Genome sequencing of two Bacillus anthracis strains: a virulent strain and a vaccinal strain

Anthrax is a strategic and important disease because it is a zoonotic disease which caused death and difficult to eradicate because it produces spores. The aim of the study was to identify anthrax in endemic areas in South Sulawesi Province. Soil samples obtained from Gowa and Pinrang regency were examined by bacterial culture test and then positive results from bacterial culture followed by multiplex PCR. Based on the results of the isolation of 52 soil samples show 35 samples from Pinrang regency show all negative anthrax, while from 17 soil samples from Gowa regency show 2 positive samples of anthrax. Positive samples of anthrax were from Timbuseng village, Patallasang district, Gowa regency. The results continued with PCR technique using Bacillus anthracis colonies derived from positive soil samples. It showed the virulent strain of Bacillus anthracis. A virulent strain is indicated by a DNA fragment of a pXO1 plasmid encoding a toxin lethal factor (Lef) of 385 bp and a DNA fragment of a pXO2 plasmid encoding a capsule of anthrax (Cap) of 264 bp and also common bacterial markers of the Bacillus genome chromosome (Ba183) of 152 bp.

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Overlapping of independent SARS-CoV-2 nosocomial transmissions in a complex outbreak

10.21203/rs.3.rs-305824/v1 ◽

2021 ◽

Author(s):

Laura Pérez-Lago ◽

Helena Martinez Lozano ◽

Jose Antonio pajares Diaz ◽

Arantxa Diaz Gomez ◽

Marina Machado ◽

...

Keyword(s):

Whole Genome Sequencing ◽

Genome Sequencing ◽

Healthcare Workers ◽

Virulent Strain ◽

Control Measures ◽

Protective Equipment ◽

Whole Genome ◽

Single Patient ◽

High Fatality Rate ◽

Depth Analysis

Abstract SARS-CoV-2 nosocomial outbreaks in the first COVID-19 wave were likely associated to a shortage of personal protective equipment and scare indications on control measures. Having covered these limitations, updates on current SARS-CoV-2 nosocomial outbreaks are required. We carried out an in-depth analysis of a 27-day nosocomial outbreak in a gastroenterology ward in our hospital, potentially involving 15 patients and three healthcare workers. Patients had stayed in one of three neighbouring rooms in the ward. The severity of the infections in six of the cases and a high fatality rate suggested the possible involvement of a single virulent strain persisting in those rooms. Whole genome sequencing of the strains from 12 patients and one healthcare worker revealed an unexpected complexity. Five different SARS-CoV-2 strains were identified, two infecting a single patient each, ruling out their relationship with the outbreak; the remaining three strains were involved in three independent overlapping limited transmission clusters with three, three, and five cases. Whole genome sequencing was key to understand the complexity of this outbreak.

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Whole Genome-Sequencing and Phylogenetic Analysis of a Historical Collection of Bacillus anthracis Strains from Danish Cattle

PLoS ONE ◽

10.1371/journal.pone.0134699 ◽

2015 ◽

Vol 10 (8) ◽

pp. e0134699 ◽

Cited By ~ 13

Author(s):

Sylviane Derzelle ◽

Guillaume Girault ◽

Branko Kokotovic ◽

Øystein Angen

Keyword(s):

Phylogenetic Analysis ◽

Bacillus Anthracis ◽

Whole Genome Sequencing ◽

Genome Sequencing ◽

Whole Genome

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Comparative Genome Sequencing for Discovery of Novel Polymorphisms in Bacillus anthracis

Science ◽

10.1126/science.1071837 ◽

2002 ◽

Vol 296 (5575) ◽

pp. 2028-2033 ◽

Cited By ~ 285

Author(s):

T. D. Read

Keyword(s):

Bacillus Anthracis ◽

Genome Sequencing ◽

Comparative Genome

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Bovine abortion by a vaccine strain of Bacillus anthracis

Ciência Rural ◽

10.1590/0103-8478cr20200264 ◽

2020 ◽

Vol 50 (12) ◽

Author(s):

Daniele Mariath Bassuino ◽

Franciele Maboni Siqueira ◽

Guilherme Konradt ◽

Andréia Vielmo ◽

Verônica Machado Rolim ◽

...

