Development of a novel selective agar for the isolation and detection of
            <i>Bacillus anthracis</i>

Rohde, Anna M.; Papp, Stefanie; Feige, P.; Grunow, Roland; Kaspari, Oliver

doi:10.1111/jam.14615

Cited by 9 publications

(9 citation statements)

References 27 publications

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“…Possibly, future progress in further developing B. anthracis -specific agar media may ameliorate this issue. Recently, for instance, a new selective agar medium for B. anthracis has been introduced [ 34 ]. The colony lift and blot ELPRA may be applied in conjunction with this improved agar medium for complex environmental samples bearing only low contamination of B. anthracis spores.…”

Section: Discussionmentioning

confidence: 99%

Enzyme-Linked Phage Receptor Binding Protein Assays (ELPRA) Enable Identification of Bacillus anthracis Colonies

Braun¹,

Rupprich²,

Neif³

et al. 2021

Viruses

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Bacteriophage receptor binding proteins (RBPs) are employed by viruses to recognize specific surface structures on bacterial host cells. Recombinant RBPs have been utilized for detection of several pathogens, typically as fusions with reporter enzymes or fluorescent proteins. Identification of Bacillus anthracis, the etiological agent of anthrax, can be difficult because of the bacterium’s close relationship with other species of the Bacillus cereussensu lato group. Here, we facilitated the identification of B. anthracis using two implementations of enzyme-linked phage receptor binding protein assays (ELPRA). We developed a single-tube centrifugation assay simplifying the rapid analysis of suspect colonies. A second assay enables identification of suspect colonies from mixed overgrown solid (agar) media derived from the complex matrix soil. Thus, these tests identified vegetative cells of B. anthracis with little processing time and may support or confirm pathogen detection by molecular methods such as polymerase chain reaction.

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Section: Discussionmentioning

confidence: 99%

Enzyme-Linked Phage Receptor Binding Protein Assays (ELPRA) Enable Identification of Bacillus anthracis Colonies

Braun¹,

Rupprich²,

Neif³

et al. 2021

Viruses

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“…swarmed across the agar plate in only one sample. Although several chromogenic and selective agars are known for the detection of B. anthracis [ 12 , 13 ], maintaining their (including PLET agars) supply is difficult and expensive in laboratories that cover only a few anthrax cases every few years. Another issue for laboratories faced with the diagnosis of sporadic anthrax is the use of M’Fadyean alternative commercial methylene blue stains, which often give mixed results and lead to diagnostic failures [ 14 ].…”

Section: Discussionmentioning

confidence: 99%

A Suggested Diagnostic Approach for Sporadic Anthrax in Cattle to Protect Public Health

et al. 2021

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The repeated occurrence of anthrax in grazing animals should be a reminder of a widespread presence of Bacillus anthracis spores in the environment. Its rapid diagnosis is critical to protect public health. Here, we report a case of anthrax in cattle that was investigated using conventional and molecular methods. In 2015, six cows suddenly died within three days and the number of dead animals increased to a total of 12 within two weeks. At necropsy, anthrax was suspected. Therefore, spleen tissue samples were collected (from 6/12 animals) and laboratory tests (microscopy, cultivation, and real-time PCR) performed. The results of tissue staining for microscopy and cultivation were in congruence, while B. anthracis real-time PCR outperformed both. Spleen tissues from all six animals were real-time PCR-positive, while B. anthracis was successfully cultivated and detected by microscopy from the spleen of only three animals. Additionally, the ear tissue from another (1/12) cow tested positive by real-time PCR, supporting the suitability of ear clippings for molecular confirmation of B. anthracis. Genotyping of the isolates using multiple-locus variable-number tandem repeat analysis (MLVA) revealed a common source of infection as all three typed isolates had an indistinguishable MLVA genotype, which has not been observed previously in Europe. The results indicate that molecular testing should be selected as the first-line tool for confirming anthrax outbreaks in animals to ensure timely protection of public health.

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“…DNA from a picked colony was tested by PCR for B. anthracis -specific markers as described in reference 1 . Additional enrichment of B. anthracis from soil samples was achieved by culturing on semiselective CEFOMA ( Bacillus ce reus sensu lato group-specific antibiotics, fo sfomycin, m acrolides a gar) as described in reference 22 .…”

Section: Methodsmentioning

confidence: 99%

Reoccurring Bovine Anthrax in Germany on the Same Pasture after 12 Years

Braun¹,

Beyer

Hanczaruk³

et al. 2022

J Clin Microbiol

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The zoonotic disease anthrax caused by the endospore-forming bacterium Bacillus anthracis is very rare in Germany. In the state of Bavaria, the last case occurred in July of 2009 resulting in four dead cows. In August of 2021, the disease reemerged after heavy rains, killing one gestating cow. Notably, both outbreaks affected the same pasture, suggesting a close epidemiological connection. B. anthracis could be grown from blood culture and the presence of both virulence plasmids (pXO1 and pXO2) was confirmed by PCR. Also, recently developed diagnostic tools enabled rapid detection of B. anthracis cells and nucleic acids directly in clinical samples. The complete genome of the strain isolated from blood, designated BF-5, was DNA-sequenced and phylogenetically grouped within the B.Br.CNEVA clade that is typical for European B. anthracis strains. The genome was almost identical to BF-1, the isolate of 2009, separated only by three single nucleotide polymorphisms on the chromosome, one on plasmid pXO2 and three indel-regions. Further, B. anthracis DNA was detected by PCR from soil-samples taken from spots, where the cow had fallen onto the pasture. New tools based on phage receptor binding proteins enabled the microscopic detection and isolation of B. anthracis directly from soil-samples. These environmental isolates were genotyped and found to be SNP-identical to BF-5. Therefore, it seems that the BF-5 genotype is currently the prevalent one at the affected premises. The contaminated area at the cadaver was subsequently disinfected with formaldehyde.

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Development of a novel selective agar for the isolation and detection of Bacillus anthracis

Cited by 9 publications

References 27 publications

Enzyme-Linked Phage Receptor Binding Protein Assays (ELPRA) Enable Identification of Bacillus anthracis Colonies

Enzyme-Linked Phage Receptor Binding Protein Assays (ELPRA) Enable Identification of Bacillus anthracis Colonies

A Suggested Diagnostic Approach for Sporadic Anthrax in Cattle to Protect Public Health

Reoccurring Bovine Anthrax in Germany on the Same Pasture after 12 Years

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