoniasis, is caused by microbiome breakdown. Döderlein's bacillus are gram-positive bacillus that form a microbiome, reproduce in the female vagina after gaining sexual maturity, secrete lactic acid, and prevent the growth of other vaginitis-causing bacteria. Clue cell are squamous epithelial cells with Gardnerella sp. attached to their cell surface. The presence of clue cell is one of the diagnostic criteria for nonspecific vaginitis. Additionally, although macrophages are reported to protect against candidal vaginitis, there are no reports of studies examining the association between macrophages and clue cell. MATERIALS AND METHODS: After re-staining 300 class I specimens by cervical cancer screening with Papanicolaou staining, the appearance of Döderlein's bacillus, macrophages, and clue cell was observed. RESULT: Age group and appearance of Döderlein's bacillus were negatively correlated. The rate of appearance of macrophages was positively correlated with the age group. In people aged 50 years or more, the appearance rate of clue cells was significantly lower in the macrophage appearance group than that in the non-appearance group. CONCLUSION: This study suggested that macrophages, and not Döderlein's bacillus, may play an important role in defense against nonspecific vaginitis.
Asunto(s)
Macrófagos/patología , Vagina/patología , Vaginitis/diagnóstico , Vaginitis/patología , Adulto , Anciano , Anciano de 80 o más Años , Bacillus/patogenicidad , Detección Precoz del Cáncer/métodos , Femenino , Humanos , Lactobacillus acidophilus/patogenicidad , Persona de Mediana Edad , Prueba de Papanicolaou/métodos , Neoplasias del Cuello Uterino/patología , Vagina/microbiología , Adulto JovenRESUMEN
A dramatic increase in global antimicrobial resistance (AMR) has been well documented. Of particular concern is the dearth of information regarding the spectrum and prevalence of AMR within Category A Select Agents. Here, we performed a survey of horizontally and vertically transferred AMR determinants among Category A agents and their near neighbors. Microarrays provided broad spectrum screening of 127 Francisella spp., Yersinia spp., and Bacillus spp. strains for the presence/absence of 500+ AMR genes (or families of genes). Detecting a broad variety of AMR genes in each genus, microarray analysis also picked up the presence of an engineered plasmid in a Y. pestis strain. High resolution melt analysis (HRMA) was also used to assess the presence of quinolone resistance-associated mutations in 100 of these strains. Though HRMA was able to detect resistance-causing point mutations in B. anthracis strains, it was not capable of discriminating these point mutations from other nucleotide substitutions (e.g., arising from sequence differences in near neighbors). Though these technologies are well-established, to our knowledge, this is the largest survey of Category A agents and their near-neighbor species for genes covering multiple mechanisms of AMR.
Asunto(s)
Infecciones Bacterianas/genética , Farmacorresistencia Bacteriana/genética , Quinolonas/uso terapéutico , Bacillus/efectos de los fármacos , Bacillus/genética , Bacillus/patogenicidad , Infecciones Bacterianas/tratamiento farmacológico , Infecciones Bacterianas/microbiología , Francisella/efectos de los fármacos , Francisella/genética , Francisella/patogenicidad , Regulación Bacteriana de la Expresión Génica/efectos de los fármacos , Humanos , Mutación/genética , Plásmidos/genética , Yersinia/efectos de los fármacos , Yersinia/genética , Yersinia/patogenicidadRESUMEN
Seed germination is a critical parameter for the successful development of sustainable agricultural practices. While seed germination is impaired by environmental constraints emerging from the climate change scenario, several types of simple procedures, known as priming, can be used to enhance it. Seed priming is defined as the process of regulating seed germination by managing a series of parameters during the initial stages of germination. Hydropriming is a highly accessible and economic technique that involves soaking of seeds in water followed by drying. Biopriming refers to the inoculation of seeds with beneficial microorganism. The present study aims to investigate whether hydropriming and biopriming could enhance seed germination. Thereby, the germination of Medicago truncatula seeds exposed to hydropriming and/or Bacillus spp. isolates was monitored for two-weeks. The seeds were sown in trays containing two types of in situ agricultural soils collected from Northern India (Karsara, Varanasi). This region is believed to be contaminated by solid waste from a nearby power plant. Phenotypic parameters had been monitored and compared to find the most appropriate combination of treatments. Additionally, qRT-PCR was used to evaluate the expression levels of specific genes used as molecular indicators of seed quality. The results show that, while hydropriming significantly enhanced seed germination percentage, biopriming resulted in improved seedling development, represented by increased biomass rather than seedling length. At a molecular level, this is reflected by the upregulation of genes involved in DNA damage repair and antioxidant defence. In conclusion, hydropriming and biopriming are efficient to improve seed germination and seedling establishment in soils collected from damaged sites of Northern India; this is reflected by morphological parameters and molecular hallmarks of seed quality.
