Susceptibility of Ixodes scapularis (Acari: Ixodidae) to Metarhizium brunneum F52 (Hypocreales: Clavicipitaceae) using three exposure assays in the laboratory

An emulsifiable concentrate (EC) and granular (G) formulation of the entomopathogenic fungus, Metarhizium brunneum strain F52 (formerly Metarhizium anisopliae strain F52) were tested against unfed adults and nymphs of Ixodes scapularis Say in the laboratory. Three exposure methods; dip, surface contact, and direct spray application, and three exposure time intervals (3, 30, and 300 min) were used to evaluate the EC formulation. Application rates ranged from 2.6 x 10(2) to 2.6 x 10(8) conidia/cm2. The surface treatment was used for granular formulation with concentrations ranging from 2.3 x 10(5) to 2.3 x 10(7) conidia/cm2 for same three exposure times. Both the EC and G formulations of this fungus were highly pathogenic against I. scapularis adults and nymphs. Logistic regression analysis found formulation, spore concentration, time of exposure, and observation period were significant or highly significant factors influencing tick mortality. For adult I. scapularis, the spray application with the EC formulation of M. brunneum F52 resulted in a lower LC50 (5.9 x 10(4) conidia/cm2) at 30 min than surface exposure to the EC (LC50 = 1.3 x 10(6) conidia/cm2) or G formulation (LC50 = 8.1 x 10(5) conidia/cm2). At higher concentrations, fungal activity was evident in adult I. scapularis held at 5 degrees C suggesting the fungus may provide control in the cooler fall season. While the observed pathogenicity of a fungus against ticks can be dependent upon the bioassay assessment, we found nymphs and adults of I. scapularis to be highly susceptible to M. brunneum F52, regardless of the exposure method used.     

Potential of Tenebrio molitor (Coleoptera: Tenebrionidae) as a bioassay probe for Metarhizium brunneum (Hypocreales: Clavicipitaceae) activity against Ixodes scapularis (Acari: Ixodidae)

The yellow mealworm, Tenebrio molitor L., has been used to indicate qualitatively the presence of entomopathogenic fungi in the soil or as a model for evaluating stress and other factors on fungal activity. Although this beetle appears highly susceptible to many of these fungi, little quantitative information is available on the sensitivity of T. molitor to a specific fungus and, therefore, fungal presence or as an indicator for pathogenicity to other species. The purpose of this study was to establish the suitability of T. molitor larvae as a bioassay probe for Metarhizium brunneum for comparison against the blacklegged tick, Ixodes scapularis. Nine concentrations of M. brunneum strain F52 ranging from 1.0 x 10(1) to 8.4 x 10(8) conidial/ml were simultaneously tested against T. molitor larvae and I. scapularis adults. Larvae of yellow mealworm were less sensitive to M. brunneum than I. scapularis adults (LC50's 4.4 x 10(7) and 1.7 x 10(5) conidia/ml, respectively, 4-wk post-treatment). The greater sensitivity of I. scapularis to the fungus suggests that the detection of fungal mycosis in mealworms would indicate sufficient inoculum to be pathogenic to I. scapularis and make this insect a suitable probe for evaluation of the presence and activity of M. brunneum against the blacklegged tick in field applications.     

Laboratory evaluation of methanolic extract of Atlantia monophylla (Family: Rutaceae) against immature stages of mosquitoes and non-target organisms

Methanolic extracts of the leaves of Atlantia monophylla (Rutaceae) were evaluated for mosquitocidal activity against immature stages of three mosquito species, Culex quinquefasciatus, Anopheles stephensi, and Aedes aegypti in the laboratory.Larvae of Cx. quinquefasciatus and pupae of An. stephensi were found more susceptible, with LC50 values of 0.14 mg/l and 0.05 mg/l, respectively. Insect growth regulating activity of this extract was more pronounced against Ae. aegypti, with EI50 value 0.002 mg/l. The extract was found safe to aquatic mosquito predators Gambusia affinis, Poecilia reticulata, and Diplonychus indicus, with the respective LC50 values of 23.4, 21.3, and 5.7 mg/l. The results indicate that the mosquitocidal effects of the extract of this plant were comparable to neem extract and certain synthetic chemical larvicides like fenthion, methoprene, etc.     

