Biological risk in a pharmaceutical dump: Brucella abortus strain viable after 20years of disposal

Abstract number: P1671

Koncan R., Bianchi S., Amendola A., Canuti M., Tridente G., Cornaglia G.

Objective: To evaluate the viability of the Brucella abortus B19 vaccine strain, recovered 20 years after storage in sealed vials

Methods: Material of potential biological risk have remained buried for years in the soil of the 12,000 square metres wasteyard of a pharmaceutical research institute in Milan (Istituto Sieroterapico Milanese, ISM). During the ISM reclamation, huge quantities of live vaccine against B. abortus were discovered in hermetically sealed vials. The procedure of reclamation of the ISM waste pit was carried out under strict safety conditions, using individual and environmental protection.

Two vaccine types of B. abortus (in liquid and in lyophilised form) have been unearthed during the process of reclamation and subsequently analyzed. A review of documentation revealed that all these vaccines had been produced by the ISM prior to 1975.

To evaluate the viability of the bacterial strains, vial contents were first inoculated into Brain Heart Infusion (Oxoid). After visible growth the colonies were subcultured onto Columbia and MacConkey agar plates (Oxoid). Brucella species were confirmed by PCR using specific primers based on the AMOS test for Brucella melitensis and B. abortus (JCM, Nov 1994).

Results: Both the liquid and the lyophilised vaccines demonstrated growth on Brain Heart Infusion. After 24 hours of incubation in 5% CO2, non-pigmented non-haemolytic colonies were observed on Columbia agar plates. The lyophilised strain also grew in MacConkey agar on the third day after seeding.

Confirmatory identification was carried out by molecular methods.

Fragments obtained with the AMOS test were sequenced and the BLAST analysis performed revealed that both lyophilised and liquid samples contained B. abortus biovar 1, i.e. the Buck 19 vaccinal strain.

Conclusion: Our results reveal the existence of a biological risk associated with the uncontrolled burial of pharmaceutical industry waste, such as live vaccines. Humans may inadvertently be exposed to products derived from vaccine manufacture by means of unintentional inoculation or other routes of exposure (aerosol, oral, injection).

It is noteworthy that Brucella is classified a class-3 organism and is a potential agent for bioterrorism according to the WHO guidelines.

Session Details

Date: 16/05/2009
Time: 00:00-00:00
Session name: 19th European Congress of Clinical Microbiology and Infectious Diseases
Location: Helsinki, Finland, 16 - 19 May 2009
Presentation type:
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