Biosafety Manual

Purpose and Scope

The purpose of this program is to define the biological safety policies and procedures pertaining to research operations at Wellesley College. These policies and procedures are designed to safeguard personnel and the environment from biologically hazardous materials and to comply with federal and state regulatory requirements. All Faculty and laboratory employees must adhere to the biological safety policies and procedures in the conduct of their research and the management of their laboratories.

Biological agents include all infectious microorganisms (bacteria, fungi, parasites, prions, rickettsias, viruses, etc.) that can cause disease in humans or pose significant environmental or agricultural impact, as well as the toxins derived from such organisms. Additionally, recombinant or synthetic nucleic acid molecules; human or non-human primate tissues, fluids, cells, or cell cultures; transgenic plants or animals; and any work with animals and their tissues, which are known to be reservoirs of zoonotic diseases, are wholly or partly covered by the procedures and policies in this manual.

For information about specific biological safety programs for operations not covered in this manual, contact a member of the Institutional Biosafety Committee.


Rules and Regulations

The following is a summary of federal and state regulations and guidelines that either regulate or provide guidelines covering the use of biological agents:

  • Centers for Disease Controls and Prevention and National Institutes of Health: Biosafety in Microbiological & Biomedical Laboratories (BMBL), 6th Edition June 2020. This document contains guidelines for microbiological practices, safety equipment, and facilities that constitute the four established biosafety levels. The BMBL is generally considered the standard for biosafety and is the basis for this manual.
  • National Institutes of Health: Guidelines for Research Involving Recombinant or Synthetic Nucleic Acid Molecules (NIH Guidelines). This document provides guidelines for constructing and handling recombinant or synthetic nucleic acid molecules and organisms containing these molecules. Although these guidelines are not subject to regulatory enforcement, institutions that receive any NIH funding for recombinant or synthetic nucleic acid molecules research are required to comply with these guidelines as a condition of funding. This document requires that institutions establish an Institutional Biosafety Committee with the authority to approve proposed research using the NIH guidelines as the minimum standard.
  • Occupational Safety and Health Administration: Bloodborne Pathogens. This regulation covers occupational exposure to human blood and other potentially infectious materials, including human tissue and cells. OSHA specifies a combination of engineering controls, work practices, and training to reduce the risk of infection. Personnel potentially exposed to human blood and other potentially infectious material must be offered immunization against hepatitis B and receive annual training. Personnel who work with HIV or hepatitis B in a research laboratory must receive additional training and demonstrate proficiency in working with human pathogens (29 CFR 1030).
  • Commonwealth of Massachusetts Department of Public Health: The Center for Environmental Health regulates the storage and disposal of potentially infectious material, and includes requirements for labeling and recordkeeping (105 CMR 480).

Select Agent Rule

The following regulations require institutions that possess, use, or transfer certain biological agents and toxins (“select agents”) to be registered and approved by DHHS and/or APHIS:

  • Department of Health and Human Services: Possession, Use, and Transfer of Select Agents and Toxins (42 CFR Part 73);
  • Department of Agriculture: Animal and Plant Health Inspection Service (7 CFR Part 331);
  • Department of Agriculture: Possession, Use, and Transfer of Biological Agents and Toxins (9 CFR Part 121).

Other Regulatory Requirements

  • U.S. Department of Transportation and the International Air Transportation Authority: These organizations have strict requirements governing the shipment and transportation of hazardous materials, including biological agents.
  • Centers for Disease Control and Prevention: The CDC has established specific regulatory requirements for importation or transportation of etiologic agents, which include a permit application that must be submitted and approved prior to any such importations. The federal regulation governing the importation of etiologic agents is 42 CFR 71.54.
  • U.S. Department of Agriculture, Animal and Plant Health Inspection Service, and Veterinary Services regulates the importation of animals and animal-derived materials to ensure that exotic animal and poultry diseases are not introduced into the United States. Generally, a USDA veterinary permit is needed for materials derived from animals or exposed to animal-source materials. Materials that require a permit include animal tissues, blood, cells, or cell lines of livestock or poultry origin, RNA/DNA extracts, hormones, enzymes, monoclonal antibodies for in vivo use in non-human species, certain polyclonal antibodies, antisera, bulk shipments of test kit reagents, and microorganisms, including bacteria, viruses, protozoa, and fungi. Exceptions to this requirement are human and nonhuman primate tissues, serum, and blood.
  • U.S. Department of Commerce has specific regulatory requirements for exportation of biological materials. These regulations are both agent and country specific and must be followed strictly.
  • Wellesley College Institutional Biosafety Committee: The IBC has promulgated a number of specific policies and procedures that are incorporated into this document as requirements or have been included as appendices.


