Sand flies (Diptera: Psychodidae) are natural vectors of Leishmania. For the initiation of sand fly experimental infections either Leishmania amastigotes or promastigotes can be used. In order to obtain comparable results, it is necessary to adjust and standardize procedures. During this study, we conducted promastigote- and amastigote-initiated infections of Leishmania infantum Nicolle, 1908 parasites in Phlebotomus (Larroussius) perniciosus Newstead, 1911 in two laboratories with different levels of biosafety protection. Protocol originally designed for a biosafety level 2 facility was modified for biosafety level 3 facility and infection parameters were compared. Particularly, specially designed plastic containers were used for blood feeding; feeders were placed outside the sand fly cage, on the top of the mesh; feeding was performed inside the climatic chamber; separation of engorged females was done in Petri dishes kept on ice; engorged females were kept in the cardboard containers until dissection. All experiments, conducted in both laboratories, resulted in fully developed late stage infections with high number of parasites and colonization of the stomodeal valve. We demonstrated that protocol originally designed for biosafety level 2 facilities can be successfully modified for other biosafety facilities, depending on the special requirements of the individual institution/laboratory.

Department of Parasitology Faculty of Science Charles University Viničná 7 Prague Czech Republic

UMR MIVEGEC IRD center 911 avenue Agropolis Montpellier France

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Alvar J., Vélez I. D., Bern C., Herrero M., Desjeux P., Cano J., Jannin J., and den Boer M.; WHO Leishmaniasis Control Team 2012. Leishmaniasis worldwide and global estimates of its incidence. PLoS One. 7: e35671. PubMed PMC

Bates P. A. 2008. Leishmania sand fly interaction: progress and challenges. Curr. Opin. Microbiol. 11: 340–344. PubMed PMC

Bongiorno G., Di Muccio T., Bianchi R., Gramiccia M., and Gradoni L.. . 2019. Laboratory transmission of an Asian strain of Leishmania tropica by the bite of the southern European sand fly Phlebotomus perniciosus. Int. J. Parasitol. 49: 417–421. PubMed

Dostálová A., and Volf P.. . 2012. Leishmania development in sand flies: parasite-vector interactions overview. Parasit. Vectors. 5: 276. PubMed PMC

Kamhawi S. 2006. Phlebotomine sand flies and Leishmania parasites: friends or foes? Trends Parasitol. 22: 439–445. PubMed

Lawyer P., Killick-Kendrick M., Rowland T., Rowton E., and Volf P.. . 2017. Laboratory colonization and mass rearing of phlebotomine sand flies (Diptera, Psychodidae). Parasite. 24: 42. PubMed PMC

Maroli M., Feliciangeli M. D., Bichaud L., Charrel R. N., and Gradoni L.. . 2013. Phlebotomine sandflies and the spreading of leishmaniases and other diseases of public health concern. Med. Vet. Entomol. 27: 123–147. PubMed

Myskova J., Votypka J., and Volf. P.. 2015. Leishmania in sand flies: comparison of quantitative polymerase chain reaction with other techniques to determine the intensity of infection. J. Med. Entomol. 45: 133–138. PubMed

R Development Core Team. 2018. R: a language and environment for statistical computing. R foundation for statistical computing, Vienna, Austria: http://www.r-project.org/.

Sadlova J., Myskova J., Lestinova T., Votypka J., Yeo M., and Volf P.. . 2017. Leishmania donovani development in Phlebotomus argentipes: comparison of promastigote- and amastigote-initiated infections. Parasitology. 144: 403–410. PubMed PMC

Volf P., and Myskova J.. . 2007. Sand flies and Leishmania: specific versus permissive vectors. Trends Parasitol. 23: 91–92. PubMed PMC

Volf P., and Volfova V.. . 2011. Establishment and maintenance of sand fly colonies. J. Vector Ecol. 36 (Suppl 1): S1–S9. PubMed