David F. Hoel

Rift Valley Fever Virus Epidemic in Kenya, 2006/2007: The Entomologic Investigations

In December 2006, Rift Valley fever (RVF) was diagnosed in humans in Garissa Hospital, Kenya and an outbreak reported affecting 11 districts. Entomologic surveillance was performed in four districts to determine the epidemic/epizootic vectors of RVF virus (RVFV). Approximately 297,000 mosquitoes were collected, 164,626 identified to species, 72,058 sorted into 3,003 pools and tested for RVFV by reverse transcription-polymerase chain reaction. Seventy-seven pools representing 10 species tested positive for RVFV, including Aedes mcintoshi/circumluteolus (26 pools), Aedes ochraceus (23 pools), Mansonia uniformis (15 pools); Culex poicilipes, Culex bitaeniorhynchus (3 pools each); Anopheles squamosus, Mansonia africana (2 pools each); Culex quinquefasciatus, Culex univittatus, Aedes pembaensis (1 pool each). Positive Ae. pembaensis, Cx. univittatus, and Cx. bitaeniorhynchus was a first time observation. Species composition, densities, and infection varied among districts supporting hypothesis that different mosquito species serve as epizootic/epidemic vectors of RVFV in diverse ecologies, creating a complex epidemiologic pattern in East Africa.

Evaluation of six mosquito traps for collection of Aedes albopictus and associated mosquito species in a suburban setting in north central Florida.

Abstract We compared 6 adult mosquito traps for effectiveness in collecting Aedes albopictus from suburban backyards with the goal of finding a more suitable surveillance replacement for the Centers for Disease Control and Prevention (CDC) light trap. Trap selection included 2 commercial propane traps (Mosquito Magnet™ Professional trap and Mosquito Magnet Liberty trap), 2 Aedes-specific traps (Fay-Prince Omnidirectional trap and Wilton trap), 1 experimental trap (Mosquito Magnet-X trap), and a standard surveillance CDC light trap that served as a control. Traps that did not generate carbon dioxide were provided with bottled CO2 at a flow rate of 500 ml/min. Those traps designed for use with chemical attractants (Mosquito Magnet traps) were baited with Lurex™ (L-lactic acid) and octenol (1-octen-3-ol) commercial baits, known attractants to Ae. albopictus. Three repetitions of a 6 × 6 Latin square test yielded a total of 37,237 mosquitoes, of which 5,280 (14.2%) were Ae. albopictus. Significantly more (P < 0.05) Ae. albopictus were collected from the experimental and commercial traps (4,244/5,280; 80.3%) than from the CDC light trap and Aedes-specific traps. The Mosquito Magnet Liberty collected the most Ae. albopictus (1,591), accounting for 30.1% of the total take, followed closely by the Mosquito Magnet-X (1,468) and the Mosquito Magnet Pro (1,185). The omnidirectional Fay-Prince trap performed better than the CDC or Wilton trap. Twenty-seven mosquito species were collected during these trials, 9 species in large enough numbers for meaningful analysis. Aedes albopictus was the second most common mosquito trapped. The results of these trials indicate that propane-powered commercial traps would serve as useful substitutes in lieu of CDC traps in Ae. albopictus surveillance efforts. Trap features advantageous for collecting Ae. albopictus and other mosquito species are discussed.

EVALUATION OF CARBON DIOXIDE, 1-OCTEN-3-OL, AND LACTIC ACID AS BAITS IN MOSQUITO MAGNETTM PRO TRAPS FOR AEDES ALBOPICTUS IN NORTH CENTRAL FLORIDA1