Roles and Responsibilities

The Principal Investigator (PI) is directly and primarily responsible for the safe operation of the laboratory, including providing application-specific training to students and researchers under their supervision. The PI's knowledge and judgment are critical in assessing risks and appropriately applying the recommendations in this Plan. However, safety is a shared responsibility among all laboratory and support staff. Many resources exist to assist the PI with these responsibilities, including the Institutional Biosafety Committee (IBC) and EHS.


Registration and Approval Process

Faculty and researchers at Wellesley College planning to carry out research using recombinant or synthetic nucleic acid molecules and/or biologically hazardous/infectious materials that pose a potential risk to the health of humans or animals, either directly through infection or indirectly through damage to the environment, must submit proposals for review and approval by the IBC prior to starting work.

When working with potentially infectious agents and human subjects or experimental animals, IBC review is necessary in addition to review by the Institutional Animal Care and Use Committee (IACUC) or the appropriate Institutional Review Board (IRB).

New Applications and Renewals

  • A new registration form shall be submitted to the IBC Chair. IBC approval is good for 3 years. After three years, another application must be resubmitted for IBC approval.
  • Information included on the renewal form must include: all proposed deviations from the protocol as initially approved (or since the last renewal notice); changes in laboratory location; changes in laboratory staff working on the project; any project titles to be added.
  • If there are significant deviations from the protocol, especially deviations that affect the containment level (e.g., new study organisms, a new host-vector-donor system, or any other modifications that may affect the containment level), the IBC may ask the PI to seek an additional approval to cover the additional experiments.


  • All changes should be detailed on the Amendment Form for review and approval. Title additions approval may be applied to several different granting agencies, but all grant titles must be registered with the IBC. Lab space additions approval applies only to work performed in registered lab space. For personnel changes, individuals must be trained in lab techniques and have completed necessary trainings.
  • If technical changes are extensive, the IBC may require that the PI submit a completely new application. A change in PI also requires full committee review. The new PI must supply a current CV in NIH format as part of the amendment.


Institutional Biosafety Committee

Wellesley College's Biosafety Program serves to protect faculty, staff, and students from exposure to biohazardous materials, to guard against the release of biohazardous materials that may harm humans, animals, plants or the environment, and to protect the integrity of experimental materials. Responsibility for oversight of the program resides with the Environmental Health and Safety Office, the Science Center Office, and the Institutional Biosafety Committee (IBC). All recombinant DNA studies, pathogenic organisms, and bloodborne pathogens must be registered with the IBC.

IBC Mission Statement


In-House Transportation and External Shipping and Receiving

If you transport any research materials including biological materials on the Wellesley College campus, you must utilize a secondary container that is labeled and leakproof. This includes transporting materials from one laboratory to another laboratory, or to/from cold/warm rooms and equipment rooms.

Shipping of materials to locations external to Wellesley College will require training if the shipment contains dry ice, batteries, radioactive materials, hazardous chemicals, or biological materials. The United States Department of Transportation (DOT) and the International Air Transport Association (IATA) have stringent requirements that include specialized training. If you believe you may need such training, email Anyone who ships a hazardous material improperly and/or without proper training may be subject to serious penalties including civil and criminal punishment.

Note that if you receive a shipment of biohazardous material, you must be approved to work with the material prior to receipt. Contact a member of the IBC in advance of ordering or making arrangements to receive a biohazardous material.

