Transovarial transmission of a core virome in the Chagas disease vector Rhodnius prolixus

Triatomine assassin bugs comprise hematophagous insect vectors of Trypanosoma cruzi, the causative agent of Chagas disease. Although the microbiome of these species has been investigated to some extent, only one virus infecting Triatoma infestans has been identified to date. Here, we describe for the first time seven (+) single-strand RNA viruses (RpV1-7) infecting Rhodnius prolixus, a primary vector of Chagas disease in Central and South America. We show that the RpVs belong to the Iflaviridae, Permutotetraviridae and Solemoviridae and are vertically transmitted from the mothers to the progeny via transovarial transmission. Consistent with this, all the RpVs, except RpV2 that is related to the entomopathogenic Slow bee paralysis virus, established persistent infections in our R. prolixus colony. Furthermore, we show that R. prolixus ovaries express 22-nucleotide viral siRNAs (vsiRNAs), but not viral piRNAs, that originate from the processing of dsRNA intermediates during viral replication of the RpVs. Interestingly, the permutotetraviruses and sobemoviruses display shared pools of vsiRNAs that might provide the basis for a cross-immunity system. The vsiRNAs are maternally deposited in the eggs, where they likely contribute to reduce the viral load and protect the developing embryos. Our results unveil for the first time a complex core virome in R. prolixus and begin to shed light on the RNAi-based antiviral defenses in triatomines.

The role of sheep ked (Melophagus ovinus) as potential vector of protozoa and bacterial pathogens

The sheep ked (Melophagus ovinus) hematophagous insect may act as a potential vector of vector-borne pathogens. The aim of this study was to detect the presence of Trypanosoma spp., Bartonella spp., Anaplasma phagocytophilum and Borrelia burgdorferi sensu lato in sheep ked collected from sheep in Poland. In total, Trypanosoma spp. was detected in 58.91% of M. ovinus, whereas Bartonella spp. and B. burgdorferi s.l. were found in 86.82% and 1.55% of the studied insects, respectively. A. phagocytophilum was not detected in the studied material. In turn, co-infection by Trypanosoma spp. and Bartonella spp. was detected in 50.39%, while co-infection with Trypanosoma spp. and Bartonella spp. and B. burgdorferi s.l. was found in 1.55% of the studied insects. The conducted study showed for the first time the presence of B. burgdorferi s. l. in M. ovinus, as well as for the first time in Poland the presence of Trypanosoma spp. and Bartonella spp. The obtained results suggest that these insects may be a potential vector for these pathogens, but further-more detailed studies are required.

Diversity and infectivity of the RNA virome among different cryptic species of an agriculturally important insect vector: whitefly Bemisia tabaci

A large number of insect-specific viruses (ISVs) have recently been discovered, mostly from hematophagous insect vectors because of their medical importance, but little attention has been paid to important plant virus vectors such as the whitefly Bemisia tabaci, which exists as a complex of cryptic species. Public SRA datasets of B. tabaci and newly generated transcriptomes of three Chinese populations are here comprehensively investigated to characterize the whitefly viromes of different cryptic species. Twenty novel ISVs were confidently identified, mostly associated with a particular cryptic species while different cryptic species harbored one or more core ISVs. Microinjection experiments showed that some ISVs might cross-infect between the two invasive whitefly cryptic species, Middle East Asia Minor 1 (MEAM1) and Mediterranean (MED), but others appeared to have a more restricted host range, reflecting the possibility of distinct long-term coevolution of these ISVs and whitefly hosts. Moreover, analysis of the profiles of virus-derived small-interfering RNAs indicated that some of the ISVs can successfully replicate in whitefly and the antiviral RNAi pathway of B. tabaci is actively involved in response to ISV infections. Our study provides a comprehensive analysis of the RNA virome, the distinct relationships and cross-cryptic species infectivity of ISVs in an agriculturally important insect vector.

A simplified protocol for in vitro rearing of human body lice

Human body lice (Pediculus humanus) are neglected ectoparasites and pathogen vectors. Difficulties in raising and maintaining colonies of body lice in a laboratory setting remain a barrier to fundamental studies of physiology and vector-pathogen interactions in these insects. Several in vivo and in vitro rearing systems have been previously described and used by multiple research groups. However, these methods suffer from drawbacks that still complicate the rearing of body lice relative to many other commonly studied hematophagous insects. Here, a simplified protocol for raising and maintaining body lice in vitro using the commercially available Hemotek apparatus is described. This protocol draws from published methods for rearing body lice as well as other hematophagous insect species to further reduce labor, time, costs, and regulatory requirements typically associated with keeping human body lice in the laboratory. Using this protocol, the insects consistently fed on commercially available rabbit blood with little mortality, reached adulthood at a high rate, and produced a significant number of viable eggs, resulting in a 4.8-fold increase in population over a period of 40 days. The data suggest that the process described here can propagate modest populations for ongoing laboratory experiments and is a useful alternative to existing methods. The use and further optimization of in vitro rearing systems may facilitate dynamic studies of body lice by a wider range of investigators, enabling new progress in combating lice infestations, and louse-borne infections.

