Plant-feeding phlebotomine sand flies, vectors of leishmaniasis, prefer Cannabis sativa.

Blood-sucking phlebotomine sand flies (Diptera: Psychodidae) transmit leishmaniasis as well as arboviral diseases and bartonellosis. Sand fly females become infected with Leishmania parasites and transmit them while imbibing vertebrates' blood, required as a source of protein for maturation of eggs. In addition, both females and males consume plant-derived sugar meals as a source of energy. Plant meals may comprise sugary solutions such as nectar or honeydew (secreted by plant-sucking homopteran insects), as well as phloem sap that sand flies obtain by piercing leaves and stems with their needle-like mouthparts. Hence, the structure of plant communities can influence the distribution and epidemiology of leishmaniasis. We designed a next-generation sequencing (NGS)-based assay for determining the source of sand fly plant meals, based upon the chloroplast DNA gene ribulose bisphosphate carboxylase large chain (rbcL). Here, we report on the predilection of several sand fly species, vectors of leishmaniasis in different parts of the world, for feeding on Cannabis sativa We infer this preference based on the substantial percentage of sand flies that had fed on C. sativa plants despite the apparent "absence" of these plants from most of the field sites. We discuss the conceivable implications of the affinity of sand flies for C. sativa on their vectorial capacity for Leishmania and the putative exploitation of their attraction to C. sativa for the control of sand fly-borne diseases.

Treatment of livestock with systemic insecticides for control of Anopheles arabiensis in western Kenya.

BACKGROUND: Despite the implementation of vector control strategies, including insecticide-treated bed nets (ITN) and indoor residual spraying (IRS) in western Kenya, this area still experiences high level of malaria transmission. Novel vector control tools are required which target such vector species, such as Anopheles arabiensis, that feed outdoors and have minimal contact with ITNs and IRS. METHODS: To address this need, ivermectin, eprinomectin, and fipronil were evaluated in Zebu cattle under semi-field conditions to evaluate the potential of these compounds to reduce the survival of blood feeding An. arabiensis. Over the course of four experiments, lactating cattle received doses of oral ivermectin at 0.1 or 0.2 mg/kg, oral eprinomectin at 0.2 or 0.5 mg/kg, topical eprinomectin at 0.5, 0.75, or 1.5 mg/kg, or oral fipronil at 0.25, 0.5, 1.0, or 1.5 mg/kg. On days 1, 3, 5, 7, 14, and 21 days post-treatment, cattle were exposed to An. arabiensis, and mosquito mortality post-blood feeding was monitored. For the analysis of survival data, the Kaplan-Meier estimator and Mantel-Haenszel test was used to contrast the treatment and control survival functions. RESULTS: All three compounds significantly reduced the survival time of An. arabiensis. Twenty-one days post-treatment, mortality of mosquitoes fed on cattle dosed orally with 0.2 or 0.5 mg/kg eprinomectin, topically with eprinomectin at 0.5 mg/kg, or orally with either 1.0 or 1.5 mg/kg fipronil was still significantly higher than control mortality. CONCLUSIONS: These data demonstrate the effectiveness of three insecticidal compounds administered systemically to cattle for controlling the cattle-feeding mosquito An. arabiensis. Eprinomectin and fipronil provided the longest-lasting control. Such endectocidal treatments in cattle are a promising new strategy for control of residual, outdoor malaria transmission and could effectively augment current interventions which target more endophilic vector species.

A rodent and tick bait for controlling white-footed mice (Peromyscus leucopus) and blacklegged ticks (Ixodes scapularis), the respective pathogen host and vector of the Lyme disease spirochetes.

