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1.
Arch Microbiol ; 203(6): 3209-3217, 2021 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-33830284

RESUMEN

The diversity of 61 Bacillus cereus strains isolated from different clinical specimens, food including raw milk and milk products, and water was evaluated. PFGE analysis could discriminate 61 distinct pulsotypes with similarity levels from 25 to 82%, which were divided into 13 clonal complexes. The similarity between clonal complexes was at least 40%. Clinical strains were divided into 10 clonal complexes, while the strains, isolated from milk, food and water were included in 9, 6 and 6 clonal complexes, respectively. Three clonal complexes were dominated by clinical isolates, while they were absent in two complexes. Bacterial isolates from food, being a probable source of alimentary toxoinfection, showed low similarity to isolates from stool specimens. The isolates from both sources were classified together in only 4 out of 13 clonal complexes. The large circular and linear plasmids with the sizes between 50 and 200 kb were detected in 24 (39.3%) and 14 (23%) B. cereus strains, respectively. Thirteen (21.3%) strains contained only one plasmid, two plasmids were found in 6 (9.8%) of strains, and three or more plasmids were obtained in 5 (8.2%) of tested strains. The plasmids were confirmed in 30.8% and 40% of isolates from clinical specimens and food and milk samples, respectively. No clear correlation between the PFGE profiles, the source as well as plasmid content among all tested strains was observed.


Asunto(s)
Bacillus cereus , Variación Genética , Plásmidos , Animales , Bacillus cereus/genética , Bacillus cereus/aislamiento & purificación , Ambiente , Heces/microbiología , Microbiología de Alimentos , Tracto Gastrointestinal/microbiología , Leche/microbiología , Plásmidos/genética , Microbiología del Agua
2.
Artículo en Inglés | MEDLINE | ID: mdl-34494947

RESUMEN

Three Gram-positive bacterial strains, BML-BC004, BML-BC017 and BML-BC059, isolated from blood samples from three inpatients in Japan, were identified as members of Bacillus cereus using matrix-assisted laser desorption ionization time-of-flight MS. The 16S rRNA gene sequences of these three strains were more than 97.1 % similar to 18 type strains belonging to the B. cereus group. Whole-genome comparisons, using average nucleotide identity (ANI) and digital DNA-DNA hybridization (dDDH), confirmed that the three strains represented three individual distinct species belonging to the B. cereus group. A phylogenetic tree showed that BML-BC004, BML-BC017 and BML-BC059 were located close to B. luti, B. mobilis and B. paramycoides, respectively. Based on these phylogenetic and phenotypic data, including values below the threshold for ANI and dDDH, the three strains should be classified as representing three different novel species of the B. cereus group: Bacillus sanguinis sp. nov., with type strain BML-BC004T (=DSM 111102T=JCM 34122T), Bacillus paramobilis sp. nov., with type strain BML-BC017T (=DSM 111100T=JCM 34124T) and Bacillus hominis sp. nov., with type strain BML-BC059T (=DSM 111101T=JCM 34125T).


Asunto(s)
Bacillus cereus/clasificación , Sangre/microbiología , Filogenia , Bacillus cereus/aislamiento & purificación , Técnicas de Tipificación Bacteriana , Composición de Base , ADN Bacteriano/genética , Ácidos Grasos/química , Humanos , Japón , Hibridación de Ácido Nucleico , ARN Ribosómico 16S/genética , Análisis de Secuencia de ADN
3.
Arch Virol ; 166(11): 3183-3188, 2021 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-34505918

RESUMEN

A new Bacillus cereus phage, SWEP1, was isolated from black soil. The host lysis activity of phage SWEP1 has a relatively short latent time (20 min) and a small burst size of 83 PFU. The genome of SWEP1 consists of 162,461 bp with 37.77% G+C content. The phage encodes 278 predicted proteins, 103 of which were assigned functionally. No tRNA genes were found. Comparative genomics analysis indicated that SWEP1 is related to Bacillus phage B4 (86.91% identity, 90% query coverage). Phenotypic and genotypic characterization suggested that SWEP1 is a new member of a new species in the genus Bequatrovirus, family Herelleviridae.


