Large scale systemic control short-circuits pathogen transmission by interrupting the sand rat (Psammomys obesus)-to-sand fly (Phlebotomus papatasi) Leishmania major transmission cycle.

Systemic control uses the vertebrate hosts of zoonotic pathogens as "Trojan horses," killing blood-feeding female vectors and short-circuiting host-to-vector pathogen transmission. Previous studies focused only on the effect of systemic control on vector abundance at small spatial scales. None were conducted at a spatial scale relevant for vector control and none on the effect of systemic control on pathogen transmission rates. We tested the application of systemic control, using Fipronil-impregnated rodent baits, in reducing Leishmania major (Kinetoplastida: Trypanosomatidae; Yakimoff & Schokhor, 1914) infection levels within the vector, Phlebotomus papatasi (Diptera: Psychodidae; Scopoli, 1786) population, at the town-scale. We provided Fipronil-impregnated food-baits to all Psammomys obesus (Mammalia:Muridae; Cretzschmar, 1828), the main L. major reservoir, burrows along the southern perimeter of the town of Yeruham, Israel, and compared sand fly abundance and infection levels with a non-treated control area. We found a significant and substantial treatment effect on L. major infection levels in the female sand fly population. Sand fly abundance was not affected. Our results demonstrate, for the first time, the potential of systemic control in reducing pathogen transmission rates at a large, epidemiologically relevant, spatial scale.

The Potential of Systemic Control of Sand Flies Using Fipronil in the Novel Leishmania major (Kinetoplastida: Trypanosomatidae) Reservoirs Meriones tristrami (Rodentia: Muridea) and Meriones crassus (Rodentia: Muridea).

Leishmania major (Yakimoff & Schokhor, 1914), an important causative agent of Old World Cutaneous Leishmaniasis (CL), is transmitted by sand flies among a limited number of gerbilline reservoir-species. We can take advantage of this strong dependency to break the pathogen transmission cycle by using systemic insecticides that render the host toxic to the blood-feeding vector. We evaluated the potential of this approach with two novel reservoir species, incriminated for CL expansion in several sites in the Middle East. Specifically, we evaluated: 1) the residuality of the systemic insecticide fipronil in Meriones tristrami (Thomas, 1892) fed on fipronil-treated baits and 2) the treatments' adulticide effect on sand flies that blood fed on treated and untreated M. tristrami and M. crassus (Sundevall, 1842). We fed M. tristrami with food pellets containing 0.1 g/kg fipronil and used gas chromatograph-mass spectrometery analysis and bioassays to examine its residual toxicity to blood-feeding female sand flies. In M. tristrami, fipronil was rapidly metabolized to fipronil sulfone, found in the blood, urine, and feces for ≥31 d after fipronil admission. The survival of sand flies that blood fed on fipronil-treated M. tristrami and M. crassus was significantly reduced for at least 15 and 9 d respectively, after fipronil admission. These results hold promise for the potential contribution of systemic control approaches to CL integrated management strategies against novel CL (due to L. major) outbreaks in Israel and elsewhere.

Systemic Control of Cutaneous Leishmaniasis Sand-Fly Vectors: Fipronil-Treated Rodent Bait Is Effective in Reducing Phlebotomus papatasi (Diptera: Psychodidae) Female Emergence Rate From Rodent Burrows.

The strong dependency of some vectors on their host as a source of habitat can be viewed as a weak link in pathogen's transmission cycles using the vertebrate host as a 'Trojan horse' to deliver insecticides directly to the vector-host point of contact (hereafter 'systemic control'). This could, simultaneously, affect the survival of blood-feeding females and coprophagic larvae. Sand-flies, vectors of leishmaniasis worldwide, are often dependent on their bloodmeal host as a source of habitat and may therefore be good candidates for systemic control. In the present study, we field-tested this methodology by baiting Meriones crassus (Sundevall, 1842) (Rodentia:Muridea) with Fipronil-treated food pellets and evaluated its effect on reducing sand-fly emergence rate, in general, and of that of blood-fed females, in particular. We demonstrated 86% reduction in the abundance of female sand-flies that exit burrows of Fipronil-treated jirds, whereas male abundance was unaffected. Furthermore, whereas in control burrows 20% of the females were blood-fed, in treatment burrows no blood-fed females were detected. Sand-fly abundance outside the burrows was not affected by burrow treatment. This highlights the focal specificity of this method: affecting female sand-flies that feed on the reservoir host. This should result in the reduction of the pathogen transmission rate in the vicinity of the treated area by reducing the prevalence of leishmania-infected sand-fly females. These results hold promise for the potential of the systemic control approach in this system. Our next-step goal is to test this methodology at a large-scale cutaneous leishmaniasis control program.

