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Atopic dermatitis (AD) is a chronic inflammatory skin disorder with a complex pathogenesis involving epidermal barrier dysfunction and aberrant lipid composition, particularly ceramides and fatty acids (FA). Conventional management options, such as topical glucocorticoids (GC), often lead to adverse effects upon prolonged usage, prompting the exploration of alternative therapeutic strategies. The lipid extract from Black Soldier Fly larvae (BSFL) biomass, holding a rich blend of FA, holds substantial potential as a novel ingredient to tackle skin barrier impairment. This study aimed to achieve proof-of-concept validation of innovative nanotechnology-based formulations tailored to enhance the topical management of AD. Specifically, solid lipid nanoparticles (SLNs) with BSFL lipid extract were developed to perform both as a carrier for dexamethasone (DEX), a representative GC, and as skin barrier repair adjuvants. Through systematic optimization using Box-Behnken Design, BSFL lipid extract-based SLNs demonstrated favorable physicochemical properties for topical application and satisfactory stability over 2 months. Notably, these SLNs exhibited favorable drug release kinetics, delivering the total DEX payload within a therapeutically relevant timeframe. Furthermore, these nanocarriers showed the ability to permeate human keratinocytes without pronounced toxicity, suggesting their potential utility in enhancing drug delivery and cellular uptake. Overall, these findings suggest that BSFL lipid extract is a promising natural and sustainable ingredient for the development of nanotechnology-driven approaches to AD management, offering a potential avenue for addressing the unmet needs in this challenging dermatologic condition., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Author(s). Published by Elsevier B.V. All rights reserved.)

Introduction: Melanoma is known as a highly lethal cancer. In melanoma cells, apoptosis signalling which relies heavily on the acute activity of mitochondria and reactive oxygen species (ROS) formation is suppressed. Our previous studies on natural compounds on melanoma suggested that mitochondria are a potential target for the melanoma treatment by selective cytotoxic effect of them. The black soldier fly is an important environmental protectant insect that based on recent studies induces apoptosis in liver and colorectal carcinoma cells through the activation of caspase 3, 8, and 9 and ultimately inhibits the growth of cancer cells., Purpose: This study was designed to evaluate the selective apoptotic effect of the n-hexane BSFL extract (BSFLE) on skin mitochondria., Materials and Methods: The mitochondria isolated from melanoma cells were treated with various concentrations (1500, 3000, and 6000 µg/ml) of n-hexane BSFLE Then MTT viability assay, ROS determination, Mitochondrial Membrane Potential (MMP), mitochondrial swelling, cytochrome c release determination, and % apoptosis were performed., Results: MTT assay showed that different concentrations of n-hexane BSFLE significantly ( P  < 0.05) decreased the SDH activity in cancerous skin mitochondria with the IC50. The ROS production and mitochondrial swelling results also showed that all concentrations of BSFL extracts significantly increased. MMP decline and the release of cytochrome c in cancer groups mitochondria. BSFLE increased apoptosis on melanoma cells., Discussion and Conclusion: It is suggested that n-hexane BSFLE compounds selectively induce a cascade of proapoptotic events that are probably defective in cancer cells. Most of these compounds target the mitochondrial transient pore caused by disruption of the mitochondrial respiratory chain. These events lead to disruption of the temporary permeability of mitochondria, swelling of mitochondria and finally the formation of apoptosome in the cytosol.

Antibacterial peptides from black soldier fly larvae extract were prepared using Flash column chromatography. Three out of five fractions (F 2 , F 3 and F 4 ) showed antibacterial activity against Listeria monocytogenes DMST 17303 with a minimum inhibitory concentration (MIC) of 1 mM, followed by Salmonella enterica Enteritidis DMST 15679 and Escherichia coli O157:H7 DMST 12743 with a MIC ranging from 4 to 8 mM. Due to the higher yield, F 2 and F 3 were further analyzed on their mode of action against L. monocytogenes DMST 17303. Both fractions, particularly F 2 , exerted antibacterial activity through inducing bacterial cell membrane disintegration and interaction with intracellular compounds including fatty acids, proteins, and nucleic acids. The F 3 did not show significant hemolytic activity up to 4 mM, while F 2 showed lower than 5% hemolysis up to 8 mM. Time-to-kill analysis revealed that F 2 was more effective and exerted a sustainable killing effect after 2 and 4 h, depending on the concentration of 1 and 2×MIC, respectively, while the F 3 at 2×MIC could completely kill the test bacteria within 24 h. Among the identified peptides in the fractions, those with charged, either positively or negatively, and moderate hydrophobicity of ranging 6.68-15.70, namely CGPPRQGPFPR, HLEEELK, LEEAEERAD, TEELEEAKKK, and KGNSELEEAKKK, are potential antimicrobial peptides., (© 2024. The Author(s).)

The black soldier fly (BSF, Hermetia illucens) is a resource insect that can utilize livestock and poultry feces. However, BSFs may also increase the risk of transmission of antibiotic resistance genes (AGRs) that are widespread in livestock and poultry farm environments. Therefore, we aimed to evaluate the biosecurity risks of different BSF treatments in the laying chicken food chain using the "chicken manure-BSF-laying hens" model. Our results indicated that different BSF treatments significantly affected antibiotic residue, ARGs, MGEs, bacterial antibiotic resistance, and bacterial microbial community composition in the food chain of laying hens fed BSFs. These risks can be effectively reduced through starvation treatment and high-temperature grinding treatment. Comprehensive risk assessment analysis revealed that starvation combined with high-temperature milling (Group H) had the greatest effect., Competing Interests: DISCLOSURES The authors declare no conflicts of interest., (Copyright © 2024. Published by Elsevier Inc.)

Background Objectives: At present, use of synthetic pesticides to manage insects and other arthropods creates a number of issues that are related to the environment and public health. The goal of the present study is to find repellent activity of Vitex negundo and Curcuma longa herbal extracts against the wild species Simulium (blackfly) bite., Methods: We have studied Simulium biting behavior and tested the repellency of herbal extract obtained from Vitex negundo L. (Lamiales: Lamiaceae) and Curcuma longa L. (Zingiberal: Zingiberaceae) along with their essential oils in three locations in Arunachal Pradesh, India on human volunteers' resistance to Simulium (blackflies). The reported herbal extracts were incorporated to topical drug delivery vehicle as a cream and gel., Results: The methanolic extract of Vitex negundo cream and gel formulation showed >2 h safety at 5% concentration and >5 h safety at 10% concentration in all the testing sites followed by ethyl acetate extract. Whereas, chloroform extract of Curcuma longa cream and gel formulation provided >1 h safety at 5% concentration and >4 h safety at 10% concentration., Interpretation Conclusion: At an optimum concentration of 10%, the methanolic extract of Vitex negundo cream provided complete protection times (CPTs) 320.4, 358.6, and 346.4 min, respectively. These findings offer confirmation of the methanolic extract and chloroform extract potential for use in new blackfly repellents., (Copyright © 2024 Copyright: © 2024 Journal of Vector Borne Diseases.)