Keyword(s):

Bacillus Anthracis ◽

Vaccine Strain ◽

Chromosomal Marker ◽

Commercial Vaccine ◽

Gram Staining ◽

Microbiological Examination ◽

Vaccinal Strain ◽

High Level ◽

Bovine Abortion ◽

Residual Virulence

ABSTRACT: This paper reports the abortion of a male Aberdeen Angus bovine by a vaccine strain of Bacillus anthracis, describing the pathological and microbiological findings and the genome sequence. Necropsy findings included multifocal areas of hemorrhage in different organs. Histologically, various organs showed hemorrhage, fibrin exudation, necrosis associated with countless bacillary bacterial clumps and severe neutrophilic inflammatory infiltrate. In the microbiological examination, numerous rough, nonhemolytic, gray and dry colonies with irregular edges were isolated from liver, lung and abomasum content samples. Gram staining revealed square-ended Gram-positive rods arranged in chains. B. anthracis identification was confirmed by detection of the molecular chromosomal marker Ba813. The genomes from the isolated B. anthracis (named SPV842_15) and from the isolated vaccinal strain (Brazilian vaccinal strain), which was recovered from a commercial vaccine used in the pregnant cow, were sequenced. Genomic comparisons displayed a high level of nucleotide identity in the comparisons between B. anthracis SPV842_15 and the B. anthracis Brazilian vaccinal strain (98,2%). Furthermore, in both strains, only the plasmid pX01 sequence was detected. Although, vaccination against anthrax is characterized by an elevated protective profile and very low residual virulence, immunization with Sterne strains can cause abortion in cattle, presumably by the plasmid pX01 toxins in rare or special situations.

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Brucella abortus Cyclic β-1,2-Glucan Mutants Have Reduced Virulence in Mice and Are Defective in Intracellular Replication in HeLa Cells

Infection and Immunity ◽

10.1128/iai.69.7.4528-4535.2001 ◽

2001 ◽

Vol 69 (7) ◽

pp. 4528-4535 ◽

Cited By ~ 91

Author(s):

Gabriel Briones ◽

Nora Iñón de Iannino ◽

Mara Roset ◽

Ana Vigliocco ◽

Patricia Silva Paulo ◽

...

Keyword(s):

Hela Cells ◽

Brucella Abortus ◽

Virulent Strain ◽

Sodium Dodecyl ◽

Wild Type ◽

Log Reduction ◽

Vaccinal Strain ◽

Intracellular Multiplication ◽

Glucan Synthetase ◽

Residual Virulence

ABSTRACT Null cyclic β-1,2-glucan synthetase mutants (cgsmutants) were obtained from Brucella abortus virulent strain 2308 and from B. abortus attenuated vaccinal strain S19. Both mutants show greater sensitivity to surfactants like deoxycholic acid, sodium dodecyl sulfate, and Zwittergent than the parental strains, suggesting cell surface alterations. Although not to the same extent, both mutants display reduced virulence in mice and defective intracellular multiplication in HeLa cells. The B. abortus S19 cgs mutant was completely cleared from the spleens of mice after 4 weeks, while the 2308 mutant showed a 1.5-log reduction of the number of brucellae isolated from the spleens after 12 weeks. These results suggest that cyclic β-1,2-glucan plays an important role in the residual virulence of the attenuatedB. abortus S19 strain. Although the cgsmutant was cleared from the spleens earlier than the wild-type parental strain (B. abortus S19) and produced less inflammatory response, its ability to confer protection against the virulent strain B. abortus 2308 was fully retained. Equivalent levels of induction of spleen gamma interferon mRNA and anti-lipopolysaccharide (LPS) of immunoglobulin G2a (IgG2a) subtype antibodies were observed in mice injected withB. abortus S19 or the cgs mutant. However, the titer of anti-LPS antibodies of the IgG1 subtype induced by thecgs mutant was lower than that observed with the parental S19 strain, thus suggesting that the cgs mutant induces a relatively exclusive Th1 response.