When shipping internally or externally, NEVER put dry ice in a container that is not able to vent as this may lead to an explosion and serious injury. The dry ice sublimates to form carbon dioxide, which exerts pressure on the wall of an airtight container and may cause an explosion.


Biohazard Assessment and Communication

The Wellesley College Institutional Biosafety Committee (IBC) is responsible for conducting risk assessments of work with biohazardous materials including recombinant DNA/RNA, human materials, microorganisms, and biological toxins. Before acquiring these materials and beginning any work, Principal Investigators must contact the IBC to review the planned work and determine whether further review and approvals may be required.

Communication of risks involved with work with biohazardous materials is done through a number of ways.

  1. The Universal Biohazard Symbol is used to denote equipment and rooms where biohazardous materials are used and stored.
  2. Biosafety training is provided by Wellesley College EHS for employees and students who work with biohazardous materials or have responsibilities that involve laboratory areas where biohazardous materials are used or stored.
  3. The Principal Investigator (PI) is responsible for ensuring risks involved in handling biohazardous materials are communicated to their laboratory group.


Risk Assessment

Risk assessment is a process used to identify the hazardous characteristics of a known biological agent or material, the activities that can result in a person’s exposure to the agent or material, the likelihood that such exposure will cause a laboratory acquired infection, and the probable consequences of such an infection. The information identified by risk assessment will provide a guide for the selection of appropriate biosafety levels and microbiological practices, safety equipment, and facility safeguards that can prevent laboratory acquired infection. The risk assessment process is the responsibility of the laboratory principal investigators and laboratory supervisors, with assistance from the Wellesley College IBC.

Principal investigators and laboratory supervisors should use risk assessment to alert their students and employees to the hazards of working with infectious agents and to the need to develop proficiency in the use of selected safe practices and containment equipment. Successful control of hazards in the laboratory also protects persons not directly associated with the laboratory, such as other occupants of the same building, and the public. The primary factors to consider in risk assessment and selection of precautions fall into two broad categories: agent hazards and laboratory procedure hazards. In addition, the capability of the laboratory staff to control hazards must be considered. This capability will depend on the training, technical proficiency, and good habits of all members of the laboratory, and the operational integrity of containment equipment and facility safeguards.


Biosafety Levels and Risk Groups

The World Health Organization (WHO) has recommended an agent risk group classification for laboratory use that describes four general risk groups based on principal characteristics and the route of transmission of the natural disease. The four groups address the risk to both the laboratory worker and the community. The NIH Guidelines established a comparable classification and assigned human etiological agents into four risk groups on the basis of hazard. The descriptions of the WHO and NIH risk group classifications are presented below in Table 1. They correlate with but do not necessarily equate to biosafety levels. The risk assessment will determine the degree of correlation between an agent’s risk group classification and biosafety level.

Wellesley College is equipped for work at Biosafety Level One (BSL-1) and Biosafety Level Two (BSL-2). The requirements for these biosafety levels are summarized in Table 2.

A fundamental objective of any biosafety program is the containment of potentially harmful biological agents. The term “containment” is used in describing safe methods, facilities, and equipment for managing infectious materials in the laboratory environment where they are being handled or maintained. The purpose of containment is to reduce or eliminate exposure of laboratory workers, other persons, and the outside environment to potentially hazardous agents. The use of vaccines may provide an increased level of personal protection. The risk assessment of the work to be done with a specific agent will determine the appropriate combination of these elements.