Black soldier fly larvae (Hermetia illucens) receive growing interest as a potential alternative animal feed source. These insects may be exposed to insecticide residues in the rearing substrate. This study aimed to investigate the effects of six different pyrethroid and organophosphate insecticides on this insect species' performance. The toxicity of two "model" substances for each of these classes (cypermethrin; pirimiphos-methyl) was quantified, with and without the synergist piperonyl butoxide (PBO). Critical effect doses corresponding to 10% yield (CED10) for cypermethrin (0.4 mg/kg) and pirimiphos-methyl (4.8 mg/kg) were determined. The addition of PBO to cypermethrin enhanced its relative potency with a factor 2.6. These data were compared against the relative toxicity of two analogue substances in each class (permethrin, deltamethrin; chlorpyrifos-methyl, malathion). Results suggest that exposure to concentrations complying with legal limits can cause significant reductions in yield. Exposure to multiple substances at lower concentrations resulted in negative additive and synergistic effects. Of the tested substances, deltamethrin was most toxic, causing 94% yield at 0.5 mg/kg. Analytical results suggested that transfer of tested substances to the larval biomass was substance- and concentration-specific, but appeared to be correlated to reduced yields and the presence of PBO. Transfer of organophosphates was overall low (<2%), but ranged from 8% to 75% for pyrethroids. Due to very low limits in insect biomass (∼0.01 mg/kg), high transfer may result in noncompliance. It is recommended that rearing companies implement lower contractual thresholds, and that policymakers consider adjusting legally allowed maximum residue levels in insect feed., (© 2023 The Authors. Insect Science published by John Wiley & Sons Australia, Ltd on behalf of Institute of Zoology, Chinese Academy of Sciences.)

Background: The onchocerciasis focus surrounding the lower Mbam and Sanaga rivers, where Onchocerca volvulus is transmitted by Simulium damnosum s.l. (Diptera: Simuliidae), was historically the largest in the southern regions of Cameroon. Annual community-directed treatment with ivermectin (CDTI) has been taking place since 2000, but recent studies have shown that new infections are occurring in children. We aimed to investigate blackfly biting and O. volvulus transmission rates along the lower Mbam river 16 years after the formal onset of annual CDTI., Methods: Black flies were collected for three consecutive days each month between July 2016 and June 2017 at two riverside villages and two inland sites situated 4.9 km and 7.9 km from the riverside. Specimens collected at each site were dissected on one of the three collection days each month to estimate parity rates and O. volvulus infection rates, while the remaining samples were preserved for pool screening., Results: In total, 93,573 S. damnosum s.l. black flies were recorded biting humans and 9281 were dissected. Annual biting rates of up to 606,370 were estimated at the riverside, decreasing to 20,540 at 7.9 km, while, based on dissections, annual transmission potentials of up to 4488 were estimated at the riverside, decreasing to 102 and 0 at 4.9 km and 7.9 km, respectively. However, pool screening showed evidence of infection in black flies at the furthest distance from the river. Results of both methods demonstrated the percentage of infective flies to be relatively low (0.10-0.36%), but above the WHO threshold for interruption of transmission. In addition, a small number of larvae collected during the dry season revealed the presence of Simulium squamosum E. This is the first time S. squamosum E has been found east of Lake Volta in Ghana, but our material was chromosomally distinctive, and we call it S. squamosum E2., Conclusions: Relatively low O. volvulus infection rates appear to be offset by extremely high densities of biting black flies which are sustaining transmission along the banks of the lower Mbam river., (© 2021. The Author(s).)

The Controlled Current Toxicity Test (CCTT) is a protocol used by the University of Georgia Black Fly Rearing and Bioassay Laboratory to evaluate the efficacy of larvicides based on Bacillus thuringiensis subsp. israelensis ( Bti ) against Simulium vittatum larvae. A standard CCTT provides a larval medium with suspended organic nutrients, a sustained current, and a clean surface for black fly larval attachment in each exposure vessel. In an effort to streamline the CCTT and eliminate a potential source of variability, 3 types of larval medium were evaluated: deionized water, distilled water, and moderately hard water, which were compared to the currently used protocol. A statistical significance in mean lethal concentration (LC 50 ) rates was observed between the CCTT (deionized water with suspended organic nutrients) medium and the other 3 experimental media. The presence of suspended organic nutrients in the CCTT significantly improved the efficacy of the Bti- based larvicide. The interaction of the suspended organic nutrient particles and the Bti- based particles in the larvicide appears to produce a mixture that is more efficiently captured and ingested by the black fly larvae than the larvicide formulation particles alone.

Background: Great strides have been made toward onchocerciasis elimination by mass drug administration (MDA) of ivermectin. Focusing on MDA-eligible areas, we investigated where the elimination goal can be achieved by 2025 by continuation of current practice (annual MDA with ivermectin) and where intensification or additional vector control is required. We did not consider areas hypoendemic for onchocerciasis with loiasis coendemicity where MDA is contraindicated., Methods: We used 2 previously published mathematical models, ONCHOSIM and EPIONCHO, to simulate future trends in microfilarial prevalence for 80 different settings (defined by precontrol endemicity and past MDA frequency and coverage) under different future treatment scenarios (annual, biannual, or quarterly MDA with different treatment coverage through 2025, with or without vector control strategies), assessing for each strategy whether it eventually leads to elimination., Results: Areas with 40%-50% precontrol microfilarial prevalence and ≥10 years of annual MDA may achieve elimination with a further 7 years of annual MDA, if not achieved already, according to both models. For most areas with 70%-80% precontrol prevalence, ONCHOSIM predicts that either annual or biannual MDA is sufficient to achieve elimination by 2025, whereas EPIONCHO predicts that elimination will not be achieved even with complementary vector control., Conclusions: Whether elimination will be reached by 2025 depends on precontrol endemicity, control history, and strategies chosen from now until 2025. Biannual or quarterly MDA will accelerate progress toward elimination but cannot guarantee it by 2025 in high-endemicity areas. Long-term concomitant MDA and vector control for high-endemicity areas might be useful.

Background: In 2012, the World Health Organization set goals for the elimination of onchocerciasis transmission by 2020 in selected African countries. Epidemiological data and mathematical modelling have indicated that elimination may not be achieved with annual ivermectin distribution in all endemic foci. Complementary and alternative treatment strategies (ATS), including vector control, will be necessary. Implementation of vector control will require that the ecology and population dynamics of Simulium damnosum (sensu lato) be carefully considered., Methods: We adapted our previous SIMuliid POPulation dynamics (SIMPOP) model to explore the impact of larvicidal insecticides on S. damnosum (s.l.) biting rates in different ecological contexts and to identify how frequently and for how long vector control should be continued to sustain substantive reductions in vector biting. SIMPOP was fitted to data from large-scale aerial larviciding trials in savannah sites (Ghana) and small-scale ground larviciding trials in forest areas (Cameroon). The model was validated against independent data from Burkina Faso/Côte d'Ivoire (savannah) and Bioko (forest). Scenario analysis explored the effects of ecological and programmatic factors such as pre-control daily biting rate (DBR) and larviciding scheme design on reductions and resurgences in biting rates., Results: The estimated efficacy of large-scale aerial larviciding in the savannah was greater than that of ground-based larviciding in the forest. Small changes in larvicidal efficacy can have large impacts on intervention success. At 93% larvicidal efficacy (a realistic value based on field trials), 10 consecutive weekly larvicidal treatments would reduce DBRs by 96% (e.g. from 400 to 16 bites/person/day). At 70% efficacy, and for 10 weekly applications, the DBR would decrease by 67% (e.g. from 400 to 132 bites/person/day). Larviciding is more likely to succeed in areas with lower water temperatures and where blackfly species have longer gonotrophic cycles., Conclusions: Focal vector control can reduce vector biting rates in settings where a high larvicidal efficacy can be achieved and an appropriate duration and frequency of larviciding can be ensured. Future work linking SIMPOP with onchocerciasis transmission models will permit evaluation of the impact of combined anti-vectorial and anti-parasitic interventions on accelerating elimination of the disease.