Actin bundles play a different role in shaping scales compared to bristles in the mosquito Aedes aegypti.

Insect epithelial cells contain cellular extensions such as bristles, hairs, and scales. These cellular extensions are homologous structures that differ in morphology and function. They contain actin bundles that dictate their cellular morphology. While the organization, function, and identity of the major actin-bundling proteins in bristles and hairs are known, this information on scales is unknown. In this study, we characterized the development of scales and the role of actin bundles in the mosquito, Aedes aegypti. We show that scales undergo drastic morphological changes during development, from a cylindrical to flat shape with longer membrane invagination. Scale actin-bundle distribution changes from the symmetrical organization of actin bundles located throughout the bristle membrane to an asymmetrical organization. By chemically inhibiting actin polymerization and by knocking out the forked gene in the mosquito (Ae-Forked; a known actin-bundling protein) by CRISPR-Cas9 gene editing, we showed that actin bundles are required for shaping bristle, hair, and scale morphology. We demonstrated that actin bundles and Ae-Forked are required for bristle elongation, but not for that of scales. In scales, actin bundles are required for width formation. In summary, our results reveal, for the first time, the developmental process of mosquito scale formation and also the role of actin bundles and actin-bundle proteins in scale morphogenesis. Moreover, our results reveal that although scale and bristle are thought to be homologous structures, actin bundles have a differential requirement in shaping mosquito scales compared to bristles.

Modelling the effects of spatial heterogeneity and temporal variation in extinction probability on mosquito populations.

Spatial synchrony plays an important role in dictating the dynamics of spatial and stage-structured populations. Here we argue that, unlike the Moran effect where spatial synchrony is driven by exogenous factors, spatial correlation in intrinsic/local-scale processes can affect the level of spatial synchrony among distinct sub-populations, and therefore the persistence of the entire population. To explore this mechanism, we modelled the consequences of spatial heterogeneity in aquatic habitat quality, and that of temporal variation in local extinction probability, on the persistence of stage-structured mosquito populations. As a model system, we used two widely distributed mosquito species, Aedes albopictus and Culex pipiens, both key vectors of a range of infectious diseases. Spatial heterogeneity in aquatic habitat quality led to increased population persistence, and this pattern was more pronounced at intermediate dispersal rates, and in the long-dispersing species (C. pipiens). The highest regional persistence was obtained at high dispersal rates. This is probably because dispersal, in our model, did not carry any additional costs. Population persistence of both species was negatively correlated with increased temporal variation in local extinction probability. These differences were stronger in the short-dispersing species (A. albopictus), especially at intermediate dispersal rates. The dispersal of A. albopictus adults in each time step was limited to the nearest habitat patches, weakening the positive effect of spatial heterogeneity in aquatic habitat quality on population persistence. In contrast, C. pipiens adults could disperse into more remote sub-populations, resulting in much higher recolonization rates. Hence, the negative effect of temporal variation in local extinction probability on patch occupancy disappeared at intermediate dispersal rates. We suggest that effectively controlling these two mosquito species requires making few spatially synchronized control efforts (i.e., generating high temporal variation in local extinction probability), rather than many asynchronized local control efforts. Finally, our model can be easily fitted to other organisms characterized by complex life cycles, and it can be also used to examine alternative scenarios, including the effect of spatial configuration of local habitat patches and dispersal kernel shape on population persistence.

Effects of fish cues on mosquito larvae development.

