Oral treatment of rodents with fipronil for feed-through and systemic control of sand flies (Diptera: Psychodidae).

The sand fly Phlebotomus papatasi Scopoli is the vector of Leishmania major (Yakimoff & Schokhor), which is maintained in populations of burrowing rodents. The purpose of this study was to conduct a laboratory study to determine the efficacy of oral treatment of rodents with fipronil for control of sand flies that feed on rodent feces as larvae or on rodent blood as adults. We determined through larval bioassays that fipronil was eliminated in feces of orally-treated hamsters at a level that was significantly toxic to sand fly larvae for 21 d after the hamsters had been withdrawn from a fipronil-treated diet. Through bloodfeeding bioassays, we also found that fipronil was present in the peripheral blood of hamsters at a concentration that was significantly toxic to bloodfeeding adult female sand flies for 49 d after the hamsters had been withdrawn from their treated diet. The results of this study suggest that fipronil acts as well as or better than feed-through or systemic insecticides that previously have been measured against sand flies, and is particularly promising because this single compound acts against both larvae and bloodfeeding adults. An area-wide approach using rodent baits containing a fipronil could suppress vector populations that originate in the vicinity of rodent reservoirs, and could be used to eliminate the most epidemiologically important part of the vector population: female sand flies that take bloodmeals on rodent reservoirs.

Laboratory evaluation of rubidium as a long-lasting marker for bloodfeeding sand flies (Diptera: Psychodidae).

The objective of this study was to evaluate the use of the trace element rubidium (Rb) as a long-lasting systemic biomarker for bloodfeeding females of the sand fly Phlebotomus papatasi Scopoli. Baits containing Rb chloride were found to be palatable to hamsters in this study. We were able to detect Rb using a portable X-ray fluorescence analyzer in all sand flies that fed on Rb-treated hamsters for at least 14 d postbloodmeal. We also detected Rb in sand flies that took a bloodmeal from hamsters up to 10 d after the hamsters were withdrawn from a Rb-treated diet. Results of this study constitute proof of concept for the incorporation of Rb chloride into rodent baits for marking bloodfeeding sand flies, and suggest that Rb marking could be used as a technique for evaluating rodent-targeted sand fly control methods and in ecological studies on sand flies.

Evaluation of three feed-through insecticides using two rodent and two sand fly species as models.

The efficacy of 3 rodent feed-through insecticides (novaluron, pyriproxyfen, and ivermectin) was determined against larvae of the sand flies Phlebotomus duboscqi and P. papatasi using Syrian hamsters (Mesocricetus auratus) and Mongolian gerbils (Meriones unguiculatus) as laboratory models. For each insecticide, there were no significant differences between the longevity or percentage survival of sand fly larvae that had been fed feces of treated rodents for each sand fly or rodent species pairing. The results of this study suggest that larvae of P. duboscqi and P. papatasi are equally susceptible to the concentrations of the rodent feed-through insecticides tested in this study and that these insecticides are pharmacologically compatible with different rodent/sand fly interactions.

Oral treatment of rodents with insecticides for control of sand flies (Diptera: Psychodidae) and the fluorescent tracer technique (FTT) as a tool to evaluate potential sand fly control methods.

In laboratory studies, insecticides (diflubenzuron, novaluron, methoprene and, pyriproxyfen) that have been incorporated into rodent diets were effective as feed-throughs against sand fly larvae. Novaluron also was effective against sand fly larvae at low concentrations and under simulated field conditions. Ivermectin has been shown to be effective as a systemic insecticide, killing 100% of blood-feeding sand flies for up to seven d after rodents were treated. The fluorescent tracer technique (FTT) is the use of certain fluorescent dyes (rhodamine B or uranine O) as feed-through transtadial biomarkers for phlebotomine sand flies, systemic biomarkers for blood-feeding sand flies, and permanent markers for nectar-feeding sand flies. The results of these laboratory studies provide proof of concept for the FTT and indicate that the FTT could be used to delineate specific foci with rodent/sand fly associations that would be susceptible to control by using feed-through or systemic insecticides, or foci where insecticide-treated sugar baits could be used against sand flies.

Evaluation of juvenile hormone analogues as rodent feed-through insecticides for control of immature phlebotomine sandflies.

The juvenile hormone analogues methoprene and pyriproxyfen were evaluated as rodent feed-through insecticides for control of immature stages of the sandfly Phlebotomus papatasi Scopoli (Diptera: Psychodidae). The development and survival of P. papatasi second-instar larvae fed faeces from Syrian hamsters, Mesocricetus auratus, that had been fed a diet containing methoprene (0, 9.788, 97.88 or 978.8 p.p.m.) or pyriproxyfen (0, 9.82, 98.2 or 982 p.p.m.) were evaluated. The faeces of methoprene-treated hamsters greatly reduced the percentage of larvae that pupated at all concentrations tested and prevented adult emergence at all but the lowest concentration (9.788 p.p.m.). Pyriproxyfen prevented both pupation and adult emergence at all concentrations tested. The results of this study suggest that a control strategy using rodent baits containing juvenile hormone analogues to control phlebotomine sandflies that live in rodent burrows and feed on rodent faeces may be possible. As rodent reservoirs and vectors of Leishmania major live in close association in many parts of the Middle East, control of the transmission of the agent of zoonotic cutaneous leishmaniasis may also be possible.

