Dynamics of Hen Flea Ceratophyllus gallinae Subpopulations in Blue Tit Nests

The hen flea is a common parasite in bird nests, in particular, in tit species, and imposes considerable fitness costs for the host. These costs are expected to lead to selection for increased host defense, which in turn should select for better-adapted parasites. Our understanding of the coevolution of this host-parasite system is currently limited by the insufficient knowledge of both the timing of flea generations and their reproductive behavior within the nesting period of their hosts. In the present study we (1) followed the demography of experimental flea subpopulations during the host's breeding cycle, (2) assessed the importance of time-temperature effects in the nest by recording temperatures within the nest material, and (3) investigated the influence of variation in host timing and duration of the breeding period on flea development. We found the following. (1) Fleas completed either one or two generations within the birds' nesting cycle, leading to two well-defined periods of cocoon formation. (2) Within-nest temperatures during the warm period of the host breeding cycle—i.e., the incubation and nestling periods—depended on both outdoor temperatures and heat production from the breeding birds. Day-degree availability, a measure of physiological time, during the host incubation was significantly explained by the duration of incubation period and its timing in the season. Similarly, day-degrees during the warmer nestling period were significantly explained by its duration and its timing in the season. (3) The number of flea larvae found in the nests correlated with the host's timing and duration of the warm period available for their development; this was not the case, however, for the number of adult fleas. These results underline the importance of time-temperature effects as determinants of flea demography within the nests. The life-cycle and time-temperature effects are discussed in the light of potential host selection on parasite behavior and life historie

Hydric stress-dependent effects of Plasmodium falciparum infection on the survival of wild-caught Anopheles gambiae female mosquitoes

<p>Abstract</p> <p>Background</p> <p>Whether <it>Plasmodium falciparum</it>, the agent of human malaria responsible for over a million deaths per year, causes fitness costs in its mosquito vectors is a burning question that has not yet been adequately resolved. Understanding the evolutionary forces responsible for the maintenance of susceptibility and refractory alleles in natural mosquito populations is critical for understanding malaria transmission dynamics.</p> <p>Methods</p> <p>In natural mosquito populations, <it>Plasmodium </it>fitness costs may only be expressed in combination with other environmental stress factors hence this hypothesis was tested experimentally. Wild-caught blood-fed <it>Anopheles gambiae </it>s.s. females of the M and S molecular form from an area endemic for malaria in Mali, West Africa, were brought to the laboratory and submitted to a 7-day period of mild hydric stress or kept with water ad-libitum. At the end of this experiment all females were submitted to intense desiccation until death. The survival of all females throughout both stress episodes, as well as their body size and infection status was recorded. The importance of stress, body size and molecular form on infection prevalence and female survival was investigated using Logistic Regression and Proportional-Hazard analysis.</p> <p>Results</p> <p>Females subjected to mild stress exhibited patterns of survival and prevalence of infection compatible with increased parasite-induced mortality compared to non-stressed females. Fitness costs seemed to be linked to ookinetes and early oocyst development but not the presence of sporozoites. In addition, when females were subjected to intense desiccation stress, those carrying oocysts exhibited drastically reduced survival but those carrying sporozoites were unaffected. No significant differences in prevalence of infection and infection-induced mortality were found between the M and S molecular forms of <it>Anopheles gambiae</it>.</p> <p>Conclusions</p> <p>Because these results suggest that infected mosquitoes may incur fitness costs under natural-like conditions, they are particularly relevant to vector control strategies aiming at boosting naturally occurring refractoriness or spreading natural or foreign genes for refractoriness using genetic drive systems in vector populations.</p

Hydric stress-dependent effects of Plasmodium falciparum infection on the survival of wild-caught Anopheles gambiae female mosquitoes