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Antioxidantes/metabolismo , Germinación/genética , Medicago truncatula/genética , Semillas/genética , Bacillus/patogenicidad , Reparación del ADN/genética , Regulación de la Expresión Génica de las Plantas/genética , Medicago truncatula/crecimiento & desarrollo , Medicago truncatula/microbiología , Plantones/genética , Plantones/crecimiento & desarrollo , Semillas/crecimiento & desarrolloRESUMEN
Bacillus velezensis FZB42 is able to activate induced systemic resistance (ISR) to enhance plant defense response against pathogen infections. Though the roles of microRNAs (miRNAs) in Bacillus-triggered ISR have been reported in Arabidopsis, the maize miRNAs responsible for the Bacillus-activated ISR process have not been discovered. To explore the maize miRNAs involved in ISR, maize miRNAs in response to FZB42 (ISR activating), FZB42â³sfpâ³alss (deficient in triggering ISR), and a control for 12 h were sequenced. A total of 146 known miRNAs belonging to 30 miRNA families and 217 novel miRNAs were identified. Four miRNAs specifically repressed in FZB42-treatment were selected as candidate ISR-associated miRNAs. All of them contained at least one defense response-related cis-element, suggesting their potential roles in activating the ISR process. Interestingly, three of the four candidate ISR-associated miRNAs belong to the conserved miR169 family, which has previously been confirmed to play roles in abiotic stress response. Moreover, 52 mRNAs were predicted as potential targets of these candidate ISR-associated miRNAs through TargetFinder software and degradome sequencing. Gene Ontology (GO) and network analyses of target genes showed that these differentially expressed miRNA might participate in the ISR process by regulating nuclear factor Y transcription factor. This study is helpful in better understanding the regulatory roles of maize miRNAs in the Bacillus-activated ISR process.
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Resistencia a la Enfermedad , MicroARNs/genética , Zea mays/genética , Bacillus/patogenicidad , MicroARNs/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Factores de Transcripción/genética , Factores de Transcripción/metabolismo , Zea mays/inmunología , Zea mays/microbiologíaRESUMEN
Background and objectives: The objective of this study was to investigate the clinical significance of isolates from blood stream infection known to be blood culture contaminants in pediatric patients. Materials and Methods: Microbiological reports and medical records of all blood culture tests issued from 2002 to 2012 (n = 76,331) were retrospectively reviewed. Evaluation for potential contaminants were done by reviewing medical records of patients with the following isolates: coagulase-negative Staphylococcus, viridans group Streptococcus, Bacillus, Corynebacterium, Micrococcus, Aerococcus, and Proprionibacterium species. Repeated cultures with same isolates were considered as a single case. Cases were evaluated for their status as a pathogen. Results: Coagulase-negative Staphylococcus had clinical significance in 23.8% of all cases. Its rate of being a true pathogen was particularly high in patients with malignancy (43.7%). Viridans group Streptococcus showed clinical significance in 46.2% of all cases. Its rate of being a true pathogen was similar regardless of the underlying morbidity of the patient. The rate of being a true pathogens for remaining isolates was 27.7% for Bacillus and 19.0% for Corynebacterium species. Conclusions: Coagulase-negative Staphylococcus and viridans group Streptococcus isolates showed high probability of being true pathogens in the pediatric population, especially in patients with underlying malignancy.
