Lecture 3 - Biosafety Levels and Guidelines | Biotechnology (Zoology) by ILLL, DU - Biotechnology Engineering (BT) PDF Download

 Page 1


Biosafety levels and guidelines 
 
Institute of Lifelong Learning, University of Delhi Page 1 
 
 
 
 
 
 
 
 
 
 
Biotechnology 
Lesson Name: Biosafety levels and guidelines 
Lesson Developer: Dr. Rashmi Tandon 
College/Dept: Department of Microbiology 
V. P. Chest Institute 
Delhi University 
 
 
 
 
 
 
 
 
 
 
 
Page 2


Biosafety levels and guidelines 
 
Institute of Lifelong Learning, University of Delhi Page 1 
 
 
 
 
 
 
 
 
 
 
Biotechnology 
Lesson Name: Biosafety levels and guidelines 
Lesson Developer: Dr. Rashmi Tandon 
College/Dept: Department of Microbiology 
V. P. Chest Institute 
Delhi University 
 
 
 
 
 
 
 
 
 
 
 
Biosafety levels and guidelines 
 
Institute of Lifelong Learning, University of Delhi Page 2 
 
 
Table of Contents        
? Introduction                                               
o Biosafety 
o  Laboratory Acquired Infections (LAIs)                              
? Biosafety Levels             
Laboratory Design and Construction           
o Laboratory Design and Construction – BSL 3        
o Laboratory Design and Construction – BSL 4        
? Safety Equipments              
o Biological Safety Cabinets          
? Laboratory Practices            
o Personal Protection           
o Procedure             
o Lab Working Area           
o Additional Requirement for BSL – 3 Laboratory     
Additional Requirement for BSL – 4 Laboratory    
? Biomedical waste and its management                                     
o Spillage                                                                               
o Radioactive waste and its disposal                                            
o Radioactivity                                                                       
o Radioactive waste                                                              
o Purpose of waste management                                         
o Radioactive waste management techniques                    
o Spillage of radioactive material                                        
? Cartagena protocol on biosafety                                                
? Summary                      
? Practice Questions            
? Glossary              
? References                                                                                  
 
 
 
 
 
 
Page 3


Biosafety levels and guidelines 
 
Institute of Lifelong Learning, University of Delhi Page 1 
 
 
 
 
 
 
 
 
 
 
Biotechnology 
Lesson Name: Biosafety levels and guidelines 
Lesson Developer: Dr. Rashmi Tandon 
College/Dept: Department of Microbiology 
V. P. Chest Institute 
Delhi University 
 
 
 
 
 
 
 
 
 
 
 
Biosafety levels and guidelines 
 
Institute of Lifelong Learning, University of Delhi Page 2 
 
 
Table of Contents        
? Introduction                                               
o Biosafety 
o  Laboratory Acquired Infections (LAIs)                              
? Biosafety Levels             
Laboratory Design and Construction           
o Laboratory Design and Construction – BSL 3        
o Laboratory Design and Construction – BSL 4        
? Safety Equipments              
o Biological Safety Cabinets          
? Laboratory Practices            
o Personal Protection           
o Procedure             
o Lab Working Area           
o Additional Requirement for BSL – 3 Laboratory     
Additional Requirement for BSL – 4 Laboratory    
? Biomedical waste and its management                                     
o Spillage                                                                               
o Radioactive waste and its disposal                                            
o Radioactivity                                                                       
o Radioactive waste                                                              
o Purpose of waste management                                         
o Radioactive waste management techniques                    
o Spillage of radioactive material                                        
? Cartagena protocol on biosafety                                                
? Summary                      
? Practice Questions            
? Glossary              
? References                                                                                  
 
 
 
 
 
 
Biosafety levels and guidelines 
 
Institute of Lifelong Learning, University of Delhi Page 3 
 
 
INTRODUCTION 
Bio-Safety 
The term „bio-safety? comprises of various techniques and practices that must be employed to 
prevent exposure of human beings, animals and the environment to harmful infectious biological 
agents. The concept of biosafety is there to ensure the containment of potentially harmful 
biological agents. Often „biosafety? is confused with „biosecurity? but they are different from each 
other. 
 
