Biological Safety Manual - Chapter 13: Arthropod Containment Guidelines (ACG)


Biological Safety Manual - Chapter 13: Arthropod Containment Guidelines (ACG)


Risk assessments for arthropod research are multifaceted and focus on risks to individual researchers (harm, injury, disease) and impacts that a breach in containment may have on the community (increased/new vector reservoir, improved vector characteristics). Specific containment recommendations for arthropods deemed to be influential on public health can be found in the "Arthropod Containment Guidelines." (ACG) The ACG is intended to supplement recommendations in the BMBL and NIH Guidelines for recombinant DNA research.

The ACG were published in hard copy in the March 2019 issue of Vector-Borne Zoonotic Diseases1 and are freely downloadable from the Mary Ann Liebert, Inc. website. The ACG guidelines is tailored specifically to arthropods that transmit pathogens, including insects (Diptera - mosquitoes, tsetse flies, black flies, sand flies, midges; Hemiptera - kissing bugs; Phthiraptera - lice; Siphonaptera - fleas) and arachnids (Acari - ticks, mites). Arthropods that cause myiasis, infestation, biting, and stinging are not included in the ACG, and the ACG also specifically excludes most uses of Drosophila spp.

The ACG recommend biosafety measures specific for arthropods of public health importance considering that:

  1. Arthropods present unique containment challenges not encountered with microbial pathogens.
  2. The diversity of arthropods often means that appropriate containment must be determined on a case-by-case basis, thus a qualitative risk assessment will require consulting multiple references, including the BMBL, the NIH Guidelines, the Canadian Laboratory Biosafety Guidelines (Public Health Agency of Canada 2004), the WHO Biosafety Guidelines (World Health Organization (WHO) 2004), and the ACAV Catalogue of Arboviruses (Centers for Disease Control and Prevention [CDC] 1985).

The ACG contain two sections of greatest interest to most researchers:

  1. The Principles of Risk Assessment that discusses arthropods in the usual context (e.g., those known to contain a pathogenic agent, those with uncertain pathogens, and those with no agent).. Arthropod risk assessment is primarily a qualitative judgment that cannot be based on a prescribed algorithm. Several factors must be considered in combination: the agents transmitted, whether the arthropod is or may be infected, the mobility and longevity of the arthropod, its reproductive potential, biological containment, and epidemiological factors influencing transmission in the proposed location or region at risk.
  2. They also consider the following:
    • biological containment is a significant factor that reduces the hazards associated with accidental escape of arthropods;
    • epidemiological context alters the risks of an escape and its impact on the location or site in which the work is performed;
    • the phenotype of the vector, such as insecticide resistance; and
    • genetically modified arthropods with an emphasis on phenotypic change.

Four Arthropod Containment Levels (ACL 1-4) add increasingly stringent measures and are similar to biosafety levels. When arthropods are known (or reasonably assumed) to be infected with another pathogen, ACLs should directly correlate with the BSL dictated for the agents that the arthropods are infected with (see BMBL Section VI). ACL-2 is the most flexible level of containment and covers most exotic and transgenic arthropods and those infected with pathogens requiring BSL-2 containment. Like BMBL, each level has the following form:

  • standard practices;
  • special practices;
  • equipment (primary barriers);
  • facilities (secondary barriers).

Table 1

Table 1. Summary of Arthropod Containment Levels [from ACG]
Anthropod containment level (ACL) 1 2 3a 4a
Arthropods free of specific pathogens Indigenous/no change in local fauna Exotic/
or transient only
Exotic with establishment potential or transgenic n/a n/a
Infection status Up to BSL-1 Up to BSL-2 Up to BSL-3 Up to BSL-4
Practices ACL-1 standard handling practices ACL-2 and BSL-2 limited access, training, signage, containment, and disposal ACL-3 and BSL-3 restricted access, training, appropriate PPE, signage, containment, disposal, record-keepinga ACL-4 with BSL-4 isolation, training, appropriate PPE, signage, containment, disposal, record-keepinga
Primary barriers Species-appropriate containers Appropriate PPE, escape-proof containers Appropriate PPE, escape-proof containers, pesticide available for emergency usea Appropriate PPE, escape-proof containers, pesticide available for emergency usea
Secondary barriers   BSL-2 facilities, breeding sites, and harborage minimized, pest control BSL-3 facilities, biological safety cabinets, other physical containment devices, pest controla BSL-4 and facility-specific procedures and equipment for arthropod handling while wearing positive pressure containment suita

General guidelines for best laboratory containment practices are shown for vector species of arthropod that are uninfected (above the bold line) or infected (below the bold line) according to biosafety and ACLs. Indigenous species are those species whose current range includes the research location. All others are considered exotic.

For uninfected arthropods, containment guidelines take into account the consequences of accidental escape from a laboratory, in which the arthropod would be:

  1. inviable as a result of exposure to unfavorable conditions;
  2. transient because conditions vary such that the arthropod would die during typical year climate cycle; or
  3. has potential for establishment because escaped arthropods could reasonably be expected to persist through a typical climatic year.

Arthropod containment specifics for each BSL should always be reviewed in the context of a laboratory-, vector-, and pathogen-specific risk assessment that is based on consultation between the investigator and the appropriate institutional oversight committee(s) and according to the constraints of the infrastructure available.

a. Additional restrictions apply for work with arthropods in association with Select Agents.

ACL, arthropod containment level; BSL, biosafety level; PPE, personal protective equipment.


  1. American Committee of Medical Entomology; American Society of Tropical Medicine and Hygiene. Vector-Borne and Zoonotic Diseases. New Rochelle (NY): Mary Ann Liebert, Inc.; 2019.

Back to Chapter Twelve

Proceed to Chapter Fourteen

Print Article


Article ID: 131897
Thu 4/8/21 9:19 PM
Mon 9/12/22 9:28 AM
Responsible Unit
School, Department, or other organizational unit issuing this document.
Environment, Health and Safety
Issuing Officer
Name of the document Issuing Officer. This is the individual whose organizational authority covers the policy scope and who is primarily responsible for the policy.
Issuing Officer Title
Title of the person who is primarily responsible for issuing this policy.
Executive Director
Next Review
Date on which the next document review is due.
12/01/2023 12:00 AM
Last Review
Date on which the most recent document review was completed.
09/12/2022 12:00 AM
Last Revised
Date on which the most recent changes to this document were approved.
09/12/2022 12:00 AM
Effective Date
If the date on which this document became/becomes enforceable differs from the Origination or Last Revision, this attribute reflects the date on which it is/was enforcable.
09/12/2022 12:00 AM
Date on which the original version of this document was first made official.
05/01/2015 12:00 AM