Asunto(s)
Fagos de Bacillus/fisiología , Bacillus cereus/virología , Genoma Viral , Fagos de Bacillus/genética , Fagos de Bacillus/aislamiento & purificación , Bacillus cereus/aislamiento & purificación , Composición de Base , Replicación del ADN/genética , Genómica , Genotipo , Microscopía Electrónica de Transmisión , Filogenia , ARN de Transferencia/genética , Microbiología del Suelo , Temperatura
4.
Anal Bioanal Chem ; 413(26): 6661-6669, 2021 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-34476520

RESUMEN

Bacillus cereus is one of the most common foodborne pathogens found in various kinds of staple foods such as rice and wheat. A rapid and accurate detection method for this pathogen is highly desirable for the sustainable production of relevant food products. While several classical and molecular-based detection methods are available for the identification of B. cereus, they suffered one or more limitations such as the requirement for a tedious and time-consuming process, less than ideal specificity, and the lack of portability. Herein, we developed the first paper-based sensing device that exhibits high species specificity with sufficiently low limit of detection for the visual detection of specific DNA sequences of B. cereus. The success is attributed to the strategic planning of fabrication in various dimensions including thorough bioinformatics search for highly specific genes, the use of the pyrrolidinyl peptide nucleic acid (PNA) probe whose selectivity advantage is well documented, and an effective PNA immobilization and DNA-binding visualization method with an internal cross-checking system for validating the results. Testing in rice matrices indicates that the sensor is capable of detecting and distinguishing B. cereus from other bacterial species. Hence, this paper-based sensor has potential to be adopted as a practical means to detect B. cereus in food industries.


Asunto(s)
Bacillus cereus/aislamiento & purificación , Técnicas Biosensibles/métodos , Microbiología de Alimentos , Ácidos Nucleicos de Péptidos/química , Pirrolidinas/química , Oryza/microbiología , Papel
5.
J Appl Microbiol ; 130(4): 1130-1141, 2021 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-32780461

RESUMEN

AIMS: To determine if antibiotics associated with conventional pig farming have a direct role in altering the populations of key soil micro-organisms isolated from piggery environments with and without exposure to antibiotics. METHODS AND RESULTS: Fluorescent Pseudomonas sp. and the Bacillus cereus group from soils adjacent to four conventional piggeries (use of antibiotics) exposed to effluent (via irrigation) and two organic piggeries (non-use of antibiotics) were assessed against nine relevant antibiotics using disc diffusion. The focus of the study was not to determine antibiotic resistance (or sensitivity) of isolates based on the manufacturer-defined sensitive break point, instead this point was used as the interpretation point to compare the populations (i.e. farm/organism combination) for the antibiotics tested. Each population was statistically analysed to determine whether the mean diameters were significantly above this selected interpretation point. Bacterial species from both environments did not show a distinct population pattern linked to the antibiotics. CONCLUSIONS: Antibiotics associated with conventional pig farming do not have a direct role in altering the environmental populations of Pseudomonas and Bacillus sp. when assessed by population shifts. SIGNIFICANCE AND IMPACT OF THE STUDY: This study confirms that an understanding of the resident soil microbiota, as compared to the transient bacteria of pig origin, is important in addressing the impact of antibiotic usage on the food-chain as a consequence of effluent re-use in, and around, pig farms.


Asunto(s)
Agricultura/métodos , Antibacterianos/farmacología , Bacillus cereus/efectos de los fármacos , Pseudomonas/efectos de los fármacos , Microbiología del Suelo , Animales , Bacillus cereus/aislamiento & purificación , Granjas , Pseudomonas/aislamiento & purificación , Suelo/química , Porcinos
6.
J Appl Microbiol ; 130(2): 370-381, 2021 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-32692860

RESUMEN

AIMS: Bacillus cereus sensu lato is a complex group of closely related bacteria, which are commonly present in the natural environment and food products. These organisms may cause food poisoning and spoilage as well as opportunistic infections. Thus far, their resistance to selected antibiotics has been explored only in part, especially in the context of strain source. Therefore, our goal was to compare the resistance of B. cereus sl from milk (environment with the potential impact of antibiotics) with strains from soil and pepper (environment without contact with antibiotics) in relation to their origin, toxicity and phylogenetic relationships. METHODS AND RESULTS: Antibiotic resistance of B. cereus sl was assessed by determining their minimal inhibitory concentrations (MICs) followed by statistical analyses. The phylogeny of the bacteria was investigated by multilocus sequence typing, and toxicity was determined with quantitative reverse-transcription real-time PCR. We found that the isolates from milk were more often multiresistant and exhibited a common resistance pattern to ß-lactams but a varied sensitivity to the tested macrolides, clindamycin, tetracycline and vancomycin. Moreover they displayed often significantly higher average MICs; however, their resistance did not correlate with phylogeny, toxicity, or in most cases, with taxonomic affiliation. CONCLUSION: We conclude that mainly food matrices may serve as an important reservoir of resistant isolates of B. cereus sl and that the use of antibiotics for the treatment of animal diseases must be carefully monitored as it strongly promotes natural selection for multiresistant strains, even among opportunist pathogens. SIGNIFICANCE AND IMPACT OF THE STUDY: The fact that compared to the isolates from natural habitats, nonpathogenic B. cereus sl isolated from food acquire antibiotic resistance faster, should increase producers and consumers awareness and result in protection of public health.


