Workflow of Lotmaria passim isolation: Experimental infection with a low-passage strain causes higher honeybee mortality rates than the PRA-403 reference strain

María Buendía-Abad; Mariano Higes; Raquel Martín-Hernández; Laura Barrios; Aranzazu Meana; Alberto Fernández Fernández; Antonio Osuna; Luis Miguel De Pablos

doi:10.1016/j.ijppaw.2020.12.003

Workflow of Lotmaria passim isolation: Experimental infection with a low-passage strain causes higher honeybee mortality rates than the PRA-403 reference strain

Int J Parasitol Parasites Wildl. 2020 Dec 31:14:68-74. doi: 10.1016/j.ijppaw.2020.12.003. eCollection 2021 Apr.

Authors

María Buendía-Abad¹, Mariano Higes¹, Raquel Martín-Hernández^{1

2}, Laura Barrios³, Aranzazu Meana⁴, Alberto Fernández Fernández⁵, Antonio Osuna^{5

6}, Luis Miguel De Pablos^{5

6}

Affiliations

¹ IRIAF - Regional Institute for Agrifood and Forestry Research and Development, Laboratory of Bee Pathology, Center for Beekeeping and Agro-environmental Research (CIAPA), Community of Castilla-La Mancha, Marchamalo, Spain.
² Institute of Human Resources for Science and Technology (INCRECYT-FEDER), Science and Technology Park Foundation from Castilla - La Mancha, Albacete, Spain.
³ Statistics Department, Computing Center SGAI-CSIC, 28006, Madrid, Spain.
⁴ Department of Animal Health, Faculty of Veterinary Medicine, Complutense University of Madrid, 28040, Madrid, Spain.
⁵ Department of Parasitology, Biochemical and Molecular Parasitology Group CTS-183, University of Granada, 18071, Granada, Spain.
⁶ Institute of Biotechnology, University of Granada, Granada, Spain.

Abstract

The impact of trypanosomatid parasites on honeybee health may represent a major threat to bee colonies worldwide. However, few axenic isolates have been generated to date and with no details on cell culture passages, a parameter that could influence parasite virulence. To address this question, a trypanosomatid isolation protocol was developed and a new strain was obtained, named L. passim C1. Using experimental infection of worker honeybees, we compared the virulence and mortality rates of the ATCC PRA-403 reference strain and C1 strain, the latter showing higher virulence from 10 days post-infection onward. This study highlights the impact of cell culture passages on the pathogenicity of L. passim in honeybees, providing new evidence of its negative effects on honeybee health.

Keywords: Experimental infection; Honeybee; Isolation; Lotmaria passim; Low-passage; Trypanosomatid.