Technical Note: Simple, scalable, and sensitive protocol for retrieving Bacillus anthracis (and other live bacteria) from heroin. (February 2016)

• Record Type: Journal Article
• Authors: Grass, Gregor; Ahrens, Bjoern; Schleenbecker, Uwe; Dobrzykowski, Linda; Wagner, Matthias; Krüger, Christian; Wölfel, Roman
• Is Part Of: Forensic science international. Volume 259(2016)
• Page Start: 32
• Page End: 35
• Publication Date: 2016-02
• Subjects: Heroin bacteria -- Endospores -- Bacillus anthracis -- Anthrax -- Cultivation -- Method; Medical jurisprudence -- Periodicals; Chemistry, Forensic -- Periodicals; Forensic Medicine -- Periodicals; Médecine légale -- Périodiques; Chimie légale -- Périodiques; Gerechtelijke geneeskunde; Gerechtelijke chemie; Gerechtelijke psychiatrie; Chemistry, Forensic; Medical jurisprudence; Electronic journals; Periodicals; Electronic journals; 614.1
• DOI: 10.1016/j.forsciint.2015.11.017
• Languages: English
• ISSNs: 0379-0738

Abstract

Highlights: Bacillus anthracis endospores spiked into heroin can be retrieved at 84-98% recovery rate. An aqueous solution of detergent works best for recovery of live bacteria from heroin. A simple, scalable, and sensitive culture-based recovery-method for B. anthracis from heroin was developed. Abstract: We describe a culture-based method suitable for isolating Bacillus anthracis and other live bacteria from heroin. This protocol was developed as a consequence of the bioforensic need to retrieve bacteria from batches of the drug associated with cases of injectional anthrax among heroin-consumers in Europe. This uncommon manifestation of infection with the notorious pathogen B. anthracis has resulted in 26 deaths between the years 2000 to 2013. Thus far, no life disease agent has been isolated from heroin during forensic investigations surrounding these incidences. Because of the conjectured very small number of disease-causing endospores in the contaminated drug it is likely that too few target sequences are available for molecular genetic analysis. Therefore, a direct culture-based approach was chosen here. Endospores of attenuated B. anthracis artificially spiked into heroin were successfully retrieved at 84-98% recovery rates using a wash solution consisting of 0.5% Tween 20 in water. Using this approach, 82 samples of un-cut heroin originating from the German Federal Criminal Police Office's heroin analysis program seized during the period between 2000 and 2014 were tested and found to be surprisingly poor in retrievable bacteria. Notably, while no B. anthracis was isolated from the drug batches, other bacteria were successfully cultured. The resulting methodical protocol is therefore suitable for analyzing un-cut heroin which can be anticipated to comprise the original microbiota from the drug's original source without interference from contaminations introduced by cutting.