A promising alternative approach to conventional vector and rodent control practices is the use of a bait containing a rodenticide and acaricide in controlling vectors and pathogen reservoirs concurrently. In the United States, Lyme disease continues to be the most prevalent vector-borne disease with approximately 500,000 Lyme disease cases estimated each year. Previous research has demonstrated the usefulness of a low dose fipronil bait in controlling Ixodes scapularis larvae feeding on white-footed mice. However, considering white-footed mice can be an unwanted species because of their association with tick-borne disease and hantaviruses, a combination rodent and tick bait (RTB) might provide a useful alternative to encourage additional community participation in integrated tick management (ITM) efforts. The purpose of this research was to evaluate the use of RTB (0.025 % warfarin, 0.005 % fipronil) in controlling white-footed mice and I. scapularis larvae. Studies were designed in part based on Environmental Protection Agency (EPA) guidelines. A laboratory choice test was conducted to evaluate the use of RTB in controlling white-footed mice over 15-day exposure when they were exposed to an alternative diet. Mice were observed every day for mortality and signs of warfarin toxicity. A simulated field test was conducted to evaluate the use of RTB, presented in the presence of an alternative diet, in controlling I. scapularis parasitizing white-footed mice over 4-day exposure. Mice were fitted with capsules and manually infested with I. scapularis larvae. The inside of each capsule was observed to evaluate tick attachment. Replete larvae detaching from each mouse were collected. Blood was collected from all treatment group mice via cardiac puncture to determine the fipronil sulfone concentration in plasma for each animal. Results indicated that RTB would be adequately consumed in the presence of an alternative diet under laboratory and simulated field conditions. Treatment with RTB resulted in 100 % mortality of white-footed mice during 15-day exposure and prevented 100 % larvae from feeding to repletion during 4-day exposure. All mice succumbing to RTB showed signs of warfarin toxicity. All mice parasitized with ticks that were exposed to RTB had fipronil sulfone detectable in plasma, with even the lowest concentration detected (8.1 parts per billion) controlling 100 % parasitizing I. scapularis larvae. The results suggest that RTB could be a useful means of rodent and tick control for use in ITM programs.

Evaluation of fipronil oral dosing to cattle for control of adult and larval sand flies under controlled conditions.

Visceral Leishmaniasis (VL) is a vector-borne disease endemic to the Indian subcontinent. The Phlebotomus genus of sand flies is the vector for VL in the Old World, with the vector on the Indian Subcontinent being Phlebotomus argentipes. Cattle are a commodity in this region and a frequent host source of P. argentipes bloodmeals. The purpose of this study was to determine the efficacy of a single oral dose fipronil against adult and larval P. argentipes. Ten Bos indicus cattle were used during the study in a controlled environment. The study was conducted in Bihar, India, and involved adult and larval bioassays using laboratory-reared P. argentipes. The results were positive in that they led to up to 100% mortality in both adult and larval sand flies over a 21-d period after a single dose of fipronil.

Efficacy of a federally approved flea bait, orally administered to white-footed mice (Peromyscus leucopus), against blood feeding Ixodes scapularis larvae under simulated field conditions.

A promising alternative approach to conventional vector control practices is the use of systemic insecticides/acaricides orally administered to relevant mammalian host species to control blood feeding disease vectors. In the United States, Lyme disease continues to be the most prevalent vector-borne disease with the Centers for Disease Control and Prevention estimating approximately 500,000 Lyme disease infections each year. Previous research has demonstrated the potential usefulness of a low dose fipronil bait in controlling Ixodes scapularis larvae feeding on white-footed mice. However, no such acaricide-only product is approved for use in treating white-footed mice to control I. scapularis. The purpose of the study was to evaluate the use of a federally approved fipronil flea control bait (Grain Bait) in controlling I. scapularis parasitizing white-footed mice (Peromyscus leucopus). A simulated field trial was conducted in which Grain Bait was presented to grouped white-footed mice alongside an alternative diet for 168 h. Mice were fitted with capsules and manually parasitized with I. scapularis larvae. Replete larvae detaching from each mouse were collected and monitored for molting to nymphs. The inside of each capsule was observed to evaluate tick attachment. Blood was collected from all Treatment group mice via cardiac puncture to determine the fipronil sulfone concentration in plasma (CP) for each animal. Results indicated that Grain Bait would be consumed in the presence of an alternative diet and that bait acceptance was greater for males, relative to females. Treatment with Grain Bait prevented 100% larvae from feeding to repletion at Day 7 post-exposure and prevented 80% of larvae from feeding to repletion and 84% from molting at Day 21 post-exposure, relative to Control groups. Molted nymphs were not recovered from mice that had CP detectable ≥18.4 ng/ml. The results suggest that this federally approved flea product could be utilized for tick control and that other medically important vector-host relationships should be considered.

Development of a low-dose fipronil deer feed: Bait-screening and range-finding to determine the optimal formulation to control blacklegged ticks (Ixodes scapularis) feeding on white-tailed deer (Odocoileus virginianus).