We investigated the effects of predator-released kairomones on life history traits of larval Culex pipiens (Linnaeus). We compared the development time and survival of sibling larvae, reared in either water conditioned by the presence of Gambusia affinis (Baird and Girard) or fishless control-water. Our results indicate that larvae developing in fish-conditioned water (FCW) pupated faster than larvae in fishless-control water. The effect of FCW on larval survival was evident only in females. Surprisingly, FCW increased female survival. In both development-time and survival, boiling the water eliminated the FCW effect, supporting our hypothesis that fish conditioning is based on kairomones. Accelerated metamorphosis in response to predator released kairomones, evident in our results, is a rarely described phenomenon. Intuitively, when exposed to predator associated signals, aquatic larvae should metamorphose earlier to escape the higher risk of predation. However, theoretical models predict this outcome only under specific conditions. Indeed, longer - rather than shorter - time to metamorphosis is usually observed in response to predation risk. We argue that the response of larval mosquitoes to predation risk is context-dependent. Shortening larval development time may not be an exceptional response, but rather represents a part of a response spectrum that depends on the level of predation risk and resource abundance.

Interactive effects of salinity and a predator on mosquito oviposition and larval performance.

Oviposition habitat selection (OHS) is increasingly being recognized as playing a large role in explaining mosquito distributions and community assemblages. Most studies have assessed the role of single factors affecting OHS, while in nature, oviposition patterns are most likely explained by multiple, interacting biotic and abiotic factors. Determining how various factors interact to affect OHS is important for understanding metapopulation and metacommunity dynamics. We investigated the individual and interactive effects of three water salinities (0, 15 and 30 p.p.t. NaCl added) and the aquatic predator Anisops debilis Perplexa (Hemiptera: Notonectidae) on OHS and larval performance of the mosquitoes Ochlerotatus caspius Pallas and Culiseta longiareolata Macquart (Diptera: Culicidae) in outdoor-artificial-pool and laboratory experiments. C. longiareolata inhabited only freshwater pools, strongly avoided pools containing A. debilis, and larvae experienced lower survival in the presence of A. debilis. Salinity concentration interacted strongly with the predator in affecting OHS and larval survival of O. caspius; oviposition increased with increasing salinity in the absence of the predator and decreased with increasing salinity in the presence of the predator. O. caspius larval survival in predator-free pools was lowest in freshwater and highest at intermediate salinity. In predator pools, survival was highest at high salinity, where predation rate was shown to be lowest in the laboratory. Our results highlight that assessing the role of single factors in affecting mosquito distributions can be misleading. Instead, multiple factors may interact to affect oviposition patterns and larval performance.

Species-specific non-physical interference competition among mosquito larvae.

Individuals of different sex, size or developmental stage can compete differently and hence contribute distinctively to population dynamics. In species with complex life cycles such as insects, competitive ability is often positively correlated with larval developmental stage. Yet, little is known on how the development and survival of early-instars is influenced by interference from late-instar larvae, especially at low densities when exploitative competition is expected to be negligible. Furthermore, the specificity and mechanisms by which interference competition operates are largely unknown. We performed two complementary experiments aiming to quantify the competitive effects of late instar Ochlerotatus caspius on early instar larvae at low densities and under high resource supply rate. The first experiment examined the net effect of interference by 4(th) on 1(st) instar O. caspius larvae, relative to the effect of 1(st) instars on themselves. The second experiment examined the effect of species-specific, non-physical interference competition (i.e., cage larvae) by 4(th) on 1(st) instar O. caspius larvae at low or high densities. Specifically, we compared the responses of O. caspius larvae raised in the presence of caged con- or hetero-specific, Culiseta longiareolata, with that of larvae in the empty-cage control group. As expected, interference from late instar larvae had a net negative effect on the development rate of first instars. In contrast, the presence of caged con-specifics (non-physical interference) accelerated the development rate of O. caspius, however, this pattern was only evident at the low density. Notably, no such pattern was detected in the presence of caged hetero-specifics. These results strongly suggest the existence of species-specific growth regulating semiochemicals.

Inter- and intra-specific density-dependent effects on life history and development strategies of larval mosquitoes.