Asunto(s)
Bacillus cereus/aislamiento & purificación , Farmacorresistencia Bacteriana , Microbiología Ambiental , Leche/microbiología , Verduras/microbiología , Animales , Antibacterianos/farmacología , Bacillus cereus/clasificación , Bacillus cereus/efectos de los fármacos , Bacillus cereus/genética , Toxinas Bacterianas/genética , Farmacorresistencia Bacteriana/efectos de los fármacos , Farmacorresistencia Bacteriana/genética , Variación Genética , Pruebas de Sensibilidad Microbiana , Tipificación de Secuencias Multilocus , Filogenia
7.
Pediatr Dev Pathol ; 24(4): 361-365, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-33729850

RESUMEN

Bacillus cereus is a gram-positive, rod-shaped bacterium that is commonly implicated in foodborne illness but has also become increasingly recognized as a source of serious non-gastrointestinal infections, including sepsis, meningitis, and pneumonia. Non-gastrointestinal B. cereus infections have been identified in children, especially in neonates; however, there are no previously described cases of fetal demise associated with B. cereus placental infection. We present a case of acute chorioamnionitis-related intrauterine fetal demise of twin A at 17 weeks gestation, noted two days after selective termination of twin B. Histological examination revealed numerous gram-positive bacilli in placental tissue, as well as fetal vasculature, in the setting of severe acute necrotizing chorioamnionitis and subchorionitis, intervillous abscesses, acute villitis, and peripheral acute funisitis. Cultures of maternal blood and placental tissue both yielded growth of B. cereus. This case underscores the importance of B. cereus as a human pathogen, and specifically demonstrates its potential as an agent of severe intraamniotic and placental infection with poor outcomes for the fetus.


Asunto(s)
Bacillus cereus/aislamiento & purificación , Corioamnionitis/diagnóstico , Muerte Fetal/etiología , Infecciones por Bacterias Grampositivas/diagnóstico , Placenta/microbiología , Complicaciones Infecciosas del Embarazo/diagnóstico , Adulto , Corioamnionitis/microbiología , Corioamnionitis/patología , Femenino , Infecciones por Bacterias Grampositivas/microbiología , Infecciones por Bacterias Grampositivas/patología , Humanos , Masculino , Placenta/patología , Embarazo , Complicaciones Infecciosas del Embarazo/microbiología , Complicaciones Infecciosas del Embarazo/patología , Embarazo Gemelar
8.
Lett Appl Microbiol ; 72(1): 53-59, 2021 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-32955742

RESUMEN

A new trans-4-hydroxy-l-proline (trans-Hyp) producing Bacillus cereus HBL-AI, was isolated from the air, which was screened just using l-proline as carbon and energy sources. This strain exhibited 73·4% bioconversion rate from initial l-proline (3 g l-1 ) to trans-Hyp. By sequencing the genome of this bacterium, 6244 coding sequences were obtained. Genome annotation analysis and functional expression were used to identify the proline-4-hydroxylase (BP4H) in HBL-AI. This enzyme belonged to a family of 2-oxoglutarate-related dioxygenases, which required 2-oxoglutarate and O2 as co-substrates for the reaction. Homologous modelling indicated that the enzyme had two monomers and contained conserved motifs, which included a distorted 'jelly roll' ß strand core and the residues (HXDXnH and RXS). The engineering Escherichia coli 3 Δ W3110/pTrc99a-proba-bp4h was constructed using BP4H, which transformed glucose to trans-Hyp in one step with high concentration of 46·2 g l-1 . This strategy provides a green and efficient method for synthesis of trans-Hyp and thus has a great potential in industrial application.


Asunto(s)
Bacillus cereus/enzimología , Genoma Bacteriano/genética , Hidroxiprolina/biosíntesis , Prolil Hidroxilasas/metabolismo , Bacillus cereus/genética , Bacillus cereus/aislamiento & purificación , Bacillus cereus/metabolismo , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Escherichia coli/genética , Escherichia coli/metabolismo , Glucosa/metabolismo , Ácidos Cetoglutáricos/metabolismo , Anotación de Secuencia Molecular , Prolina/metabolismo , Prolil Hidroxilasas/genética
9.
Biotechnol Lett ; 43(7): 1349-1355, 2021 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-33694018