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Bacteriemia/diagnóstico , Cultivo de Sangre/normas , Pediatría/normas , Aerococcus/aislamiento & purificación , Aerococcus/patogenicidad , Bacillus/aislamiento & purificación , Bacillus/patogenicidad , Bacteriemia/sangre , Cultivo de Sangre/estadística & datos numéricos , Preescolar , Corynebacterium/aislamiento & purificación , Corynebacterium/patogenicidad , Femenino , Humanos , Lactante , Recién Nacido , Masculino , Micrococcus/aislamiento & purificación , Micrococcus/patogenicidad , Pediatría/métodos , Estudios Retrospectivos , Staphylococcus/aislamiento & purificación , Staphylococcus/patogenicidad , Estreptococos Viridans/aislamiento & purificación , Estreptococos Viridans/patogenicidadRESUMEN
Bacillus Calmette-Guérin (BCG)-activated macrophages (BAMs) have anti-tumor effects, especially on fibrosarcoma cells. However, the mechanism governing this process has not been elucidated to date. TRIM59 is an up-regulated membrane protein expressed on the surface of BAMs. In this study, we found that up-regulated TRIM59 macrophages exhibited excellent growth inhibition on MCA207 fibrosarcoma and induced tumor apoptosis. Moreover, TRIM59 enhanced macrophage infiltration and increased the M1 phenotype macrophages inside the tumor. Furthermore, the cytotoxic T cells and B cells in the spleen and lymphnode have not been affected by TRIM59. These results showed that macrophages expressing TRIM59 exhibited the main cytotoxic effect on tumors. In vitro, we co-cultured TRIM59 up-regulated macrophages fixed with 1% paraformaldehyde or cell culture supernatant and tumor cells. We found that the killing activities of macrophages decreased after treatment with anti-TRIM59 antibody, and the supernatant of TRIM59 up-regulated macrophages had no tumoricidal effect on fibrosarcoma cells, which demonstrated that TRIM59 may be involved in tumoricidal effects via cell-cell contact. In addition, the PI3K-Akt pathway of MCA207 co-cultured with macrophages highly expressing TRIM59 was significantly inhibited, whereas the activation of the PI3K-Akt pathway in MCA207 was not affected after co-culture with TRIM59-CKO macrophages. These results define a vital role of TRIM59 as an anti-tumor effector molecule of BAMs and suggest a new therapeutic target for the treatment of fibrosarcoma.
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Apoptosis/fisiología , Bacillus/patogenicidad , Fibrosarcoma/metabolismo , Péptidos y Proteínas de Señalización Intracelular/metabolismo , Activación de Macrófagos/fisiología , Macrófagos/metabolismo , Proteínas de la Membrana/metabolismo , Proteínas de Motivos Tripartitos/metabolismo , Animales , Línea Celular Tumoral , Femenino , Melanoma Experimental/metabolismo , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Ratones Transgénicos , Mycobacterium bovis/patogenicidad , Fosfatidilinositol 3-Quinasas/metabolismo , Células RAW 264.7 , Transducción de Señal/fisiología , Regulación hacia Arriba/fisiologíaRESUMEN
BACKGROUND: The study of the mechanisms by which larvae of the Culex quinquefasciatus mosquito survive exposure to the entomopathogen Lysinibacillus sphaericus has benefited substantially from the generation of laboratory-selected colonies resistant to this bacterium. One such colony, RIAB59, was selected after regular long-term exposure of larvae to the L. sphaericus IAB59 strain. This strain is characterized by its ability to produce the well known Binary (Bin) toxin, and the recently characterized Cry48Aa/Cry49Aa toxin, able to kill Bin-resistant larvae. Resistance to Bin is associated with the depletion of its receptor, Cqm1 α-glucosidase, from the larvae midgut. This study aimed to identify novel molecules and pathways associated with survival of the RIAB59 larvae and the resistance phenotype. METHODS: A transcriptomic approach and bioinformatic tools were used to compare the profiles derived from the midguts of larvae resistant and susceptible to L. sphaericus IAB59. RESULTS: The RNA-seq profiles identified 1355 differentially expressed genes (DEGs), with 673 down- and 682 upregulated transcripts. One of the most downregulated DEGs was cqm1, which validates the approach. Other strongly downregulated mRNAs encode the enzyme pantetheinase, apolipoprotein D, lipases, heat-shock proteins and a number of lesser known and hypothetical polypeptides. Among the upregulated DEGs, the top most encodes a peroxisomal enzyme involved in lipid metabolism, while others encode enzymes associated with juvenile hormone synthesis, ion channels, DNA binding proteins and defense polypeptides. Further analyses confirmed a strong downregulation of several enzymes involved in lipid catabolism while the assignment of DEGs into metabolic pathways highlighted the upregulation of those related to DNA synthesis and maintenance, confirmed by their clustering into related protein networks. Several other pathways were also identified with mixed profiles of down- and upregulated transcripts. Quantitative RT-PCR confirmed the changes in levels seen for selected mRNAs. CONCLUSIONS: Our transcriptome-wide dataset revealed that the RIAB59 colony, found to be substantially more resistant to Bin than to the Cry48Aa/Cry49Aa toxin, developed a differential expression profile as well as metabolic features co-selected during the long-term adaptation to IAB59 and that are most likely linked to Bin resistance.