Nutritional ecology of the formosan subterranean termite (Isoptera: Rhinotermitidae): feeding response to commercial wood species

The feeding preferences of the Formosan subterranean termite, Coptotermes formosanus Shiraki, were tested in three separate experiments on 28 different wood species. Experiment 1 was a multiple-choice test designed to test relative preferences among 24 wood species commercially available in New Orleans, LA. Experiment 2 was a similar study designed to test relative preferences among 21 wood species shown or reported to be unpalatable to the Formosan subterranean termite. Experiment 3 was a no-choice test to examine the feeding deterrence of the 10 least preferred wood species. Preference was determined by consumption rates. Birch (Betula alleghaniensis Britton), red gum (Liquidambar styraciflua L.), Parana pine [Araucaria angustifolia (Bert.) 1, sugar maple (Acer saccharum Marsh.), pecan (Carya illinoensis Wangenh.), and northern red oak (Quercus rubra L.) were the most preferred species by C. formosanus in order of consumption rate. All of these species were significantly more preferred than southern yellow pine (Pinus taeda L.), widely used for monitoring. Sinker cypress [ = old growth bald cypress, Taxodium distichum (L.)], western red cedar (Thuja plicata Donn), Alaskan yellow cedar (Chamaecyparis nootkatensis D. Don), eastern red cedar (Juniperus virginiana L.), sassafras [Sassafras albidum (Nutt.)], Spanish cedar (Cedrella odorata L.), Honduras mahogany (Swietenia macrophyla King), Indian rosewood (Dalbergia latifolia Roxb.), Honduras rosewood (D. stevensonii Standl.), and morado (Machaerium sp.) induced significant feeding deterrence and mortality to C. formosanus. The last eight species produced 100% mortality after 3 mo.     

Chemical composition and anti-mosquito potential of rhizome extract and volatile oil derived from Curcuma aromatica against Aedes aegypti (Diptera: Culicidae).

Crude rhizome extracts and volatile oils of Curcuma aromatica were evaluated for chemical composition and anti-mosquito potential, including larvicidal, adulticidal, and repellent activities against the Aedes aegypti mosquito. Chemical identification achieved by GC/MS analysis revealed that xanthorrhizol, 1H-3a, 7-methanoazulene and curcumene at 35.08 and 13.65%, 21.81 and 30.02%, and 13.75 and 25.71%, were the main constituents in hexane extracts and volatile oils, respectively. Volatile oil of Cu. aromatica possessed a significantly higher larvicidal activity against the 4th instar larvae of Ae. aegypti than that of hexane extracts, with LC50 values of 36.30 and 57.15 ppm, respectively. In testing for adulticidal activity, on the other hand, hexane-extracted Cu. aromatica (LC50: 1.60 microg/mg female) was found to be slightly more effective against female Ae. aegypti than volatile oil (LC50: 2.86 microg/mg female). However, the repellency of these two products against Ae. aegypti adult females differed significantly. The hexane-extracted Cu. aromatica, with a median complete protection time of 1 h (range = 1-1.5 h) when applied at a concentration of 25%, appeared to have significantly higher repellency than that of distillate oil (0.5 h, range = 0-0.5 h). The different results obtained from both products of Cu. aromatica were probably due to variety in quantity and type of active ingredients as well as the biological and physiological characteristics that differed between both developmental stages of mosquitoes, larvae, and adults.     

Ovicidal and larvicidal activity against Aedes aegypti and Anopheles gambiae complex mosquitoes of essential oils extracted from three spontaneous plants of Burkina Faso

Essential oils extracted from dried leaves of three spontaneous plants naturally growing in Burkina Faso, i.e. Cymbopogon proximus, Lippia multiflora and Ocimum canum, exhibited larvicidal activity by the WHO standard protocol against 3rd and 4th instar F1-larvae of field-collected mosquitoes vectors of human disease, namely Aedes aegypti and members of the Anopheles gambiae complex, An. arabiensis and An. gambiae. The median lethal concentration (LC50) for Ae. aegypti and An. gambiae s.l. larvae ranged between 53.5-258.5 ppm and 61.9-301.6 ppm, respectively. The LC90 estimates ranged 74.8-334.8 ppm for Ae. aegypti, and 121.6-582.9 ppm for An. gambiae s.l. Ovicidal activity against eggs of An. gambiae s.l. was also demonstrated. The LC50 values for An. gambiae s.l. eggs ranged between 17.1-188.7 ppm, while LC90 values ranged between 33.5-488 ppm. Lippia multiflora showed the highest activity against An. gambiae s.l. eggs and Ae. aegypti larvae, whereas no difference was found among C. proximus and L. multiflora in their activity against An. gambiae s.l. larvae. Of the three plants, essential oils from O. canum had the lowest activity against both eggs and larvae. Eggs were more susceptible than larvae. Ae. aegypti larvae were more susceptible than larvae of An. gambiae s.l.     