ABSTRACT The impact of the attractants 1-octen-3-ol (octenol) and l-lactic acid (LurexTM) on collection of Aedes albopictus in suburban backyards was assessed in Mosquito MagnetTM Pro traps. These carbon dioxide-producing traps were additionally baited with commercial formulated lures with octenol, lactic acid, octenol + lactic acid, or no attractant (control) and evaluated in 4 residential sites. Three repetitions of the study resulted in the total collection of 1,321 Ae. albopictus. Significantly more Ae. albopictus were captured in traps baited with octenol + lactic acid than in traps baited only with octenol. Lactic acid-baited and control trap captures were not significantly different from octenol + lactic acid- or octenol-baited trap totals. Octenol- + lactic acid-baited traps collected 36.2% and 52.0% more Ae. albopictus than lactic acid-baited and control traps, respectively. Male Ae. albopictus accounted for 26.7% of the total capture. Other mosquito species collected in sufficient numbers for analysis included Cx. nigripalpus, Ochlerotatus infirmatus, Psorophora ferox, and Cx. erraticus. Larger numbers of these species were collected in traps that were unbaited or baited with only octenol than in traps baited with lactic acid or octenol + lactic acid.

Efficacy of Ovitrap Colors and Patterns for Attracting Aedes albopictus at Suburban Field Sites in North-Central Florida1

Abstract We sought to visually enhance the attractiveness of a standard black ovitrap routinely used in surveillance of the Asian tiger mosquito, Aedes albopictus, and now being used as lethal ovitraps in Ae. aegypti dengue control programs. Black plastic drinking cups (ovitraps) were visually altered to offer field populations of gravid female Ae. albopictus 6 different oviposition site choices. Trials were conducted at 3 field locations in Gainesville, Orange Park, and Jacksonville, FL, during July–August 2009. A black glossy cup served as the control and was tested against 5 cup choices consisting of white, blue, orange, or black-and-white contrasting patterns (checkered or vertically striped). Means (SE) of eggs collected over 6 wk for each choice were: black 122.53 (9.63) > blue 116.74 (10.74) > checkered 101.84 (9.53) > orange 97.15 (7.95) > striped 84.62 (8.17) > white 81.84 (8.74). Black ovitraps outperformed competing colored and contrasting patterned ovicups with respect to choice from gravid Ae. albopictus seeking artificial oviposition sites.

Effects of Ivermectin on Blood-Feeding Phlebotomus papatasi, and the Promastigote Stage of Leishmania major

Ivermectin (IVM) is a chemically modified macrocyclic lactone of Streptomyces avermitilis that acts as a potent neurotoxin against many nematodes and arthropods. Little is known of IVMs effect against either blood-feeding Phlebotomus sand flies, or the infective promastigote stage of Leishmania transmitted by these flies. We injected hamsters subcutaneously with two standard IVM treatments (200 and 400 μg/kg body weight) and allowed cohorts of Leishmania major-infected Phlebotomus papatasi to blood-feed on these animals at various posttreatment time points (4 h, 1, 2, 6, and 10 days). Infected and uninfected sand flies that bit treated and untreated hamsters served as controls. Serum levels of IVM in low- and high-dose-treated hamsters were determined at the five time points. Sand fly mortality following blood feeding was recorded at 24-h intervals and, in relation to IVM treatment, was time and dose dependent. Mortality was most rapid and greatest among infected flies that fed nearest to time of dosing. Mean survival of infected sand flies after feeding on untreated hamsters was 11.5 days, whereas that of infected sand flies that fed 4 h, 1 day, or 2 days posttreatment on high-dose-treated hamsters (400 μg/kg) was 1.6, 2.1, and 2.7 days, respectively. Infected and uninfected sand flies that blood fed 6 days following low-dose IVM treatment (200 μg/kg) still experienced significantly greater mortality (p < 0.02) than controls. Promastigotes dissected out of surviving flies that fed on IVM-treated hamsters showed typical motility and survival. Moreover, 21.7% of IVM-treated hamsters developed lesions after being fed upon by infected sand flies. L. major promastigotes appeared to be tolerant to ng/mL blood levels of IVM that caused significant mortality for up to 10 days posttreatment in blood-feeding P. papatasi.

Profiling of Human Acquired Immunity Against the Salivary Proteins of Phlebotomus papatasi Reveals Clusters of Differential Immunoreactivity

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.