RESUMEN

BACKGROUND: All RecJ proteins are known to date only perform exonuclease activity. The present study reports that a novel RecJ protein obtained from Bacillus cereus isolated from marine sediments has both endonuclease and exonuclease activities. METHODS: Analysis of the BcRecJ expression induction in E. coli BL21 revealed that the BcRecJ protein cleaved plasmids and genomic DNA in the host cell, and led to cell death and decreased the DNA content. Further, the BcRecJ protein had the ability to degrade supercoiled plasmid DNA into circular or linear forms in vitro. Meanwhile, the BcRecJ protein loaded with an S-modified guide facilitated plasmid linearization and reduced smear formation. RESULTS: The results suggested that this novel BcRecJ protein was different from any reported RecJs and had a longer C-terminus. Testing the BcRecJ mutants indicated that the endonuclease activity was affected by two residues of BcRecJ (D561, E637) after testing the BcRecJ mutants. CONCLUSION: The discovery of the type of protein is a new breakthrough for the RecJ proteins, which has both endonuclease and exonuclease activities.


Asunto(s)
Bacillus cereus/enzimología , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Escherichia coli/crecimiento & desarrollo , Exodesoxirribonucleasas/genética , Exodesoxirribonucleasas/metabolismo , Bacillus cereus/aislamiento & purificación , Cromosomas Bacterianos/metabolismo , Clonación Molecular , Endonucleasas/genética , Endonucleasas/metabolismo , Escherichia coli/genética , Exonucleasas/genética , Exonucleasas/metabolismo , Regulación Bacteriana de la Expresión Génica , Regulación Enzimológica de la Expresión Génica , Sedimentos Geológicos/microbiología , Viabilidad Microbiana , Plásmidos/metabolismo , Transformación Bacteriana
10.
Food Microbiol ; 93: 103604, 2021 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-32912579

RESUMEN

This study addresses the biodiversity of Bacillus cereus group population present along the value chain of milk for consumption. The B. cereus population did not grow and remained mainly unaltered during storage of milk at 4 °C while storage at a suboptimal temperature at 8 °C (representative of a broken cold chain) caused a major shift in its composition. Mesophilic strains dominated the B. cereus population in raw milk and after storage at 4 °C, while psycrotrophic strains dominated after storage at 8 °C. All psycrotrophic and mesophilic isolates (n = 368) demonstrated high spoilage potentials of the milk components. Fifteen out of 20 mesophilic isolates but only two out of 40 psychrotrophic isolates, exhibited vero cell toxicity. No genes encoding the emetic toxin cereulide were detected in the genomes of 100 milk isolates while 14 of them harbored the enterotoxin genes cytK1/cytK2. Both psycrotrophic and mesophilic isolates carried the enterotoxin genes nheA and hblA. Together, the results provide insight into the composition and properties, of the B. cereus population present in milk along the value chain and during storage at optimal refrigerated temperature and at suboptimal temperature. This knowledge is useful in the dairy industry's work to assure high quality products and for risk assessment.


Asunto(s)
Bacillus cereus/clasificación , Bacillus cereus/genética , Bacillus cereus/aislamiento & purificación , Microbiología de Alimentos , Leche/microbiología , Animales , Proteínas Bacterianas/genética , Toxinas Bacterianas/genética , Biodiversidad , ADN Bacteriano/genética , Depsipéptidos , Enterotoxinas/genética , Fermentación , Contaminación de Alimentos/análisis , Proteínas Hemolisinas/genética , Filogenia , Temperatura
11.
Food Microbiol ; 98: 103759, 2021 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-33875199

RESUMEN

B. cereus is a human pathogen associated with food poisoning leading to gastrointestinal disorders, as well as local and severe systemic infections. The pathogenic spectrum of B. cereus ranges from strains used as probiotics in humans to lethal highly toxic strains. In this study, we gathered a collection of 100 strains representative of the pathological diversity of B. cereus in humans, and characterized these strains for their cytotoxic potential towards human cells. We analyzed the correlation between cytotoxicity to epithelial and macrophage cells and the combination of 10 genes suspected to play a role during B. cereus virulence. We highlight genetic differences among isolates and studied correlations between genetic signature, cytotoxicity and strain pathological status. We hope that our findings will improve our understanding of the pathogenicity of B. cereus, thereby making it possible to improve both clinical diagnosis and food safety.