 
 
 
Laboratory Acquired Infections (LAIs) 
LAIs are the reason why biosafety rules and practices are needed. It was at the start of 20
th
 
century, when cases of LAIs first got reported. By 1978, 4079 cases of LAIs were identified by 
Pike and Sulkin. Out of these, 168 patients died.
1-4
 The ten most common infective agents that 
were identified were- Brucella spp, Coxiella burnetii, hepatitis B virus (HBV), Salmonella typhi, 
Francisella tularensis, Mycobacterium tuberculosis, Blastomyces dermatitidis, Venezuelan equine 
encephalitis virus, Chlamydia psittaci and Coccidioides immitis. 
Twenty years later, Harding and Byers reported 1267 infections with 22 deaths.
5 
Out of these, 
1074 infections were caused due to M. tuberculosis, Coxiella burnetii, hantavirus, arboviruses, 
HBV, Brucella spp, Salmonella spp., Shigella spp., hepatitis C virus and Cryptosporidium spp. 
Also, research labs are known to contribute more towards LAIs than clinical (diagnostics) 
settings.
6
 All this necessitates the formulation of clear biosafety guidelines to prevent such 
incidences. 
 
BIOSAFETY LEVELS
7
 
There are 4 biosafety levels (BSL-1 through BSL-4) each having its own set of precautions and 
practices to contain the biological agents handled therein. The biosafety levels are nothing but 
designations given to laboratory facilities. It is a sum total of: 
a. laboratory design and construction,  
b. safety equipments and  
c. laboratory practices. 
Before going into the details of each component, it is required to know about the various classes 
of microorganisms that can be handled in each laboratory type. The microorganism are 
 
„Biosecurity? refers to the security measures that are employed to prevent 
theft, loss, intentional misuse of dangerous pathogens and toxins. 
Page 4


Biosafety levels and guidelines 
 
Institute of Lifelong Learning, University of Delhi Page 1 
 
 
 
 
 
 
 
 
 
 
Biotechnology 
Lesson Name: Biosafety levels and guidelines 
Lesson Developer: Dr. Rashmi Tandon 
College/Dept: Department of Microbiology 
V. P. Chest Institute 
Delhi University 
 
 
 
 
 
 
 
 
 
 
 
Biosafety levels and guidelines 
 
Institute of Lifelong Learning, University of Delhi Page 2 
 
 
Table of Contents        
? Introduction                                               
o Biosafety 
o  Laboratory Acquired Infections (LAIs)                              
? Biosafety Levels             
Laboratory Design and Construction           
o Laboratory Design and Construction – BSL 3        
o Laboratory Design and Construction – BSL 4        
? Safety Equipments              
o Biological Safety Cabinets          
? Laboratory Practices            
o Personal Protection           
o Procedure             
o Lab Working Area           
o Additional Requirement for BSL – 3 Laboratory     
Additional Requirement for BSL – 4 Laboratory    
? Biomedical waste and its management                                     
o Spillage                                                                               
o Radioactive waste and its disposal                                            
o Radioactivity                                                                       
o Radioactive waste                                                              
o Purpose of waste management                                         
o Radioactive waste management techniques                    
o Spillage of radioactive material                                        
? Cartagena protocol on biosafety                                                
? Summary                      
? Practice Questions            
? Glossary              
? References                                                                                  
 
 
 
 
 
 
Biosafety levels and guidelines 
 
Institute of Lifelong Learning, University of Delhi Page 3 
 
 
INTRODUCTION 
Bio-Safety 
The term „bio-safety? comprises of various techniques and practices that must be employed to 
prevent exposure of human beings, animals and the environment to harmful infectious biological 
agents. The concept of biosafety is there to ensure the containment of potentially harmful 
biological agents. Often „biosafety? is confused with „biosecurity? but they are different from each 
other. 
 
