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Phlebotomus papatasi sand flies inject their hosts with a myriad of pharmacologically active salivary proteins to assist with blood feeding and to modulate host defenses. In addition, salivary proteins can influence cutaneous leishmaniasis disease outcome, highlighting the potential of the salivary components to be used as a vaccine. Variability of vaccine targets in natural populations influences antigen choice for vaccine development. Therefore, the objective of this study was to investigate the variability in the predicted protein sequences of nine of the most abundantly expressed salivary proteins from field populations, testing the hypothesis that salivary proteins appropriate to target for vaccination strategies will be possible. PpSP12, PpSP14, PpSP28, PpSP29, PpSP30, PpSP32, PpSP36, PpSP42, and PpSP44 mature cDNAs from field collected P. papatasi from three distinct ecotopes in the Middle East and North Africa were amplified, sequenced, and in silico translated to assess the predicted amino acid variability. Two of the predicted sequences, PpSP12 and PpSP14, demonstrated low genetic variability across the three geographic isolated sand fly populations, with conserved multiple predicted MHCII epitope binding sites suggestive of their potential application in vaccination approaches. The other seven predicted salivary proteins revealed greater allelic variation across the same sand fly populations, possibly precluding their use as vaccine targets., Competing Interests: The authors declare they have no competing interests.

The Togolese Republic has a tropical and humid climate which constitutes an ideal environment for mosquitoes to breed and transmit diseases. The Aedes mosquito is known to transmit yellow fever (YF), dengue, chikungunya, and Zika viruses in West Africa. Togo has been suffering from YF virus transmission, despite vaccination efforts. Unfortunately, there is scarcity in the data that reflect mosquito spatial distribution in Togo, specifically possible YF vectors. In the current study, mosquito surveillance efforts targeted areas with confirmed YF cases between July and August 2012. Indoor mosquitoes were collected using knockdown insecticide spraying, whereas Biogents (BG) traps were used to collect outdoor mosquito adults. Mosquito larval surveillance was conducted as well. In total, 17 species were identified. This investigation revealed the presence of medically important vectors in Togo, especially the Aedes aegypti (Linnaeus) (Diptera: Culicidae) which was collected in the four regions. Screening of all pools of female Aedes mosquitoes for YF, by real-time PCR, showed negative results. This is the first record for Coquillettidia flavocincta (Edwards) (Diptera: Culicidae) species in West Africa. This preliminary work serves as a baseline for further mosquito distribution studies in Togo., (© 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.)

Determination of the residual activity of insecticides is an essential component in the selection of an appropriate insecticide for indoor residual spraying operations. This report presents the results of a laboratory study to evaluate the residual bio-efficacy of four insecticides sprayed on the most common house-wall surfaces that occur in Egypt (wood, mud, and cement) against Phlebotomus papatasi (Scopoli, 1786) (Diptera: Psychodidae) and Culex pipiens Linnaeus, 1758 (Diptera: Culicidae). In total, 28,050 P. papatasi females and 31,275 Cx. pipiens females were subjected to the WHO cone bioassay. Effective and extended control (≥80% mortality) was produced by lambda-cyhalothrin on indoor wood and cement surfaces. Lambda-cyhalothrin effectively controlled (>80% mortality) P. papatasi and Cx. pipiens for 10 and 12 wk postspray on wood surfaces, respectively. Deltamethrin effectively controlled Cx. pipiens for 8 wk on indoor wood, mud, and cement surfaces. Indoor and outdoor-kept surfaces treated with permethrin and malathion provided negligible efficacy against P. papatasi and Cx. pipiens. Phlebotomus papatasi was better able to survive bioassay exposure than Cx. pipiens against all insecticides investigated. The role surfaces might play in inhibiting IRS-based vector control endeavors in rural areas in developing countries was highlighted in this study. The current insecticide labeling system that includes both sand flies with mosquitoes under the same dosage category should be revised periodically., (Published by Oxford University Press on behalf of Entomological Society of America 2019.)

