Specific detection of the floodwater mosquitoes Aedes sticticus and Aedes vexans DNA in predatory diving beetles

Floodwater mosquitoes (Diptera: Culicidae) are associated with periodically flooded wet meadows, marshes, and swamps in floodplains of major rivers worldwide, and their larvae are abundant in the shallow parts of flooded areas. The nuisance caused by the blood‐seeking adult female mosquitoes motivates mosquito control. Larviciding with Bacillus thuringiensis israelensis is considered the most environmentally safe method. However, some concern has been raised whether aquatic predatory insects could be indirectly affected by this reduction in a potential vital prey. Top predators in the temporary wetlands in the River Dalälven floodplains are diving beetles (Coleoptera: Dytiscidae), and Aedes sticticus and Ae. vexans are the target species for mosquito control. For detailed studies on this aquatic predator–prey system, we developed a polymerase chain reaction (PCR) assay for detection of mosquito DNA in the guts of medium‐sized diving beetles. Primers were designed for amplifying short mitochondrial DNA fragments of the cytochrome C oxidase subunit I (COI) gene in Ae. sticticus and Ae. vexans, respectively. Primer specificity was confirmed and half‐life detectability of Ae. sticticus DNA in diving beetle guts was derived from a feeding and digestion experiment. The Ae. sticticus DNA within diving beetle guts was detected up to 12 h postfeeding, and half‐life detectability was estimated to 5.6 h. In addition, field caught diving beetles were screened for Ae. sticticus and Ae. vexans DNA and in 14% of the diving beetles one or both mosquito species were detected, showing that these mosquito species are utilized as food by the diving beetles.     

Environmental predictors of the occurrence of exotic <Emphasis Type="Italic">Hydrilla verticillata</Emphasis> (L.f.) Royle and native <Emphasis Type="Italic">Egeria najas</Emphasis> Planch. in a sub-tropical river floodplain: the Upper River Paraná, Brazil

The Upper River Paraná Floodplain System comprises the rivers Ivinheima, Baía and Paraná, which with their associated waterbodies form three subsystems, each showing individual characteristics. Hydrilla verticillata recently invaded the Upper Paraná Floodplain, while Egeria najas is the native most abundant submersed macrophyte. A large flood-pulse, during January–March 2007, abruptly reduced macrophyte stands in many areas to near-zero and dispersed propagules over the entire floodplain. From April 2007 to April 2008, we conducted three surveys sampling for the presence–absence of H. verticillata and E. najas and environmental variables aiming to answer: (1) How rapid is the colonization–regeneration process for both species? (2) Which habitats seem to be more susceptible to their colonization? (3) Which environmental factors can best predict their occurrence? Neither H. verticillata nor E. najas colonized the Ivinheima subsystem. In the Baía subsystem, E. najas had only two occurrences while H. verticillata was not present. In the Paraná subsystem, E. najas predominantly occurred in river channels, but it was also common in floodplain lakes. In April 2007, it was found in 13% of the sites in the Paraná subsystem, increasing to 30% in November 2007 and reaching 34% in April 2008. H. verticillata did not successfully colonize floodplain lakes of the Paraná subsystem. In channels, it had 34% occurrence in April 2007, increasing to 62% in November 2007 and remaining at 62% in April 2008. The role of environmental variables in predicting species occurrence changed depending upon the scale of the analysis. Considering the whole Upper Paraná floodplain, water transparency followed by electrical conductivity were the strongest predictors for both species. Colonization by submersed plants seems improbable in the Ivinheima subsystem owing to its low water transparency besides frequent localized floods; in the Baía subsystem, it seems inhibited by transparency and low alkalinity. Considering just the Paraná subsystem, the proportion of organic matter in sediment, ten times higher in floodplain lakes than in channels, was the best predictor for H. verticillata occurrence (also related to water pH and transparency), while E. najas was only significantly explained by transparency.     