 
 
 
Laboratory Acquired Infections (LAIs) 
LAIs are the reason why biosafety rules and practices are needed. It was at the start of 20
th
 
century, when cases of LAIs first got reported. By 1978, 4079 cases of LAIs were identified by 
Pike and Sulkin. Out of these, 168 patients died.
1-4
 The ten most common infective agents that 
were identified were- Brucella spp, Coxiella burnetii, hepatitis B virus (HBV), Salmonella typhi, 
Francisella tularensis, Mycobacterium tuberculosis, Blastomyces dermatitidis, Venezuelan equine 
encephalitis virus, Chlamydia psittaci and Coccidioides immitis. 
Twenty years later, Harding and Byers reported 1267 infections with 22 deaths.
5 
Out of these, 
1074 infections were caused due to M. tuberculosis, Coxiella burnetii, hantavirus, arboviruses, 
HBV, Brucella spp, Salmonella spp., Shigella spp., hepatitis C virus and Cryptosporidium spp. 
Also, research labs are known to contribute more towards LAIs than clinical (diagnostics) 
settings.
6
 All this necessitates the formulation of clear biosafety guidelines to prevent such 
incidences. 
 
BIOSAFETY LEVELS
7
 
There are 4 biosafety levels (BSL-1 through BSL-4) each having its own set of precautions and 
practices to contain the biological agents handled therein. The biosafety levels are nothing but 
designations given to laboratory facilities. It is a sum total of: 
a. laboratory design and construction,  
b. safety equipments and  
c. laboratory practices. 
Before going into the details of each component, it is required to know about the various classes 
of microorganisms that can be handled in each laboratory type. The microorganism are 
 
„Biosecurity? refers to the security measures that are employed to prevent 
theft, loss, intentional misuse of dangerous pathogens and toxins. 
Biosafety levels and guidelines 
 
Institute of Lifelong Learning, University of Delhi Page 4 
 
classified into WHO risk groups 1 to 4; with 1 being that of lowest risk to 4 being the one with 
organisms of highest risk. 
The characteristic features of different risk groups are given in Table 1.  The microorganisms 
belonging to WHO risk group 1 are dealt with in a BSL-1 laboratory, risk group 2 pathogens in 
BSL-2, risk group 3 agents in BSL-3.   
For more information on biosafety levels, click on the link below: 
http://www.youtube.com/watch?v=UmP9GqEr00w 
 
Table 1: WHO classification of risk groups 
 RISK GROUP 1 RISK GROUP 2 RISK GROUP 3 RISK GROUP 4 
Causes 
human/animal 
disease 
No Yes Yes Yes 
Serious 
infection 
No May / may not Yes Yes 
Spread of 
infection 
No Limited No Yes 
Availability of 
treatment 
No Yes Yes No 
Individual risk No/very low Moderate High High 
Community risk No/very low Low Low High 
Examples
a  
of 
infectious 
agents 
Escherichia coli, 
Bacillus subtilis, 
Bacillus 
licheniformis 
C. difficile, most 
Chlamydiae, 
hepatitis A, B, 
and C, 
orthopoxviruses 
(other than 
smallpox), 
influenza A, 
Borrelia 
burgdorferi sensu 
stricto, Borrelia 
afzelii, Borrelia 
garinii, 
Salmonella, 
mumps virus 
(Rubulavirus sp.), 
measles 
(Morbillivirus), 
Methicillin-
resistant 
Staphylococcus 
aureus (MRSA), 
and Vancomycin-
resistant 
Staphylococcus 
aureus (VRSA) 
Yersinia pestis, 
Francisella 
tularensis, 
Leishmania 
donovani, 
Mycobacterium 
tuberculosis, 
Chlamydia psittaci, 
Venezuelan equine 
encephalitis virus, 
Eastern equine 
encephalitis virus, 
SARS coronavirus, 
Coxiella burnetii, 
Rift Valley fever 
virus, Rickettsia 
rickettsii, several 
species of Brucella, 
rabies virus, 
chikungunya, yellow 
fever virus, and 
West Nile virus. 
Bolivian and 
Argentine 
hemorrhagic 
fevers, Marburg 
virus, Ebola 
virus, Lassa 
virus, Crimean-
Congo 
hemorrhagic 
fever, and 
various other 
hemorrhagic 
disease 
 
a
 http://en.wikipedia.org/wiki/Biosafety_level         
It must also be noted that laboratories where in biological organisms are handled that can cause 
infection, a biohazard warning sign (Figure 1) must be put up on the laboratory doors. 
Page 5