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Bacillus/patogenicidad , Culex/genética , Culex/microbiología , Resistencia a la Enfermedad/genética , Animales , Toxinas Bacterianas/metabolismo , Biología Computacional , Sistema Digestivo/enzimología , Femenino , Perfilación de la Expresión Génica , Genes de Insecto , Larva/genética , Larva/microbiología , Fenotipo , RNA-Seq , alfa-Glucosidasas/metabolismoRESUMEN
The Bacillus cereus group includes several Bacillus species with closely related phylogeny. The most well-studied members of the group, B. anthracis, B. cereus, and B. thuringiensis, are known for their pathogenic potential. Here, we present the historical rationale for speciation and discuss shared and unique features of these bacteria. Aspects of cell morphology and physiology, and genome sequence similarity and gene synteny support close evolutionary relationships for these three species. For many strains, distinct differences in virulence factor synthesis provide facile means for species assignment. B. anthracis is the causative agent of anthrax. Some B. cereus strains are commonly recognized as food poisoning agents, but strains can also cause localized wound and eye infections as well as systemic disease. Certain B. thuringiensis strains are entomopathogens and have been commercialized for use as biopesticides, while some strains have been reported to cause infection in immunocompromised individuals. In this article we compare and contrast B. anthracis, B. cereus, and B. thuringiensis, including ecology, cell structure and development, virulence attributes, gene regulation and genetic exchange systems, and experimental models of disease.
Asunto(s)
Bacillus cereus/clasificación , Bacillus cereus/patogenicidad , Bacillus/clasificación , Bacillus/patogenicidad , Filogenia , Animales , Carbunco/terapia , Vacunas contra el Carbunco , Bacillus/genética , Bacillus/fisiología , Bacillus anthracis/clasificación , Bacillus anthracis/patogenicidad , Bacillus cereus/genética , Bacillus cereus/fisiología , Bacillus thuringiensis/clasificación , Bacillus thuringiensis/patogenicidad , Toxinas Bacterianas/química , Toxinas Bacterianas/clasificación , Vacunas Bacterianas , Agentes de Control Biológico/metabolismo , ADN Bacteriano , Modelos Animales de Enfermedad , Ecología , Enfermedades Gastrointestinales/microbiología , Regulación Bacteriana de la Expresión Génica , Genes Bacterianos/genética , Genoma Bacteriano/genética , Humanos , Infecciones/microbiología , Invertebrados , Especificidad de la Especie , Esporas Bacterianas/citología , Virulencia/genéticaRESUMEN
Colonization resistance is an important attribute for bacterial interactions with hosts, but the mechanism is still not completely clear. In this study, we found that Phytobacter sp. SCO41T can effectively inhibit the in vivo colonization of Bacillus nematocida B16 in Caenorhabditis elegans, and we revealed the colonization resistance mechanism. Three strains of colonization-resistant bacteria, SCO41T, BX15, and BC7, were isolated from the intestines of the free-living nematode C. elegans derived from rotten fruit and soil. The primary characteristics and genome map of one of the three isolates was investigated to explore the underlying mechanism of colonization resistance in C. elegans. In addition, we performed exogenous iron supplementation and gene cluster knockout experiments to validate the sequencing results. The results showed that relationship was close among the three strains, which was identified as belonging to the genus Phytobacter. The type strain is SCO41T (= CICC 24103T = KCTC 52362T). Whole genome analysis showed that csgA, csgB, csgC, csgE, csgF, and csgG were involved in the curli adhesive process and that fepA, fepB, fepC, fepD, and fepG played important roles in SCO41T against the colonization of B. nematocida B16 in C. elegans by competing for iron. Exogenous iron supplementation showed that exogenous iron can increase the colonization of B. nematocida B16, which was additionally confirmed by a deletion mutant strain. The csg gene family contributes to the colonization of SCO41T in C. elegans. Curli potentially contribute to the colonization of SCO41T in C. elegans, and enterobactin has a key role in SCO41T to resist the colonization of B. nematocida B16 by competing for iron.