Laboratory evaluation of the efficacy of fluorescent biomarkers for sugar-feeding sand flies (Diptera: Psychodidae).

The purpose of this study was to evaluate the use of four fluorescent dyes (rhodamine B, uranine O, auramine O, and erythrosin B) and two nonfluorescent dyes (carmoisine and indigotine) incorporated into sugar baits as biomarkers for phlebotomine sand flies. Each dye could be detected in sand flies fed baits with dye for 24 h when examined using bright field microscopy, although there was considerable variability in the marking produced; all sand flies that had ingested rhodamine B-treated sucrose solution were marked clearly. Sand flies that had ingested sucrose solution containing rhodamine B or uranine O at concentrations as low as 10 mg/L were consistently detected under fluorescence microscopy. None of the treatments in this study reduced the longevity of sand flies. All sand flies fed sucrose solution containing rhodamine B or uranine O were marked for at least 14 d, whereas only 20% of sand flies were marked 3 d after feeding on a carmoisine-treated solution. When rhodamine B and uranine O were combined in a single sucrose solution or when the dyes were fed sequentially to sand flies, both dyes could be detected in sand flies using fluorescence microscopy. We propose that rhodamine B- or uranine O-treated sucrose baits could be used in ecological studies or to identify portions of the adult sand fly population that could be targeted with insecticide-treated sugar baits.

Oral treatment of rodents with ivermectin for the control of Phlebotomus papatasi (Diptera: Psychodidae) under laboratory conditions.

The macrocyclic lactone ivermectin was evaluated as a rodent systemic and feed-through insecticide for control of adult and immature sand flies (Phlebotomus papatasi Scopoli). Syrian hamsters were fed diets containing 0 or 20 mg/kg ivermectin for 9 days, and sand flies were allowed to take bloodmeals from the hamsters 0, 3, 7, and 14 days after they were withdrawn from their diets. Ivermectin treatment of hamsters was 100% effective against bloodfeeding sand flies for up to 7 days after hamsters were withdrawn from ivermectin-treated diets. The survival and fecundity of sand flies that took bloodmeals from hamsters 14 days after they had been withdrawn from their ivermectin-treated diets were not significantly different from sand flies that took bloodmeals from control hamsters. Feces of the hamsters were collected 0, 3, 7, and 14 days after the hamsters had been withdrawn from their diets, and the feces were fed to 2nd instars of P. papatasi. All larvae that were fed feces of ivermectin-treated hamsters voided 0 days after being withdrawn from their diets died before pupation; larvae fed feces voided by ivermectin-treated hamsters 3 and 7 days after being withdrawn from their treated diets had significantly reduced survival. The results of this study suggest that oral ivermectin treatment of rodents could control immature and adult female sand flies that are closely associated with rodents.

Laboratory evaluation of novaluron as a rodent feed-through insecticide against sand fly larvae (Diptera: Psychodidae).

Experiments were conducted to evaluate novaluron as a feed-through larvicide to control immature phlebotomine sand flies (Phlebotomus papatasi Scopoli) (Diptera: Psychodidae). The minimum effective concentration of novaluron against sand fly larvae fed hamster feces treated with novaluron, or feces of hamsters fed a diet containing novaluron, was 9.88 and 9.88 x 10(-1) mg/kg, respectively. Feces of novaluron-treated hamsters were held under conditions simulating the inside of a rodent burrow for up to 30 d, and all larvae that consumed these feces died before pupation; a significant reduction in treated larval survival relative to control was observed when the feces were aged for up to 150 d. Novaluron was shown to be effective as a feed-though larvicide when novaluron-treated food made up only a portion of the diet of hamsters. The results of this study suggest that novaluron could be effective as a rodent feed-through insecticide in a field setting.

Laboratory evaluation of insecticide-treated sugar baits for control of phlebotomine sand flies (Diptera: Psychodidae).