Asunto(s)
Bacillus cereus/patogenicidad , Enfermedades Transmitidas por los Alimentos/microbiología , Infecciones por Bacterias Grampositivas/microbiología , Animales , Bacillus cereus/clasificación , Bacillus cereus/genética , Bacillus cereus/aislamiento & purificación , Línea Celular , Células Epiteliales/microbiología , Contaminación de Alimentos/análisis , Microbiología de Alimentos , Humanos , Macrófagos/microbiología , Filogenia , Virulencia
12.
J Dairy Sci ; 104(2): 1465-1473, 2021 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-33272582

RESUMEN

In this study, a new vancomycin (Van)-modified poly-l-lysine (PLL) magnetic bead (MB) technique was developed for isolation of gram-positive bacteria. The method combines magnetic separation with a multiplex PCR (mPCR) assay and gel electrophoresis for easy and rapid detection of Bacillus cereus. Vancomycin was used as a molecular ligand between the MB and the d-alanyl-d-alanine moieties on the cell wall surface of B. cereus. The PLL served as a flexible molecular tether between the MB and Van that reduced steric hindrance maintaining the biological activity of Van. The MB-PLL-Van capture nanoprobes exhibited excellent capture and isolation efficiency for B. cereus in spiked milk matrix samples without interference from the complex food matrix. The subsequent mPCR assay showed high specificity for the 4 target genes in B. cereus, the entFM, cesB, cer, and 16S rRNA genes, that were used to achieve efficient genotyping and detection. Under optimum conditions, the limit of detection reached 103 cfu/mL, with a dynamic range of detection at 103 to 107 cfu/mL in pure culture. Application of the MB-PLL-Van mediated mPCR assay for B. cereus in milk matrix samples achieved results similar to those of the pure culture. In addition, with a 6-h pre-enrichment of B. cereus that was spiked in milk matrix samples, the limit of detection reached 101 cfu/mL. The MB-PLL-Van mediated mPCR assay developed in this study could be used as a universal technology platform for the efficient enrichment and genotyping of gram-positive bacteria.


Asunto(s)
Bacillus cereus/aislamiento & purificación , Campos Magnéticos , Leche/microbiología , Reacción en Cadena de la Polimerasa Multiplex , Polilisina , Vancomicina , Animales , Bacillus cereus/clasificación , Bacillus cereus/genética , Microbiología de Alimentos/métodos , Técnicas de Genotipaje , Microesferas , Reacción en Cadena de la Polimerasa Multiplex/veterinaria , ARN Ribosómico 16S
13.
Foodborne Pathog Dis ; 18(1): 8-15, 2021 01.
Artículo en Inglés | MEDLINE | ID: mdl-32865425

RESUMEN

This study was conducted to reveal the genotyping, antimicrobial susceptibility, and biofilm formation of Bacillus cereus isolated from powdered food products in China. Five hundred powdered food samples were collected from five provinces in China: 100 samples each of powdered infant formula (PIF), soy milk powder (SMP), lotus root powder (LRP), walnut powder (WP), and rice flour (RF). The genotyping of isolates was analyzed using multilocus sequence typing; meanwhile, antimicrobial susceptibility, and ability of biofilms formation on stainless steel tube of isolates were evaluated. Forty-two B. cereus strains were detected with an overall contamination rate of 8.4%, as well as, the highest B. cereus contamination rate was found in SMP (10%), followed by LRP (9%), WP (9%), RF (8%), and PIF (6%). These isolates were divided into 22 sequence types (STs); among them, ST32 (4/42, 9.5%) was the predominant ST. Phylogenetic relationships showed that the 42 strains of B. cereus were divided into three groups (group I, group II, and group III). Antimicrobial susceptibility testing indicated that all isolates were susceptible to tetracycline, gentamicin, erythromycin, and chloramphenicol, while resistant to ampicillin, cefepime, oxacillin, and rifampin. The analysis of ability of biofilm formation on stainless steel tube showed optical density (OD)595 value of 66.7% of B. cereus isolates was greater than 1. The OD595 level of isolates belonging to group III was higher compared with the other two groups, and OD595 values of B. cereus HB1 and HN5 were greater than 2. These findings improved the understanding of the characteristics of B. cereus isolated from powdered food products in China, and provided a theoretical basis for the prevention and control of B. cereus in food industry.


Asunto(s)
Bacillus cereus/aislamiento & purificación , Biopelículas/crecimiento & desarrollo , Farmacorresistencia Bacteriana/fisiología , Contaminación de Alimentos/análisis , Alimentos Especializados/microbiología , Bacillus cereus/genética , Bacillus cereus/fisiología , China , Microbiología de Alimentos , Genotipo , Humanos , Pruebas de Sensibilidad Microbiana , Filogenia , Polvos
14.
Foodborne Pathog Dis ; 18(5): 306-314, 2021 05.
Artículo en Inglés | MEDLINE | ID: mdl-33769083