Treating municipal sewage sludge (MSS) sustainably and economically in China remains a challenge because of risks associated with the heavy metals it contains. In this study, black solider fly larvae (BSFL) were used for MSS treatment. The resistance of larvae to combined heavy metals and their potential use in conversion of MSS were investigated. The results indicated that seven MSS samples contained large amounts of heavy metals, with the lead and nickel contents of several samples exceeding Chinese national discharge standards. BSFL were highly tolerant to an artificial diet spiked with combined heavy metals. Principal component analysis revealed that high concentrations of lead, nickel, boron, and mercury potentially interfered with larval weight gain, while zinc, copper, chromium, cadmium, and mercury slightly reduced larval survival. The addition of chicken manure and wheat bran as co-substrates improved the conversion process, which was influenced by the nature and amount of added co-substrate and especially the quantity of nitrogen added. With the amended substrate, the BSFL accumulated heavy metals into their bodies but not into extracted larval oil. The heavy metal content of the treatment residue was lower than that considered safe for organic-inorganic compound fertilizers standards in China and the harvested larvae could be used as a source of oil for industrial application.

Many blackfly species (Diptera: Simuliidae) are economically important insect pests, both as nuisance biters and as vectors of pathogens of medical and veterinary relevance. Among the important blackfly pest species in North America is Simulium vittatum Zetterstedt sensu lato. The objective of this study was to identify compounds excreted by mammalian hosts that are attractive to host-seeking S. vittatum females. The attractiveness of putative compounds to colonized S. vittatum was tested through electrophysiological (electroantennography; n = 58 compounds) and behavioural (Y-tube assays; n = 7 compounds in three concentrations) bioassays. Five compounds were significantly attractive to host-seeking S. vittatum females: 1-octen-3-ol; 2-heptanone; acetophenone; 1-octanol, and naphthalene. These candidate compounds might be useful as attractants in traps that could be developed for use in alternative or complementary management tactics in programmes to suppress nuisance blackfly populations, or for the collection of samples in which to study the transmission ecology of pathogens transmitted by blackflies of the S. vittatum complex., (© 2016 The Royal Entomological Society.)

Uganda is the only country in sub-Saharan Africa whose onchocerciasis elimination programme extensively uses vector control and biannual treatment with ivermectin. The purpose of this study was to assess the impact of combined strategies on interrupting onchocerciasis transmission in the Kashoya-Kitomi focus. Mass Drug Administration annually (13 years) followed by biannual treatments (6 years) and ground larviciding (36 cycles in 3 years) with temephos (Abate ® , EC500) against Simulium neavei were conducted. Routine fly catches were conducted for over seven years in six catching sites and freshwater crabs Potamonautes aloysiisabaudiae were examined for immature stages of Simulium neavei. Epidemiological assessments by skin snip were performed in 2004 and 2013. Collection of dry blood spots (DBS) from children <10 years for IgG4 antibodies analysis were done in 2010 and 2013. Treatment coverage with ivermectin improved with introduction of biannual treatment strategy. Microfilaria prevalence reduced from 85% in 1991 to 62% in 2004; and to only 0.5% in 2013. Crab infestation reduced from 59% in 2007 to 0% in 2013 following ground larviciding. Comparison of total fly catches before and after ground larviciding revealed a drop from 5334 flies in 2007 to 0 flies in 2009. Serological assays conducted among 1,362 children in 2010 revealed 11 positive cases (0.8%; 95% CI: 0.4%-1.2%). However, assessment conducted on 3246 children in 2013 revealed five positives, giving point prevalence of 0.15%; 95% CI: 0.02%-0.28%. Four of the five children subjected to O-150 PCR proved negative. The data show that transmission of onchocerciasis has been interrupted based on national and WHO Guidelines of 2012 and 2016, respectively., (Copyright © 2016 Elsevier B.V. All rights reserved.)

The black soldier fly Hermetia illucens is a good candidate for waste management. The harvested insects are rich in protein and have the potential to be used in animal feed. However, people are wary about heavy metals in waste. Therefore, it is necessary to understand how the uptake of heavy metals could affect H. illucens and where and to what extent metals are accumulated by the black soldier fly. Based on these considerations, developmental parameters were investigated in the different life stages of H. illucens fed an increasing concentration gradient of cadmium (Cd) and chromium (Cr); additionally, Cd and Cr distribution in the body parts of H. illucens at the different life stages was monitored. We found that Cd and Cr have no effects on larvae survival and eclosion rate, but they do have effects on larvae duration and pupation rate. Both Cd and Cr were transferred into larvae, prepupae, and pupae. While the concentrations of Cd in larvae and prepupae were much higher than that in their diets, the opposite case was observed with Cr. The concentrations of Cd and Cr in H. illucens decreased in later development stages. In individual larva and prepupa, Cd and Cr were mainly included in the body and not in the integument. In the pupa, the puparium contained higher Cd and Cr concentrations than the pupa body. The distribution of Cd and Cr in the different life stages and body parts may present a potential strategy for how H. illucens tolerate and remove heavy metal stress.

Hematophagous insects can be vectors of pathogens and cause significant economic loss in zootechnical production. Among biting insects, many dipteran species feed on horse blood. The black fly (Diptera: Simuliidae) group, is responsible for several disorders in horses and inflicts painful bites that lead to undesirable behaviours in horses, particularly when bites occur in sensitive areas such as the inner ear. A field study was conducted in a French equestrian center during which a semiochemical was applied on horses' ears to assess repellent efficacy against simulid infestation. During the first phase of the study, efficacy was evaluated over a one hour period. Then, during the second phase of the study, persistency of the effect was tested at 8, 9 and 10h after application. The results of the study's first phase showed 90% efficacy over one hour, with 121.5 insects found in control ears and 12 insects in treated ears (p=0.001). In the second phase of the study, a total amount of 411 insects were observed on control ears whereas only 2 insects were observed on treated ears (p<0.0001); the treatment remained over 98% effective up to 10hours after application. When using a slow release excipient, this semiochemical may offer at least 10h of protection against simulids. This safe, efficient, and long lasting protection could help horses and their owners to manage simulid parasitism., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Background: Onchocerciasis is one of the most important public health problems over large areas of tropical Africa countries including Ethiopia. The African Program for Onchocerciasis Control (APOC) has been working with ultimate goal of reducing the public health and socio-economic problems of onchocerciasis through administration of the tablet for continuous 12-15 years using the strategy of yearly community-directed treatment with ivermectin (CDTI) in endemic areas of Africa to kill the microfilariae that invade the eyes and are present in the skin to be transported to another victim by the black fly. The objective of this study was to assess knowledge, attitude and practice of community drug distributors (CDDs) towards onchocerciasis and CDTI in Quara district., Result: Of all the study participating CDD 11.4% (9/79) said that they knew about the etiology of the disease, 35.4% (28/79) had good level of knowledge, 19 (24.1%) had good level of positive attitude and 18 (22.8%) had good level of positive practice about onchocerciasis. Similarly, 45.6% (36/79), 81.0% (64/79) and 29.1% (23/79) had good level of knowledge, attitude and practice about CDTIP, respectively. Being a female CDD (adjusted OR 7.246, P = 0.035, 95% CI 1.147, 45.455) and being older than 35 years (adjusted OR 8.435, P = 0.001, 95% CI 4.53, 9.003) were significantly associated with the likelihood of having good level of knowledge about the disease., Conclusion: Although onchocerciasis is endemic in Quara district, large proportion of the CDDs had misconceptions about its causation, transmission and prevention. Therefore, CDTIP for onchocerciasis control need to be supported by proper and continuous training, and health education about different aspects of the disease.

