The Absence of Abdominal Pigmentation in Livestock Associated Culicoides following Artificial Blood Feeding and the Epidemiological Implication for Arbovirus Surveillance

Culicoides midges (Diptera: Ceratopogonidae), the vectors of economically important arboviruses such as bluetongue virus and African horse sickness virus, are of global importance. In the absence of transovarial transmission, the parity rate of a Culicoides population provides imperative information regarding the risk of virus dispersal. Abdominal pigmentation, which develops after blood feeding and ovipositioning, is used as an indicator of parity in Culicoides. During oral susceptibility trials over the last three decades, a persistent proportion of blood engorged females did not develop pigment after incubation. The present study, combining a number of feeding trials and different artificial feeding methods, reports on this phenomenon, as observed in various South African and Italian Culicoides species and populations. The absence of pigmentation in artificial blood-fed females was found in at least 23 Culicoides species, including important vectors such as C. imicola, C. bolitinos, C. obsoletus, and C. scoticus. Viruses were repeatedly detected in these unpigmented females after incubation. Blood meal size seems to play a role and this phenomenon could be present in the field and requires consideration, especially regarding the detection of virus in apparent “nulliparous” females and the identification of overwintering mechanisms and seasonally free vector zones.

Influence of GST- and P450-based metabolic resistance to pyrethroids on blood feeding in the major African malaria vector Anopheles funestus

Insecticide resistance genes are often associated with pleiotropic effects on various mosquito life-history traits. However, very little information is available on the impact of insecticide resistance, especially metabolic resistance, on blood feeding process in mosquitoes. Here, using two recently detected DNA-based metabolic markers in the major malaria vector, An. funestus, we investigated how metabolic resistance genes could affect blood meal intake.After allowing both field F1 and lab F8 Anopheles funestus strains to feed on human arm for 30 minutes, we assessed the association between key parameters of blood meal process including, probing time, feeding duration, blood feeding success and blood meal size, and markers of glutathione S-transferase (L119F-GSTe2) and cytochrome P450 (CYP6P9a_R) - mediated metabolic resistance. None of the parameters of blood meal process was associated with L119F-GSTe2 genotypes. In contrast, for CYP6P9a_R, homozygote resistant mosquitoes were significantly more able to blood-feed than homozygote susceptible (OR = 3.3; CI 95%: 1.4-7.7; P =0.01) mosquitoes. Moreover, the volume of blood meal ingested by CYP6P9a-SS mosquitoes was lower than that of CYP6P9a-RS (P<0.004) and of CYP6P9a-RR (P<0.006). This suggests that CYP6P9a gene affects the feeding success and blood meal size of An. funestus. However, no correlation was found in the expression of CYP6P9a and that of genes encoding for salivary proteins involved in blood meal process.This study suggests that P450-based metabolic resistance may increase the blood feeding ability of malaria vectors and potential impacting their vectorial capacity.

Feeding performance on a novel host: no adaptation over generations and differential patterns in two flea species

To model the colonization of a novel host by fleas, Synosternus cleopatrae and Xenopsylla ramesis, we established experimental lines maintained for 15 generations on a principal or a novel host (either co-occurring with a flea or not). We compared the blood meal size and the energy expended for digestion by fleas from the 15th generation of each line on these hosts between hosts within a line and between lines within a host asking (a) whether fleas adapt to a novel host (increased blood consumption/decreased energy expended for digestion); (b) if yes, whether this adaptation leads to the loss of ability to exploit an original host, and (c) whether the success of adaptation to a novel host depends on its ecological co-occurrence with a flea. The blood consumption and digestion energetics of fleas fed on the principal host differed from those on other hosts. The effect of the principal host on feeding performance differed between fleas, with S. cleopatrae consuming less blood and expending more energy for digestion on the principal than on any other host, whereas the opposite was true for X. ramesis. No changes in feeding performance on a novel host over generations were found. We propose several explanations for the lack of adaptation to a novel host over time. We explain the poor performance of S. cleopatrae on its principal host via its immune response mounting pattern. We argue that the principal host of a parasite is not necessarily the host on which the parasite demonstrates the best performance.