Phylogeography of Aedes (Stegomyia) aegypti (L.) and Aedes (Stegomyia) albopictus (Skuse) (Diptera: Culicidae) based on mitochondrial DNA variations

Aedes (Stegomyia) aegypti (l.) and Aedes (Stegomyia) albopictus (Skuse) are the most important vectors of the dengue and yellow-fever viruses. Both took advantage of trade developments to spread throughout the tropics from their native area: A. aegypti originated from Africa and a. albopictus from South-East Asia. We investigated the relationships between A. aegypti and A. albopictus mosquitoes based on three mitochondrial-DNA genes (cytochrome b, cytochrome oxidase I and NADH dehydrogenase subunit 5). Little genetic variation was observed for a. albopictus, probably owing to the recent spreading of the species via human activities. For A. aegypti, most populations from South America were found to be genetically similar to populations from South-East Asia (Thailand and Vietnam), except for one sample from Boa Vista (northern Amazonia), which was more closely related to samples from Africa (Guinea and Ivory Coast). This suggests that African populations of A. aegypti introduced during the slave trade have persisted in Boa Vista, resisting eradication campaigns.     

Essential oils of Cupressus funebris, Juniperus communis, and J. chinensis (Cupressaceae) as repellents against ticks (Acari: Ixodidae) and mosquitoes (Diptera: Culicidae) and as toxicants against mosquitoes

Juniperus communis leaf oil, J. chinensis wood oil, and Cupressus funebris wood oil (Cupressaceae) from China were analyzed by gas chromatography and gas chromatography-mass spectrometry. We identified 104 compounds, representing 66.8-95.5% of the oils. The major components were: α-pinene (27.0%), α-terpinene (14.0%), and linalool (10.9%) for J. communis; cuparene (11.3%) and δ-cadinene (7.8%) for J. chinensis; and α-cedrene (16.9%), cedrol (7.6%), and β-cedrene (5.7%) for C. funebris. The essential oils of C. funebris, J. chinensis, and J. communis were evaluated for repellency against adult yellow fever mosquitoes, Aedes aegypti (L.), host-seeking nymphs of the lone star tick, Amblyomma americanum (L.), and the blacklegged tick, Ixodes scapularis Say, and for toxicity against Ae. aegypti larvae and adults, all in laboratory bioassays. All the oils were repellent to both species of ticks. The EC(95) values of C. funebris, J. communis, and J. chinensis against A. americanum were 0.426, 0.508, and 0.917 mg oil/cm(2) filter paper, respectively, compared to 0.683 mg deet/cm(2) filter paper. All I. scapularis nymphs were repelled by 0.103 mg oil/cm(2) filter paper of C. funebris oil. At 4 h after application, 0.827 mg oil/cm(2) filter paper, C. funebris and J. chinensis oils repelled ≥80% of A. americanum nymphs. The oils of C. funebris and J. chinensis did not prevent female Ae. aegypti from biting at the highest dosage tested (1.500 mg/cm(2) ). However, the oil of J. communis had a Minimum Effective Dosage (estimate of ED(99) ) for repellency of 0.029 ± 0.018 mg/cm(2) ; this oil was nearly as potent as deet. The oil of J. chinensis showed a mild ability to kill Ae. aegypti larvae, at 80 and 100% at 125 and 250 ppm, respectively.     