Background: Recent reports of transmission interruption of Onchocerca volvulus, the causing agent of river blindness, in former endemic foci in the Americas, and more recently in West and East Africa, raise the question whether elimination of this debilitating disease is underway after long-term treatment of the population at risk with ivermectin. The situation in Central Africa has not yet been clearly assessed., Methods and Findings: Entomologic data from two former endemic river basins in North Cameroon were generated over a period of 43 and 48 months to follow-up transmission levels in areas under prolonged ivermectin control. Moreover, epidemiologic parameters of animal-borne Onchocerca spp. transmitted by the same local black fly vectors of the Simulium damnosum complex were recorded and their impact on O. volvulus transmission success evaluated. With mitochondrial DNA markers we unambiguously confirmed the presence of infective O. volvulus larvae in vectors from the Sudan savannah region (mean Annual Transmission Potential 2009-2012: 98, range 47-221), but not from the Adamawa highland region. Transmission rates of O. ochengi, a parasite of Zebu cattle, were high in both foci., Conclusions/significance: The high cattle livestock density in conjunction with the high transmission rates of the bovine filaria O. ochengi prevents the transmission of O. volvulus on the Adamawa plateau, whereas transmission in a former hyperendemic focus was markedly reduced, but not completely interrupted after 25 years of ivermectin control. This study may be helpful to gauge the impact of the presence of animal-filariae for O. volvulus transmission in terms of the growing human and livestock populations in sub-Saharan countries.

The suite of pheromones that promote communal oviposition by Simulium vittatum, a North American black fly species, was identified and characterized using gas chromatography-mass spectrometry, electrophysiological, and behavioral bioassays. Behavioral assays demonstrated that communal oviposition was induced by egg-derived compounds that were active at short range and whose effect was enhanced through direct contact. Three compounds (cis-9-tetradecen-1-ol, 1-pentadecene, and 1-tridecene) were identified in a non-polar solvent extract of freshly deposited S. vittatum eggs that were capable of inducing the oviposition response. Electroantennography demonstrated that two of these three compounds (1-pentadecene and 1-tridecene) actively stimulated antennal neurons. Identification of the oviposition pheromones of this family may be helpful in developing control measures for nuisance black flies and for medically-important species such as Simulium damnosum sensu lato.

Background: Entomological indicators are considered key metrics to document the interruption of transmission of Onchocerca volvulus, the etiological agent of human onchocerciasis. Human landing collection is the standard employed for collection of the vectors for this parasite. Recent studies reported the development of traps that have the potential for replacing humans for surveillance of O. volvulus in the vector population. However, the key chemical components of human odor that are attractive to vector black flies have not been identified., Methodology/principal Findings: Human sweat compounds were analyzed using GC-MS analysis and compounds common to three individuals identified. These common compounds, with others previously identified as attractive to other hematophagous arthropods were evaluated for their ability to stimulate and attract the major onchocerciasis vectors in Africa (Simulium damnosum sensu lato) and Latin America (Simulium ochraceum s. l.) using electroantennography and a Y tube binary choice assay. Medium chain length carboxylic acids and aldehydes were neurostimulatory for S. damnosum s.l. while S. ochraceum s.l. was stimulated by short chain aliphatic alcohols and aldehydes. Both species were attracted to ammonium bicarbonate and acetophenone. The compounds were shown to be attractive to the relevant vector species in field studies, when incorporated into a formulation that permitted a continuous release of the compound over time and used in concert with previously developed trap platforms., Conclusions/significance: The identification of compounds attractive to the major vectors of O. volvulus will permit the development of optimized traps. Such traps may replace the use of human vector collectors for monitoring the effectiveness of onchocerciasis elimination programs and could find use as a contributing component in an integrated vector control/drug program aimed at eliminating river blindness in Africa.

Bacillus thuringiensis subsp. israelensis (Bti) is the first Bacillus thuringiensis to be found and used as an effective biological control agent against larvae of many mosquito and black fly species around the world. Its larvicidal activity resides in four major (of 134, 128, 72 and 27 kDa) and at least two minor (of 78 and 29 kDa) polypeptides encoded respectively by cry4Aa, cry4Ba, cry11Aa, cyt1Aa, cry10Aa and cyt2Ba, all mapped on the 128 kb plasmid known as pBtoxis. These six δ-endotoxins form a complex parasporal crystalline body with remarkably high, specific and different toxicities to Aedes, Culex and Anopheles larvae. Cry toxins are composed of three domains (perforating domain I and receptor binding II and III) and create cation-selective channels, whereas Cyts are composed of one domain that acts as well as a detergent-like membrane perforator. Despite the low toxicities of Cyt1Aa and Cyt2Ba alone against exposed larvae, they are highly synergistic with the Cry toxins and hence their combinations prevent emergence of resistance in the targets. The lack of significant levels of resistance in field mosquito populations treated for decades with Bti-bioinsecticide suggests that this bacterium will be an effective biocontrol agent for years to come.

The effect of insect vectors on avian exposure to infection by pathogens remains poorly studied. Here, we used an insect repellent treatment to reduce the number of blood-sucking flying insects in blue tit Cyanistes caeruleus nests and examined its effect on nestling health status measured as body mass, nestling phytohaemagglutinin (PHA) response and blood parasite prevalence. We found that (i) the insect repellent treatment significantly reduced the number of blood-sucking flying insects in nests and (ii) the number of blood-sucking flying insects had a significant effect on the prevalence of the blood parasite Trypanosoma independently of the treatment. In addition, we found support for an adverse effect of parasite infections on nestling PHA response. Nestlings infected by Trypanosoma mounted a weaker response against PHA than non-parasitized ones. In addition, the number of blowflies in the nest was negatively associated with nestling PHA response. Overall, we found support for the hypothesis that blood-sucking flying insects attacking nestlings increase their exposure to parasite infections. Our results further substantiate the adverse effect of parasites on nestling condition.

The Itwara onchocerciasis focus is located around the Itwara forest reserve in western Uganda. In 1991, annual treatments with ivermectin started in the focus. They were supplemented in 1995 by the control of the vector Simulium neavei, which was subsequently eliminated from the focus. The impact of the two interventions on the disease was assessed in 2010 by nodule palpations, examinations of skin snips by microscopy and PCR, and Ov16 recombinant ELISA. There was no evidence of any microfilaria in 688 skin snips and only 2 (0.06%) of 3316 children examined for IgG4 were slightly above the arbitrary cut off of 40. A follow up of the same children 21 months later in 2012 confirmed that both were negative for diagnostic antigen Ov-16, skin snip microscopy and PCR. Based on the World Health Organization (WHO) elimination criteria of 2001 and the Uganda onchocerciasis certification guidelines, it was concluded that the disease has disappeared from the Itwara focus after 19 years of ivermectin treatments and the elimination of the vector around 2001. Ivermectin treatments were recommended to be halted., (Copyright © 2013 Elsevier B.V. All rights reserved.)