Table 1: Risk Group Classifications for Biological Agents

Risk Group Classification NIH Guidelines WHO Laboratory Biosafety Manual
Risk Group 1
(e.g. E. coli K12)
Agents not associated with disease in healthy adult humans. No or low individual and community risk. A microorganism unlikely to cause human or animal disease.
Risk Group 2
(e.g. Streptococcus pyogenes)
Agents associated with human disease that is rarely serious and for which preventive or therapeutic interventions are often available. Moderate individual risk and low community risk. A pathogen that can cause human or animal disease but is unlikely to be a serious hazard to laboratory workers, the community, livestock or the environment. Laboratory exposures may cause serious infection, but effective treatment and preventive measures are available and the risk of spread of infection is limited.
Risk Group 3
(e.g. Francisella tularensis)
Agents associated with serious or lethal human disease for which preventative or therapeutic interventions may be available. High invidividual risk, low community risk. High individual risk and low community risk. A pathogen that usually causes serious human or animal disease but does not ordinarily spread from one infected individual to another. Effective treatment and preventive measures are available.
Risk Group 4
(e.g. Ebola Virus)
Agents likely to cause serious or lethal human disease for which preventive or therapeutic interventions are not usually available. High individual risk, high community risk. High individual and community risk. A pathogen that usually causes serious human or animal disease and can be readily transmitted from one individual to another, directly or indirectly. Effective treatment and preventive measures are not usually available.


Table 2: Biosafety Levels

BSL Agents Practices Primary Barriers and Safety Equipment Facilities (Secondary Barriers)

None known to consistently cause diseases in healthy adults

Standard microbiological practices
  • No primary barriers required
  • Lab coats, gloves, eye/face protection as needed
Lab bench and sink required
  • Agents associated with human disease
  • Routes of transmission include percutaneous injury, ingestion, mucous membrane exposure
BSL-1 practices plus:
  • Limited access
  • Biohazard warning signs
  • Sharps precautions
  • Biosafety manual defining any needed waste decontamination or medical surveillance policies
  • BSCs or other physical containment devices must be used for all manipulations that cause splashes or aerosols of infectious materials
  • Lab coats, gloves, eye/face protection as needed
BSL-1 plus autoclave



Audits and Inspections

Periodic internal audits and inspections of Wellesley College work areas including laboratories are conducted by the Environmental Health and Safety Office. They may be prearranged or unannounced. The purpose of these internal activities is to identify areas of non-compliance and remediate them quickly. In addition, local, state, and federal regulatory agencies with jurisdiction over Wellesley College may make prearranged or unannounced inspections. Therefore, it is important that anyone who observes a potential non-compliance situation or unsafe condition reports the information to the Environmental Health and Safety Office.



Biosafety training is required by a number of biosafety regulations and guidelines. All employees and students who work with human materials must have Bloodborne Pathogens Training. To schedule training, email


Labels and Signs

The Universal Biohazard Symbol is utilized on door signage, as well as stickers found on laboratory equipment, to denote a biological hazard. All rooms that contain biohazardous agents must be posted with the Universal Biohazard Symbol. The background must be red/orange in color with a black universal biohazard symbol and black lettering. All equipment (centrifuges, water baths, refrigerators/freezers, incubators, etc.) that comes in contact with biohazardous materials must be labeled with the universal biohazard symbol.


Medical Surveillance

Research with certain biological agents may require medical surveillance to help assure the health of employees and students who have potential workplace exposures to hazards including, but not limited to, animal allergens, biohazardous materials, and high noise levels. The medical surveillance program is coordinated by the Wellesley College Health Service and the Environmental Health and Safety Office. Participants typically complete a health history questionnaire and may also receive a medical examination by a licensed physician. These screenings are used to establish a baseline of the participant’s health. Periodic future assessments are compared to the baseline to monitor potential occupational exposures. In some cases, vaccinations may be recommended. For example, the Hepatitis B vaccination is recommended for all employees who have potential occupational exposures to human blood or other potentially infectious materials.


Injury Involving Biological Material

All injuries and illnesses involving biological materials must be reported immediately to EHS (x3882), no matter how minor they may appear. If the incident involved a student, the Wellesley College Health Service (x2810) must also be contacted. Suspected exposures and “near misses” must also be reported. Near misses are defined as unplanned occurrences that did not lead to an injury or exposure, but could have. For example, if a researcher is splashed with a culture of an infectious organism, but is wearing sufficient personal protective equipment that no skin or mucous membrane contact was made, this event would be considered a “near miss.”

For medical emergencies, call x5555.