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Infection with a Nonencapsulated Bacillus anthracis Strain in Rabbits—The Role of Bacterial Adhesion and the Potential for a Safe Live Attenuated Vaccine

Toxins ◽

10.3390/toxins10120506 ◽

2018 ◽

Vol 10 (12) ◽

pp. 506 ◽

Cited By ~ 2

Author(s):

Itai Glinert ◽

Shay Weiss ◽

Assa Sittner ◽

Elad Bar-David ◽

Amir Ben-Shmuel ◽

...

Keyword(s):

Bacillus Anthracis ◽

Bacterial Adhesion ◽

Virulent Strain ◽

Wild Type ◽

Live Attenuated Vaccine ◽

Adhesion Protein ◽

Brain Infection ◽

Systemic Dissemination ◽

Intravenous Inoculation

Nonencapsulated (∆pXO2) Bacillus anthracis strains are commonly used as vaccines and for anthrax research, mainly in the mouse model. Previously, we demonstrated that the infection of rabbits, intranasally or subcutaneously, with the spores of a fully virulent strain results in the systemic dissemination of the bacteria, meningitis, and death, whereas ∆pXO2 strains are fully attenuated in this animal model. We used the intravenous inoculation of rabbits to study the pathogenicity of the ∆pXO2 strain infection. Bacteremia, brain bacterial burden, and pathology were used as criteria to compare the Vollum∆pXO2 disease to the wild type Vollum infection. To test the role of adhesion in the virulence of Vollum∆pXO2, we deleted the major adhesion protein BslA and tested the virulence and immunogenicity of this mutant. We found that 50% of the rabbits succumb to Vollum∆pXO2 strain following i.v. infection, a death that was accompanied with significant neurological symptoms. Pathology revealed severe brain infection coupled with an atypical massive bacterial growth into the parenchyma. Contrary to the Vollum strain, deletion of the bslA gene fully attenuated the ∆pXO2 strain. Though the Vollum∆pXO2 cannot serve as a model for B. anthracis pathogenicity in rabbits, deletion of the bslA gene prevents central nervous system (CNS) infections, possibly leading to the generation of a safer vaccine.

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Whole Genome Sequencing for Studying Bacillus anthracis from an Outbreak in the Abruzzo Region of Italy

Microorganisms ◽

10.3390/microorganisms8010087 ◽

2020 ◽

Vol 8 (1) ◽

pp. 87

Author(s):

Alexandra Chiaverini ◽

Mostafa Y. Abdel-Glil ◽

Jörg Linde ◽

Domenico Galante ◽

Valeria Rondinone ◽

...

Keyword(s):

Bacillus Anthracis ◽

Whole Genome Sequencing ◽

Genome Sequencing ◽

Southern Italy ◽

Previous History ◽

Whole Genome ◽

Genome Sequences ◽

Abruzzo Region ◽

History Of ◽

Bacterium Bacillus

Anthrax is a serious infectious disease caused by the gram-positive and spore-forming bacterium Bacillus anthracis. In Italy, anthrax is an endemic disease with sporadic cases each year and few outbreaks, especially in Southern Italy. However, new foci have been discovered in zones without previous history of anthrax. During summer 2016, an outbreak of anthrax caused the death of four goats in the Abruzzo region, where the disease had not been reported before. In order to investigate the outbreak, we sequenced one strain and compared it to 19 Italian B. anthracis genomes. Furthermore, we downloaded 71 whole genome sequences representing the global distribution of canonical SNP lineages and used them to verify the phylogenetic positioning. To this end, we analyzed and compared the genome sequences using canonical SNPs and the whole genome SNP-based analysis. Our results demonstrate that the outbreak strain belonged to the Trans-Eurasian (TEA) group A.Br.011/009, which is the predominant clade in Central-Southern Italy. In conclusion, the high genomic relatedness of the Italian TEA strains suggests their evolution from a common ancestor, while the spread is supposedly driven by trade as well as human and transhumance activities. Here, we demonstrated the capabilities of whole genome sequencing (WGS), which can be used as a tool for outbreak analyses and surveillance activities.

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