In the present study we have evaluated the repellent activity of mixture of Curcuma longa, Zanthoxylum limonella and Pogostemon heyneanus essential oils in 1:1:2 ratio at 5%, 10% and 20% concentration against blackflies in northeastern India. Initially the essential oil mixture tested here has been found effective against Aedes albopictus mosquitoes. The average protection recorded in 20% concentration (170.56 ± 4.0; 95% CI = 162.09-179.02) was higher as compared to other two concentrations (F = 90.2; p<0.0001; df = 53). Percentage repellency and repellency index was found to be higher in 20% concentration (p ≤ 0.017). No appreciable clinical and behavioral signs were observed in the acute dermal toxicity using rat model. No changes were observed in biochemical profiles of treatment group animals. Similarly, no prominent lesions were observed in vital organs of treatment in both the sexes. The study concludes that tested repellent is safe for use and has multi-insects repellent property.

The year 2012 marks the 25th anniversary of the donation of ivermectin to fight onchocerciasis and the projected date for elimination of transmission of the disease in the Americas. This review looks at the history of onchocerciasis, from its discovery through to 2025, by which time it is projected that the disease will have been eliminated as a public health problem, except in a handful of sub-Saharan countries, where it should be well on the way towards elimination., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

Black flies, a non-target species of the insecticides used in fruit production, represent a severe medical and veterinary problem. Large increases in the level of resistance to the pyrethroids fenvalerate (more than 355-fold) and deltamethrin (162-fold) and a small increase in resistance to the organophosphate azinphos methyl (2-fold) were observed between 1996-2008 in black fly larvae under insecticide pressure. Eventually, no change or a slight variation in insecticide resistance was followed by a subsequent increase in resistance. The evolution of pesticide resistance in a field population is a complex and stepwise process that is influenced by several factors, the most significant of which is the insecticide selection pressure, such as the dose and frequency of application. The variation in insecticide susceptibility within a black fly population in the productive area may be related to changes in fruit-pest control. The frequency of individuals with esterase activities higher than the maximum value determined in the susceptible population increased consistently over the sampling period. However, the insecticide resistance was not attributed to glutathione S-transferase activity. In conclusion, esterase activity in black flies from the productive area is one mechanism underlying the high levels of resistance to pyrethroids, which have been recently used infrequently. These enzymes may be reselected by currently used pesticides and enhance the resistance to these insecticides.

Use of repellents seems to be most reliable method of personal protection against annoyance and infections associated with haematophagous insects. We have investigated the biting activity of Simulium and tested the repellency of five essential oils extracted from Homalomena aromatica Schott (Alismatales: Araceae), Pogostemon heyneanus Bentham (Lamiales: Lamiaceae), Citrus aurantifolia Swingle (Sapindales: Rutaceae), Vitex negundo L. (Lamiales: Lamiaceae), and Ageratum conzoides L. (Asterales: Asteraceae) on the human volunteers against Simulium (blackflies) in three locations of Arunachal Pradesh, India. Blackflies preferred biting legs (> 79%) as compared to hand and face with profound biting activity during 1000-1100 h (> 23%) and 1500 - 1600 h (> 28%). The essential oil extracted from Homalomena aromatica, Vitex negundo and Ageratum conizoides provided > 2 h protection at 5% concentration and > 5 h protection at 10% concentration in all the three testing locations. The repellency of Homalomena aromatica, Vitex negundo and Ageratum conizoides essential oils after 6 h application was > 50% at 5% concentration and > 90% at 10% concentration. The study provides evidence for the potential of these essential oils in developing new repellents against blackflies.

Water was collected from a site on the Susquehanna River in eastern Pennsylvania, where less-than-optimal black fly larval mortality had been occasionally observed after treatment with Bacillus thuringiensis subsp. israelensis de Barjac insecticidal crystalline proteins (Bti ICPs). A series of experiments was conducted with Simulium vittatum Zetterstedt larvae to determine the water related factors responsible for the impaired response to Bti ICPs (Vectobac 12S, strain AM 65-52). Seston in the water impaired the effectiveness of the ICPs, whereas the dissolved substances had no impact on larval mortality. Individual components of the seston then were exposed to the larvae followed by exposure to Bti ICPs. Exposure of larvae to selected minerals and nutritive organic material before ICP exposure resulted in no significant decrease in mortality. Exposure of larvae to silicon dioxide, cellulose, viable diatoms, and purified diatom frustules before Bti ICP exposure resulted in significant reductions in mortality. Exposure of larvae to purified diatom frustules from Cyclotella meneghiniana Kützing resulted in the most severe impairment of mortality after Bti ICP exposure. It is postulated that frustule-induced impairment of feeding behavior is responsible for the impairment of larval mortality.

The root and leaf extracts of four plants, Occimum gratissimum, Azadirachta indica, Pterocarpus santalinoides, and Pistia hyptis, were studied for repellent activities against the adults of Simulium damnosum sensu lato. The leaves and roots were extracted with 95% ethanol and the stocks were diluted with paraffin. The repellent activities of the extracts were investigated using human baits along the banks of River Oyan and River Ogun in southwestern Nigeria. The results showed that the root extract of O. grattissium and leaf extract of P. hyptis had highest repellent potentials with 78% and 78.1% protection against S. damnosum sensu lato, respectively, whereas the root and leaf of P. santalinoides recorded the least. Although there were significant differences in the percentage of protection of the extracts of the plants (p < 0.05), the variations in the percentage of protection of the leaf and root extracts were not statistically significant (p > 0.05). The study concludes that there exist some repellent efficacies in the extracts of the plants, most importantly O. grattissimum and P. hyptis. The plant extracts can further be developed in the prevention of man-vector contact in onchocerciasis endemic communities.

Spinosad is a naturally derived insecticide that has shown potential as a mosquito larvicide. To determine the activity of spinosad against blackflies, late-instar larvae from a community comprising Simulium triittatum (63.6%) and seven other species, including three known vectors of onchocerciasis in Mexico (S. metallicum, S. ochraceum, and S. callidum), were subjected to concentration-mortality laboratory bioassays following World Health Organization guidelines. Cephalic capsule measurements confirmed the relatively homogeneous distribution of experimental larvae. The 50% lethal concentration of spinosad was estimated at 1.48 ppm spinosad (95% confidence interval: 1.07-2.33) for a 10-min exposure period, whereas larvae treated with 0.05 ppm of the organophosphate temephos experienced 61% mortality. Immature aquatic insects were identified to genus and tested for their susceptibility to spinosad in the laboratory. After exposure to 12 ppm spinosad for 10 min, ephemeropterans, odonates, trichopterans, and hemipterans did not experience significantly increased mortality over that of untreated controls, whereas a significant increase in mortality was observed in spinosad-treated Plecoptera (P < 0.001). Tilapia and trout fry exposed to 12 ppm spinosad for 10 min did not experience increased mortality at 24-h postexposure over that of the controls. We conclude that spinosad is less toxic than temephos to these blackfly species, but is likely to have a low impact on nontarget members of the aquatic community.