Work with certain biological agents may be contraindicated if pregnant or planning to become pregnant. Student questions or concerns about biological materials and their potential impact on pregnancy should be directed to the Wellesley College Health Service for a confidential medical assessment. Employees with questions or concerns may contact EHS, who will facilitate a confidential assessment by a qualified medical professional.


OSHA 300 Log and Sharps Log

The Occupational Safety and Health Administration (OSHA) requires that employers record employee workplace injuries and illnesses. This record is known as the OSHA 300 Log and each year a new 300 Log is completed for the period of January through December. All occupational injuries and illnesses incurred by Wellesley College employees that require more than first aid treatment must be recorded in the OSHA 300 Log. The log is posted by February 1 each year for the previous year’s injuries and illnesses. Names are not included in the posted OSHA 300 Log. The OSHA 300 Log allows Wellesley College to summarize the number of occupational injuries and illnesses in a given year and calculate the injury rate based on the number of employees.

For workplace injuries involving sharps such as needles and razor blades in a laboratory or health clinic setting, a similar log known as the Sharps Log is completed. The Sharps Log is used for recording percutaneous injuries from contaminated sharps. The Sharps Log must contain, at a minimum, information about the injury, the type and brand of device involved in the injury (if known), the department or work area where the exposure occurred, and an explanation of how the incident occurred. The log must be recorded and maintained in such a manner so as to protect the confidentiality of the injured employee (e.g., removal of personal identifiers).


Bloodborne Pathogens

Bloodborne pathogens are microorganisms that may be present in human blood and other human-derived materials such as human cell lines, blood components, body fluids, tissues, and organs. All work with human materials is covered by OSHA's Bloodborne Pathogen Standard, which addresses exposure control plans, universal precautions, engineering and work practice controls, personal protective equipment, housekeeping, laboratories, hepatitis B vaccination, post-exposure follow-up, hazard communication and training, and recordkeeping.

All employees who work with human materials will be offered the Hepatitis B vaccination free of charge. They may accept or decline, and if they initially decline they may change their mind at a later time and receive the vaccination free of charge.

All employees and students that work with human materials in a laboratory setting or have the potential for occupational exposure through job duties must receive initial training and annual retraining. Contact for training.


UV Lights

Ultraviolet (UV) lights are used in a variety of laboratory applications. UV lights may be found as an accessory within a biological safety cabinet (BSC). UV lights may also be used to visualize gels or to induce DNA damage. UV light is hazardous to humans and can cause burns to skin and eyes with as little as a few seconds of exposure.

Prior to purchasing a new BSC, it is important to review the need for a UV light with EHS and only purchase a BSC that has a sash interlock. Older models may not have a sash interlock to disconnect the UV light when the sash is opened and should be reported EHS for review. A UV light in a BSC is never a substitute for thorough cleaning and disinfection with a chemical disinfectant such as 70% ethanol or a freshly prepared solution of sodium hypochlorite (bleach).

Use of UV lights for other laboratory applications must be conducted with the appropriate personal protective equipment to shield skin and eyes from UV exposure. At a minimum, a lab coat, gloves, and a UV-rated faceshield should be worn.


Autoclave Use

An autoclave is a piece of laboratory equipment that uses high pressure and high temperature steam to sterilize materials such as microbiological media, glassware, and biohazardous waste.

The use of an autoclave to sterilize biohazardous and sharps waste is regulated by the State of Massachusetts. All autoclaves used for this purpose must be periodically validated through the use of biological indicators. Biological indicators use thermophilic bacteria, typically Geobacillus stearothermophilus, to verify that cycle time and temperature are adequate to kill microorganisms that may be present in the waste.

When using an autoclave, it is important to remember that interior and exterior surfaces may be very hot. Wear heat-resistant protective equipment and allow the contents to cool prior to removal from the autoclave.