Biosafety levels and guidelines 
 
Institute of Lifelong Learning, University of Delhi Page 1 
 
 
 
 
 
 
 
 
 
 
Biotechnology 
Lesson Name: Biosafety levels and guidelines 
Lesson Developer: Dr. Rashmi Tandon 
College/Dept: Department of Microbiology 
V. P. Chest Institute 
Delhi University 
 
 
 
 
 
 
 
 
 
 
 
Biosafety levels and guidelines 
 
Institute of Lifelong Learning, University of Delhi Page 2 
 
 
Table of Contents        
? Introduction                                               
o Biosafety 
o  Laboratory Acquired Infections (LAIs)                              
? Biosafety Levels             
Laboratory Design and Construction           
o Laboratory Design and Construction – BSL 3        
o Laboratory Design and Construction – BSL 4        
? Safety Equipments              
o Biological Safety Cabinets          
? Laboratory Practices            
o Personal Protection           
o Procedure             
o Lab Working Area           
o Additional Requirement for BSL – 3 Laboratory     
Additional Requirement for BSL – 4 Laboratory    
? Biomedical waste and its management                                     
o Spillage                                                                               
o Radioactive waste and its disposal                                            
o Radioactivity                                                                       
o Radioactive waste                                                              
o Purpose of waste management                                         
o Radioactive waste management techniques                    
o Spillage of radioactive material                                        
? Cartagena protocol on biosafety                                                
? Summary                      
? Practice Questions            
? Glossary              
? References                                                                                  
 
 
 
 
 
 
Biosafety levels and guidelines 
 
Institute of Lifelong Learning, University of Delhi Page 3 
 
 
INTRODUCTION 
Bio-Safety 
The term „bio-safety? comprises of various techniques and practices that must be employed to 
prevent exposure of human beings, animals and the environment to harmful infectious biological 
agents. The concept of biosafety is there to ensure the containment of potentially harmful 
biological agents. Often „biosafety? is confused with „biosecurity? but they are different from each 
other. 
 
 
 
 
Laboratory Acquired Infections (LAIs) 
LAIs are the reason why biosafety rules and practices are needed. It was at the start of 20
th
 
century, when cases of LAIs first got reported. By 1978, 4079 cases of LAIs were identified by 
Pike and Sulkin. Out of these, 168 patients died.
1-4
 The ten most common infective agents that 
were identified were- Brucella spp, Coxiella burnetii, hepatitis B virus (HBV), Salmonella typhi, 
Francisella tularensis, Mycobacterium tuberculosis, Blastomyces dermatitidis, Venezuelan equine 
encephalitis virus, Chlamydia psittaci and Coccidioides immitis. 
Twenty years later, Harding and Byers reported 1267 infections with 22 deaths.
5 
Out of these, 
1074 infections were caused due to M. tuberculosis, Coxiella burnetii, hantavirus, arboviruses, 
HBV, Brucella spp, Salmonella spp., Shigella spp., hepatitis C virus and Cryptosporidium spp. 
Also, research labs are known to contribute more towards LAIs than clinical (diagnostics) 
settings.
6
 All this necessitates the formulation of clear biosafety guidelines to prevent such 
incidences. 
 
BIOSAFETY LEVELS
7
 
There are 4 biosafety levels (BSL-1 through BSL-4) each having its own set of precautions and 
practices to contain the biological agents handled therein. The biosafety levels are nothing but 
designations given to laboratory facilities. It is a sum total of: 
a. laboratory design and construction,  
b. safety equipments and  
c. laboratory practices. 
Before going into the details of each component, it is required to know about the various classes 
of microorganisms that can be handled in each laboratory type. The microorganism are 
 
„Biosecurity? refers to the security measures that are employed to prevent 
theft, loss, intentional misuse of dangerous pathogens and toxins. 
Biosafety levels and guidelines 
 