<p>Abstract</p> <p>Background</p> <p>Whether <it>Plasmodium falciparum</it>, the agent of human malaria responsible for over a million deaths per year, causes fitness costs in its mosquito vectors is a burning question that has not yet been adequately resolved. Understanding the evolutionary forces responsible for the maintenance of susceptibility and refractory alleles in natural mosquito populations is critical for understanding malaria transmission dynamics.</p> <p>Methods</p> <p>In natural mosquito populations, <it>Plasmodium </it>fitness costs may only be expressed in combination with other environmental stress factors hence this hypothesis was tested experimentally. Wild-caught blood-fed <it>Anopheles gambiae </it>s.s. females of the M and S molecular form from an area endemic for malaria in Mali, West Africa, were brought to the laboratory and submitted to a 7-day period of mild hydric stress or kept with water ad-libitum. At the end of this experiment all females were submitted to intense desiccation until death. The survival of all females throughout both stress episodes, as well as their body size and infection status was recorded. The importance of stress, body size and molecular form on infection prevalence and female survival was investigated using Logistic Regression and Proportional-Hazard analysis.</p> <p>Results</p> <p>Females subjected to mild stress exhibited patterns of survival and prevalence of infection compatible with increased parasite-induced mortality compared to non-stressed females. Fitness costs seemed to be linked to ookinetes and early oocyst development but not the presence of sporozoites. In addition, when females were subjected to intense desiccation stress, those carrying oocysts exhibited drastically reduced survival but those carrying sporozoites were unaffected. No significant differences in prevalence of infection and infection-induced mortality were found between the M and S molecular forms of <it>Anopheles gambiae</it>.</p> <p>Conclusions</p> <p>Because these results suggest that infected mosquitoes may incur fitness costs under natural-like conditions, they are particularly relevant to vector control strategies aiming at boosting naturally occurring refractoriness or spreading natural or foreign genes for refractoriness using genetic drive systems in vector populations.</p

Visual and olfactory associative learning in the malaria vector Anopheles gambiae sensu stricto

<p>Abstract</p> <p>Background</p> <p>Memory and learning are critical aspects of the ecology of insect vectors of human pathogens because of their potential effects on contacts between vectors and their hosts. Despite this epidemiological importance, there have been only a limited number of studies investigating associative learning in insect vector species and none on Anopheline mosquitoes.</p> <p>Methods</p> <p>A simple behavioural assays was developed to study visual and olfactory associative learning in <it>Anopheles gambiae</it>, the main vector of malaria in Africa. Two contrasted membrane qualities or levels of blood palatability were used as reinforcing stimuli for bi-directional conditioning during blood feeding.</p> <p>Results</p> <p>Under such experimental conditions <it>An. gambiae </it>females learned very rapidly to associate visual (chequered and white patterns) and olfactory cues (presence and absence of cheese or Citronella smell) with the reinforcing stimuli (bloodmeal quality) and remembered the association for up to three days. Associative learning significantly increased with the strength of the conditioning stimuli used. Importantly, learning sometimes occurred faster when a positive reinforcing stimulus (palatable blood) was associated with an innately preferred cue (such as a darker visual pattern). However, the use of too attractive a cue (e.g. Shropshire cheese smell) was counter-productive and decreased learning success.</p> <p>Conclusions</p> <p>The results address an important knowledge gap in mosquito ecology and emphasize the role of associative memory for <it>An. gambiae</it>'s host finding and blood-feeding behaviour with important potential implications for vector control.</p

Effects of larval growth condition and water availability on desiccation resistance and its physiological basis in adult Anopheles gambiae sensu stricto