A controlled current toxicity test (CCTT) was developed to evaluate the response of black fly (Simulium vittatum) larvae to insecticidal proteins following exposure to various antibiotics. The bacterium, Bacillus thuringiensis subsp. israelensis (Bti), produces proteins that are toxic to Nemotoceran Diptera, such as black flies and mosquitoes, when ingested. These insecticidal crystalline proteins (ICPs) are highly efficacious in controlling black flies; however, speculation has arisen regarding the potential for antibiotic contamination of waterways to mitigate the toxicity of these proteins. A series of experiments was conducted with the CCTT in which black fly larvae were exposed to enrofloxacin, tylosin, sulfamethoxazole, and trimethoprim followed by exposure to Bti ICPs. These antibiotics were selected based on their use in agricultural and documented anthropogenic contamination of rivers. Anthropogenic concentrations of a mixture of these four antibiotics did not affect the response of the larvae to Bti ICPs. Subsequent experiments were conducted with antibiotic concentrations 10,000 to 80,000 times higher than those found in contaminated rivers. Exposure of black fly larvae to high levels of enrofloxacin (0.5 mg/L) had no effect upon the susceptibility to Bti ICPs; however, exposure to high levels of tylosin (8 mg/L) resulted in a significant increase in the susceptibility of the larvae to Bti ICPs at 72 h of exposure, but not at 48 h. Exposure of black fly larvae to high concentrations of a mixture of sulfamethoxazole and trimethoprim resulted in a significant increase in the efficacy of the larvicide after 48 and 72 h of exposure. These results suggest that impairment of the efficacy of Bti ICPs to black fly larvae is not due to antibiotic contamination of the larval environment., (Copyright 2010 SETAC)

Sertraline is a selective serotonin reuptake inhibitor (SSRI) prescribed as an antidepressant. Although SSRIs are known to block serotonin reuptake sites on cell membranes, they also have been shown to inhibit acetylcholinesterase (AChE) activity. Thus, the interaction of these chemicals with other AChE inhibitors, namely, organophosphate and carbamate insecticides, is of interest. In addition, these insecticides have been shown to interact with serotonergic neuronal pathways creating questions as to how these chemicals might interact. In this study, the interactive effect of sertraline (SSRI) in binary combinations with carbaryl (carbamate insecticide) and diazinon (organophosphate insecticide) was assessed using a 48-h acute toxicity test with black fly larvae, Simulium vittatum IS-7. Results showed that observed mortality was bracketed by the independent action model and concentration addition model with the independent action model slightly underestimating mortality and the concentration addition model slightly overestimating mortality. Varying the concentration of the chemicals in the mixture did not indicate that sertraline was interacting with the insecticides to make them more toxic or vice versa. These results indicate that sertraline and the insecticides are likely eliciting toxicity at separate neuronal pathways since no interaction was observed.

The island of Bioko is part of the Republic of Equatorial Guinea and is the only island in the World to have endemic onchocerciasis. The disease is hyperendemic and shows a forest-type epidemiology with low levels of blindness and high levels of skin disease, and the whole population of 68,000 is estimated to be at risk. Control of onchocerciasis began in 1990 using ivermectin and this yielded significant clinical benefits but transmission was not interrupted. Feasibility and preparatory studies carried out between 1995 and 2002 confirmed the probable isolation of the vector on the island, the high vectorial efficiency of the Bioko form of Simulium yahense, the seasonality of river flow, blackfly breeding and biting densities, and the distribution of the vector breeding sites. It was proposed that larviciding should be carried out from January to April, when most of the island's rivers were dry or too low to support Simulium damnosum s.l., and that most rivers would not need to be treated above 500 m altitude because they were too small to support the breeding of S. damnosum s.l. Larviciding (with temephos) would need to be carried out by helicopter (because of problems of access by land), supplemented by ground-based delivery. Insecticide susceptibility trials showed that the Bioko form was highly susceptible to temephos, and insecticide carry was tested in the rivers by assessing the length of river in which S. damnosum s.l. larvae were killed below a temephos dosing point. Regular fly catching points were established in 1999 to provide pre-control biting densities, and to act as monitoring points for control efforts. An environmental impact assessment concluded that the proposed control programme could be expected to do little damage, and a large-scale larviciding trial using ground-based applications of temephos (Abate 20EC) throughout the northern (accessible) part of the island was carried out for five weeks from 12 February 2001. Following this, a first attempt to eliminate the vectors was conducted using helicopter and ground-based applications of temephos from February to May 2003, but this was not successful because some vector populations persisted and subsequently spread throughout the island. A second attempt from January to May 2005 aimed to treat all flowing watercourses and greatly increased the number of treatment points. This led to the successful elimination of the vector. The last biting S. damnosum s.l. was caught in March 2005 and none have been found since then for more than 3 years.

Mechanistic and stereoselective based in vitro metabolism assays were utlilized to gain insight into the toxic mode of action of the 1,2,4-triazole fungicide, triadimefon, with black fly (Diptera: Simuliidae) larvae. Based on results from enzyme inhibitor studies, the metabolism of triadimefon in black fly larvae microsomes was found to occur predominantly via an oxidative P450-mediated pathway; triadimenol was formed via the stereoselective reduction of the prochiral carbonyl group of triadimefon. The relatively minor contribution of carbonyl reduction suggests that triadimefon may inhibit ecdysone 20-monooxygenase and disrupt insect molting hormone biosynthesis. 48-h LC50 tests for triadimefon and triadimenol with black fly larvae yielded median values (with 95% confidence intervals) of 6.1 (5.8-6.4) and 22.3 (20.3-24.1) mg/L respectively. The exposure of black fly larvae to sublethal concentrations of triadimefon resulted in increased microsomal P450 activity and affected the microsomal rates of both triadimefon depletion and triadimenol formation. In contrast to trout, black fly larvae produced a higher fraction of the more toxic triadimenol stereoisomers, which may explain in part why triadimefon exhibited a significantly greater toxicity with black fly larvae than trout. These results illustrate that while LC50 tests conducted with commercial triadimenol would presumably expose each organism to the same relative abundance of the four triadimenol stereoisomers, LC50 tests with triadimefon ultimately expose each organism to a unique set of triadimenol stereoisomers depending upon the organism's stereoselective metabolism.

Here, we validate the use of a citronella (natural oil) based repellent to reduce the abundance of flying blood-sucking insects in avian nests. These insects are important parasites of birds affecting them as blood feeders and as vectors of a diversity of pathogens. When nestling were 10 days old, we assigned wild great tit Parus major nests to one of two treatments, control and fumigated nests. The abundance of biting midges and blackflies captured during 3 days following the treatment application were lower in fumigated nests with respect to control ones. By contrast, the abundance of blowfly pupae measured when nestlings left their nests was not affected by the treatment. Although many experimental studies modify the abundance of nest-dweller ectoparasites, to our knowledge, this is the first one describing an easy, safe, and effective method, reducing the total abundance of both biting midges and blackflies in wild avian nests. Our results could be used in future conservation projects and experimental studies on host-parasite evolution affecting the abundance of flying blood-feeder insects under natural conditions.