We explored how inter- and intra-specific competition among larvae of two temporary-pool mosquito species, Culiseta longiareolata and Ochlerotatus caspius, affect larval developmental strategy and life history traits. Given that their larvae have similar feeding habits, we expected negative reciprocal inter-specific interactions. In a microcosm experiment, we found sex-specific responses of larval survival and development to both intra- and inter-specific larval competition. C. longiareolata was the superior competitor, reducing adult size and modifying larval developmental time of O. caspius. We observed two distinct waves of adult emergence in O. caspius, with clear sex-specific responses to its inter-specific competitor. In males, this pattern was not affected by C. longiareolata, but in females, the timing and average body size of the second wave strongly varied with C. longiareolata density. Specifically, in the absence of C. longiareolata, the second wave immediately followed the first wave. However, as C. longiareolata abundance increased, the second wave was progressively delayed and the resulting females tended to be larger. This study improves our understanding of the way intra- and inter-specific competition combine to influence the life histories of species making up temporary pond communities. It also provides strong evidence that not all individuals of a cohort employ the same strategies in response to competition.

Foraging behavior of an urban bird species: molt gaps, distance to shelter, and predation risk.

Predation cost (Pc) is often regarded as a pivotal component determining foraging behavior. We hypothesized that variations in two of its major constituents, predation risk (mu) and the marginal value of energy ([see text for symbol]Fs/ [see text for symbol]e, where Fs is the survivor's fitness and e represents the amount of acquired energy), will translate into variations in patch use behavior of ground-foraging birds. We studied patch use behavior of House Sparrows (Passer domesticus), as affected by the proximity to shelter, in two large outdoor aviaries. Proximity to shelter should affect mu. We manipulated the birds' flight performance by clipping primary flight feathers from their wings to increase mu, but the clipping may also increase [see text for symbol]Fs/ [see text for symbol]e. To help distinguish between the birds' response to these confounding effects, we further augmented food in the aviaries to reduce [see text for symbol]Fs/ [see text for symbol]e. Patch use, as measured by giving-up densities (GUD, the amount of food left behind in a resource patch following exploitation) was affected by distance from shelter only slightly and mainly when the birds were feather-clipped and food was not augmented. Food augmentation had a homogenizing effect on foraging costs by increasing GUDs and washing out the effects of distance and feather clipping. We argue that mu increases with distance from shelter but that, for the highly urban House Sparrow, this increase is only slight. Feather clipping then increased mu further to the point at which patch use discernibly decreased with distance from shelter. Our experimental manipulation of feather clipping also acted to increase [see text for symbol]Fs/ [see text for symbol]e and resulted in an overall lowering of GUDs. The seed augmentation counteracted the effect of feather clipping on [see text for symbol]Fs/ [see text for symbol]e, allowing the birds to reduce their foraging efforts and washing out the qualitative effect of mu with respect to distance from shelter.

Drinking water boosts food intake rate, body mass increase and fat accumulation in migratory blackcaps (Sylvia atricapilla).

Fat accumulation by blackcaps (Sylvia atricapilla) is a prerequisite for successful migratory flight in the autumn and has recently been determined to be constrained by availability of drinking water. Birds staging in a fruit-rich Pistacia atlantica plantation that had access to water increased their body mass and fat reserves both faster and to a greater extent than birds deprived of water. We conducted a series of laboratory experiments on birds captured during the autumn migration period in which we tested the hypotheses that drinking water increases food use by easing limitations on the birds' dietary choices and, consequently, feeding and food processing rates, and that the availability of drinking water leads to improved digestion and, therefore, to higher apparent metabolizable energy. Blackcaps were trapped in autumn in the Northern Negev Desert, Israel and transferred to individual cages in the laboratory. Birds were provided with P. atlantica fruit and mealworms, and had either free access to water (controls) or were water-deprived. In experiment 1, in which mealworm availability was restricted, water-deprived birds had a fourfold lower fruit and energy intake rates and, consequently, gained less fat and total mass than control birds. Water availability did not affect food metabolizability. In experiment 2, in which mealworms were provided ad libitum, water availability influenced the birds' diet: water-restricted birds ate more mealworms, while control birds consumed mainly P. atlantica fruit. Further, in experiment 2, fat and mass gain did not differ between the two treatment groups. We conclude that water availability may have important consequences for fat accumulation in migrating birds while they fatten at stopover sites, especially when water-rich food is scarce. Restricted water availability may also impede the blackcap's dietary shift from insectivory to frugivory, a shift probably necessary for successful pre-migratory fattening.