RESUMEN

Bacillus cereus is a common foodborne pathogen that can cause both gastrointestinal and nongastrointestinal diseases. In this study, we collected 603 meat and meat products from 39 major cities in China. The positive contamination rate of B. cereus in the collected samples was 26.37% (159/603), and the contamination level in 5.03% (8/159) positive samples exceeded 1100 most probable number/g. The detection rates of virulence genes were 89.7% for the nheABC gene group, 37.1% for the hblACD gene cluster, 82.3% for cytK-2, and 2.9% for cesB. Notably, all isolates presented with multiple antibiotic resistance, and 99.43% of isolates were resistant to five classes of antibiotics. In addition, the multilocus sequence typing results indicated that all isolates were rich in genetic diversity. Collectively, we conducted a systematic investigation on the prevalence and characterization of B. cereus in meat and meat products in China, providing crucial information for assessing the risk of B. cereus occurrence in meat and meat products.


Asunto(s)
Bacillus cereus/aislamiento & purificación , Farmacorresistencia Bacteriana Múltiple/genética , Microbiología de Alimentos/estadística & datos numéricos , Productos de la Carne/microbiología , Carne/microbiología , Animales , Antibacterianos/farmacología , Bacillus cereus/genética , China/epidemiología , Tipificación de Secuencias Multilocus , Prevalencia , Factores de Virulencia/genética , Factores de Virulencia/aislamiento & purificación
15.
Environ Microbiol ; 22(12): 5248-5264, 2020 12.
Artículo en Inglés | MEDLINE | ID: mdl-32815215

RESUMEN

Bacillus cereus is a ubiquitous endospore-forming bacterium, which mainly affects humans as a food-borne pathogen. Bacillus cereus can contaminate groundwater used to irrigate food crops. Here, we examined the ability of the emetic strain B. cereus F4810/72 to survive abiotic conditions encountered in groundwater. Our results showed that vegetative B. cereus cells rapidly evolved in a mixed population composed of endospores and asporogenic variants bearing spo0A mutations. One asporogenic variant, VAR-F48, was isolated and characterized. VAR-F48 can survive in sterilized groundwater over a long period in a vegetative form and has a competitive advantage compared to its parental strain. Proteomics analysis allowed us to quantify changes to cellular and exoproteins after 24 and 72 h incubation in groundwater, for VAR-F48 compared to its parental strain. The results revealed a significant re-routing of the metabolism in the absence of Spo0A. We concluded that VAR-F48 maximizes its energy use to deal with oligotrophy, and the emergence of spo0A-mutated variants may contribute to the persistence of emetic B. cereus in natural oligotrophic environments.


Asunto(s)
Bacillus cereus/fisiología , Proteínas Bacterianas/genética , Enfermedades Transmitidas por los Alimentos/microbiología , Agua Subterránea/microbiología , Factores de Transcripción/genética , Bacillus cereus/genética , Bacillus cereus/aislamiento & purificación , Bacillus cereus/metabolismo , Proteínas Bacterianas/metabolismo , Microbiología de Alimentos , Humanos , Viabilidad Microbiana/genética , Mutación , Esporas Bacterianas/genética , Esporas Bacterianas/aislamiento & purificación , Esporas Bacterianas/metabolismo , Esporas Bacterianas/fisiología , Factores de Transcripción/metabolismo
16.
BMC Microbiol ; 20(1): 363, 2020 11 26.
Artículo en Inglés | MEDLINE | ID: mdl-33243143

RESUMEN

BACKGROUND: Bacillus thuringiensis bacteria share similar genetic, physiological, and biochemical characteristics with other members of the Bacillus cereus group. Their diversity and entomopathogenic origin are related to their mobile genetic elements. However, the effects of wide-spread application of B. thuringiensis-based pesticides on genetically related B. cereus group populations present in the environment remain poorly understood. RESULTS: We first identified pBMB76 from B. thuringiensis tenebrionis as a new conjugative plasmid. Mixed mating experiments suggested that pBMB76 may compete with pHT73, another known conjugative plasmid. Applications of single (tenebrionis 4AA1 and kurstaki HD73 carrying pBMB76 and pHT73, respectively) and mixed (4AA1 + HD73) B. thuringiensis strains were performed in confined plot habitats (soil and leaf) over two planting seasons. In total, 684 B. cereus group isolates were randomly selected from different treatment sets, and the transmissibility and occurrence rate of potential conjugative plasmids were surveyed. Results showed that the percentage of isolates with plasmid mobility was markedly enhanced in the B. thuringiensis-sprayed groups. Furthermore, we performed multi-locus sequence typing (MLST) for a subset of 291 isolates, which indicated that the dominant sequence types in the treated habitats were identical or related to the corresponding sprayed formulations. CONCLUSIONS: The application of B. thuringiensis strains with conjugal and mobilizing capability drove gene transmissibility within the B. cereus group populations in confined habitats and potentially modified the population structure.


