Guidance document written by expert members of the Infravec2 consortium, summarising the state of the art in secure insectary design at CL2 and CL3. We expect this original version to evolve as techniques change or evolve, new techniques are added, novel safety-related questions need to be addressed etc. As such, we also aim to add advice on vectors such as ticks that require adaptation of methodologies described in this first version, in the future. Comments as well as questions are encouraged to improve these guidelines. Those should be directed to: Alain Kohl.

As part of the Infravec2 project, funded by Horizon 2020, the Infravec2 project is aiming to establish guidelines for setting up and running insectaries (in the first instance, for mosquitoes) at different containment levels. These guidelines should provide information to those interested developing new insectaries but may also provide useful information to existing facilities. The idea is borne out of discussions between scientists involved in mosquito (and other arthropod vector) studies and the difficulties faced at various levels, which can impact the of experiments that are feasible but may also impact reproducibility of results between laboratories.

Version history

• • Infravec2 guidelines for the design and operation of containment level 2 and 3 insectaries in Europe. Published online in Pathogens and Global Health.
  • Version 3
  • Version 2.1
  • Version 2
  • Version 1

  Your help is requested in this endeavor by responding to the survey below to help us create the most useful guidelines possible.

  Link to survey: https://tinyurl.com/Infravec2InsectarySurvey

  Your input is of high value.

  Thank you!

Version 1 is based on protocols and reagents currently being assessed by Infravec2 partners. It will be updated with information and technical details as further reagents and data become available.

Standardization of the protocol for Leishmania spp. promastigote and amastigote-initiated infections in sand flies (Diptera, Phlebotominae) conducted in BSL2 facilities. Charles University, Faculty of Science, Department of Parasitology, Prague, Czech Republic MIVEGEC, Institute of Research for Development (IRD), Montpellier, France

Standardization of the protocol for Leishmania spp. promastigote and amastigote-initiated infections in sand flies (Diptera, Phlebotominae) conducted in BSL3 facilities. Charles University, Faculty of Science, Department of Parasitology, Prague, Czech Republic and MIVEGEC, Institute of Research for Development (IRD), Montpellier, France.

Reference: Vaselek S, Prudhomme J, Myskova J, Lestinova T, Spitzova T, Bañuls AL, Volf P. Comparative Study of Promastigote- and Amastigote-Initiated Infection of Leishmania infantum (Kinetoplastida: Trypanosomatidae) in Phlebotomus perniciosus (Diptera: Psychodidae) Conducted in Different Biosafety Level Laboratories. J Med Entomol. 2019 Nov 8. pii: tjz199. doi: 10.1093/jme/tjz199. [PubMed] (see Publications)

The document describes an experimental protocol for oral TOSV infection of sand flies. Experiments with sand flies were performed at Charles University, Faculty of Science, Department of Parasitology, Prague, Czech Republic. It includes all necessary information and procedures on how to infect sand flies with TOSV within Biosafety Level 2 (BSL2).

The document describes an experimental protocol for Anopheles coluzzii infections with Plasmodium falciparum. It uses the P. falciparum NF54 clone provided by Radboud University Medical Center (RUMC) and was drafted by the Max Planck Institute for Infection Biology (MPIIB).

To optimize laboratory breeding of Anopheles coluzzii mosquitoes for malaria research two rearing protocols from expert laboratories were compared. Differences between methods included the number of/Anopheles/larvae that were raised per tray, the available volume of water, and type of food.