Burkholderia cepacia complex :Approach for Microbiological Examination of Pharmaceutical Product

Burkholderia cepacia complex :Approach for Microbiological Examination of Pharmaceutical Product

Kingdom	Bacteria
Phylum	Proteobacteria
Class	Betaproteobacteria
Order	Burkholderiales
Family	Burkholderiaceae
Genus	Burkholderia
Species	B. cepacia complex
Binomial name	Burkholderia cepacia complex

Burkholderia cepacia complex is one of the most common bacterial species found in contaminated pharmaceutical products (water-based drug products).

Burkholderia cepacia complex can survive trough different antiseptics, preservatives and resist several antibiotics. Burkholderia cepacia complex can grow and survive in pharmaceutical products, active ingredients, and excipients.

Most probable root causes of contamination with Burkholderia cepacia complex in pharmaceutical industries:

• Inadequate cleaning procedures and lack of cleaning validation studies.
• Use of unsuitable grade of water as potable water.
• Poor water system control.
• Poor water system design.
• Insufficient microbiological controls.
• Lack of environmental floral studies.
• Using specific type of disinfectant for long period of time.

As there are currently no compendial methods for detection of Burkholderia cepacia complex, we have provided suggestions for a potential validation approach and some points to consider when designing your validation studies. However, any validated method capable of detecting Burkholderia cepacia complex organisms would be adequate. It is currently sufficient to precondition representative strain(s) of Burkholderia cepacia complex in water and/or your drug product to demonstrate that your proposed method is capable of detecting small numbers of Burkholderia cepacia complex.

SUGGESTED METHOD

Sample Preparation and Pre-Incubation: Prepare a sample using a 1 in 10 dilution of not less than 1 g of the product to be examined as described in preparation of sample and use 10 mL or the quantity corresponding to 1 g or 1 mL, to inoculate a suitable amount of Soybean-Casein Digest Broth, mix and incubate at 30°C to 35°C for 18 to 24 hours.

Selection and Subculture: Subculture on a plate of Burkholderia Cepacia Selective Agar , and incubate at 35°C to 37°C for period of 4 days.

Interpretation: Growth of colonies indicates the possible presence of Burkholderia cepacia complex. This is confirmed by Microbial identification system.

Test organisms use for Growth promotion, indicative and inhibitory test of Burkholderia Cepacia Selective Agar Media:

Growth promotion + Indicative:

Burkholderia cepacia ATCC25608

(Sage Green Colonies with Bright Pink Medium)

Inhibitory:

Pseudomonas aeruginosa ATCC 9027

*Staphylococcus aureus ATCC 6538

*Escherichia coli ATCC 8739

*Note: validation must be done for Inhibition properties with above organisms.

Manufacturer of media:

**Hi-media: Burkholderia Cepacia Agar Base M1640+Burkholderia Cepacia Selective FD232

**Growth promotion, indicative and inhibitory test must be revised accordingly if manufacturer of media is different. 

REFERENCE

• https://www.fda.gov/Drugs/DrugSafety/ucm559508.htm https://www.fda.gov/downloads/Drugs/DevelopmentApprovalProcess/SmallBusinessAssistance/UCM547287.pdf
• http://medcraveonline.com/IJVV/IJVV-03-00064.pdf
• http://www.americanpharmaceuticalreview.com/Featured-Articles/160451-Recovery-of-Stressed-Acclimated-Burkholderia-cepacia-Complex-Organisms/
• https://www.fda.gov/downloads/aboutfda/centersoffices/cder/ucm275569.pdf
• http://himedialabs.com/TD/M1640.pdf
• http://himedialabs.com/TD/FD232.pdf