Phlebotomus papatasi is one of the most medically important sand fly species in the Old World, serving as a vector of Leishmania parasites and phleboviruses. Chemical control is still considered the most effective method for rapidly reducing populations of flying insects involved in vector-borne disease transmission, but is increasingly threatened by insecticide resistance in the target insect posing significant problems for entomologists responsible for control programs. This study was conducted to determine pyrethroid resistance mechanisms and the biological, physiological, and molecular impacts of resistance in Ph. papatasi, and to compare their resistance mechanisms against those reported for mosquitoes and other intensely studied dipterans. Field-collected Ph. papatasi from Aswan, Egypt, were subjected to sublethal doses of permethrin and reared as a resistant strain under laboratory conditions through 16 generations. Biological parameter observations of resistant Ph. papatasi revealed an association of resistance with productivity cost. Physiological analysis revealed that concentrations of oxidase and esterase enzymes increased in early generations of the resistant colony, and then subsided through the F16 generation to levels similar to those in a susceptible colony. The activity levels of acetylcholinesterase were higher in field-collected Ph. papatasi than in susceptible colony flies, but decreased significantly despite subsequent exposure to permethrin. The molecular search for gene mutations in the resistant strain of Ph. papatasi failed to identify any mutations common in pyrethroid-resistant mosquitoes. Our study revealed that the mechanism of pyrethroid resistance in sand flies is different than that in mosquitoes, at least at the genetic level., (Published by Oxford University Press on behalf of Entomological Society of America 2016. This work is written by US Government employees and is in the public domain in the US.)

Mosquitoes of various species mate in swarms comprised of tens of thousands of flying males. In this study, we examined Aedes aegypti swarming behavior and identified associated chemical cues. Novel evidence is provided that Ae. aegypti females aggregate by means of olfactory cues, such as aggregation pheromones. Isolation of Ae. aegypti aggregation pheromones was achieved by aeration of confined mosquitoes and collection of associated volatiles by glass filters. The collected volatiles were identified through gas chromatography mass spectrometry (GCMS). Three aggregation pheromones were collected and identified as 2,6,6-trimethylcyclohex-2-ene-1,4-dione (ketoisophorone) (CAS# 1125-21-9, t(R) = 18.75), 2,2,6-trimethylcyclohexane-1,4-dione (the saturated analog of ketoisophorone) (CAS# 20547-99-3, t(R) = 20.05), and 1-(4-ethylphenyl) ethanone (CAS# 937-30-4, t(R) = 24.22). Our biological studies revealed that the identified compounds stimulated mosquito behavior under laboratory conditions. The mechanism of mosquito swarm formation is discussed in light of our behavioral study findings. A preliminary field trial demonstrated the potential application of the isolated aggregation pheromones in controlling Ae. aegypti., (© 2014 The Society for Vector Ecology.)

Phlebotomus papatasi sand flies are among the primary vectors of Leishmania major parasites from Morocco to the Indian subcontinent and from southern Europe to central and eastern Africa. Antibody-based immunity to sand fly salivary gland proteins in human populations remains a complex contextual problem that is not yet fully understood. We profiled the immunoreactivities of plasma antibodies to sand fly salivary gland sonicates (SGSs) from 229 human blood donors residing in different regions of sand fly endemicity throughout Jordan and Egypt as well as 69 US military personnel, who were differentially exposed to P. papatasi bites and L. major infections in Iraq. Compared with plasma from control region donors, antibodies were significantly immunoreactive to five salivary proteins (12, 26, 30, 38, and 44 kDa) among Jordanian and Egyptian donors, with immunoglobulin G4 being the dominant anti-SGS isotype. US personnel were significantly immunoreactive to only two salivary proteins (38 and 14 kDa). Using k-means clustering, donors were segregated into four clusters distinguished by unique immunoreactivity profiles to varying combinations of the significantly immunogenic salivary proteins. SGS-induced cellular proliferation was diminished among donors residing in sand fly-endemic regions. These data provide a clearer picture of human immune responses to sand fly vector salivary constituents.

