Mostafa BentahirCentre de Technologies Moléculaires Appliquées (CTMA)

Defence Laboratories (DLD)03-04 December 2018

BFREE

Safe handling and preparation of CBRN mixed samples:

Biological challenges and solutions

Presented at the Civil-Military Cooperation in CBRN-E

12th CoU Meeting

Brussels

www.eda.europa.eu2

MAIN OBJECTIVE

The aim of BFREE was to obtain an efficient sample processing and risk mitigation

method for both ensuring safe handling and preparation of mixed CBRN samples for

the following analysis of CBRN agents.

• Harmonised approach for receiving and handling CBRN

mixed samples, considering;- reception, screening, sample preparation, laboratory analysis, safety practises, choice

of equipment

• Removal of biological contamination from CBRN mixed

samples- for safe analysis of chemical and radiological agents

www.eda.europa.eu3

FINAL RESULTS

The BFREE consortium proposed a Recommended Operating Procedure (ROP)

for receiving and handling of mixed CBRN threat samples

The procedure was divided into four major steps

1. package receipt and transport container screening

2. screening of outer and inner container

3. sample screening and

4. documentation of results.

1: Package receipt and transport container screening

Performed preferably in a separated area with good security and access controls

Hold a Team of Expert (ToE) meeting

Establish/continue Chain of custody

Review corresponding documentation and interview the delivery person

If possible communicate with the sampling team

Visually inspect the transport container and make photo documentation

Check for explosive device – screen for radiation and volatile chemicals

Prepare sample for transport to BSL3 or equivalent laboratory

Document observations, complete Sample Receipt Forms

Carry out threat assessment and develop an analysis plan

www.eda.europa.eu4

FINAL RESULTS

For the removal of biological contamination the BFREE project recommends using a three step protocol/method

1) double filtration through 0.2 µm (removal of spores and bacteria)

2) a 30 kDa filter (removal of virus)

3) ultra violet (UV) irradiation (inactivation of residual microorganisms).

Simple

Easy to use

Suited for field laboratories

Quick, less than 10 minutes

Cheap

www.eda.europa.eu5

MILITARY BENEFITS

Sweden: The National CBRN Defence Centre is the Swedish Armed Forces’ knowledge centre for

protection against chemical, biological, radiation and nuclear threats. Within in their

organisation their Deployable Field Laboratories are during the fall 2018 they updating

the protocols for handling and analysing mixed CBRN samples based on the results from

the BFREE project.

Norway: The Norwegian Defence Research Establishment are working on the protocols but the

procedures have not yet been implemented.

Austria: They have since two years ago a new BSL-3 laboratory. Within this facility, there is a NBC-

isolator-line for being able to handle and analyse mixed CBRNE samples. The procedures

and protocols are based on the results of BFREE. This laboratory is sub-ordinated the

MoD for military systems. There is also an ongoing cooperation between the Public

Service and Armed Forces.

Holland: They have chosen to follow a slightly different approach for the elimination of B-agents

with extraction of samples with organic solvents followed by centrifugation.

Germany and France: No information

www.eda.europa.eu6

MILITARY BENEFITS

Belgium: The Belgium laboratory (CTMA/DLD) has continued to develop and validate

the filter methods. They are also working on procedures for powder and soil

samples.

Peer-review article of the double filtration method for safe handling and

separation of mixed CBRN samples.

Development of microfluidic filtration device

monitoring of microbial contamination of the surface of the parcel (Outer

assessment with hand-Held device

Ongoing work of inactivation of Bio-agent with high energy UVC

www.eda.europa.eu7

MILITARY BENEFITS

Peer review publication

www.eda.europa.eu8

MILITARY BENEFITS microfluidic filtration device prototype

www.eda.europa.eu9

MILITARY BENEFITS Monitoring surface contamination

www.eda.europa.eu10

MILITARY BENEFITS Monitoring surface contamination

www.eda.europa.eu11

MILITARY BENEFITS: Monitoring surface contamination

www.eda.europa.eu12

WAY FORWARD

Even though BFREE achieved good results, the conclusion and discussion

identified the need for follow up work.

• Changes in chemical composition due to oxidation caused by UV irradiation.

• Develop a closed system for performing UV irradiation.

• Extend the evaluation of the “BFREE method” to other sample matrixes (i.e., powders).

• Extend the evaluation of the “BFREE method” to real biological weapon agents (BWA).

• Extend the evaluation of the “BFREE method” to more chemicals.

• Reactivations of DNA damages caused by UV irradiation in bacterial agents.

• Evaluate more filter types in order to improve bacterial/virus removal.

• Study transfer of various biological threat agents from water to organic phase.

www.eda.europa.eu13

• Swedish Defence Research Agency (FOI)

• Bundeswehr Research Institute for Protective

Technologies and NBC‐Protection (WIS)

• Centre for Applied Molecular

Technologies/Defence Department

Laboratories (CTMA/DLD‐Bio)

• Norwegian Defence Research Establishment

(FFI)

• The Netherlands Organization For Applied

Scientific Research (TNO)

• The Armaments and Defence Technology

Agency (ARWT)

• DGA CBRN Defence

Acknowledgement