Phlebotomus papatasi SP15: mRNA expression variability and amino acid sequence polymorphisms of field populations

BackgroundThe Phlebotomus papatasi salivary protein PpSP15 was shown to protect mice against Leishmania major, suggesting that incorporation of salivary molecules in multi-component vaccines may be a viable strategy for anti-Leishmania vaccines.MethodsHere, we investigated PpSP15 predicted amino acid sequence variability and mRNA profile of P. papatasi field populations from the Middle East. In addition, predicted MHC class II T-cell epitopes were obtained and compared to areas of amino acid sequence variability within the secreted protein.ResultsThe analysis of PpSP15 expression from field populations revealed significant intra- and interpopulation variation.. In spite of the variability detected for P. papatasi populations, common epitopes for MHC class II binding are still present and may potentially be used to boost the response against Le. major infections.ConclusionsConserved epitopes of PpSP15 could potentially be used in the development of a salivary gland antigen-based vaccine.

Discovery of Diurnal Resting Sites of Phlebotomine Sand Flies in a Village in Southern Egypt

ABSTRACT 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 ♂♂ and 7 ♀♀ of Phlebotomus papatasi and 3 ♂♂ 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.

Deposition from Ultra-Low Volume Application of Public Health Insecticides in a Hot Desert Environment1

Abstract Three insecticides commonly used for mosquito and sand fly control were applied 30 min to 3 h after sunset during June and July 2010, at Camp Buehring, Kuwait, to determine the relative quantity of pesticides to height and distance traveled in a hot desert environment. A BVA dilution oil was used for the control. Oil-based adulticides were sprayed using a truck-mounted Curtis DynaFog Maxi-Pro 4 ultra-low volume (ULV) sprayer. Malathion (Fyfanon ULV, 96% active ingredient [AI]), resmethrin (Scourge 4 + 12, 4% AI), pyrethrins (ULD BP-300, 3% AI), and BVA Spray 13 (100% refined petroleum distillate) were mixed with Uvitex optical brightener fluorescent dye and applied at 2 speeds on evenings when wind speed was less than 16.1 km/h (10 mph). Collection targets using biodegradable cotton ribbons (1 m × 2.5 cm) were later read with a fluorometer to quantify the amount of insecticide deposited on targets set at heights of 15.2, 76.2, and 152.4 cm (6, 30, and 60 in.) and distances of 1.5, 6.1, 15.2, 30.5, 61.0, and 91.4 m (5, 20, 50, 100, 200, and 300 ft). Mean insecticide deposition across all distances was 31% on 76.2-cm targets and 49% on 152.4-cm targets, while 15.2-cm targets typically collected <20% of test spray. Mean ground temperatures were typically within 5°C of air temperatures at 152.4 cm and within 1 to 5°C of air at 15.2 cm or 76.2 cm. Collectively, mean insecticide deposition was 80% at or above 76.2 cm for all insecticides. This finding may explain in part why control of low-flying phlebotomine sand flies with ULV insecticides has been met with less than optimal success by US military forces deployed in the Middle East.

SR450 and Superhawk XP Applications of Bacillus Thuringiensis Israelensis Against Culex Quinquefasciatus

Abstract Sprayer comparisons and larval morality assays were conducted following SR450 backpack mist blower and Superhawk XP thermal fogger applications of Vectobac® WDG Bacillus thuringiensis israelensis (Bti) against Culex quinquefasciatus. Bacillus thuringiensis israelensis was applied at maximum label rate in a 232.26-m2 field plot located in north-central Florida with containers placed at 2 heights (ground level and 1.52 m above ground) on stakes positioned 3.04, 6.09, 9.14, 12.19, and 15.24 m from the spray line. Results indicated that there was no significant (P > 0.05) difference in 24- and 48-h larval mortality between the 2 sprayers or between the 2 heights. There was significant difference (P < 0.05) among the 5 rows, with mortality continuously decreasing with increasing distance from sprayer. Both sprayers provided on average >70% larval mortality 3.04–9.14 m from the spray line, and <60% mortality at 12.19 and 15.24 m. The data suggest that the SR450 and Superhawk XP may be comparable sprayers for use with Bti to control mosquito larvae.