Lyme disease is the most prevalent vector-borne disease in the U.S., and acaricidal feeds administered to white-tailed deer (Odocoileus virginianus) have potential to disrupt blood feeding by the blacklegged tick, Ixodes scapularis. Two studies were conducted with the aim of determining an ideal formulation to deliver oral acaricides to white-tailed deer and finding the lowest fipronil dose level to effectively control I. scapularis. During formulation screening, various commercial attractants (baits) were presented to deer in large paddocks under semi-field conditions and motion-sensitive cameras were used to monitor consumption by deer and non-target species. During dose range-finding, deer were housed individually and presented a fipronil feed at one of five dose levels for 48 h (two deer untreated). At 24 h post-exposure, the deer were anesthetized, blood was drawn, and deer were manually infested with 20 I. scapularis mating pairs in feeding capsules. Tick mortality was monitored up to nine days post-attachment. Results of formulation screening indicated that loose/granular formulations were substantially more palatable to deer, relative to block formulations, and would be more effective under field conditions. Dose range-finding concluded fipronil feeds with doses of 25 ppm and higher would eliminate 100% of I. scapularis parasitizing deer. Additionally, 10 ppm fipronil feed controlled a considerable proportion of ticks, and results suggested 100% ticks could be eliminated if fipronil sulfone was present in plasma at ≥25.1 ppb. These results were paramount in developing a low dose fipronil deer feed for tick control and should provide valuable insights prior to execution of future field trials.

Development of a low-dose fipronil deer feed: evaluation of efficacy against two medically important tick species parasitizing white-tailed deer (Odocoileus virginianus) under pen conditions.

BACKGROUND: Odocoileus virginianus (the white-tailed deer) is a key reproductive host for medically important tick species, including Ixodes scapularis and Amblyomma americanum. Orally administering a systemic acaricide to white-tailed deer has the potential to reduce tick reproduction, abundance and pathogen-infected tick bites. Prior studies have demonstrated considerable efficacy of a low-dose fipronil mouse bait in controlling larval I. scapularis parasitizing the pathogen reservoir, Peromyscus leucopus. No prior studies have evaluated the efficacy of a fipronil product in controlling ticks parasitizing white-tailed deer. METHODS: A pen study was conducted to evaluate the efficacy of a fipronil deer feed in controlling I. scapularis and A. americanum adult ticks. Individually housed deer (n = 24) were exposed to deer feed containing 0.0025% fipronil (fipronil deer feed) for 48 h and 120 h, and a control group of deer were exposed to an untreated placebo. On post-exposure day 7 and day 21, all deer were parasitized with 20 mating pairs of feeding capsule-enclosed I. scapularis and A. americanum. Post-attachment, engorgement and mortality of ticks were recorded. The concentrations of fipronil in plasma, feces and tissues from euthanized deer were estimated using liquid chromatography-mass spectrometry. RESULTS: The fipronil deer feed efficaciously controlled ticks parasitizing pen-reared white-tailed deer. Efficacy in reducing survivorship of blood-feeding female I. scapularis exceeded 90% in all instances except for when ticks parasitized 48-h treated deer at day 21 post-exposure (47.2%). Efficacy in reducing survivorship of A. americanum females exceeded 80% in all instances. In the 120-h exposure group there was 100% tick mortality at day 7 post-exposure for both tick species. A significant correlation was observed between reductions in tick survivorship and concentrations of fipronil sulfone in plasma. The results of tissue analysis suggest that a withdrawal period may be needed to allow for fipronil degradation prior to hunting season. CONCLUSIONS: The results provide proof-of-concept for the use of a fipronil-based oral acaricide in controlling two medically important tick species infesting a key reproductive host. A field trial is necessary to confirm the efficacy and toxicology of the product in wild deer populations. Fipronil deer feed may provide a means of controlling multiple tick species parasitizing wild ruminants to be integrated into tick management programs.

Identification of bloodmeals in wild caught blood fed Phlebotomus argentipes (Diptera: Psychodidae) using cytochrome b PCR and reverse line blotting in Bihar, India.

Identification of the source of bloodmeals in vectors plays an important role in epidemiological studies by determining the host preferences of wild sand flies in natural habitat. The anthropophilic index is a crucial component in human leishmaniasis. Bloodmeal analysis can identify the reservoir hosts of vector borne diseases. The amplification of the mitochondrial cytochrome b gene, followed by reverse line blot analysis, helps to identify the bloodmeal ingested by the wild caught sand flies. In the current study, blood fed sand flies were collected from three different villages in Bihar, India, by using Centers for Disease Control mini traps with incandescent light. Traps were placed in five different sites in the villages. Whole genome DNA was extracted from the blood fed sand flies and was amplified for the cytochrome b region, followed by reverse line blot analysis. In total, 442 blood fed sand flies were analyzed out of which 288 (65%) were positive to cytochrome b polymerase chain reaction. Humans, cattle, buffalo, and goats were the major bloodmeals identified, followed by chickens. In some of the blood fed sand flies, multiple bloodmeals were identified. In the current study, sand flies mostly fed on humans, followed by cattle, buffalo, and goats. In this regard, it is necessary to also consider cattle, buffalo, and goats when addressing vector control in Bihar, India.

