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Chiggers are larval mites that pose a significant health risk globally via the spread of scrub typhus. However, fundamental studies into the bacterial microbiome in North America have never been considered. In this investigation, chiggers were collected in the wild from two locally common rodent host species (i.e., Sigmodon hispidus and Peromyscus leucopus) in three different ecoregions of North Carolina (NC), United States to investigate the composition of their bacterial communities, including potential pathogens. DNA was extracted from the chiggers, and the V3-V4 regions of the bacterial 16S rRNA gene were sequenced using next-generation sequencing (NGS). Alpha diversity metrics revealed significant differences in bacterial diversity among different collection counties. Beta diversity metrics also revealed that bacterial communities across counties were significantly different, suggesting changes in the microbiome as the environment changed. Specifically, we saw that the two western NC collection counties had similar bacterial composition as did the two eastern collection counties. In addition, we found that the chigger microbiome bacterial diversity and composition differed between rodent host species. The 16S rRNA sequence reads were assigned to 64 phyla, 106 orders, 199 families, and 359 genera. The major bacterial phylum was Actinobacteria. The most abundant species were in the genera Corynebacterium, Propionibacterium, class ZB2, and Methylobacterium. Sequences derived from potential pathogens within the genera Orientia and Rickettsia were also detected. Our findings provide the first insights into the ecology of chigger microbiomes in the US. Further research is required to determine if the potential pathogens found detected in chiggers are a threat to humans and wildlife. [ABSTRACT FROM AUTHOR]

Scrub typhus, a rickettsial disease caused by Orientia spp., is transmitted by infected larval trombiculid mites (chiggers). We report the molecular detection of Orientia species in free-living Eutrombicula chiggers collected in an area in North Carolina, USA, to which spotted fever group rickettsiae infections are endemic. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Skip oviposition is considered as an example of spatial bet hedging and involves a tradeoff between the benefit of reducing the risk of complete reproductive failure due to stochastic loss of a breeding site and the energetic and mortality costs associated with dispersal across several oviposition sites. Skip oviposition has been studied extensively for container-breeding mosquitoes but has never been studied with sand flies. By conducting a series of bioassays using solitary gravid females exposed to varying numbers of oviposition sites and to oviposition sites of variable quality at small (oviposition jar) and medium (free-flight cage) scales, we showed that sand flies exhibited skip oviposit at both scales. Specifically, with low-quality oviposition sites, females spread their eggs across several oviposition sites with total egg clutch size remaining constant and number of eggs per oviposition site decreasing with increasing number of oviposition sites. With variable quality sites, sand flies biased their oviposition to sites containing increasing levels of organic matter (OM), but also laid eggs in poor-quality sites; a behavior consistent with spatial bet hedging. We also demonstrated that the presence of OM stimulated larger egg clutch size and increased percent skip oviposition. Skip oviposition was less frequent at the free flight cage scale. But, at this scale, females were shown to be stimulated to lay more eggs when in the presence of other females than when alone. [ABSTRACT FROM AUTHOR]

Scrub typhus, a rickettsial disease caused by Orientia spp., is transmitted by infected larval trombiculid mites (chiggers). We report the molecular detection of Orientia species in free-living Eutrombicula chiggers collected in an area in North Carolina, USA, to which spotted fever group rickettsiae infections are endemic.

In phlebotomine sand fly, the topic of the factors stimulating or inhibiting egg hatching has been largely ignored. In this study, we evaluated the hypothesis that pharate neonate sand flies are able to regulate their hatching time adaptively in response to cues indicative of the presence of food or potential competitors. In this study, we evaluated the independent and combined effects of organic matter (OM) extract and proximity to conspecific eggs on the hatching proportion of Phlebotomus papatasi eggs. In one set of larval rearing jars, we introduced 16 eggs into a small hole in the center of a plaster base of the jar. In another set, we introduced a single egg into each small hole of a 4-by-4 symmetrical array. To one set, we added an aqueous OM extract, and to the other, we added deionized water (DI). OM stimulated egg hatching while egg clustering slightly inhibited egg hatching. Results of this experiment are biologically important because they show, for the first time, that pharate sand fly neonates are able to adaptively regulate their hatching time in response to external cues.

Phlebotomine sand flies are worldwide vectors of Leishmania parasites as well as other bacterial and viral pathogens. Due to the variable impact of traditional vector control practices, a more ecologically based approach is needed. The goal of this study was to isolate bacteria from the most attractive substrate to gravid Phlebotomus papatasi Scopoli sand flies and determine the role of bacterial volatiles in the oviposition attractancy of P. papatasi using behavioral assays. We hypothesized that gravid sand flies are attracted to bacterially derived semiochemical cues associated with breeding sites. Bacteria were isolated from a larvae-conditioned rearing medium, previously shown to be highly attractive to sand flies. The isolated bacteria were identified by amplifying and sequencing 16S rDNA gene fragments, and 12 distinct bacterial species were selected for two-choice olfactometer bioassays. The mix of 12 bacterial isolates elicited strong attraction at the lower concentration of 107 cells per ml and significant repellence at a high concentration of 109 cells per ml. Three individual isolates (SSI-2, SSI-9, and SSI-11) were particularly attractive at low doses. In general, we observed dose-related effects, with some bacterial isolates stimulating negative and some positive dose-response curves in sand fly attraction. Our study confirms the important role of saprophytic bacteria, gut bacteria, or both, in guiding the oviposition-site selection behavior of sand flies. Identifying the specific attractive semiochemical cues that they produce could lead to development of an attractive lure for surveillance and control of sand flies., (© The Author(s) 2020. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

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., (© The Author(s) 2020. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

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., (© The Author(s) 2020. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Most sand flies and mosquitoes require a bloodmeal for egg production, but when blood-sources are scarce, some of them can reproduce without it, so called facultative autogeny. The evolution of autogenous reproduction is thought to involve a trade-off between the benefit of reproducing in the absence of bloodmeal hosts versus the quantitative cost of reduced fecundity and/or or qualitative effect on reduced offspring development and survivorship. We blood-fed (BF) some Phlebotomous papatasi (Scopoli) sand fly females on mice while keeping others (from the same cohort) not BF. We then compared the fecundity of BF and non-blood-fed (NBF) females and also evaluated their egg mass and hatching rate, larval development rate and survivorship, pupa mass and eclosion rates, and progeny fecundity. Among NBF females, only 55% became gravid and produced three times less mature oocytes than BF ones. Autogenous females laid 3.5 and 5.7 times fewer eggs in individual and multi-female bioassays, respectively. Egg mass and hatching rate were not affected by blood-feeding. Individual-larvae bioassays suggested reduced survival during larval stages in the autogenous group. In multi-larvae bioassays, overall and especially pupae survival was significantly reduced in the autogenous group. Development rate was slower and pupal mass was reduced in progeny from autogenous mothers. These effects were particularly apparent at high larval density. Mothers' blood-feeding history did not affect daughter's fecundity. Studies on the costs of autogeny provides insights on the evolution of blood feeding. Moreover, it also provides insights regarding potential implications of autogeny to the emergence of vector-borne diseases., (© The Author(s) 2019. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)