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Bed bugs are hematophagous insects with a long history of presence in human communities. Over the last three decades, infestations by bed bugs in human dwellings have drastically increased, leading to a rise in bed bug concerns. This study was conducted in May 2019 in the Miarinarivo district of Madagascar. Eight locations with suspected infestations were inspected. Of these, five locations were found with active bed bug infestations. Overall, 23 (19 adults and four nymphs) specimens with numbers per apartment ranging from 2 to 6 were collected and analyzed using morphological and molecular approaches. Of 19 adults collected, 18 were morphologically identified as Cimex hemipterus (Hemiptera: Cimicidae), with the final adult identified as C. lectularius. Morphological identification of adults was confirmed by sequencing 450 bp fragment of COI gene. All four nymphs were also identified molecularly as C. hemipterus. Maximum likelihood phylogenetic tree of COI gene and sequence alignment of C. hemipterus specimens revealed the presence of two groups (first group clustered with previously reported haplotype from Asian and African countries and the second group encompassed a lesser known haplotype exclusively from Africa (Kenya and Senegal). This study presents low genetic diversity among C. hemipterus specimens and, for the first time, reports the occurrence of C. lectularius, known as temperate species, in Madagascar. These results can be helpful in control management strategies against these ectoparasites in this country.

Bed bugs, Cimex lectularius and C. hemipterus, are common blood-sucking ectoparasites of humans with a large geographical distribution, worldwide. In France, little is known about the status of bed bugs’ infestation and their resistance to insecticides, particularly, pyrethroids. Here, we aimed to find mutations in the kdr gene, known to be involved in resistance to insecticides. We gathered bed bugs from various infested locations, including 17 private houses, 12 HLM building complex, 29 apartments, 2 EHPAD, and 2 immigrants’ residences. A total of 1211 bed bugs were collected and morphologically identified as C. lectularius. Two fragments of the kdr gene, encompassing codons V419L and L925I, were successfully amplified for 156 specimens. We recorded sense mutation in the first amplified fragment (kdr1) in 89 out of 156 (57%) samples, in which in 61 out of 89 (68.5%) sequences, a change of valine (V) into leucine (L) V419L was observed. Within the second fragment (kdr2), a homozygous mutation was recorded in 73 out of 156 (46.7%) specimens at the codon 925. At this position, 43 out of 73 (58.9%) specimens had a sense mutation leading to the replacement of leucine (L) by isoleucine (I). Among 162 mutant sequences analyzed (89 for the kdr1 fragment and 73 for the kdr2 one), we detected single point mutation in 26.6%, while 73.4% presented the mutation in both kdr1 and kdr2 fragments. All modifications recorded in bed bug populations of Paris are described to be involved in the knockdown resistance (kdr) against pyrethroids.