This study evaluated the toxicity and duration of 3 residual insecticides against the Old World sand fly, Phlebotomus papatasi, an important vector of cutaneous leishmaniasis, on 2 types of tent material used by the US military in Afghanistan and the Middle East. Vinyl and cotton duck tent surfaces were treated at maximum labeled rates of lambda-cyhalothrin (Demand CS, Zeneca Inc, Wilmington, DE), bifenthrin (Talstar P Professional, FMC Corporation, Philadelphia, PA) and permethrin (Insect Repellent, Clothing Application, 40%), then subsequently stored in indoor, shaded spaces at room temperature (60%-70% relative humidity (RH), 22°C-25°C), and under sunlight and ambient air temperatures outdoors (20%-30% RH, 29°C-44°C). Insecticide susceptible colony flies (F110) obtained from the insectary of US Naval Medical Research Unit No. 3, Cairo, Egypt, were exposed to treated tent surfaces for 30-minute periods twice monthly for up to 5 months, then once monthly thereafter, using the World Health Organization cone assay. Lambda-cyhalothrin treated cotton duck tent material stored indoors killed P. papatasi for 8 months, while the complementary sun-exposed cotton duck material killed adult flies for 1 month before the efficacy dropped to less than 80%. Sand fly mortality on permethrin- and bifenthrin-treated cotton duck decreased below 80% after 2 weeks exposure to sunlight. Shade-stored permethrin and bifenthrin cotton duck material killed more than 80% of test flies through 5 months before mortality rates decreased substantially. Vinyl tent material provided limited control (less than 50% mortality) for less than 1 month with all treatment and storage regimes. Lambda-cyhalothrin-treated cotton duck tent material provided the longest control and produced the highest overall mortalities (100% for 8 months (shaded), more than 90% for 1 month (sunlight-exposed)) of both cotton duck and vinyl tents.

Background: Sand fly saliva can drive the outcome of Leishmania infection in animal models, and salivary components have been postulated as vaccine candidates against leishmaniasis. In the sand fly Phlebotomus papatasi, natural sugar-sources modulate the activity of proteins involved in meal digestion, and possibly influence vectorial capacity. However, only a handful of studies have assessed the variability of salivary components in sand flies, focusing on the effects of environmental factors in natural habitats. In order to better understand such interactions, we compared the expression profiles of nine P. papatasi salivary gland genes of specimens inhabiting different ecological habitats in Egypt and Jordan and throughout the sand fly season in each habitat., Results: The majority of investigated genes were up-regulated in specimens from Swaymeh late in the season, when the availability of sugar sources is reduced due to water deprivation. On the other hand, these genes were not up-regulated in specimens collected from Aswan, an irrigated area less susceptible to drought effects., Conclusion: Expression plasticity of genes involved with vectorial capacity in disease vectors may play an important epidemiological role in the establishment of diseases in natural habitats.

Lighted Centers for Disease Control and Prevention (CDC) light traps were baited with carbon dioxide (CO2) produced from three different sources to compare the efficacy of each in collecting phlebotomine sand flies in Bahrif village, Aswan Governorate, Egypt. Treatments consisted of compressed CO2 gas released at a rate of 250 ml/min, 1.5 kg of dry ice (replaced daily) sublimating from an insulated plastic container, CO2 gas produced from a prototype FASTGAS (FG) CO2 generator system (APTIV Inc., Portland, OR), and a CDC light trap without a CO2 source. Carbon dioxide was released above each treatment trap's catch opening. Traps were placed in a 4 x 4 Latin square designed study with three replications completed after four consecutive nights in August 2007. During the study, 1,842 phlebotomine sand flies were collected from two genera and five species. Traps collected 1,739 (94.4%) Phlebotomus papatasi (Scopoli), 19 (1.0%) Phlebotomus sergenti, 64 (3.5%) Sergentomyia schwetzi, 16 (0.9%) Sergentomyia palestinensis, and four (0.2%) Sergentomyia tiberiadis. Overall treatment results were dry ice (541) > FG (504) > compressed gas (454) > no CO2 (343). Total catches of P. papatasi were not significantly different between treatments, although CO2-baited traps collected 23-34% more sand flies than the unbaited (control) trap. Results indicate that the traps baited with a prototype CO2 generator were as attractive as traps supplied with CO2 sources traditionally used in sand fly surveillance efforts. Field-deployable CO2 generators are particularly advantageous in remote areas where dry ice or compressed gas is difficult to obtain.