Snail predation by larvae of Sepedon scapularis Adams (Diptera: Sciomyzidae), a potential biocontrol agent of snail intermediate hosts of schistosomiasis in South Africa

Malacophagous larvae of the fly Sepedon scapularis Adams were shown experimentally to be effective predators of three species of aquatic pulmonate snails tested as prey: Bulinus africanus (Krauss) an important intermediate host of Schistosoma haematobium (Bilharz), Bulinus tropicus (Krauss) and the invasive species Physa acuta Draparnaud. Survival of S. scapularis larvae from instar to instar was negatively affected by the size of prey snails, since larvae tended to be asphyxiated by the mucous secretions of the snails, or by the larval hydrofuge hairs becoming entangled in snail faeces. In experiments to test the choice of S.scapularis larvae for different species and sizes of snails, B.africanus was significantly disfavoured compared with the other two snail species. Small snails (< 3 mm) were more frequently killed by all three larval instars of S.scapularis. First instars killed few, if any, large snails (> 7 mm), whereas second and third instars preyed effectively on all sizes of snails. Third instars killed significantly more snails than younger instars. For larvae offered only one species of snail as prey, the mean total number of snails killed per larva during its entire development was 49 B.africanus, 45 B.tropicus or 34 P.acuta. It is concluded that S.scapularis is potentially useful as a biological control agent for use against indigenous Bulinus and exotic Physa snails.     

Larvicidal activity of Tagetes patula essential oil against three mosquito species

Larvicidal activity of Tagetes patula essential oil was tested against the fourth instar larvae of Aedes aegypti, Anopheles stephensi, and Culex quinquefaciatus. Five different concentrations of essential oil were studied and the results were compared with that of synthetic insecticide, malathion. A. aegypti (LC(50) 13.57, LC(90) 37.91) was most susceptible followed by An. stephensi (LC(50) 12.08, LC(90) 57.62) and C. quinquefaciatus (LC(50) 22.33, LC(90) 71.89).     

Introduction of Culex toxicity into Bacillus thuringiensis Cry4Ba by protein engineering

Bacillus thuringiensis mosquitocidal toxin Cry4Ba has no significant natural activity against Culex quinquefasciatus or Culex pipiens (50% lethal concentrations [LC(50)], >80,000 and >20,000 ng/ml, respectively). We introduced amino acid substitutions in three putative loops of domain II of Cry4Ba. The mutant proteins were tested on four different species of mosquitoes, Aedes aegypti, Anopheles quadrimaculatus, C. quinquefasciatus, and C. pipiens. Putative loop 1 and 2 exchanges eliminated activity towards A. aegypti and A. quadrimaculatus. Mutations in a putative loop 3 resulted in a final increase in toxicity of >700-fold and >285-fold against C. quinquefasciatus (LC(50) congruent with 114 ng/ml) and C. pipiens (LC(50) 37 ng/ml), respectively. The enhanced protein (mutein) has very little negative effect on the activity against Anopheles or AEDES: These results suggest that the introduction of short variable sequences of the loop regions from one toxin into another might provide a general rational design approach to enhancing B. thuringiensis Cry toxins.     

Impacts of granivorous and frugivorous arthropods on pre-dispersal seed production of western juniper (<Emphasis Type="Italic">Juniperus occidentalis</Emphasis>)

Arthropods impact seed production in various juniper species, but effects of pre-dispersal seed predation are generally unknown for arthropods that feed on western juniper (Juniperus occidentalis). From 2009 to 2013, we quantified impacts of three arthropod granivores and a frugivorous insect on western juniper seed production at two N. California field sites—Madeline and Shinn Peak. Insect larvae were genetically identified using the COI barcode region. Seed damage by all arthropod taxa varied both spatially and temporally. Juniper berry mites (Trisetacus quadricetus) had the greatest effect on reducing seed production at Madeline, and granivorous moths (Periploca spp. and Argyresthia spp.) had the greatest effect at Shinn Peak. Three findings supported the predator satiation hypothesis, which suggests that unpredictable masting events overwhelm responses of seed predators. First, estimated berry production had significant negative effects on the proportion of seeds damaged across sites by a chalcidoid wasp (Eurytoma juniperina) and by granivorous moths at Shinn Peak. Second, seed damage by granivorous moths was significantly reduced in a mast year of juniper berry production. Third, number of seeds per berry positively affected seed damage across sites by granivorous moths in all years except the mast year as well as damage by a frugivorous tortricid moth (Henricus infernalis) at Madeline. Distance to neighboring trees had positive, site-specific effects on damage by granivorous moths and Henricus, perhaps due to limited movement of parasitoids that attack these insects. Our results suggest that arthropod damage can significantly impact seed production of western juniper.     

