Detection of Arthropod-Borne Bacteria and Assessment of MALDI-TOF MS for the Identification of Field-Collected Immature Bed Bugs from Mauritania

SIMPLE SUMMARY: Bed bugs are hematophagous insects that have been described as a cause of nuisance since ancient times. Two species are known to parasitize humans: Cimex lectularius, prevalent in temperate areas, and C. hemipterus, also known as the tropical bed bug. A global increase in infestation has been observed in recent decades. However, data is lacking concerning species distribution, which may be affected by population movements (e.g., travelers, migrants). Moreover, although these insects are not known to harbor and transmit infectious agents to humans, continuous monitoring of their infection status is part of active surveillance. We analyzed bed bug specimens collected from beddings in Mauritania, and assessed MALDI-TOF MS as an identification tool for immature specimens. Moreover, we also conducted bacterial detection in each specimen. Overall, our findings supported MALDI-TOF MS as being a robust tool allowing species level identification of immature Cimex specimens. ABSTRACT: Human infestations by bed bugs have upsurged globally in recent decades, including in African countries, where recent reports pointed out an increase in infestation. Sympatric dwelling has been described for two species of bed bug parasitizing humans: Cimex hemipterus (the tropical bed bug) and C. lectularius. Identification of these two species is based on morphological characteristics, and gene sequencing, and may also rely on Matrix-Assisted Laser Desorption Ionization Time-Of-Flight Mass Spectrometry (MALDI-TOF MS). The present work aimed to assess whether MALDI-TOF MS was applicable for species level identification of immature stages of Cimex. Arthropods were collected in domestic settings in Nouakchott, Mauritania. Identification used morphological keys and MALDI-TOF MS identification was assessed for immature stages. Quantitative PCR and sequencing assays were used to detect arthropod-associated bacteria in each specimen. A total of 92 arthropods were collected, all morphologically identified as C. hemipterus (32 males, 14 females and 45 immature stages). A total of 35/45 specimens produced good quality MALDI-TOF MS spectra. Analysis allowed species level identification of all immature C. hemipterus after their spectra were entered into our in-house MALDI-TOF MS arthropod spectra database. Molecular screening allowed detection of Wolbachia DNA in each specimen. These results suggested that MALDI-TOF MS is a reliable tool for species level identification of Cimex specimens, including immature specimens. Future studies should assess this approach on larger panels of immature specimens for different Cimex species and focus on the precise staging of their different immature developmental stages.