Mosquito-only flaviviruses,isolated from <Emphasis Type="Italic">Aedes albopictus</Emphasis> in Slovenia: results of a pilot mosquito monitoring program

Autochthonous transmissions of mosquito-borne diseases were described in close proximity to Slovenia where the knowledge of mosquito species and the viruses they transmit remains very scarce. Therefore we aimed to assess the burden mosquitoes bring to the local community and to test mosquitoes for the presence of the selected zoonotic arboviruses, i. e. Dengue (DENV), West Nile (WNV) and Chikungunya (CHIKV). A pilot mosquito monitoring was conducted in 2012 within two municipalities based on adult mosquito trapping. Altogether 2007 adult mosquitoes were caught. Females of Aedes albopictus and Culex pipiens represented the majority of the total mosquito catch. In sites with more households and nearby greenery, catches of Ae. albopictus were more abundant. Highest numbers of Cx. pipiens females were caught close to a nature reserve, where many bird species stop during their migrations. Trapping results are consistent with both species’ feeding behaviour. Female mosquitoes were grouped into 58 pools according to species, location and date of catch. Real time RT-PCRs for WNV, DENV and CHIKV were performed, but no arboviral genomes were detected in any of the tested mosquito pools. However, amplicons were obtained in three pools when using generic RT-PCR markers for flaviviruses. The viruses detected were propagated and isolated in mosquito cell line C3/36 and identified with sequencing as mosquito-only flaviviruses (MOFs). The important contributions of this study are a successful first isolation of MOFs and confirmation of the presence of the invasive mosquito Ae. albopictus in Slovenia.     

Molecular analysis of the phylogenetic relationships among the diploid <Emphasis Type="Italic">Aegilops</Emphasis> species of the section <Emphasis Type="Italic">Sitopsis</Emphasis>

RAPD analysis was used to study the intraspecific variation and phylogenetic relationships of Sgenome diploid Aegilops species regarded as potential donors of the B genome of cultivated wheat. In total, 21 DNA specimens from six S-genome diploid species were examined. On a dendrogram, Ae. speltoides and Ae. aucheri formed the most isolated cluster. Among the other species, Ae. searsii was the most distant while Ae. longissima and Ae. sharonensis were the closest species. The maximum difference between individual accessions within one species was approximately the same (0.18–0.22) in Ae. bicornis, Ae. longissima, Ae. sharonensis, and Ae. searsii. The difference between the clusters of questionable species Ae. speltoides and Ae. aucheri corresponded to the intraspecific level; the difference between closely related Ae. longissima and Ae. sharonensis corresponded to the interspecific level.     

Diving beetle assemblages of flooded wetlands in relation to time,wetland type and Bti-based mosquito control

We investigated the abundance and taxonomic composition of the aquatic predatory insect fauna, with focus on adult diving beetles (Coleoptera: Dytiscidae), in eight temporary flooded wet meadows and two alder swamps in the River Dalälven floodplains, central Sweden from 2002 to 2006. Diving beetles are generalist predators and often abundant in various waters, including temporary wetlands. In the River Dalälven floodplains, recurrent floods induce massive hatching of flood-water mosquitoes (Diptera: Culicidae), which constitute a superabundant patchy and irregular food resource for aquatic predatory insects. Our aims were (1) to characterize the assemblage of adult diving beetles occurring in the wetlands during floods in relation to time and wetland type and (2) to evaluate the effect on the aquatic predator assemblage of strongly reducing the abundance of a potential prey, flood-water mosquito larvae with Bacillus thuringiensis var. israelensis (Bti) during floods. We found diving beetles to be the dominating aquatic predatory insect taxa in all 10 wetlands. There was a difference in Dytiscidae species richness but not in diversity between wet meadows and alder swamps after rarefaction. The cluster analysis based on dytiscid species and abundances showed very high similarities between the wetlands. The variance component analysis was unable to distinguish any factor that could explain more than 7.4% of the variation in the dytiscid species assemblages. The only effect of Bti-treatment against flood-water mosquito larvae, potential food for the predatory dytiscids, was a slight increase in abundance of the medium-sized dytiscid species. Our results are in accordance with previous studies, suggesting that irregular and recurrent flood dynamic structure the dytiscid fauna more than food limitations and environmental factors.     

AFLP,RAPD, and ISSR analysis of intraspecific polymorphism and interspecific differences of allotetraploid species <Emphasis Type="Italic">Aegilops kotschyi</Emphasis> Boiss. and <Emphasis Type="Italic">Aegilops variabilis</Emphasis> Eig