Asunto(s)
Bacillus cereus/genética , Bacillus thuringiensis/genética , Conjugación Genética , Bacillus cereus/clasificación , Bacillus cereus/aislamiento & purificación , Agentes de Control Biológico , Ecosistema , Genes Bacterianos/genética , Hojas de la Planta/microbiología , Plásmidos/genética , Microbiología del Suelo
17.
BMC Microbiol ; 20(1): 310, 2020 10 14.
Artículo en Inglés | MEDLINE | ID: mdl-33054711

RESUMEN

BACKGROUND: Bacillus cereus is a foodborne pathogen commonly found in nature and food and can cause food spoilage and health issues. Although the prevalence of B. cereus in foods has been reported worldwide, the extent of contamination in edible fungi, which has become increasingly popular as traditional or functional food, is largely unknown. Here we investigated the prevalence, toxin genes' distribution, antibiotic resistance, and genetic diversity of B. cereus isolated from edible fungi in China. RESULTS: Six hundred and ninety-nine edible fungi samples were collected across China, with 198 (28.3%) samples found to be contaminated by B. cereus, with an average contamination level of 55.4 most probable number (MPN)/g. Two hundred and forty-seven B. cereus strains were isolated from the contaminated samples. Seven enterotoxin genes and one cereulide synthetase gene were detected. The detection frequencies of all enterotoxin genes were ≥ 80%, whereas the positive rate of the cesB gene in B. cereus was 3%. Most isolates were resistant to penicillins, ß-lactam/ß-lactamase inhibitor combinations, cephems, and ansamycins, but were susceptible to penems, aminoglycosides, macrolides, ketolide, glycopeptides, quinolones, phenylpropanol, tetracyclines, lincosamides, streptogramins, and nitrofurans. Meanwhile, 99.6% of all isolates displayed multiple antimicrobial resistance to three or more classes of antimicrobials. Using genetic diversity analysis, all isolates were defined in 171 sequence types (STs), of which 83 isolates were assigned to 78 new STs. CONCLUSIONS: This study provides large-scale insight into the prevalence and potential risk of B. cereus in edible fungi in China. Approximately one-third of the samples were contaminated with B. cereus, and almost all isolates showed multiple antimicrobial resistance. Detection frequencies of all seven enterotoxin genes were equal to or more than 80%. These new findings may indicate a need for proper pre-/post-processing of edible fungi to eliminate B. cereus, thereby preventing the potential risk to public health.


Asunto(s)
Agaricales , Bacillus cereus/aislamiento & purificación , Enterotoxinas/análisis , Contaminación de Alimentos/análisis , Antibacterianos/farmacología , Bacillus cereus/efectos de los fármacos , Bacillus cereus/genética , China , Recuento de Colonia Microbiana , Farmacorresistencia Microbiana , Microbiología de Alimentos
18.
BMC Microbiol ; 20(1): 175, 2020 06 22.
Artículo en Inglés | MEDLINE | ID: mdl-32571217

RESUMEN

BACKGROUND: Incidences of heat stress due to the changing global climate can negatively affect the growth and yield of temperature-sensitive crops such as soybean variety, Pungsannamul. Increased temperatures decrease crop productivity by affecting biochemical, physiological, molecular, and morphological factors either individually or in combination with other abiotic stresses. The application of plant growth-promoting endophytic bacteria (PGPEB) offers an ecofriendly approach for improving agriculture crop production and counteracting the negative effects of heat stress. RESULTS: We isolated, screened and identified thermotolerant B. cereus SA1 as a bacterium that could produce biologically active metabolites, such as gibberellin, indole-3-acetic acid, and organic acids. SA1 inoculation improved the biomass, chlorophyll content, and chlorophyll fluorescence of soybean plants under normal and heat stress conditions for 5 and 10 days. Heat stress increased abscisic acid (ABA) and reduced salicylic acid (SA); however, SA1 inoculation markedly reduced ABA and increased SA. Antioxidant analysis results showed that SA1 increased the ascorbic acid peroxidase, superoxide dismutase, and glutathione contents in soybean plants. In addition, heat stress markedly decreased amino acid contents; however, they were increased with SA1 inoculation. Heat stress for 5 days increased heat shock protein (HSP) expression, and a decrease in GmHSP expression was observed after 10 days; however, SA1 inoculation augmented the heat stress response and increased HSP expression. The stress-responsive GmLAX3 and GmAKT2 were overexpressed in SA1-inoculated plants and may be associated with decreased reactive oxygen species generation, altered auxin and ABA stimuli, and enhanced potassium gradients, which are critical in plants under heat stress. CONCLUSION: The current findings suggest that B. cereus SA1 could be used as a thermotolerant bacterium for the mitigation of heat stress damage in soybean plants and could be commercialized as a biofertilizer only in case found non-pathogenic.