Institute of Lifelong Learning, University of Delhi Page 4 
 
classified into WHO risk groups 1 to 4; with 1 being that of lowest risk to 4 being the one with 
organisms of highest risk. 
The characteristic features of different risk groups are given in Table 1.  The microorganisms 
belonging to WHO risk group 1 are dealt with in a BSL-1 laboratory, risk group 2 pathogens in 
BSL-2, risk group 3 agents in BSL-3.   
For more information on biosafety levels, click on the link below: 
http://www.youtube.com/watch?v=UmP9GqEr00w 
 
Table 1: WHO classification of risk groups 
 RISK GROUP 1 RISK GROUP 2 RISK GROUP 3 RISK GROUP 4 
Causes 
human/animal 
disease 
No Yes Yes Yes 
Serious 
infection 
No May / may not Yes Yes 
Spread of 
infection 
No Limited No Yes 
Availability of 
treatment 
No Yes Yes No 
Individual risk No/very low Moderate High High 
Community risk No/very low Low Low High 
Examples
a  
of 
infectious 
agents 
Escherichia coli, 
Bacillus subtilis, 
Bacillus 
licheniformis 
C. difficile, most 
Chlamydiae, 
hepatitis A, B, 
and C, 
orthopoxviruses 
(other than 
smallpox), 
influenza A, 
Borrelia 
burgdorferi sensu 
stricto, Borrelia 
afzelii, Borrelia 
garinii, 
Salmonella, 
mumps virus 
(Rubulavirus sp.), 
measles 
(Morbillivirus), 
Methicillin-
resistant 
Staphylococcus 
aureus (MRSA), 
and Vancomycin-
resistant 
Staphylococcus 
aureus (VRSA) 
Yersinia pestis, 
Francisella 
tularensis, 
Leishmania 
donovani, 
Mycobacterium 
tuberculosis, 
Chlamydia psittaci, 
Venezuelan equine 
encephalitis virus, 
Eastern equine 
encephalitis virus, 
SARS coronavirus, 
Coxiella burnetii, 
Rift Valley fever 
virus, Rickettsia 
rickettsii, several 
species of Brucella, 
rabies virus, 
chikungunya, yellow 
fever virus, and 
West Nile virus. 
Bolivian and 
Argentine 
hemorrhagic 
fevers, Marburg 
virus, Ebola 
virus, Lassa 
virus, Crimean-
Congo 
hemorrhagic 
fever, and 
various other 
hemorrhagic 
disease 
 
a
 http://en.wikipedia.org/wiki/Biosafety_level         
It must also be noted that laboratories where in biological organisms are handled that can cause 
infection, a biohazard warning sign (Figure 1) must be put up on the laboratory doors. 
Biosafety levels and guidelines 
 
Institute of Lifelong Learning, University of Delhi Page 5 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Figure 1: Biohazard warning sign 
Source: http://www.who.int/csr/resources/publications/biosafety/en/Biosafety7.pdf 
A.  Laboratory Design and Construction 
The laboratories must be designed keeping in mind the kind of work to be carried out. The 
designs for typical biosafety levels 1 and 2 laboratories are as given in figures 2 and 3. The 
laboratories are exactly similar to each other except for the fact that BSL-2 laboratory has an 
autoclave for treating all the contaminated waste prior to disposal. In order to qualify as a BSL 1 
or 2 laboratory, a lab must have the following features: 
? It should be spacious enough to provide for smooth working of the lab, carrying out 
routine jobs along with easy cleaning and maintenance of lab. Overcrowding of 
equipments or other materials should not be there. 
? Floor shouldn?t be slippery. 
? Bench tops (working platform), floor and walls must be impervious to liquids and 
resistant to various disinfectants that are used in the laboratory. 
? Labs should be well lit. 
? Basin for washing hands must be there, preferably near the exit of the lab. 
? Sufficient storage space must be there. 
? Gas connections must be proper. 
? Self closing gates with a vision panel as shown in the figures 3 and 4. 
? First aid facility must be available in close vicinity. 
One of the important features of a biosafety lab is the biosafety cabinets which shall be 
discussed in the following section.