Ground Anthrax Bacillus Refined Isolation (GABRI) method for analyzing environmental samples with low levels of Bacillus anthracis contamination

Fasanella, Antonio; Taranto, Pietro Di; Garofolo, Giuliano; Colao, Valeriana; Mariño, Leonardo; Buonavoglia, Domenico; Pedarra, Carmine; Adone, Rosanna; Hugh‐Jones, Martin

doi:10.1186/1471-2180-13-167

Cited by 28 publications

(36 citation statements)

References 14 publications

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“…The Ground Anthrax Bacillus Refined Isolation (GABRI) was performed as described by [ 36 ]. After incubation at 37°C for 24 hours the suspected colonies appearing whitish and without hemolysis were picked and spread on Columbia blood agar.…”

Section: Methodsmentioning

confidence: 99%

Microevolution of Anthrax from a Young Ancestor (M.A.Y.A.) Suggests a Soil-Borne Life Cycle of Bacillus anthracis

et al. 2015

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During an anthrax outbreak at the Pollino National Park (Basilicata, Italy) in 2004, diseased cattle were buried and from these anthrax-foci Bacillus anthracis endospores still diffuse to the surface resulting in local accumulations. Recent data suggest that B. anthracis multiplies in soil outside the animal-host body. This notion is supported by the frequent isolation of B. anthracis from soil lacking one or both virulence plasmids. Such strains represent an evolutionary dead end, as they are likely no longer able to successfully infect new hosts. This loss of virulence plasmids is explained most simply by postulating a soil-borne life cycle of the pathogen. To test this hypothesis we investigated possible microevolution at two natural anthrax foci from the 2004 outbreak. If valid, then genotypes of strains isolated from near the surface at these foci should be on a different evolutionary trajectory from those below residing in deeper-laying horizons close to the carcass. Thus, the genetic diversity of B. anthracis isolates was compared conducting Progressive Hierarchical Resolving Assays using Nucleic Acids (PHRANA) and next generation Whole Genome Sequencing (WGS). PHRANA was not discriminatory enough to resolve the fine genetic relationships between the isolates. Conversely, WGS of nine isolates from near-surface and nine from near-carcass revealed five isolate specific SNPs, four of which were found only in different near-surface isolates. In support of our hypothesis, one surface-isolate lacked plasmid pXO1 and also harbored one of the unique SNPs. Taken together, our results suggest a limited soil-borne life cycle of B. anthracis.

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Section: Methodsmentioning

confidence: 99%

Microevolution of Anthrax from a Young Ancestor (M.A.Y.A.) Suggests a Soil-Borne Life Cycle of Bacillus anthracis

et al. 2015

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“…Biopsy samples from skin lesions of each patient were directly cultured on 5% sheep blood agar at 37˚C overnight and a single colony was isolated as an independent strain. The colonies were identified using biochemical tests and PCR [18].…”

Section: Bacterial Strains and Dna Extractionmentioning

confidence: 99%

Epidemiological and comparative genomic analysis of Bacillus anthracis isolated from northern Vietnam

et al. 2020

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To understand the epidemiological and genetic background of anthrax cases occurring in Vietnam from 2011 to 2015, we surveilled and genetically analyzed Bacillus anthracis isolated in the north of the country. Epidemiological surveillance showed that most human cutaneous anthrax cases occurred in association with animal dissection. Whole-genome sequences were obtained from six B. anthracis strains from human patients with cutaneous anthrax in the endemic area. Comparative genomic analysis showed that the genetic homogeneity among Vietnamese B. anthracis strains was very high. All Vietnamese B. anthracis strains belonged to the canSNP lineage of A.Br.011/009, which mostly consists of strains of the trans-Eurasian (TEA) group, including the most closely related strain, Carbosap. To clarify the genetic diversity of Vietnamese strains and strains belonging to A.Br.011/009 and A. Br.008/011 canSNP lineages, we applied a reference genome-based single-nucleotide polymorphism (SNP) and gene-by-gene genomic analysis (whole-genome MLST) strategy. The phylogeny from core genome SNPs revealed that the Vietnamese strains were positioned close to each other; moreover, several SNPs specific to Vietnamese B. anthracis were identified. Whole-genome MLST analysis revealed the differences in the number of SNPs between Vietnamese strains, which could enable discrimination at the strain level. OPEN ACCESS Citation: Hoang TTH, Dang DA, Pham TH, Luong MH, Tran ND, Nguyen TH, et al. (2020) Epidemiological and comparative genomic analysis of Bacillus anthracis isolated from northern Vietnam. PLoS ONE 15(2): e0228116. https://doi.

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“…To detect B. anthracis from samples, the Ground Anthrax Bacillus Refined Isolation (GABRI) method was used (Fasanella et al, 2013a). The GABRI method is a sensitive method for isolation of B. anthracis spores from soil particles as it disrupts hydrophobic interactions between spores and the adhering solid matrices.…”

Section: Isolation Of B Anthracis From Soil Samples and Inactivationmentioning

confidence: 99%

“…All non-hemolytic candidate strains retrieved by the modified GABRI method (Fasanella et al, 2013a) from Bangladesh samples were identified as B. anthracis by diagnostic real time PCR on three markers dhp61 (chromosomal), pagA (pXO1) and capC (pXO2), harboring both virulence plasmids (pXO1 and pXO2) (data not shown). The geographic attribution of these isolates within Bangladesh is presented in Table 1.…”

Section: B Anthracis Isolation From Environmental Samples and Initiamentioning

confidence: 99%

Unexpected genomic relationships between Bacillus anthracis strains from Bangladesh and Central Europe

Rume

Ahsan

Biswas

et al. 2016

Infection, Genetics and Evolution

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The zoonosis anthrax caused by the bacterium Bacillus anthracis has a broad geographical distribution. Active enzootic areas are typically located away from central and northern Europe where cases of the disease occur only sporadically and in limited numbers. In contrast, a few out of the 64 districts of Bangladesh are hyper-endemic for anthrax and there the disease causes major losses in live-stock. In this study we genotyped eight strains of B. anthracis collected from the districts of Sirajganj and Tangail in 2013. All these strains belonged to canSNP group A.Br.001/002 Sterne differing only in a few of 31 tandem-repeat (MLVA)-markers. Whole genome sequences were obtained from five of these strains and compared with genomic information of B. anthracis strains originating from various geographical locations. Characteristic signatures were detected defining two "Bangladesh" clusters potentially useful for rapid molecular epidemiology. From this data high-resolution PCR assays were developed and subsequently tested on additional isolates from Bangladesh and Central Europe. Remarkably, this comparative genomic analysis focusing on SNP-discovery revealed a close genetic relationship between these strains from Bangladesh and historic strains collected between 1991 and 2008 in The Netherlands and Germany, respectively. Possible explanations for these phylogenetic relationships are discussed.

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Ground Anthrax Bacillus Refined Isolation (GABRI) method for analyzing environmental samples with low levels of Bacillus anthracis contamination

Cited by 28 publications

References 14 publications

Microevolution of Anthrax from a Young Ancestor (M.A.Y.A.) Suggests a Soil-Borne Life Cycle of Bacillus anthracis

Microevolution of Anthrax from a Young Ancestor (M.A.Y.A.) Suggests a Soil-Borne Life Cycle of Bacillus anthracis

Epidemiological and comparative genomic analysis of Bacillus anthracis isolated from northern Vietnam

Unexpected genomic relationships between Bacillus anthracis strains from Bangladesh and Central Europe

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