What's New in Biosafety

By: Leslie Hofherr, MS, MPH, CBSP

January 1o, 2011

As a service to my clients and colleagues, I am very pleased to present a summary of important recent biosafety changes prepared by Leslie Hofherr, MS, MPH, CBSP.  Leslie has rich experience in biosafety and has assisted a number of my clients in biosafety matters over the years.


Biosafety compliance guidance and regulatory requirements have changed significantly in the last 3 years. Cal/OSHA has promulgated new regulations covering aerosol transmissible diseases (ATDs) and the primary biosafety compliance guidelines have been updated. In the future we can expect an increase in regulatory oversight of biohazardous materials due to concerns of the general public and congress for the spread of infectious diseases and bioterrorism, and changes in technology.


Below is a summary of some important changes and reminders for biosafety compliance in the areas of:

Cal/OSHA Aerosol Transmissible Disease Standards (ATD)

Biosafety in Microbiological and Biomedical Laboratories (BMBL) 5th edition

Cell Line Use and Safety

NIH Guidelines for Research Involving Recombinant DNA Molecules


ATDCal/OSHA Aerosol Transmissible Disease Standards (ATD):

Effective August 5, 2009.


http://www.dir.ca.gov/title8/5199.html   http://www.dir.ca.gov/title8/5199-1.html

The new standards focus primarily, but not entirely, on employers working in the health care industry. There are two new Cal/OSHA ATD standards.

The more narrow "zoonotics" standard, 8 CCR 5199.1, applies only to employers that handle animals or animal by-products, and is intended to prevent the transmission of diseases between animals and humans.These regulations require compliance by services that capture, sample, transport or dispose of birds and other wildlife; farms producing animals or animal products (including untreated animal products, byproducts or wastes); slaughterhouses; veterinary animal inspection; importers of live or untreated animals or animal products; zoos; animal parks; pet stores; and laboratory operations.

These covered facilities and services must follow the standard whenever an employee is involved in capturing or sampling of animals to detect the presence of infection with zoonotic ATDs or is involved in the transportation or disposal of animals suspected to be infected with zoonotic ATD or covered under a quarantine or alert issued by Centers for Disease Control, California Department of Food and Agriculture, California Department of Fish and Game, California Department Public Health, U.S. Department of Agriculture or U.S. Department of the Interior.


The much broader ATD standard, 8 CCR 5199, covers, but is not limited to, health care facilities including hospitals, nursing facilities, clinics, medical offices, long-term care providers, and emergency services and transport providers. The standard also covers other defined "high risk" workplaces where the exposure of ATDs is deemed more likely, including homeless shelters, drug treatment programs, correctional facilities, and air-handling machinery maintenance or repair operations where contamination might be expected. The standard further covers certain laboratories and police services.


There are four classes of employers identified that have different compliance requirements: Covered, Referring, Laboratory, or Exempt employers. Below is a summary of the Laboratory Employer requirements.


 Aerosol Transmissible Diseases - Laboratories

Workers in clinical and research biological laboratories are at risk of contracting aerosol transmissible diseases (ATDs) due to laboratory procedures that generate aerosols. Laboratory workers have contracted serious diseases including tuberculosis (TB), SARS, and meningococcal disease. Pathogens such as brucellosis, which are not generally transmitted between people can also be transmitted by laboratory aerosols. The Aerosol Transmissible Disease (ATD) standard (Section 5199) requires laboratories to adopt standard biosafety practices to protect laboratory workers when handling materials containing pathogens that may be spread through aerosols and which can cause serious disease. A minimum list of these pathogens is included in Appendix D of the ATD Standard. The standard requires that laboratories implement control measures that are consistent with the recommendations of the U.S. Centers for Disease Controls and Prevention (CDC)for handling these materials. When laboratory workers have direct contact with patients the employer is required to comply with other requirements of the standard applicable to patient contact.

The ATD standard requires laboratory employers to use feasible engineering and work practice controls to limit exposure to aerosols, and to provide personal protective equipment and respirators when that equipment is necessary to control exposures. In addition, the employer is required to develop, implement, and annually review, a written Biosafety Plan (BSP)that includes:

1. Identification of qualified biosafety officer(s) who will be responsible for implementing, reviewing and updating the BSP, and for reviewing plans to modify the facility.

2. A list of job classifications, tasks and procedures in which employees may be exposed to ATPs-L.

3. A list of the ATPs-L that are present or anticipated to be present in materials in the laboratory.

4. Safe handling procedures and a list of prohibited practices, such as sniffing in vitro cultures, that may increase employee exposure to infectious agents.

5. Engineering controls, including containment facilities such as biosafety cabinets.

6. Procedures requiring the use of personal protective equipment and/or respirators.

7. Effective decontamination and disinfection procedures for laboratory surfaces and equipment.

8. A requirement that all incoming materials containing ATPs-L be treated as containing the virulent or wild-type pathogen, until procedures conducted at the laboratory verify they are deactivated or attenuated.