Host feeding responses of Aedes aegypti (L.) exposed to deltamethrin

Escape responses of mated and unmated nulliparous Aedes aegypti mosquitoes were compared using three different concentrations of deltamethrin in the presence or absence of a live animal host using an excito-repellency (ER) test system. Both insecticide contact (excitation) and non-contact (repellency) test configurations were compared. For contact trials, mated mosquitoes showed similar escape movements among the three concentrations when host stimuli were absent. Significant differences in responses were seen between the lower concentrations of (LC(50) and LC(75) ) deltamethrin with and without hosts present (P<0.05). Presence or absence of host stimuli produced no significant differences in escape response for unmated females when exposed to the highest concentration (LC(90) ) of deltamethrin. Our findings indicate that as deltamethrin concentrations decrease to sublethal levels, mating status and host cues play a more significant role in escape behavior. Therefore, insemination can influence the outcome of feeding success and flight movement of nulliparous female Ae. aegypti in contact with deltamethrin and in the presence of live host stimuli. The ER assay system serves as a useful tool for observing excitation and repellency responses of Ae. aegypti to insecticides in the presence or absence of other environmental and biological cues that can affect mosquito behavior.     

Larvicide and oviposition deterrent effects of fruit and leaf extracts from Melia azedarach L. on Aedes aegypti (L.) (Diptera: Culicidae)

Aedes aegypti (L.) (Diptera: Culicidae), the main urban vector of dengue, has developed resistance to various insecticides, making its control increasingly difficult. We explored the effects of Argentine Melia azedarach L. (Meliaceae) fruit and senescent leaf extracts on Ae. aegypti larval development and survival, by rearing cohorts of first instar mosquitoes in water with different extract concentrations. We also analysed oviposition deterrent activity in choice tests with extract-treated ovitraps. The leaf extract showed a strong larvicide activity, with all larvae dying before pupation, and significantly delayed development time. It strongly inhibited oviposition by Ae. aegypti females. The fruit extract showed much weaker effects. This first report of highly effective larvicidal, growth regulating and oviposition deterrent activity of a senescent leaf extract of M. azedarach against Ae. aegypti, suggests that such extract could represent a promising tool in the management of this mosquito pest.     

Derris (Lonchocarpus) urucu (Leguminosae) extract modifies the peritrophic matrix structure of Aedes aegypti (Diptera:Culicidae)

Aqueous suspension of ethanol extracts of Derris (Lonchocarpus) urucu (Leguminosae), collected in the state of Amazonas, Brazil, were tested for larvicidal activity against the mosquito Aedes aegypti (Diptera:Culicidae). The aim of this study was to observe the alterations of peritrophic matrix in Ae. aegypti larvae treated with an aqueous suspension of D. urucu extract. Different concentrations of D. urucu root extract were tested against fourth instar larvae. One hundred percent mortality was observed at 150 microg/ml (LC(50) 17.6 microg/ml) 24 h following treatment. In response to D. urucu feeding, larvae excreted a large amount of amorphous feces, while control larvae did not produce feces during the assay period. Ultrastructural studies showed tha larvae fed with 150 microg/ml of D. urucu extract for 4 h have an imperfect peritrophic matrix and extensive damage of the midgut epithelium. Data indicate a protective role for the peritrophic matrix. The structural modification of the peritrophic matrix is intrinsically associated with larval mortality.     

Intercanopy community structure across a heterogeneous landscape in a western juniper‐encroached ecosystem

Questions: How does landscape position influence biotic and abiotic attributes of western juniper (Juniperus occidentalis) encroached ecosystems? How does intercanopy plant community structure respond to changes in soil moisture and temperature based on juniper cover and topographic position? Location: Steens Mountain, southeast Oregon, USA. Objectives: Competition with western juniper modifies plant community composition, alters soil hydrology, and reduces plant productivity. Research is needed to understand these influences across heterogeneous landscapes. This study characterizes the relationship between juniper encroachment and soil water, soil temperature, topographic position, and intercanopy plant community structure. Methods: Using a completely randomized block design, plant density and cover, percent bare ground, percent soil moisture, soil temperature, heat accumulation, and elevation were sampled in 10 m² plots representing low (<1%), moderate (～14%), and high (～27%) juniper cover at four aspects. The relationship and difference between vegetation patterns and environmental variables were analyzed using AOV, NMS, and MRPP (α=0.1). Indicator species analysis tested for shifts in dominant species along ecological gradients. Results: Soil moisture remained higher in low juniper cover sites than moderate and high juniper cover sites. North‐facing sites had highest soil moisture at 5 cm depth with low and moderate juniper cover levels. With increasing soil temperature from May to June, soil moisture declined by 19.7% at 5 cm depth. Achnatherum lemmonii and Pseudoreogneria spicata occurred in closed juniper stands while Achnatherum occidentale and Leymus cinereus were common when encroachment was limited. Application: This approach can be used to predict ecosystem response to western juniper encroachment across heterogeneous landscapes.     