Efficacy of low-dose fipronil bait against blacklegged tick (Ixodes scapularis) larvae feeding on white-footed mice (Peromyscus leucopus) under simulated field conditions.

BACKGROUND: Lyme disease, caused primarily by Borrelia burgdorferi sensu stricto, is the most prevalent vector-borne disease in the United States. Treatment of rodent pathogen reservoirs with an oral acaricide may suppress the production of infected host-seeking ticks posing a risk for human infection. A previous study showed that an oral fipronil bait effectively controlled larval Ixodes scapularis ticks on white-footed mice (Peromyscus leucopus) up to 15 days post-bait exposure. The present study expands upon this finding by exposing group-housed white-footed mice to fipronil bait under simulated field conditions prior to tick infestation. METHODS: Mice (n = 80) were housed in groups of 10 within large enclosures and offered a choice between fipronil bait within a commercial bait station and an alternative diet. The mice were assigned to two treatment groups and two control groups to undergo bait exposure durations of either 24 h (reduced) or 168 h (extended). Groups were further differentiated by the time point post-bait exposure when larval ticks were applied to mice within feeding capsules (reduced day 1, day 15; extended day 21, day 35). For 4 days post-tick introduction, attached larvae were observed by microscopy and replete larvae were recovered. Replete larvae were monitored for molting success. Plasma was collected from all treatment group mice to obtain fipronil plasma concentrations (CP). RESULTS: The fipronil bait (0.005% fipronil) was palatable and controlled larval ticks on white-footed mice when presented under simulated field conditions. Efficacy in preventing attached larvae from feeding to repletion was 100% (day 1), 89.0% (day 15), 85.8% (day 21), and 75.2% (day 35). When also considering molting success, the fipronil bait prevented 100% (day 1), 91.1% (day 15), 91.7% (day 21), and 82.5% (day 35) of larvae attaching to mice from molting. The mean CP per mouse was 191.5 ng/ml (day 1), 29.4 ng/ml (day 15), 10.6 ng/ml (day 21), and 1.0 ng/ml (day 35). CONCLUSIONS: The results suggest that fipronil bait will be consumed by white-footed mice in the presence of an alternative diet, and effectively control larval ticks on treated mice. A field trial is needed to confirm the results of this study. Low-dose fipronil bait may provide a cost-effective means of controlling blacklegged ticks to be integrated into tick management programs.

Evaluating the use of a low dose fipronil bait in reducing black-tailed prairie dog (Cynomys ludovicianus) fleas at reduced application rates.

Plague is a virulent zoonosis, vectored by fleas, posing danger to black-tailed prairie dogs (BTPDs) (Cynomys ludovicianus), black-footed ferrets (Mustela nigripes), and humans in North America. During prior research, a fipronil grain bait (0.005%) applied at rates of 1-½ cup/burrow, reduced flea abundance by > 95-100% when applied three times February-March in northern Colorado. The objective of the current study was to determine the efficacy of fipronil bait against fleas in northern Colorado at reduced application rates (½ cup/burrow) and frequencies (1-2 applications). The field study was conducted in Larimer county, Colorado USA between June-November 2018. Three test plots were selected: two treatment plots (1 vs. 2 fipronil bait applications) and one untreated control. Fipronil was applied at a rate of ½ cup (~95 g)/burrow. Fleas were collected from captured BTPDs and swabs of active burrows prior to bait application and up to 134-days post-treatment. A total of 203 BTPDs and 210 active burrows were sampled. Within the treatment plots, no fleas were collected from BTPDs up to 134-days post-treatment (100% efficacy). Five fleas were recovered from burrows within the one-application plot (<40-days post-application) with efficacy ranging from 97.1 to 100%. No fleas were recovered from burrows within the two-application plot. We caution that while fleas were present within the control plot throughout the study, abundances were low. The efficacy results are supported by those of prior field research conducted in South Dakota and suggest that fipronil bait may be applied at lower rates and frequencies than initially proposed, with potential to sustain flea removal >4-months.