To evaluate genetic variation, 27 accessions of allotetraploid species Aegilops kotschyi and Ae. variabilis with the US genome were analyzed using the AFLP, RAPD, and ISSR methods. A total of 316 polymorphic RAPD fragments, 750 polymorphic AFLP fragments, and 234 polymorphic ISSR fragments were obtained. It was demonstrated that the analyzed species were characterized by a considerable level of nuclear genome variation. According to the data of ISSR and RAPD analysis, the average value of the Jaccard similarity coefficient for the accessions of Ae. variabilis from different geographical regions was slightly lower than that for the accessions of Ae. kotschyi. At the same time, AFLP analysis showed no considerable differences in the levels of intraspecific variation of the studied species. Analysis of the summarized RAPD, ISSR, and AFLP marking data in the Structure software program showed that most of the analyzed accessions with high degree of probability could be assigned to one of two groups, the first of which corresponded to Ae. kotschyi and the second corresponded to Ae. variabilis, thereby confirming the species independence of Ae. kotschyi and Ae. variabilis. Accessions k900, k907, k908, and v90 could not with a sufficiently high degree of probability be assigned to one of the species, which possibly was the result of interspecific hybridization. Analysis of the species diversity using different molecular markers made it possible to identify the accessions that were notably different from other accessions of its species.     

Half a century of changes in the riverine landscape of Limay River: the origin of a riparian neoecosystem in Patagonia (Argentina)

Our study aims to determine the dynamic that led to the spread of exotic Salicaceae on the Limay River floodplain and its implications in shaping the current neoecosystem. We used images obtained by the HRG sensor on board the SPOT-5 satellite. We selected two images dates allowed a comparison of the floodplain under different flooding regimes programmed by the Interjuridiccional Basin Authority, at a rate of 1290 m³/s in Spring and a flow rate of less than 400 m³/s in Summer. To characterize the vegetation cover, the Normalized Difference Vegetation Index (NDVI) was used to compare images of September and December, also the Normalized Difference Water Index (NDWI) was applied to delineate the riverbed and floodplain in September. To evaluate the influence of flooding regime on the detected patches a Principal Component Analysis (PCA), was performed; and biotic and abiotic factors on the composition of the dominant tree species in each patch by Multiple Factor Analysis (MFA) were analized. A 58.4 % coverage of forest patches had developed on the Limay River floodplain. Patches of older trees grew on surfaces at 1.5 m above the water level. However, the surfaces not reached by floods >1.5 m have a very low rocky coverage (1 %). The analysis of the age of the trees downstream of the dam system showed that the vegetation, often exposed to high floods before the dams were built (1971), was composed of the native willow (Salix humboldtiana) and the exotic Salix alba. As the river regime was attenuated and extraordinary floods disappeared with the operation of the dam Arroyito in 1980, the first patches of Populus nigra spread.     

Diversity of fish species in suichuan river and shushui river and conservation value,China

Based on the data collected from the Suichuan and Shushui Rivers, both tributaries of the Ganjiang River between April and July of 2015, the fish species identified were classified into 68 and 46 species, 14 and 12 families in the Suichuan River and Shushui River respectively. Cyprinidae is the most common family that accounts for 52.9% and 58.7% in the total number of fish species in the Suichuan River and Shushui River, respectively. The dominant species were Pseudohemiculter dispar, Squalidus argentatus, Silurus asotus and Leptobotia elongate for the Suichuan River and Squalidus argentatus, Acrossocheilus parallens, Pseudohemiculter dispar, Silurus asotus, Pelteobagrus fulvidraco, Acrossocheilus fasciatus and Zacco platypus for the Shushui River. The diversity of fish species showed more abundant and diverse for the Suichuan River. The composition of ecotype of fish indicated the rich diversities of ecotype in both the Suichuan and Shushui Rivers. The tributary (Suichuan River) and main stream of the Ganjiang River demonstrated a highly fauna similarity and the fish resources indicated its significance to maintain the fish diversity in the middle of the Ganjiang River and its benefits to the existence of the fish species in mountain streams. Dam construction, sand excavation and heavy metal pollution are the most significant threat to fish diversity and ecosystem functioning in Ganjiang River basin. In order to protect fish diversity and fisheries more effectively, relevant laws should be strengthened and conservation areas should be established for the survival of freshwater fish species.     

Odonata larvae of Kaliningrad oblast: Species composition and distributional features

In this study, a total of 27 species of dragonfly and damselfly larvae were identified, including 14 species from suborder Anisoptera and 13 from Zygoptera. The dominant species are Libellula quadrimaculata, Enallagma cyathigerum, and Erythromma najas. Frequently occurring species are Coenagrion hastulatum, C. lunulatum, C. pulchellum, Ischnura elegans, Platycnemis pennipes, Lestes virens, and Libellula depressa. Rare species are Lestes sponsa, Libellula fulva, Gomphus flavipes, Gomphus vulgatissimus, Sympetrum flaveolum, Aeshna viridis, Ae. cyanea, Ae. grandis, Ae. mixta, Calopteryx splendens, C. virgo, and Ophiogomphus cecilia.     