Asunto(s)
Bacillus cereus/fisiología , Endófitos/fisiología , Glycine max/crecimiento & desarrollo , Proteínas de Choque Térmico/genética , Ácido Abscísico/metabolismo , Bacillus cereus/aislamiento & purificación , Bacillus cereus/metabolismo , Productos Agrícolas/genética , Productos Agrícolas/crecimiento & desarrollo , Productos Agrícolas/microbiología , Regulación de la Expresión Génica de las Plantas/efectos de los fármacos , Respuesta al Choque Térmico , Proteínas de Plantas/genética , Ácido Salicílico/metabolismo , Microbiología del Suelo , Glycine max/genética , Glycine max/microbiología , Termotolerancia
19.
Microb Pathog ; 140: 103966, 2020 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-31911192

RESUMEN

Replacing agrochemicals with plant growth promoting bacteria (PGPB) may offset some of the environmental impacts of food production. The objectives of this study were to (1) isolate and characterize bacterial strains from tomato rhizosphere, including root, shoot and leaf, (2) select and identify the most promising PGPB strains, (3) verify the phytostimulatory activity and mineral uptake potential of selected strains. Bacterial strains isolated from tomato rhizosphere, were screened for phosphorous (P) solubilization, production of indole acetic acid (IAA), amylase activity, antibiotic resistance, and quick test strip (QTS) for biochemical characterization. The tested strains, positive for all five of these assays were selected for molecular identification and subjected to greenhouse growth trails with tomato and mung bean. Two strains were selected and identified as Bacillus cereus (B. cereus) isolated from rhizosphere and Klebsiella variicola (K. variicola) isolated from root endosphere using 16s rRNA sequences. Both strains produced IAA, gibberellic acid (GA3) and kinetin, however B. cereus showed potential GA3 and IAA production as compared to K. variicola. In tomato, only one growth variable (shoot length) was increased over the control by one of the selected bacterial strains (B. cereus). In mung bean, inoculation with either strain B. cereus or K. variicola increased shoot length and dry weight. Moreover, our results showed that the use of PGPB significantly increased plant growth and Fe, Zn, Ca, Mg, Cu, Na and K contents of plants. It seems that evaluated strains had a higher ability in boosting plant growth and higher yield.


Asunto(s)
Bacterias/aislamiento & purificación , Microbiología del Suelo , Solanum lycopersicum/crecimiento & desarrollo , Solanum lycopersicum/microbiología , Bacillus cereus/clasificación , Bacillus cereus/genética , Bacillus cereus/aislamiento & purificación , Bacillus cereus/metabolismo , Bacterias/clasificación , Bacterias/genética , Bacterias/metabolismo , Giberelinas/metabolismo , Ácidos Indolacéticos/metabolismo , Klebsiella/clasificación , Klebsiella/genética , Klebsiella/aislamiento & purificación , Klebsiella/metabolismo , Filogenia , Hojas de la Planta/microbiología , Raíces de Plantas/microbiología , Rizosfera
20.
Exp Eye Res ; 193: 107959, 2020 04.
Artículo en Inglés | MEDLINE | ID: mdl-32032628

RESUMEN

Bacillus cereus (B. cereus) endophthalmitis is a devastating intraocular infection primarily associated with post-traumatic injuries. The majority of these infections result in substantial vision loss, if not loss of the eye itself, within 12-48 h. Multifactorial mechanisms that lead to the innate intraocular inflammatory response during this disease include the combination of robust bacterial replication, migration of the organism throughout the eye, and toxin production by the organism. Therefore, the window of therapeutic intervention in B. cereus endophthalmitis is quite narrow compared to that of other pathogens which cause this disease. Understanding the interaction of bacterial and host factors is critical in understanding the disease and formulating more rational therapeutics for salvaging vision. In this review, we will discuss clinical and research findings related to B. cereus endophthalmitis in terms of the organism's virulence and inflammogenic potential, and strategies for improving of current therapeutic regimens for this blinding disease.


Asunto(s)
Antibacterianos/uso terapéutico , Bacillus cereus/aislamiento & purificación , Endoftalmitis/microbiología , Infecciones Bacterianas del Ojo/microbiología , Endoftalmitis/tratamiento farmacológico , Infecciones Bacterianas del Ojo/tratamiento farmacológico , Humanos
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