9. Inspection procedures including an audit of biosafety procedures, to be performed at least annually.

10. Emergency procedures for uncontrolled releases within the laboratory facility and untreated releases outside the laboratory facility including reporting such incidents to the local health officer.

11. Procedures for medical surveillance, including:

a. vaccinations as recommended by the CDC or California Department of Public Health

b. annual tests for latent TB infection in clinical laboratories and in laboratories where materials containing M. tuberculosis may be present.

c. medical follow-up for employees who have had a significant exposure to an ATP-L without benefit of applicable required control measures.

12. Procedures for initial and annual employee training.

13. Procedures to involve employees in the evaluation of the effectiveness of the BSP in their work areas.

The employer is required to keep records of training, medical surveillance and exposure incidents, inspections, and evaluation of engineering controls and other control measures.


BMBLUpdated CDC/NIH  "Biosafety in Microbiological and Biomedical Laboratories" (BMBL) to the 5th edition, 2007


First introduced in 1984, BMBL is an advisory document recommending best practices for the safe conduct of work in biomedical and clinical laboratories. Since its inception, it has become one of the most frequently used codes of practice in biosafety, and an authoritative reference for: the development of laboratory policies and procedures, the construction of new laboratories, and the renovation of existing laboratories. Periodic updates have been made to the BMBL to "refine guidance based on new knowledge and experiences and to address contemporary issues that present new risks that confront laboratory workers and the public health".


General revisions include update of Biosafety Level 1-4 containment criteria, agent summary statements, risk assessment section, biosecurity section, agricultural Biosafety Level 3 section, and expanded guidance on a number of topics, including decontamination, sterilization, occupational medicine, and immunization.  Below are listed the major revisions for Biosafety Level 1 (BSL-1) and BSL-2 compliance.


Biosafety Level 1 (BSL-1) Laboratory updates:

Standard Microbiological Practice

  • Access control to laboratory is enforced by lab supervisor.
  • Sharps use policy and controls must be developed and implemented to reduce sharps injury.
  • Biohazard door card is required when infectious agents are present in the lab.
  • Agent infomation is posted on door card as per the institutional policy.

Safety Equipment

  • Gloves must be worn to protect against exposure and selected based on a risk assessment.
  • Persons who wear contact lenses in the labs should also wear eye protection.

Laboratory Facility

  • Chairs used in lab work must be covered with non-porous materials that can be easily cleaned and decontaminated with appropriate disinfectant.

Biosafety Level 2 (BSL-2) Laboratory updates (including BSL-1 updates above):


  • Provide all lab personnel with information regarding immune competence, conditions that may predispose them to infection, and encouragement to self identify to your healthcare provider.
  • A lab-specific biosafety manual is required.
  • Document lab personnel proficiency in standard and special microbiological practices before they work with BSL-2 agents.
  • Spills involving infectious materials are contained, decontaminated, and cleaned up by staff properly trained and equipped to work with infectious material.
  • All procedures involving the manipulation of infectious materials that may generate an aerosol should be conducted within a BSC or other physical containment devices.

Safety Equipment

  • Eye and face protection, and gloves must be disposed of with other contaminated lab waste or decontaminated before reuse if reusable.
  • Eye, face, and respiratory protection should be used in rooms containing infected animals as determined by the risk assessment.

Laboratory Facility

  • Lab doors should be self-closing and have locks as described in the institutional policies.
  • The sink should be located near the exit door.
  • Lab windows that open to the exterior are not recommended.
  • Vacuum lines should be protected with HEPA filters or their equivalent and liquid disinfectant traps as needed. 
  • At least annual certification of Class II Type A BSC required if BSC air is re-circulated back into the lab. BSC must be operated according to manufacturer's recommendations. Institute provisions to assure proper safety cabinet performance and BSC air system operation must be verified.
  • A method for decontaminating all lab wastes should be available in the facility (e.g., autoclave, chemical disinfection, incineration, or other validated decontamination method).

cellsCell line use hazards and safety


The risk of a laboratory infection from working with cell cultures in general is low, however, risk increases when working with human and other primate cells, and primary cells from other mammalian species. There are reports of infection of laboratory workers handling primary rhesus monkey kidney cells and the bloodborne pathogen risks from working with primary human cells, tissues and body fluids are widely recognized.


Potential laboratory hazards associated with human cells and tissues include the bloodborne pathogens HBV, HIV, HCV, HTLV, EBV, HPV and CMV as well as agents such as Mycobacterium tuberculosis that may be present in human lung tissue. Other primate cells and tissues also present risks to laboratory workers.Cells immortalized with viral agents such as SV-40, EBV adenovirus or HPV, as well as cells carrying viral genomic material also present potential hazards to laboratory workers. Tumorigenic human cells also are potential hazards.There has been one reported case of development of a tumor from an accidental needle-stick.