Visceral leishmaniasis on the Indian Subcontinent: Efficacy of fipronil-based cattle treatment in controlling sand fly populations is dependent on specific aspects of sand fly ecology.

BACKGROUND: Visceral leishmaniasis (VL) is a deadly disease transmitted by the sand fly Phlebotomus argentipes on the Indian subcontinent, with a promising means of vector control being orally treating cattle with fipronil-based drugs. While prior research investigating the dynamic relationship between timing of fipronil-based control schemes and the seasonality of sand flies provides insights into potential of treatment on a large scale, ecological uncertainties remain. We investigated how uncertainties associated with sand fly ecology might affect our ability to assess efficacy of fipronil-based control schemes. To do this, we used a previously-described, individual-based, stochastic sand fly model to quantify how uncertainties associated with 1) the percentage of female sand flies taking blood meals from cattle, and 2) the percentage of female sand flies ovipositing in organic matter containing feces from treated cattle might impact the efficacy of fipronil-based sand fly control schemes. PRINCIPAL FINDINGS: Assuming no prior knowledge of sand fly blood meal and oviposition sites, the probabilities of achieving effective sand fly population reduction with treatments performed 3, 6 and 12 times per year were ≈5-22%, ≈27-36%, and ≈46-54%, respectively. Assuming ≥50% of sand flies feed on cattle, probabilities of achieving efficacious control increased to ≈8-31%, ≈15-42%, and ≈52-65%. Assuming also that ≥50% of sand flies oviposit in cattle feces, the above probabilities increased further to ≈14-53%, ≈31-81%, and ≈89-97%. CONCLUSIONS: Our assessments of the efficacy of fipronil-based cattle treatments in controlling sand fly populations depend on our assumptions regarding key aspects of sand fly ecology. Assessments are most sensitive to assumptions concerning the percentage of sand flies ovipositing in feces of treated cattle, thus emphasizing the importance of identifying sand fly oviposition sites. Our results place the evaluation of fipronil-based cattle treatment within a broader ecological context, which could aid in the planning and execution of a largescale field trial.

Efficacy of a low dose fipronil bait against blacklegged tick (Ixodes scapularis) larvae feeding on white-footed mice (Peromyscus leucopus) under laboratory conditions.

BACKGROUND: Lyme disease is the most prevalent vector-borne disease in the USA with cases continuing to increase. Current control measures have not been shown to be impactful, and therefore alternatives are needed. Treating pathogen reservoirs with low dose systemic acaricides in endemic areas may provide a useful tool for disrupting the cycle of the vector and pathogen. The purpose of this study was to determine the efficacy of a 0.005% fipronil bait, presented orally to white-footed mice, in controlling blacklegged tick larvae (larvae). METHODS: Sixty mice were assigned to 3 treatment groups and three untreated control groups. All individually housed mice in treatment groups were exposed to 0.005% fipronil bait for 48 hours. Larvae were manually applied to mice within feeding capsules at one of three timepoints: Day 1, Day 9 and Day 15 post-exposure. For 4-days post-tick attachment, replete larvae were collected from water moats underneath each cage and attached larvae were observed by microscopy. Plasma from 4 treated mice at Day-1, Day 13 and Day 19, and 4 control mice (n = 16) was collected to obtain fipronil plasma concentrations (CP). RESULTS: Fipronil bait did not appear to produce neophobia in mice, as the amount of bait eaten at 24- and 48-hours exposure did not differ significantly. The 48-hour fipronil bait exposure prevented 100% of larvae from feeding to repletion at Day 1, Day 9 and Day 15 post-treatment. Within the treatment groups, all larvae observable within the capsules expired and were prevented from detaching by Day 4. In contrast, within the control groups a total of 502 replete larvae were collected from moats and 348 larvae observable within the capsules successfully detached. CP averaged 948.9, 101.2 and 79.4 ng/ml for mice euthanized at Day 1, Day 9 and Day 15, respectively. No fipronil was detected in control mice. CONCLUSIONS: We provide early indication that low dose fipronil bait, orally presented to white-footed mice, can effectively control blacklegged tick larvae. Future research should modify the exposure duration and post-exposure tick attachment timepoints to simulate various field scenarios under which successful efficacy might be obtained. Low dose fipronil bait could provide a cost-effective, practical means of controlling blacklegged ticks and other arthropod vectors.

Short-term movement of Phlebotomus argentipes (Diptera: Psychodidae) in a visceral leishmaniasis-endemic village in Bihar, India.