<p>Abstract</p> <p>Background</p> <p>Natural populations of the malaria mosquito <it>Anopheles gambiae </it>s.s. are exposed to large seasonal and daily fluctuations in relative humidity and temperature, which makes coping with drought a crucial aspect of their ecology.</p> <p>Methods</p> <p>To better understand natural variation in desiccation resistance in this species, the effects of variation in larval food availability and access to water as an adult on subsequent phenotypic quality and desiccation resistance of adult females of the Mopti chromosomal form were tested experimentally.</p> <p>Results</p> <p>It was found that, under normal conditions, larval food availability and adult access to water had only small direct effects on female wet mass, dry mass, and water, glycogen and body lipid contents corrected for body size. In contrast, when females subsequently faced a strong desiccation challenge, larval food availability and adult access to water had strong carry-over effects on most measured physiological and metabolic parameters, and affected female survival. Glycogen and water content were the most used physiological reserves in relative terms, but their usage significantly depended on female phenotypic quality. Adult access to water significantly influenced the use of water and body lipid reserves, which subsequently affected desiccation resistance.</p> <p>Conclusions</p> <p>These results demonstrate the importance of growth conditions and water availability on adult physiological status and subsequent resistance to desiccation.</p

Impact of three years of large scale Indoor Residual Spraying (IRS) and Insecticide Treated Nets (ITNs) interventions on insecticide resistance in Anopheles gambiae s.l. in Benin

<p>Abstract</p> <p>Background</p> <p>In Benin, Indoor Residual Spraying (IRS) and long-lasting insecticidal nets (LLINs) are the cornerstones of malaria prevention. In the context of high resistance of <it>Anopheles gambiae </it>to pyrethroids, The National Malaria Control Program (NMCP) has undertaken a full coverage of IRS in a no-flood zone in the Oueme region, coupled with the distribution of LLINs in a flood zone. We assessed the impact of this campaign on phenotypic resistance, <it>kdr </it>(knock-down resistance) and <it>ace-1^R</it>(insensitive acetylcholinesterase) mutations.</p> <p>Methods</p> <p>Insecticides used for malaria vector control interventions were bendiocarb WP (0.4 g/m²) and deltamethrin (55 mg/m²), respectively for IRS and LLINs. Susceptibility status of <it>An. gambiae </it>was assessed using World Health Organization bioassay tests to DDT, permethrin, deltamethrin and bendiocarb in the Oueme region before intervention (2007) and after interventions in 2008 and 2010. <it>An. gambiae </it>specimens were screened for identification of species, molecular M and S forms and for the detection of the West African <it>kdr </it>(L1014F) as well as <it>ace-1^R</it>mutations using PCR techniques.</p> <p>Results</p> <p>The univariate logistic regression performed showed that <it>kdr </it>frequency has increased significantly during the three years in the intervention area and in the control area. Several factors (LLINs, IRS, mosquito coils, aerosols, use of pesticides for crop protection) could explain the selection of individual resistant <it>An. gambiae</it>. The <it>Kdr </it>resistance gene could not be the only mechanism of resistance observed in the Oueme region. The high susceptibility to bendiocarb is in agreement with a previous study conducted in Benin. However, the occurrence of <it>ace-1^R</it>heterozygous individuals even on sites far from IRS areas, suggests other factors may contribute to the selection of resistance other than those exerted by the vector control program.</p> <p>Conclusion</p> <p>The results of this study have confirmed that <it>An.gambiae </it>have maintained and developed the resistance to pyrethroids, but are still susceptible to bendiocarb. Our data clearly shows that selection of resistant individuals was caused by other insecticides than those used by the IRS and LLINs.</p

Multilevel Analyses of Genetic Differentiation in Anopheles gambiae s.s. Reveal Patterns of Gene Flow Important for Malaria-Fighting Mosquito Projects

Malaria control projects based on the introduction and spread of transgenes into mosquito populations depend on the extent of isolation between those populations. On the basis of the distribution of paracentric inversions, Anopheles gambiae has been subdivided into five subspecific chromosomal forms. Estimating gene flow between and within these forms of An. gambiae presents a number of challenges. We compared patterns of genetic divergence (F(ST)) between sympatric populations of the Bamako and Mopti forms at five sites. We used microsatellite loci within the j inversion on chromosome 2, which is fixed in the Bamako form but absent in the Mopti form, and microsatellites on chromosome 3, a region void of inversions. Estimates of genetic diversity and F(ST)'s suggest genetic exchanges between forms for the third chromosome but little for the j inversion. These results suggest a role for the inversion in speciation. Extensive gene flow within forms among sites resulted in populations clustering according to form despite substantial gene flow between forms. These patterns underscore the low levels of current gene flow between chromosomal forms in this area of sympatry. Introducing refractoriness genes in areas of the genome void of inversions may facilitate their spread within forms but their passage between forms may prove more difficult than previously thought