The essential oil of catmint, Nepeta cataria L., was hydrogenated to yield an oil enriched in dihydronepetalactone (DHN) diastereomers, termed. This material was used for the preparation of liquid alcohol-based and lotion formulations. The efficacy of these formulations as repellents was tested after application to human test subjects at two locations in the United States: Maine and Florida. In Maine, data on repellency of the hydrogenated catmint oil formulations toward black flies (Simulium decorum Walker) and mosquitoes (primarily Aedes intrudens Dyar) were obtained. In these tests, protection from black flies was conferred for 6 h or more with all formulations, and both liquid and lotion formulations at 15 wt% active ingredient gave complete protection for 7.5 h. All formulations conferred protection from mosquitoes for >4 h, with the best (15 wt% lotion) giving >8 h of complete protection. In Florida, data on repellency toward a mixed population of mosquitoes indicated that all formulations conferred protection for >4 h, with the 15 wt% lotion giving >6 h complete protection from bites.

Cytotaxonomic identifications of larvae of members of the Simulium damnosum Theobald (Diptera: Simuliidae) complex collected in forest zones of southeast Ghana and southwest Togo between 1977 and 1996 showed that the Djodji form of Simulium sanctipauli Vajime & Dunbar, a vector of onchocerciasis, was eliminated in 1988 by larvicide operations conducted by the World Health Organization (WHO) Onchocerciasis Control Programme (OCP) in West Africa. No members of the form were identified amongst 997 larvae collected up to 8 years after systematic control operations began in February 1988. The results are discussed in relation to estimates of the numbers of samples required to certify elimination and the possibility that other members of the S. damnosum complex were also eliminated by the OCP.

Objective: To assess the phytochemical properties of Chromoleana odorata in the control of blackflies to help check the problem of environmental pollution from the use of chemical insecticides., Methods: Dried pulverized leaves of Chromolaena odorata (100 g) were soaked in distilled water for 24 hours and the extract was filtered, marc pressed and evaporated over water bath. Stock solution of the dried mass (1 g) was concentrated by dissolving in 100 mL of water. Serial dilutions (100 mg/mL, 10 mg/mL, 1 mg/mL, 0.1 mg/mL, 0.01 mg/L, and 0.001 mg/mL) of both the extract and Chlopyrifos (organo phosphorus) were prepared and toxicity was tested on the larvae of Simulium species., Results: The chemical compound recorded 100% larval mortality at all concentrations while the plant extract recorded 100% larval mortality in three (100, 10, and 1 mg/mL) of its diluent concentration. The percentage of larval mortality by chemical compound and plant extract was statistically insignificant (t=0.2456, P>0.05), but the mortality rate was significant within 30 min interval of exposure time (t=3.756, P<0.05). The LC50 of the plant extract was determined at about 0.001 mg/mL concentration. This showed that the plant extract had an appreciable larvicidal property close to that of the chemical compound., Conclusion: This result indicates the usefulness of plant materials in the control of obnoxious insect vectors of mn. It also opens additional approach to the management of environment from toxic chemicals that are non-biodegradable and cause environmental imbalance in the ecosystem.

Fipronil is a phenylpyrazole insecticide used in agricultural and domestic settings for controlling various insect pests in crops, lawns, and residential structures. Fipronil is chiral; however, it is released into the environment as a racemic mixture of two enantiomers. In this study, the acute toxicity of the (S,+) and (R,-) enantiomers and the racemic mixture of fipronil were assessed using Simulium vittatum IS-7 (black fly), Xenopus laevis (African clawed frog), Procambarus clarkii (crayfish), Palaemonetes pugio (grass shrimp), Mercenaria mercenaria (hardshell clam), and Dunaliella tertiolecta (phytoplankton). Results showed that S. vittatum IS-7 was the most sensitive freshwater species to the racemic mixture of fipronil (LC50 = 0.65 microg/L) while P. pugio was the most sensitive marine species (LC50 = 0.32 microg/L). Procambarus clarkii were significantly more sensitive to the (S,+) enantiomer while larval P. pugio were significantly more sensitive to the (R,-) enantiomer. Enantioselective toxicity was not observed in the other organisms tested. Increased mortality and minimal recovery was observed in all species tested for recovery from fipronil exposure. These results indicate that the most toxic isomer of fipronil is organism-specific and that enantioselective toxicity may be more common in crustaceans than in other aquatic organisms.

The transmission of Onchocerca volvulus Leuckart (Spirudida: Onchocercidae) and the prospects of Simulium neavei Roubaud (Diptera: Simuliidae) vector elimination through ground larviciding were investigated in the Mpamba-Nkusi focus, western Uganda. Transmission levels and the initiated vector elimination activities were assessed to supplement the ongoing ivermectin mass distribution programme. Searches for breeding sites, adult fly catches, dissection of flies, river treatment with temephos (Abate) and a review of annual ivermectin treatment data were conducted. High levels of crab infestation with S. neavei sensu stricto immature stages were recorded; 57.9% and 100% for the Mpamba and Nyabugando river systems, respectively. The mean numbers of larvae/pupae per crab were 3.6 +/- 0.5 in the Mpamba and 20.6 +/- 1.8 in the Nyabugando systems. Pre-intervention mean biting densities were 39 and 32 flies/(man day) in 2001 and 2002, respectively, and an annual biting rate in 2001 of > 14 000. The bimodal biting pattern of S. neavei s.s. consisted of two peaks; one in the morning (09.00-10.00 hours) and one in the afternoon (14.00-15.00 hours) with a mid-day lull in biting. The infection/infective rates were 13.3%/2.8% and 16.6%/2.9% in the dissected parous flies from the Mpamba and Nyabugando river systems, respectively. Out of approximately 1000 parous flies, 129 and 109 were found to be harbouring infective larvae of Onchocerca volvulus in their heads from the Mpamba and Nyabugando river systems, respectively. In spite of the > 10 years of ivermectin treatment, at a mean coverage of 71.3%, infection remained relatively high. Ground larviciding with temephos (Abate) initiated in June and October 2002 had a significant impact. In the Mpamba river system there was a significant (P < 0.001) reduction in positive crabs from 57.9% in 2001 to 0.06% in 2003 and a decrease in the mean number of larvae/pupae per crab from 3.6 +/- 0.5 in 2001 to 0.0007 +/- 0.0001 (P < 0.002) in 2003. Similarly, in the Nyabugando river system, a significant (P < 0.001) reduction in crab infestation from 100% in 2001 to 0.06% in 2003 and a decrease in the mean number of larvae/pupae per crab from 20.6 +/- 1.8 in 2001 to 0.06 +/- 0.03 in 2003. Drastic reductions were observed in the mean number of biting flies from 3 flies/h in 2001 to 0 flies/h in 2003 and the annual biting rates fell from 14,235 flies/year in 2001 to only 730 flies/year in 2003. These data suggest that substantial progress towards the goal of S. neavei s.s. vector elimination has been made and this will enhance the ongoing ivermectin treatment in this isolated focus.

In the Lower Cross River basin in Nigeria, no pre-control entomologic profile of Onchocerca volvulus infection in the local Simulium damnosum population was available prior to the initiation of an ivermectin control program in 1995. A longitudinal entomologic study was therefore carried out over a 12-month period (January-December 2001) at the Agbokim waterfalls and Afi River, which are breeding sites of S. damnosum in the river basin. A total of 9,287 adult S. damnosum were caught on human bait; 9,048 (97.43%) were dissected, of which 313 (3.46%) were infected. Annual biting rates (ABRs) of 42,419 and 28,346 bites per persons per year were recorded at the Agbokim Waterfalls and Afi River, respectively. The annual transmission potential (ATP) was 419 infective larvae per person per year at the Agbokim Waterfalls and 427 at the Afi River. Monthly biting rate and monthly transmission potential varied significantly (P < 0.05) at the two sites. Transmission was highly seasonal from April to September, corresponding to the peak biting period of the vector. The high ATP and ABR values are a measure of the mesoendemicity of onchocerciasis in the river basin. There was a significant F(0).05 (1, 10) (P < 0.05) variation in the relative fly abundance from both sites. It was observed that human activities such as farming, fishing, timber cutting, and hunting are done in the early morning and late afternoon, which corresponds to the peak diurnal biting period of the vector. Changes in these practices and attitudes may markedly affect the disease intensity and transmission.