Four types of commercial mosquito control traps, the Mosquito Magnet Pro (MMP), the Sentinel 360 (S360), the BG-Sentinel (BGS), and the Mega-Catch Ultra (MCU), were compared with a standard Centers for Disease Control and Prevention (CDC) light trap for efficacy in collecting phlebotomine sand flies (Diptera: Psychodidae) in a small farming village in the Nile River Valley 10 km north of Aswan, Egypt. Each trap was baited with either carbon dioxide (CO2) from combustion of butane gas (MMP), dry ice (CDC and BGS traps), light (MCU and S360), or dry ice and light (CDC). Traps were rotated through five sites in a5 x 5 Latin square design, repeated four times during the height of the sand fly season (June, August, and September 2007) at a site where 94% of sand flies in past collections were Phlebotomus papatasi (Scopoli). A total of 6,440 sand flies was collected, of which 6,037 (93.7%) were P. papatasi. Of the CO2-baited traps, the BGS trap collected twice as many P. papatasi as the MMP and CDC light traps, and at least three times more P. papatasi than the light-only MCU and S360 traps (P < 0.05). Mean numbers (+/- SE) of P. papatasi captured per trap night were as follows: BGS 142.1 (+/- 45.8) > MMP 56.8 (+/- 9.0) > CDC 52.3 (+/- 6.1) > MCU 38.2 (+/- 6.4) > S360 12.6 (+/- 1.8). Results indicate that several types of commercial traps are suitable substitutes for the CDC light trap in sand fly surveillance programs.

In an attempt to find diurnal resting sites of adult phlebotomine sand flies, potential phlebotomine adult habitats were aspirated in the village of Bahrif in Aswan, Egypt. During this survey, sand flies were aspirated from low (30-45 cm high) irregular piles of mud bricks found under high date palm canopies between the village and the Nile River. There were 5 males and 7 females of Phlebotomus papatasi and 3 males of Sergentomyia schwetzi. Six of the 7 aspirated females were engorged with blood. A total of 78 sand flies was captured on 3 glue boards placed overnight on the ground next to the mud bricks. Attempts to aspirate sand flies from adjacent walls and plants were unsuccessful. The identification of diurnal resting sites in less structured habitats may ultimately lead to more effective adult sand fly control.

Centers For Disease Control and Prevention (CDC) light traps were modified for use with light-emitting diodes (LED) and compared against a control trap (incandescent light) to determine the effectiveness of blue, green, and red lights against standard incandescent light routinely used for sand fly surveillance. Light traps were baited with dry ice and rotated through a 4 x 4 Latin square design during May, June, and July, 2006. Trapping over 12 trap nights yielded a total of 2,298 sand flies in the village of Bahrif, 6 km north of Aswan on the east bank of the Nile River in southern Egypt. Phlebotomus papatasi comprised 94.4% of trap collections with five other species collected in small numbers. Over half (55.13%) of all sand flies were collected from red light traps and significantly more sand flies (P < 0.05) were collected from red light traps than from blue, green, or incandescent light traps. Red light traps collected more than twice as many sand flies as control (incandescent) traps and > 4 x more than blue and green light traps. Results indicate that LED red light is a more effective substitute for standard incandescent light when surveying in areas where P. papatasi is the predominant sand fly species. Each LED uses approximately 15% of the energy that a standard CDC lamp consumes, extending battery life and effective operating time of traps. Our prototype LED-modified traps performed well in this hot, arid environment with no trap failures.