Bio burden & its implication in pharmaceutical sciences

2. BIO BURDEN
Submitted to: Sir Umair
Submitted by: Faiza Nasar
CMS# 12397
ROLL# 112

3. CONTENTS:
 Definition
 Difference b/w bio burden & biofilm
 Where do biofilm & bio burden occurs?
 Impact of biofilm & bio burden on patients safety
 ATP Bioluminescence test
 Turbidimetric method
 Agar plate count method
 Quantifying bacterial bio burden in Negative Pressure Wound Therapy
 Formulation of disinfectants
 Monitoring of bio burden in pharmaceutical clean rooms
 Microbial bio burden on OSDF
 Bio burden in endoscopy
 Future Horizon

4. BIO BURDEN:
 The term bio burden is used to define the number of
microorganisms on a device or a drug prior to sterilization

5. BIO BURDEN & BIOFILM:
 The number of microorganisms with which an object is
contaminated is referred to as bio burden.
 Biofilm is an assemblage of microbes that are irreversibly associated
with a surface & is enclosed in a matrix primarily of polysaccharide
material.
These two terms are distinct & should not be confused…

6. WHERE DO BIO BURDEN & BIOFILM
OCCURS?
 Biofilm & Bio burden can be found on any
surface that has been exposed to organic
material and/or microbial contaminants.

7. IMPACT OF BIOFILM & BIOBURDEN ON
PATIENTS HEALTH:
 Bacteria within the biofilm can have up to 1000 times
higher resistance to microbes.
 Biofilm is impenetrable to host immunes system
 Is impenetrable to antibiotics
 Account for over 80% of microbial infections.

8. Biofilm & bio burden formation on a
catheters surface after incubation for 24
hours

9. TESTING OF BIO BURDEN:
 ATP Bioluminescence test
 Turbidimetric test
 Agar plate count

10. ATP BIOLUMINESCENCE TEST:
Swab of microbial contaminants + Luciferin + ATP + O2
Oxyluciferin + AMP +Ppi +
Light is directly proportional to the amount of the microbial count

11. TURBIDIMETRIC METHOD:
 Specimen incubated with LAL (limulus amebocyte lysate)
 Rate of in turbidity is measured spectrophotpometrically &
compared with standard test.
Rate of turbidity is directly proportional to microbial count.

12. PLATE COUNT AGAR:
Plate Count Agar (PCA), also called Standard Methods Agar (SMA), is a
microbiological growth medium commonly used to assess
or to monitor "total" or viable bacterial growth of a sample. PCA is not a selective
medium.
The composition of plate count agar may vary, but typically it contains (w/v):
0.5% peptone
0.25% yeast extract
0.1% glucose
1.5% agar
pH adjusted to neutral at 25 C.

13. QUANTIFYING BACTERIAL BIO BURDEN
DURING NEGATIVE PRESSURE WOUND
THERAPY:
 Harmful effects can be observed if microbial count is >10^3 colonies of
bacteria/gram of tissues
 Excellent healing rate is observed
 Clinical levels of bio burden is not yet determined
 Negative pressure causes bacterial clearance
that leads to bio burden levels

14. FORMULATION OF DISINFECTANTS:
 6 categories of disinfectants (alcohols, chlorine, iodophore etc…) were
tested in the absence of bio burden
 Chemicals with best results were retested in the presence of bio burden
 RESULTS: only products containing high ethyl alcohol concentration has
increased the antimicrobial activity…
Better disinfectant formulations were prepared…

15. MONITORING OF BIO BURDEN IN
PHARMACEUTICAL CLEANROOMS:
 FDA has specific guidelines of bio burden for pharmaceutical cleanrooms
 Enhanced electrical filtration(EEF) has reduced bio burden levels in
pharmaceutical cleanrooms.
 EEF HEPA traps & kills S.epidermidis & E.coli

16. MICROBIAL BIO BURDEN IN OSDF:
 Most pharmaceutical companies test oral solid dosage form (OSDF) for
microbial bio burden on a routine basis but it is not necessary.
 More than 70% of microbial bio burden is killed during drying process.

17. ENDOSCOPY BIO BURDEN:
 Endoscopy is a source of nosocomial infection
 Bio burden for upper GIT is ranging from 10^6 – 10^7 CFU
 Bio burden for lower GIT is ranging from 10^8- 10^9 CFU
 Cleaning of endoscope from bio burden reduces the nosocomial
outbreaks.
Cleaning of endoscopy should be observed with the proper disinfectant
before using it

18. FUTURE HORIZON:
 Gamma radiation plant will start running next year in Portugal to sterilize
wine cork stoppers… that leads to decreased bio burden level in wine cork
stopper.
 Portugal is the biggest world producer of wine cork stopper
 Initial bio burden on cork stopper is 10^4 cfu/cork stopper