Sub-lethal insecticide exposure affects host biting efficiency of Kdr-resistant Anopheles gambiae

The massive use of insecticide-treated nets (ITNs) has drastically changed the environment for malaria vector mosquitoes, challenging their host-seeking behaviour and biting success. Here, we investigated the effect of a brief exposure to an ITN on the biting behaviour of Anopheles mambiae mosquitoes and the interaction between such behaviour and the kdr mutation that confers resistance to pyrethroids. To this aim, we developed a video assay to study the biting behaviour of mosquitoes with similar genetic background, but different kdr locus genotypes (SS i.e. homozygous susceptible, RS i.e. heterozygous and RR i.e. homozygous resistant), after a brief exposure to either control untreated nets or one of two types of pyrethroid-treated nets (deltamethrin or permethrin). In presence of untreated nets, the kdr mutation did not influence mosquito blood feeding success but caused differences in feeding and prediuresis durations and blood meal size. Exposure to deltamethrin ITN decreased the blood feeding success rate of RR and RS mosquitoes, whereas in presence of permethrin ITN, the kdr mutation increased the blood-feeding success of mosquitoes. Exposure to the two types of pyrethroid-treated nets reduced feeding duration, prediuresis duration and blood meal size of all three genotypes. Our study demonstrates a complex interaction between insecticide exposure and the kdr mutation on the biting behavior of mosquitoes, which may substantially impact malaria vector fitness and disease transmission.

Effects of Bartonella spp. on Flea Feeding and Reproductive Performance

ABSTRACTNumerous pathogens are transmitted from one host to another by hematophagous insect vectors. The interactions between a vector-borne organism and its vector vary in many ways, most of which are yet to be explored and identified. These interactions may play a role in the dynamics of the infection cycle. One way to evaluate these interactions is by studying the effects of the tested organism on the vector. In this study, we tested the effects of infection withBartonellaspecies on fitness-related variables of fleas by usingBartonellasp. strain OE 1-1,Xenopsylla ramesisfleas, andMeriones crassusjirds as a model system. Feeding parameters, including blood meal size and metabolic rate during digestion, as well as reproductive parameters, including fecundity, fertility, and life span, were compared between fleas experimentally infected withBartonellaand uninfected fleas. In addition, the developmental time, sex ratio, and body size of F1offspring fleas were compared between the two groups. Most tested parameters did not differ between infected and uninfected fleas. However, F1males produced byBartonella-positive females were significantly smaller than F1males produced byBartonella-negative female fleas. The findings in this study suggest that bartonellae are well adapted to their flea vectors, and by minimally affecting their fitness they have evolved to better spread themselves in the natural environment.

Ingestive behaviour and physiology of the medicinal leech

Ingestion lasts 25 min in Hirudo medicinalis and is characterized by pharyngeal peristalsis which fills the crop. This peristalsis has an initial rate of 2.4 Hz which decays smoothly to 1.2 Hz at termination of ingestion. During ingestion, the leech body wall undergoes peristalsis which appears to aid in filling the crop diverticula. Body peristalsis begins at a rate of 10 min-1 and decreases linearly to 2 min-1 at termination. The body also undergoes dorsoventral flexions when blood flow is occluded. Blood meal size increases slightly with leech size: 8.4 g for 1-g leeches and 9.7 g for 2-g leeches. However, relative meal size decreases markedly with increasing animal size; from 8.15 times body mass for 1-g to 4.80 times for 2-g leeches. When intact leeches were exposed to micromolar concentrations of serotonin, there was an increase in the rate of pharyngeal peristalsis and the size of the blood meals. Leeches excrete the plasma from their ingested blood meals. Excretion is activated during ingestion, which increases feeding efficiency by increasing the proportion of blood cells in the ingestate. Excretion continues for 4–6 days following ingestion, removing all the remaining plasma from the ingestate. Leech ingestion comprises stereotyped muscular movements, secretion of saliva and excretion of plasma. A strikingly similar feeding physiology is seen in the blood-sucking insect Rhodnius, and we suggest that efficient sanguivory may require the convergent evolution of similar ingestive mechanisms.