Susceptibility of two different strains of Aedes aegypti (Diptera: Culicidae) to plant oils

The toxicity of 11 oils extracted from plants commonly grown in the Himalayan region was studied using larvae of two Aedes aegypti (L.) strains. A strain from Liverpool, England, was highly susceptible to these oils. The LC50 values were much higher in a local laboratory strain. Daucus carota L. oil was highly toxic in both strains. Differences in the susceptibility of these strains to the action of the test oils and their potential use in integrated pest management are discussed.     

Bruchid (Coleoptera: Bruchidae) ovicidal phenylbutanoid from Zingiber purpureum

The larvicidal activity of the dichloromethane extract of Zingiber purpureum Roscoe (Zingiberaceae) rhizome against the second instar of Aedes aegypti (L.) (Diptera: Culicidae) is shown to be due to 4-(3',4'-dimethoxyphenyl)buta-1,3-diene. The diene also showed ovicidal activity against the bruchid Callosobruchus maculatus (F.) (Coleoptera: Bruchidae). Most of the eggs laid by bruchids on treated cowpea seeds were transparent, and very few of them contained developing embryos. The few larvae produced from these embryos were unable to penetrate the seed coat and enter the seed. Similar effects were seen when adults were exposed to the compound and then placed on untreated cowpea seeds, suggesting that a new type of maternally mediated ovicidal effect was involved. Coated and impregnated granular formulations of the extract were evaluated for use in the control of bruchid infestation of stored cowpea seeds. Coated granules showed activity similar to that of the crude extract but were found to lose activity rapidly. Impregnated granules were found to be less active than the crude extract.     

Importance of juniper to birds nesting in piñon–juniper woodlands in northwest New Mexico

Piñon–juniper (Pinus spp.–Juniperus spp.) woodlands are common throughout western North America, yet relatively little is known about the habitat use and requirements for many members of its avian community. During summer 2005–2007, we assessed avian nesting substrates within piñon (Pinus edulis)–juniper (Juniperus osteosperma) woodlands in northwestern New Mexico. Of all nests in live trees, 86% were in junipers. The selection of juniper as a nest tree was significantly higher than expected from the region's piñon–juniper ratio (1:1.06) for the community as a whole, for both open cup and cavity nesting species, and for 8 species (of which 6 are piñon–juniper obligate or semi-obligate species). Nest survival, however, was not higher in juniper than in piñon for the nesting community as a whole or for chipping sparrows (Spizella passerina), the single species that was well represented nesting in piñon. The high use of juniper as a nesting substrate differs from previous studies, which have suggested that a presence of piñon is among the most important habitat features for many piñon–juniper species. Because of their importance to nesting birds, managers should avoid preferential thinning of junipers within piñon–juniper woodlands. © 2011 The Wildlife Society.     

Larvicidal and repellent potential of Albizzia amara Boivin and Ocimum basilicum Linn against dengue vector, Aedes aegypti (Insecta:Diptera:Culicidae)

Investigations were made to test the larval toxicity and smoke repellent potential of Albizzia amara and Ocimum basilicum at different concentration (2%, 4%, 6%, 8% and 10%) against the different instar (I, II, III and IV) larvae and pupae of Aedes aegypti. The LC50 values of A. amara and O. basilicum for I instar larvae was 5.412 and 3.734, II instar 6.480 and 4.154, III instar 7.106 and 4.664, IV instar 7.515 and 5.124, respectively. The LC50 and LC90 values of pupae were 6.792%, 5.449% and 16.925%, 15.474%. The smoke toxicity of A. amara was more effective against A. aegypti than the O. basilicum.