Visceral leishmaniasis (VL), transmitted by the sand fly, Phlebotomus argentipes, is frequently reported on the Indian subcontinent where its basic ecology is largely unknown. Our objective was to evaluate the effectiveness of sugar solution (10%), containing colored food dye (0.5%) applied to peridomestic vegetation, to mark P. argentipes and subsequently estimate horizontal movement by capturing dye-marked specimens in CDC light traps in a village in Bihar, India. From September 30 to November 27, 2016, a total of 667 captured sand flies were successfully marked using sugar baits, of which 608 were P. argentipes (~91.2%). Although the majority of P. argentipes were captured <100 m from the respective marking sites, a significantly greater proportion of females (15.7%) was captured >100 m from marking sites when compared to males (3.1%). Sand flies that ingested sugar bait were only collected from areas containing >eight vegetation types and mature banana plants. The average number of marked P. argentipes captured per trap-night (±SD) <100 m from respective marking sites was greatest in peridomestic vegetation (Males: 0.9 ± 1.97; Females: 0.63 ± 1.44), followed by areas with livestock (Males: 0.66 ± 2.75; Females: 0.24 ± 0.69), and areas with humans only (Males: 0.1 ± 0.36; Females: 0.11 ± 0.31). To our knowledge, this is the only study in Bihar in which sand flies have been marked with food dyes, and the results demonstrate the potential usefulness of food dyes in estimating short-term movement of P. argentipes. Limitations of this experiment are that the number of each trap location type, vegetation composition at marking sites, and distance of all trap sites from marking sites were not homogenous, and the total number of marked sand flies collected were relatively low. In spite of the above limitations, these data should prove useful in developing a large-scale study addressing the caveats. Results of such a study could provide important information regarding the dynamics of VL transmission and inspire managers to pursue alternative means of sand fly control on the Indian subcontinent.

Field evaluation of low-dose warfarin baits to control wild pigs (Sus scrofa) in North Texas.

Wild pigs (Sus scrofa) are a highly detrimental invasive species that occupy a rapidly expanding range within the United States. In Australia, field trials evaluating baits containing 0.09% warfarin resulted in wild pig population reduction >95%. The objective of this study was to conduct an EPA-approved field trial to evaluate the use of bait containing low-dose warfarin (0.01% and 0.005%) in reducing wild pig numbers in Texas. An 8-week field test was conducted in the panhandle ~100 km southeast of Amarillo. Two ~8 km2 treatment plots were selected and each presented with either 0.01% or 0.005% warfarin baits. One control plot (~8 km2) was presented placebo. The baits were delivered using 30 species-specific feeders per plot (n = 90) that kept wildlife from accessing the toxicant. Pig movements and feed consumption were monitored during pre-treatment, treatment, and post-treatment periods. All pigs with VHF transmitters within the 0.005% warfarin-treated plot (n = 14) succumbed to the warfarin (100% mortality). Overall, 35 wild pigs were found dead from warfarin toxicosis, within both treatment plots. Total feed consumption by wild pigs was reduced by ~97.8% and ~96.2% for the 0.005% and 0.01% warfarin baited plots, respectively, indicating the absence of pigs was a result of toxic bait consumption. Results of 97 systematic searches of the treatment plots indicated no warfarin-induced non-target wildlife fatalities. Warfarin residues in wild pig livers averaged 3.69 mg/kg (n = 13) and 2.89 mg/kg (n = 9) for pigs recovered within the 0.005% plot and 0.01% warfarin plot, respectively. This study is the first efficacy field evaluation of a wild pig toxicant conducted in the US. The results suggest low-dose warfarin bait, presented in species-specific feeders, can effectively reduce wild pig numbers and pose minimal risk to non-target wildlife and domestic animals. A product containing warfarin may provide another management tool in reducing wild pig problems.

Field evaluation of a 0.005% fipronil bait, orally administered to Rhombomys opimus, for control of fleas (Siphonaptera: Pulicidae) and phlebotomine sand flies (Diptera: Psychodidae) in the Central Asian Republic of Kazakhstan.