All primary human cell cultures (explants) and subsequent in vitro passages fall under the OSHA BBP standard. To be exempted from the BBP requirements, primary cell cultures and lines must undergo testing and characterization (documented) for blood-borne pathogens such as HBV, HCV and HIV and others.

Safety precautions

  • Laboratory workers should never handle autologous cells or tissues
  • Conduct a risk assessment based on the origin of the cells or tissues (species and tissue type), as well as the source (recently isolated or well characterized
  • Human source cells require compliance with the Cal/OSHA Bloodborne Pathogens (BBP) Standard and BSL-2
  • Use "Universal Precautions" for all human and primate cell cultures
  • Perform work involving human and primate cells following BSL-2 containment
  • Perform all work in a biosafety cabinet (BSC)
  • Autoclave or disinfect all cell contaminated materials prior to discarding
  • Wear PPE while performing work with cells - lab coat, gloves, eye protection at a minimum
  • Follow occupational medicine program for BBP Standard
  • Follow company specific BBP Exposure Control Plan policies and procedures




NIHNIH Guidelines for Research Involving Recombinant DNA Molecules Update September 2009



The "NIH Guidelines" provides biosafety guidance covering uses of recombinant DNA. In September 2009 the "NIH Guidelines" were updated to better cover recombinant influenza viruses uses. With the advent of new recombinant technology "designer" influenza viruses can be created including the 1918 H1N1 strain. The amendments clarify and augment the current guidance for research with potentially pandemic influenza virus and harmonize with the CDC/NIH Biosafety in Microbiological and Biomedical Laboratories (5th edition) and other regulatory policies. Section III-D-7 provides guidance regarding the biosafety level containment for research with influenza viruses generated by recombinant methods. In Appendix B, the potentially pandemic influenza viruses human H2N2 (1957-1968), 1918 H1N1, and HPAI H5N1 are classified as Risk Group 3 agents. Appendix G-II-C-5 provides additional biosafety guidance for research with these influenza viruses including Biosafety Level 3 enhanced containment, practices and training, animal containment and occupational health.


Some other actions at the NIH Office of Biotechnology (OBA) in 2010:

  • At its December 7-8, 2010 meeting, the NIH Recombinant DNA Advisory Committee (RAC) discussed a proposal to certify Kluyveromyces lactis as a new host-vector system under the NIH Guidelines for Research Involving Recombinant DNA Molecules.
  • In July 2010, the NIH Office of Biotechnology Activities (OBA) published a proposal to revise the NIH Guidelines to exempt the breeding of well-characterized transgenic rodents that can be maintained under Biosafety Level (BL) 1 conditions. This exemption would not apply, however, to the breeding of rodents that have a gene encoding more than fifty percent of an exogenous eukaryotic virus or those transgenic rodents in which the transgene is under the control of a gammaretroviral promoter. Currently these proposed changes have not been incorporated into the "NIH Guidelines".
  • In 2010, OBA sent a memorandum to inform OBA-registered investigators who are using or propose to use lentiviral or retroviral vectors in hematopoietic or other stem cells that a "relative clonal dominance" was detected during follow-up of a subject who is participating in a French human gene transfer trial being conducted for individuals with β-Thalassemia Major and Sickle Cell Anemia.
  • In March and May 2010, OBA proposed to broadening the scope of the NIH Guidelines, to encompass nucleic acids that are synthesized chemically or by other means without the use of recombinant technology, changing the review level for recombinant or synthetic experiments involving more than half but less than two-thirds of the genome of certain viruses in tissue culture, as described in Section III-E-1 of the NIH Guidelines, and Revising the criteria for determining when introduction of a drug resistance trait into a microorganism must be reviewed and approved by the NIH Director. Currently these proposed changes have not been incorporated into the "NIH Guidelines".

 Synthetic biology has advanced to the point where some pathogens can be manufactured from scratch. This technical leap has beneficent implications for medical research and vaccine design, but it also raises concerns that the technology could be used to produce a deadly pathogen for nefarious use. Federal government agencies have reviewed the advances in synthetic biology but have not moved to regulate its use yet." 

Services I Provide 
Contact me and I would be happy to discuss ways I can assist you and your company meet your obligations and maintain a safe work place.  Some of the related services I provide include:
artServed by an Expert
Arthur Mahoney, Principal Consultant

For 17 years, he has been assisting public and private companies to comply with safety, hazardous material management, transportation and safety concerns. He also provides chemical inventory database services.


Leslie Hofherr, MS, MPH, CBSP
For over 20 years she has been assisting companies and educational  institutions in biosafety and health and safety matters.  
Feel free to contact us at 650-347-0417 or by email

Arthur Mahoney
Hazard Solutions LLC

Disclaimer:  The information presented above should not be construed in any way as legal advice or an interpretation of regulations.  It is meant to provide basic information about topics that may affect clients and colleagues.



Home | Training | Services | Onsite EHS Services | Chemical Storage & Risk Management Assistance
Database Solutions | Resources | About | Contact