Disinfection and Decontamination

Disinfection is the process of applying a product directly to a surface or object to destroy or irreversibly inactivate most pathogenic microorganisms, but not usually bacterial or fungal spores. Disinfection reduces the level of microbial contamination to an acceptably safe level. Decontamination renders an area, device, item or material safe to handle, and may be accomplished through the process of cleaning with soap and water, disinfection, or sterilization.

All work areas and equipment where biological materials have been used should be wiped down with an effective disinfectant after use and after any spills or contamination incidents. Appropriate laboratory disinfectants include 70% ethanol (except for bacterial or fungal spore-forming agents) and a freshly prepared 10% sodium hypochlorite (bleach) solution consisting of 1 part bleach in 9 parts water. Label all ethanol and bleach solution containers and bottles.


Spill Management

Spill kits should be readily available in all areas where biohazardous materials are used.

For BL-1; BL-2; rDNA; and blood spills:

  1. Alert lab staff and students that a spill has occurred, and to avoid the area.
  2. If spill is of a BL-2 substance:
    1. Avoid inhaling airborne material and evacuate room.
    2. Close door and post “DO NOT ENTER” signage.
    3. Wait 30 minutes for aerosols to settle.
    4. Re-enter space and follow instructions below.
  3. Don the Personal Protective Equipment (PPE) supplied in the spill kit, which includes eye protection, gloves, and boot covers. Wear your lab coat.
  4. Use the red biological waste bags provided as the collection receptacle.
  5. Place the spill pads over the spilled area to begin soaking up any liquid contamination. Place pads over entire affected areas to soak up all the liquid waste.
  6. Spray down the contaminated spill pads with the bleach, let sit for at least 20 minutes.
  7. Place soiled pads/towels into a red biological bag as waste.
  8. Collect any solid waste such as glass or plastic and dispose in biological sharps container. Use scoops/scrapers/forceps within spill kit to pick up broken glass or plastic.
  9. Disinfect spill area again with bleach spray to include nearby splash zone. Wipe up with paper towels or spill pads and place into the red biological waste bag.
  10. When complete, tie off bag, and place inside another red biological waste bag and tie off. This ensures that if a leak were to happen, there is a secondary containment for the hazard.
  11. Dispose of soiled PPE into a biological waste container in the lab for pick up and disposal.
  12. Contact EHS at extension x3882 to report the incident and fill out an incident report.



Emergency Eyewash and Drench Shower

Emergency eyewash and drench showers are located in areas where hazardous materials are used or stored. They must be kept accessible at all times. Do not store materials under or around the eyewash and drench shower as they may block access during an emergency situation. The emergency eyewash and drench showers are checked periodically by EHS or their designee. Promptly report any malfunctions to the Science Center Office (x3136) for repair.

If a hazardous material exposure occurs, quickly move to the eyewash or shower area. Inform others of the situation, ask for assistance, and have someone report the event to Public Safety (x5555). If there is an exposure to the eye, activate the eyewash and rinse for 15 minutes. If there is an exposure to the body, activate the shower and rinse for 15 minutes, removing clothing as necessary.


Personal Protective Equipment

Personal protective equipment (PPE) is an essential element of primary containment and laboratory safety. PPE provided to Wellesley College students, staff and faculty includes, but is not limited to:

  • Gloves
  • Laboratory coats (impervious)
  • Side shields (for glasses)
  • Face shields/masks
  • Safety glasses
  • Goggles
  • Hoods
  • Shoe covers
  • Respiratory protection – only if approved and in the Respiratory Protection Program
  • Other site-specific PPE

At a minimum, laboratory personnel should wear gloves and a laboratory coat whenever handling biological agents or cells and tissues. Safety glasses with side shields, goggles, or face shield shall be worn when these materials could potentially be splashed in the face. Laboratory personnel should wear other personal protective equipment (apron, face shield, mask, etc.) as needed or required to prevent potentially infectious materials from reaching their clothes, skin, eyes, mouth, or other mucous membranes. PPE must be removed prior to leaving the work area and placed in designated areas. PPE must be treated as medical waste when discarded. If PPE is not disposed, PPE shall be cleaned with disinfectant before and after use.


Waste Disposal


Revised June 2024