Effect of water source and feed regime on development and phenotypic quality in Anopheles gambiae (s.l.): prospects for improved mass-rearing techniques towards release programmes

Abstract Background In many malaria-endemic sub-Saharan countries, insecticide resistance poses a threat to existing mosquito control measures, underscoring the need for complementary control methods such as sterile and/or genetically-modified mosquito release programmes. The sibling species Anopheles gambiae and An. coluzzii are responsible for malaria transmission in most of this region. In their natural habitat, these species generally breed in clean, soft water and it is believed that divergent preference in their larval breeding sites have played a role in their speciation process. Mosquito release programmes rely on the rearing of mosquitoes at high larval densities. Current rearing protocols often make use of deionised water regardless of the strain reared. They also depend on a delicate balance between the need for adequate feeding and the negative effect of toxic ammonia and food waste build-up on mosquito development, making managing and improving water quality in the insectary imperative. Methods Here, we investigated the impact of water source and feed regimes on emergence rate and phenotypic quality of mosquitoes in the insectary. First-instar larvae of An. gambiae (Kisumu strain) and An. coluzzii (Mopti and VK3 strains) were reared in three water sources with varying degrees of hardness (deionised, mineral and a mix of the two), with a daily water change. Larvae were fed daily using two standardised feeding regimes, solution and powder feed. Results Water source had a significant impact on mosquito size and development time for all strains. Earlier emergence of significantly larger mosquitoes was observed in mineral water with the smallest mosquitoes developing later from deionised water. Wing-length was significantly longer in mineral, mixed water and in powder feed, irrespective of sex, strains or water types. Deionised water was the least favourable for mosquito quality across all strains. Conclusions Mineral water and powder feed should be used in rearing protocols to improve mosquito quality where the optimal quality of mosquitoes is desired. Although results obtained were not significant for improved mosquito numbers, the phenotypic quality of mosquitoes reared was significantly improved in mineral water and mix water. Further studies are recommended on the impact mineral water has on other fitness traits such as longevity, fecundity and mating competitiveness

Cooperative Blood-feeding and the Function and Implications of Feeding Aggregations in the Sand Fly, Lutzomyia longipalpis (Diptera: Psychodidae)

Given the importance that the evolution of cooperation bears in evolutionary biology and the social sciences, extensive theoretical work has focused on identifying conditions that promote cooperation among individuals. In insects, cooperative or altruistic interactions typically occur amongst social insects and are thus explained by kin selection. Here we provide evidence that in Lutzomia longipalpis, a small biting fly and an important vector of leishmaniasis in the New World, cooperative blood-feeding in groups of non-kin individuals results in a strong decrease in saliva expenditure. Feeding in groups also strongly affected the time taken to initiate a bloodmeal and its duration and ultimately resulted in greater fecundity. The benefits of feeding aggregations were particularly strong when flies fed on older hosts pre-exposed to sand fly bites, suggesting that flies feeding in groups may be better able to overcome their stronger immune response. These results demonstrate that, in L. longipalpis, feeding cooperatively maximizes the effects of salivary components injected into hosts to facilitate blood intake and to counteract the host immune defences. As a result, cooperating sand flies enjoy enormous fitness gains. This constitutes, to our knowledge, the first functional explanation for feeding aggregations in this species and potentially in other hematophagous insects and a rare example of cooperation amongst individuals of a nonsocial insects species. The evolution of cooperative group feeding in sand flies may have important implications for th