Determination of the most sensitive and susceptible organismal life stage is important for use of the organism in toxicity assessments of environmental pollutants. In the present study, the sensitivities and susceptibilities of larval developmental stages of two black fly sibling species, Simulium vittatum Zetterstedt cytospecies IS-7 and S. vittatum Zetterstedt cytospecies IIIL-1, were determined using the organophosphate-insecticide chlorpyrifos. Differences in sensitivity and susceptibility were determined through analysis of slopes and median lethal concentration values produced from 24-h orbital shaker toxicity tests, respectively. The results showed no difference in sensitivity or susceptibility between sibling species. However, early instar (second and third) as well as mid-instar (fourth and fifth) groupings of S. vittatum IS-7 were significantly more susceptible than late-instar (sixth and seventh) larval groupings. Likewise, mid-instar groupings of S. vittatum IIIL-1 were more susceptible than late-instar larval groupings. However, neither species showed significant differences in sensitivity among instar groupings. The results of the present study indicate that the mid-instar groupings of black fly larvae are the best choice for use in toxicity tests, both because of their increased susceptibility compared to late-instar groupings and because of their ease in handling and manipulating in the laboratory compared to early instar groupings.

The bacterium Bacillus thuringiensis (Bt) produces parasporal crystals containing delta-endotoxins responsible for selective insecticidal activity on larvae. Upon ingestion, these crystals are solubilized in the midgut lumen and converted into active toxins that bind to receptors present on the microvilli causing serious damage to the epithelial columnar cells. We investigated the effect of these endotoxins on larvae of the Simulium pertinax, a common black fly in Brazil, using several concentrations during 4 h of the serovar israelensis strain IPS-82 (LFB-FIOCRUZ 584), serotype H-14 type strain of the Institute Pasteur, Paris. Light and electron microscope observations revealed, by time and endotoxin concentration, increasing damages of the larvae midgut epithelium. The most characteristic effects were midgut columnar cell vacuolization, microvilli damages, epithelium cell contents passing into the midgut lumen and finally the cell death. This article is the first report of the histopathological effects of the Bti endotoxins in the midgut of S. pertinax larvae and the data obtained may contribute to a better understanding of the mode of action of this bacterial strain used as bioinsecticide against black fly larvae.

The study was set up to evaluate the impact of two commercial larvicide formulations, Bacillus thuringiensis serovar israelensis base (Bti) at 15 ppm/1 min and temephos at 0.03 ppm of active ingredient, used to control Simulium pertinax populations, on associated non-target entomofauna occupying the same breeding sites. The experiments were carried out on the Pedra Branca and Muricana rivers, on the slopes of Serra do Mar massif, municipality of Paraty, state of Rio de Janeiro, Brazil. Bti was applied to the river Pedra Branca and temephosto theriver Muricana. On both rivers, treatment and control sections were labeled as such, each one with two observation posts: slow moving water and fast water regions respectively. Artificial substrata was used to evaluate the abundance of associated entomofauna. Attached immature stages of arthropods were removed from both of its surfaces fortnightly. Were collected, from the two rivers, 28 477 specimens of the entomofauna associated with S. pertinax. The families Hydropsychidae, Chironomidae, Bactidae, Simuliidae, Blephariceridae and Megapodagrionidae were represented. These was an impact of temephos on the entomofauna associated with S. pertinax only in Simuliidae and Chironomidae, and to Bti only in Simuliidae. However, the reduction in their numbers was not statistically significant.

Mixed populations of the pest blackflies Simulium bonaerense Coscarón & Wygodzinsky, S. wolffhuegeli (Enderlein) and S. nigristrigatum Wygodzinsky & Coscarón (Diptera: Simuliidae) are highly resistant to DDT and pyrethroids in the Neuquén Valley, a fruit-growing area of northern Patagonia, Argentina. As these insecticides have not been used for blackfly control, resistance is attributed to exposure to agricultural insecticides. Pre-treatment with the synergist piperonyl butoxide (PBO) reduced both DDT and fenvalerate resistance, indicating that resistance was partly due to monooxygenase inhibition. Pre-treatment with the synergist tribufos to inhibit esterases slightly increased fenvalerate toxicity in the resistant population. Even so, biochemical studies indicated almost three-fold higher esterase activity in the resistant population, compared to the susceptible. Starch gel electrophoresis confirmed higher frequency and staining intensity of esterase electromorphs in the resistant population. Incomplete synergism against metabolic resistance indicates additional involvement of a non-metabolic resistance mechanism, such as target site insensitivity, assumed to be kdr-like in this case. Glutathione S-transferase activities were low and inconsistent, indicating no role in Simulium resistance. Knowing these spectra of insecticide activity and resistance mechanisms facilitates the choice of more effective products for Simulium control and permits better coordination with agrochemical operations.

Populations of Simulium (Chirostilbia) pertinax Kollar, 1832 from Southern and Southeastern States of Brazil were analyzed for temephos susceptibility considering control historical information and possible resistance. In situ bioassays were carried out for populations from the states of Paraná (Tibaji and Rolândia), Rio de Janeiro (Muriqui) and São Paulo (Barra do Una, Ilhabela and Morungaba). The populations were characterized as susceptible (S) or resistant (R) by submitting larvae in the last instars to an operational concentration (0.1ppm a.i./10min) of temephos (Abate 500E) as diagnostic. The possible mechanisms for the organophosphorus resistance development are discussed considering old and new control strategies.

Chromosomal studies were carried on six larval populations of Simulium (Chirostilbia) pertinax from different locations in Brazil. Larvae were collected in the states of Paraná, Rio Grande do Sul, Rio de Janeiro and São Paulo. Polytene chromosome map comparisons within and among populations showed no differences in banding pattern, except for some limited polymorphism (secondary NOR and four band polymorphisms). There were no chromosomal variations associated with the resistance or susceptibility of the larvae to temephos. The chromosomal homosequentiality found among the six populations suggests that S. pertinax may be a monomorphic species.

The relative toxicity of leaf litter to nematocerous dipteran larvae characteristic of mosquito developmental sites was investigated. Culicidae, Chironomidae, and Simuliidae taxa originating from alpine hydrosystems were tested together with two laboratory nonindigenous culicid taxa. Bioassays indicate that ingestion of 10-month-old decaying leaves from Alnus glutinosa, Populus nigra, and Quercus robur by larvae is more deleterious for Aedes aegypti, A. albopictus, Culex pipiens, Simulium variegatum, and Chironomus annularius than for A. rusticus. Histopathological observations reveal that the midgut epithelium is the main target organ of the toxic effect of dietary leaf litter, which appears to be stronger than that of previously reported tannic acid. There is a general response of the nematocerous larval midgut epithelium to dietary tannins-phenolic compounds: clear cells of the anterior midgut showing symptoms of intoxication before dark cells of the posterior midgut., (Copyright 2000 Academic Press.)