The purpose of this study was to evaluate the use of boric acid, imidacloprid, ivermectin, or abamectin incorporated into sugar baits as oral toxicants for adult phlebotomine sand flies. Variable toxicity of insecticide-sugar bait solutions to adult male and female sand flies was demonstrated, based on male female median lethal concentration values of 0.10-0.08, 6.13-9.53, and 9.03-18.11 mg/liter of imidacloprid, ivermectin, and abamectin, respectively. Complete control of sand flies could not be achieved with as high as 40 g/liter of boric acid in sugar bait solution; concentrations >40 g/liter were found repellent to the sand flies. Uranine O (a fluorescent tracer dye that can be used to measure the ingestion of sugar baits by sand flies) did not interact negatively with imidacloprid, ivermectin, or abamectin when it was combined with the insecticides in a sugar bait. Also, incorporation of imidacloprid, ivermectin, or abamectin into sugar baits did not reduce the effect whether adult male and female sand flies fed on these sugar baits. We propose that imidacloprid, ivermectin, or abamectin could be used to control adult sand fly populations with targeted use of insecticide-treated sugar baits.

Evaluation of rhodamine B as an orally delivered biomarker for rodents and a feed-through transtadial biomarker for phlebotomine sand flies (Diptera: Psychodidae).

The purpose of this study was to evaluate the use of rhodamine B as an orally delivered biomarker for rodents and a feed-through transtadial biomarker for phlebotomine sand flies (Diptera: Psychodidae). Rhodamine B-treated hamsters were visibly marked for up to 8 wk, and their feces were fluorescent when examined under a fluorescence microscope. The development and survival of sand fly larvae fed feces of rhodamine B-treated hamsters were not significantly different from control sand flies. Adult male and female sand flies, that had been fed as larvae the feces of rhodamine B-treated hamsters, were fluorescent when examined using fluorescent microscopy and could be distinguished from control sand flies. Adult female sand flies that took bloodmeals from rhodamine B-treated hamsters were fluorescent when examined immediately after feeding. Rhodamine B incorporated rodent baits could be used to detect adult male and female sand flies that fed on the feces of baited rodents as larvae, or adult female sand flies that have taken a bloodmeal from bait-fed rodents. This would allow the delineation of specific foci with rodent-sand fly associations that would be susceptible to control by using feed-through or systemic insecticides.

Ivermectin as a rodent feed-through insecticide for control of immature sand flies (Diptera: Psychodidae).

Ivermectin was evaluated as a potential rodent feed-through for the control of immature stages of Phlebotomus papatasi. The survival of sand fly larvae fed feces of Syrian hamsters (Mesocricetus auratus) that had been fed a diet containing 0, 2, 6, 10, 20, 60, or 100 ppm ivermectin was measured. Sand fly larvae fed the feces of ivermectin-treated hamsters had significantly reduced survival, with 100% mortality of larvae fed feces of hamsters fed a diet containing 20, 60, and 100 ppm ivermectin. The results of this study suggest that a control strategy using rodent baits containing ivermectin to control phlebotomine sand flies may be possible. Because rodent reservoirs and sand fly vectors of Leishmania major live in close association in many parts of the Middle East, the control of transmission of the agent of zoonotic cutaneous leishmaniasis also may be possible.

Evaluation of novaluron as a feed-through insecticide for control of immature sand flies (Diptera: Psychodidae).

The development and survival of sand fly Phlebotomus papatasi Scopoli (Diptera: Psychodidae) larvae fed feces of Syrian hamsters, Mesocricetus auratus, that had been fed a diet containing novaluron were evaluated. In total, six larval diets were used in sand fly larval bioassays. Four groups of larvae were fed feces of hamsters that had been maintained on a diet containing either 0, 9.88, 98.8, or 988 ppm novaluron. Two additional groups were fed a larval diet composed of equal parts composted rabbit feces and rabbit chow containing either 0 or 988 ppm novaluron. No pupation, hence no adult emergence, occurred when larvae were fed feces of hamsters that were fed diets containing novaluron. The mortality of sand flies fed feces of treated hamsters occurred during larval molts. The results of this study suggest that a control strategy using rodent baits containing novaluron to control phlebotomine sand flies and zoonotic cutaneous leishmaniasis may be possible.

Laboratory evaluation of diflubenzuron as a feed-through for control of immature sand flies (Diptera: Psychodidae).

The benzoylurea chitin synthesis inhibitor diflubenzuron was evaluated as a rodent feed-through for the control of immature stages of Phlebotomus papatasi Scopoli (Diptera: Psychodidae). The development and survival of second instars of P. papatasi larvae that were fed feces from Syrian hamsters, Mesocricetus auratus, that had been fed a diet containing 0, 8.97, 89.7, or 897 ppm diflubenzuron was evaluated. No pupation or adult emergence occurred when larvae were fed feces from hamsters that were fed diets containing diflubenzuron. The mortality of sand flies fed feces from treated hamsters was coincident with pupation of the controls, suggesting a specific effect on the larval-to-pupal molt. The results of this study suggest that a control strategy using rodent baits containing diflubenzuron for phlebotomine sand flies and zoonotic cutaneous leishmaniasis may be possible.