Influence of Trypanosoma cruzi (Kinetoplastida: Trypanosomatidae) Infection on Mortality of the Sylvatic Triatomine Vector, Mepraia spinolai (Heteroptera: Reuviidae), Under Fasting

The etiologic agent of Chagas disease, Trypanosoma cruzi, is transmitted by hematophagous insect vectors that subsist on repeated blood meals over their lives separated by periods of fasting. Using naturally infected Mepraia spinolai, we measured the influence of parasite infection on this host vector’s mortality during regular feeding and after fasting. After their capture, the insects were fed twice with uninfected mice to evaluate parasitic infection in their fecal samples by microscopic observation and PCR. Then the insects were subjected to a fasting period, followed by a third (final) feeding. After each feeding, a fecal sample was obtained to evaluate T. cruzi infection. To determine its progress through ontogeny, mortality and ecdysis of the infected and uninfected nymphs and adults were recorded on three occasions, over 140 d, and analyzed. Detections of infection by T. cruzi between the two first feedings increased, but this detection level was generally reduced after final feeding unless reinfected. For nymphs (stages III–V), their mortality was highest when infected after the fasting period, whereas adults were equally resistant to death after fasting when infected with T. cruzi. Metacyclic trypomastigotes were principally excreted in the fecal samples. Our results confirm that T. cruzi is pathogenic to its invertebrate hosts under nutritional stress conditions, when nymphs’ mortality is higher while infected than uninfected when they were hungry. These results are epidemiologically important because T. cruzi harms the fasting vector M. spinolai, reducing its lifespan and competence as a disease vector, and thereby its rates of parasite transmission.

Insects found in birds’ nests from Argentina. Furnarius rufus (Gmelin, 1788) (Aves: Furnariidae) and their inquiline birds, the true hosts of Acanthocrios furnarii (Cordero & Vogelsang, 1928) (Hemiptera: Heteroptera: Cimicidae)

The nest of Furnarius rufus (Gmelin, 1788) [Aves: Furnariidae] is a domed mud structure, with a partition separating the breeding chamber from the outside. In general each couple builds one nest per year, but each nest is used for one clutch or two consecutive clutches in the same breeding season. Some nests can remain in the field for 2 or 3 years, but some had a longer permanence (up to more than 8 years). Thus the nests are widely used by other vertebrate inquilines. Insects found in nests of F. rufus mentioned in the literature belong to Coleoptera [Chrysomelidae (1 sp.), Histeridae (1 sp.)], Hemiptera [Cimicidae (1 sp.), Reduviidae: Triatominae (1 sp.)], Hymenoptera [Apidae: Meliponinae (1 sp.)], and Diptera [Muscidae (2 spp.)]. A total of 9 orders, 35 families (two unidentified), and 67 species of insects (including 14 identified to genus and 10 undetermined), were found in 251 nests of F. rufus sampled in Argentina (Chaco [6], Córdoba [14], Santa Fe [2], Entre Ríos [7], San Luis [1], La Pampa [12], and Buenos Aires [209]). The most abundant species was one hematophagous insect, Acanthocrios furnarii (Cordero & Vogelsang, 1928) [Hemiptera: Cimicidae], an exclusive parasite on the inquiline birds. Agelaioides badius badius (Vieillot, 1819) [Aves: Icteridae], Sicalis flaveola pelzelni Sclater, 1872 [Aves: Emberizidae], and Troglodytes aedon Vieillot, 1809 [Aves: Troglodytidae] are new hosts of A. furnarii. Progne chalybea (Gmelin, 1789) [Aves: Hirundinidae] was found for the first time infested by A. furnarii inside the nests of F. rufus. The insects in the nests of Furnarius cristatus Burmeister, 1888, and those of P. domesticus, Progne sp., S. f. pelzelni, and T. aedon outside the nests of F. rufus were also investigated. Some ticks (Acarina: Argasidae, Ixodidae) found in the nests of F. rufus and P. domesticus are mentioned.

Occurrences of Ornithocoris toledoi (Cimicidae) on a free-range chicken farm in the state of Rio de Janeiro, Brazil

Ornithocoris toledoi is a hematophagous insect that parasites birds, particularly, galliformes. Although the occurrence of this arthropod is relatively low in Brazil, this is an important ectoparasite associated with backyarding poultry. The objective of this study was to report the occurrence of O. toledoi in a free-range chicken farm in the state of Rio de Janeiro, Brazil, including aspects of its taxonomic identification, biology and epidemiology.