Plague (Yersinia pestis) and zoonotic cutaneous leishmaniasis (Leishmania major) are two rodent-associated diseases which are vectored by fleas and phlebotomine sand flies, respectively. In Central Asia, the great gerbil (Rhombomys opimus) serves as the primary reservoir for both diseases in most natural foci. The systemic insecticide fipronil has been previously shown to be highly effective in controlling fleas and sand flies. However, the impact of a fipronil-based rodent bait, on flea and sand fly abundance, has never been reported in Central Asia. A field trial was conducted in southeastern Kazakhstan to evaluate the efficacy of a 0.005% fipronil bait, applied to gerbil burrows for oral uptake, in reducing Xenopsylla spp. flea and Phlebotomus spp. sand fly abundance. All active gerbil burrows within the treated area were presented with ~120 g of 0.005% fipronil grain bait twice during late spring/early summer (June 16, June 21). In total, 120 occupied and 14 visited gerbil colonies were surveyed and treated, and the resulting application rate was minimal (~0.006 mg fipronil/m2). The bait resulted in 100% reduction in Xenopsylla spp. flea abundance at 80-days post-treatment. Gravid sand flies were reduced ~72% and 100% during treatment and at week-3 post-treatment, respectively. However, noticeable sand fly reduction did not occur after week-3 and results suggest environmental factors also influenced abundance significantly. In conclusion, fipronil bait, applied in southeastern Kazakhstan, has the potential to reduce or potentially eliminate Xenopsylla spp. fleas if applied at least every 80-days, but may need to be applied at higher frequency to significantly reduce the oviposition rate of Phlebotomus spp. sand flies. Fipronil-based bait may provide a means of controlling blood-feeding vectors, subsequently reducing disease risk, in Central Asia and other affected regions globally.

Bionomics of Phlebotomus argentipes in villages in Bihar, India with insights into efficacy of IRS-based control measures.

BACKGROUND: Visceral leishmaniasis (VL) is a deadly vector-borne disease. Approximately 90% of Indian VL cases occur in Bihar, where the sand fly, Phlebotomus argentipes, is the principal vector. Sand fly control in Bihar consists of indoor residual spraying (IRS), the practice of spraying the inner walls of village dwellings with insecticides. Prior researchers have evaluated success of IRS-control by estimating vector abundance in village houses, but the number of sampling periods (n = 2-3) were minimal, and outdoor-resting P. argentipes were neglected. We describe a large-scale field study, performed in 24 villages within two Bihari districts, during which P. argentipes were collected biweekly over 47-weeks, in cattle enclosures, houses, and outdoors in peri-domestic vegetation. The objectives of this study were to provide updated P. argentipes ecological field data, and determine if program-initiated IRS-treatment had led to noticeable differences in vector abundance. PRINCIPAL FINDINGS: P. argentipes (n = 126,901) relative abundance was greatest during the summer months (June-August) when minimum temperatures were highest. P. argentipes were most frequently collected from cattle enclosures (~46% total; ~56% blood fed). Many sand flies were found to have taken blood from multiple sources, with ~81% having blood fed on humans and ~60% blood feeding on bovines. Nonparametric statistical tests were determined most appropriate for evaluating IRS-treatment. Differences in P. argentipes abundance in houses, cattle enclosures and vegetation were detected between IRS-treated and untreated villages in only ~9% of evaluation periods occurring during the peak period of human-vector exposure (June-August) and in ~8% of the total observations. No significant differences were detected between the numbers of P. argentipes collected in vegetation close to the experimental villages. CONCLUSION: The results of this study provide updated data regarding P. argentipes seasonal abundance, spatial distribution, and host preferances, and suggest vector abundance has not significantly declined in IRS-treated villages. We suggest that IRS be supplemented with vector control strategies targeting exophagic, exophilic P. argentipes, and that disease surveillance be accompanied by rigorous vector population monitoring.

Efficacy of a fipronil bait in reducing the number of fleas (Oropsylla spp.) infesting wild black-tailed prairie dogs.

Bubonic plague (Yersinia pestis) is a deadly zoonosis with black-tailed prairie dogs (Cynomys ludovicianus) as a reservoir host in the United States. Systemic insecticides are a promising means of controlling the vectors, Oropsylla spp. fleas, infesting these prairie dogs, subsequently disrupting the Y. pestis cycle. The objective of this study was to conduct a field trial evaluating the efficacy of a grain rodent bait containing fipronil (0.005%) against fleas infesting prairie dogs. The study was performed in Larimer County, CO, where bait was applied to a treatment area containing a dense prairie dog population, three times over a three-week period. Prairie dogs were captured and combed for fleas during four study periods (pre-, mid-, 1st post-, and 2nd post-treatment). Results indicated the use of bait containing fipronil significantly reduced flea burden. The bait containing fipronil was determined to reduce the mean number of fleas per prairie dog >95% for a minimum of 52 days post-initial treatment application and 31 days post-final treatment application. These results suggest the potential for this form of treatment to reduce flea population density on prairie dogs, and subsequently plague transmission, among mammalian hosts across the United States and beyond.