Asunto(s)
Caenorhabditis elegans/microbiología , Gammaproteobacteria/genética , Animales , Bacillus/patogenicidad , Caenorhabditis elegans/genética , Gammaproteobacteria/aislamiento & purificación , Gammaproteobacteria/patogenicidad , Microbioma Gastrointestinal/fisiología , Técnicas de Inactivación de Genes , Intestinos/microbiología , Virulencia , Secuenciación Completa del Genoma/métodosRESUMEN
Due to the critical role played by autophagy in pathogen clearance, pathogens have developed diverse strategies to subvert it. Despite previous key findings of bacteria-autophagy interplay, asystems-level insight into selective targeting by the host and autophagy modulation by the pathogens is lacking. We predicted potential interactions between human autophagy proteins and effector proteins from 56 pathogenic bacterial species by identifying bacterial proteins predicted to have recognition motifs for selective autophagy receptors SQSTM1/p62, CALCOCO2/NDP52 and MAP1LC3/LC3. Using structure-based interaction prediction, we identified bacterial proteins capable to modify core autophagy components. Our analysis revealed that autophagy receptors in general potentially target mostly genus-specific proteins, and not those present in multiple genera. The complementarity between the predicted SQSTM1/p62 and CALCOCO2/NDP52 targets, which has been shown for Salmonella, Listeria and Shigella, could be observed across other pathogens. This complementarity potentially leaves the host more susceptible to chronic infections upon the mutation of autophagy receptors. Proteins derived from enterotoxigenic and non-toxigenic Bacillus outer membrane vesicles indicated that autophagy targets pathogenic proteins rather than non-pathogenic ones. We also observed apathogen-specific pattern as to which autophagy phase could be modulated by specific genera. We found intriguing examples of bacterial proteins that could modulate autophagy, and in turn being targeted by autophagy as ahost defense mechanism. We confirmed experimentally an interplay between a Salmonella protease, YhjJ and autophagy. Our comparative meta-analysis points out key commonalities and differences in how pathogens could affect autophagy and how autophagy potentially recognizes these pathogenic effectors. Abbreviations: ATG5: autophagy related 5; CALCOCO2/NDP52: calcium binding and coiled-coil domain 2; GST: glutathione S-transferase; LIR: MAP1LC3/LC3-interacting region; MAP1LC3/LC3: microtubule associated protein 1 light chain 3 alpha; OMV: outer membrane vesicles; SQSTM1/p62: sequestosome 1; SCV: Salmonella containing vesicle; TECPR1: tectonin beta-propeller repeat containing 1; YhjJ: hypothetical zinc-protease.
Asunto(s)
Autofagia/genética , Proteínas Bacterianas/metabolismo , Proteínas Asociadas a Microtúbulos/metabolismo , Proteínas Nucleares/metabolismo , Proteína Sequestosoma-1/metabolismo , Secuencias de Aminoácidos , Bacillus/metabolismo , Bacillus/patogenicidad , Análisis por Conglomerados , Biología Computacional , Evolución Molecular , Interacciones Huésped-Patógeno/genética , Humanos , Listeria/metabolismo , Listeria/patogenicidad , Proteínas de la Membrana/metabolismo , Proteínas Asociadas a Microtúbulos/genética , Proteínas Nucleares/genética , Unión Proteica/genética , Mapas de Interacción de Proteínas , Salmonella/metabolismo , Salmonella/patogenicidad , Proteína Sequestosoma-1/genética , Shigella/metabolismo , Shigella/patogenicidad , Factores de Virulencia/metabolismoRESUMEN
BACKGROUND: Biotic and abiotic factors have been reported to affect the larvicidal efficacy of Bacillus thuringiensis israelensis (Bti) and Bacillus sphaericus (Bs), although the extent to which they are affected has been poorly documented. This paper studies the effect of sunlight exposure on the efficacy of a new larvicide formulation based on both Bti and Bs, herein after referred to as BTBSWAX, applied against two different larval stages. METHODS: The emergence of inhibition exhibited by BTBSWAX at three different dosages (1 g/m2, 1.5 g/m2, and 2 g/m2) was monitored under semi-field conditions using a total of 32 containers comprising 16 that were covered and 16 that were uncovered. Two experiments were conducted using first- and second-instar larvae of Anopheles gambiae, respectively. RESULTS: BTBSWAX at 2 g/m2 in covered containers exhibited high emergence inhibition (> 80%) when larvae were exposed from 1st instar on day-6 post-treatment, whereas the emergence inhibition was only 28% in uncovered containers. For larvae exposed from 1st instar on day-12 post-treatment, the emergence inhibition was moderate (70%) in covered containers but was low (< 20%) in uncovered containers. For larvae exposed from 2nd instar on day-10 post-treatment, the emergence inhibition was moderate (31%) in covered containers but was very low (< 10%) in uncovered containers. Moreover, the residual efficacy of BTBSWAX was markedly affected by environmental stresses, including sunlight exposure (Hazard ratio (HR) = 0.12, p < 0.001 and HR = 0.63, p = 0.033 for BTBSWAX at 2 g/m2 against 1st and 2nd instar larvae, respectively). CONCLUSION: These findings emphasize the impact of environmental variables (e.g., sunlight exposure) on the residual efficacy of Bti and Bs biolarvicides in the field. They hence highlight the need to take these factors into account for larvicide formulation development processes. Moreover, studies of the ecology of Anopheles larvae in targeted areas are also crucial for the integration of larval control strategies into malaria transmission plans devised by national malaria control programmes of endemic countries.
Asunto(s)
Anopheles/fisiología , Bacillus/patogenicidad , Bacillus/efectos de la radiación , Insecticidas/farmacología , Control de Mosquitos/métodos , Luz Solar , Animales , Anopheles/microbiología , Bioensayo , Femenino , Larva/microbiología , Larva/fisiologíaRESUMEN
Bacillus cereus sensu lato also known as B. cereus group is composed of an ecologically diverse bacterial group with an increasing number of related species, some of which are medically or agriculturally important. Numerous eï¬orts have been undertaken to allow presumptive diï¬erentiation of B. cereus group species from one another. FCC41 is a Bacillus sp. strain toxic against mosquito species like Aedes aegypti, Aedes (Ochlerotatus) albifasciatus, Culex pipiens, Culex quinquefasciatus, and Culex apicinus, some of them responsible for the transmission of vector-borne diseases. Here, we report the complete genome sequence of FCC41 strain, which consists of one circular chromosome and eight circular plasmids ranging in size from 8 to 490 kb. This strain harbors six crystal protein genes, including cry24Ca, two cry4-like and two cry52-like, a cry41-like parasporin gene and multiple virulence factors. The phylogenetic analysis of the whole-genome sequence of this strain with molecular approaches places this strain into the Bacillus wiedmannii cluster. However, according with phenotypical characteristics such as the mosquitocidal activity due to the presence of Cry proteins found in the parasporal body and cry genes encoded in plasmids of different sizes, indicate that this strain could be renamed as B. wiedmannii biovar thuringiensis strain FCC41.