Visceral Leishmaniasis on the Indian Subcontinent: Modelling the Dynamic Relationship between Vector Control Schemes and Vector Life Cycles.

BACKGROUND: Visceral leishmaniasis (VL) is a disease caused by two known vector-borne parasite species (Leishmania donovani, L. infantum), transmitted to man by phlebotomine sand flies (species: Phlebotomus and Lutzomyia), resulting in ≈50,000 human fatalities annually, ≈67% occurring on the Indian subcontinent. Indoor residual spraying is the current method of sand fly control in India, but alternative means of vector control, such as the treatment of livestock with systemic insecticide-based drugs, are being evaluated. We describe an individual-based, stochastic, life-stage-structured model that represents a sand fly vector population within a village in India and simulates the effects of vector control via fipronil-based drugs orally administered to cattle, which target both blood-feeding adults and larvae that feed on host feces. PRINCIPLE FINDINGS: Simulation results indicated efficacy of fipronil-based control schemes in reducing sand fly abundance depended on timing of drug applications relative to seasonality of the sand fly life cycle. Taking into account cost-effectiveness and logistical feasibility, two of the most efficacious treatment schemes reduced population peaks occurring from April through August by ≈90% (applications 3 times per year at 2-month intervals initiated in March) and >95% (applications 6 times per year at 2-month intervals initiated in January) relative to no control, with the cumulative number of sand fly days occurring April-August reduced by ≈83% and ≈97%, respectively, and more specifically during the summer months of peak human exposure (June-August) by ≈85% and ≈97%, respectively. CONCLUSIONS: Our model should prove useful in a priori evaluation of the efficacy of fipronil-based drugs in controlling leishmaniasis on the Indian subcontinent and beyond.

Visceral leishmaniasis prevalence and associated risk factors in the saran district of Bihar, India, from 2009 to July of 2011.

India is one of three countries that account for an estimated 300,000 of 500,000 cases of visceral leishmaniasis (VL) occurring annually. Bihar State is the most affected area of India, with more than 90% of the cases. Surveys were conducted in two villages within the Saran district of Bihar, India, from 2009 to July of 2011 to assess risk factors associated with VL. Forty-five cases were identified, and individuals were given an oral survey. The results indicated that men contracted the disease more than women (58%), and cases over the age of 21 years accounted for 42% of the total VL cases. April to June showed the highest number of new cases. Of 135 households surveyed for sleeping conditions, 95% reported sleeping outside, and 98% slept in beds. Proximity to VL cases was the greatest risk factor (cluster 1 relative risk = 11.89 and cluster 2 relative risk = 138.34). The VL case clustering observed in this study can be incorporated in disease prevention strategies to more efficiently and effectively target VL control efforts.

The role of Palmyra palm trees (Borassus flabellifer) and sand fly distribution in northeastern India.

Visceral leishmaniasis (VL), known as Kala-azar in India, is a parasite transmitted by the bite of the sand fly vector Phlebotomus argentipes. Published information on the species indicates it is a poor flyer, mainly hopping and gliding. This study describes the vector as more arboreal than previously documented. Data collected indicate the ability of P. argentipes and Sergentomyia spp to attain vertical heights in Palmyra palm trees Borassus flabellifer up to 18.4 m above ground level. To determine if sand flies were either climbing the tree trunk to rest in the canopy or flying, sticky traps were set around the tree trunk and checked for captures overnight. CDC traps set in the palm tree canopy resulted in the capture of 5,067 sand flies, 3,990 of which were P. argentipes. Traps were set during daylight hours to determine if sand flies remained and rested in the canopy. A total of 128 sand flies were trapped over 29 trap days in the palm trees. With the CDC traps, 130 P. argentipes and no Sergentomyia spp were captured. The converse was true for the sticky traps set around tree trunks 3 m below the CDC traps. Of the 105 sand flies collected, only one was P. argentipes and 104 were Sergentomyia spp. As reported elsewhere, this indicates Sergentomyia spp tend to climb and hop, wheareas P. argentipes are capable of longer and more sustained flight. Data presented herein suggest that P. argentipes is more exophylic and exophagic than previously reported. These findings have implications for sand fly control.