Cover thresholds for impacts of an exotic grass on the structure and assembly of a wet prairie community

Potential impacts of an exotic grass, Hemarthria altissima, on restoration of wet prairie community structure (species richness and cover of indicator species) and assembly processes (temporal turnover rates of plant species) on the Kissimmee River floodplain in Central Florida, USA, were evaluated over a 12-year period before and after restoration of hydrologic regimes (2001), and implementation of herbicide treatments (2006–2007) to control its spread. Thresholds for impacts were derived from comparisons of sample sites with variable levels of H. altissima cover. Prior to herbicide treatments, cover of H. altissima exhibited a logistic increase over time, with peak colonization and expansion occurring during major flood events. Mean post-restoration cover of three native wet prairie indicator species (Polygonum punctatum, Panicum hemitomon, and Luziola fluitans) increased to 37.8 ± 3.4 % in plots in which H. altissima cover was <12 %, and did not exceed 15 % in any plots with H. altissima cover >30 %. Prior to and after herbicide treatments, these indicator species largely accounted for observed differences in wet prairie community structure (i.e., cover of wetland forbs and grasses) between heavily infested sites and plots with low or no cover of H. altissima. The cover threshold at which H. altissima began to have these community-level effects was 40–50 %, but lower species richness was found only where H. altissima cover was >80 %. Increasing cover of H. altissima led to a significant decline in temporal turnover rates of plant species (P < 0.001, r² = 0.10), but also was largely due to plots with very high (>75 %) cover of H. altissima. Mean temporal turnover rates of plant species increased significantly (P = 0.03) after herbicide treatments and subsequently were highest during an ensuing flood pulse. However, 2–3 years after herbicide treatments, regrowth of H. altissima reestablished high cover (mean = 59 ± 9.5 %) in over half of the treated plots. The ability of H. altissima to establish dominant cover in restored hydrologic conditions on the Kissimmee River floodplain, and documented regrowth following herbicide treatments, increase the potential for this exotic grass species to be a pervasive threat to successful reestablishment of wet prairie community structure and assembly processes.     

The Application of Chironomid Pupal Exuvial Technique (CPET) for Ecological Analysis in a Neotropical Large River System

This field ecological study, based on the chironomid pupal exuvial technique (CPET), is new for the Paraná River and proposes an efficient tool to be used in future ecological approaches and biomonitoring. Drifting of pupal exuviae in a river-floodplain system of the Middle Paraná River floodplain was represented by 34 Chironomidae taxa, being the characteristic association obtained from the CPET: Lopescladius, Onconeura, Paralauterborniella, Polypedilum, and Harnischia complex. Diversity, richness, dominance, total density, and density of dominant taxa were different between the longitudinal and lateral dimensions but not between hydrologic phases, with a greater diversity and richness in the main channel of the river and higher density and dominance in floodplain habitats. The species turnover is the dominant process in structuring studied assemblages in spatial and temporal analysis, increasing in the floodplain habitats and in low-water phase. The results obtained showed that drifting exuviae in the longitudinal axis were coming from different assemblages and environments of a wider area (regional), while exuviae recorded in the connections of the floodplain environments in the lateral dimension could reflect the local assemblages. We demonstrated the ecological value of CPET studies to interpret the attributes of Chironomidae assemblage in river-floodplain systems of large rivers in an integrated way.     

Use of foreign genetic variability to improve the quality of wheat grain

Foreign genetic variability, which is represented by different wild-growing relatives of wheat such as Ae. umbellulata (UU, 2n = 14), Ae. cylindrica (CCDD, 2n = 28), Ae. tauschii (DD, 2n = 14), Ae. ventricosa (DDUnUn, 2n = 28), Ae. variabilis (UUSS, 2n = 28), and T. palmovae (AADD, 2n = 28) is used in interspecies crossings with the wheat cultivar T. aestivum for the purpose of transferring exotic Gli/Glu alleles into the genome of the crop. As a result, a series of new exotic Gli/Glu alleles is introgressed into the genome of wheat cultivar. An essential negative as well as positive influence of the wild exotic alleles on the baking quality indicators of the flour and the consistency of the wheat endosperm is discovered in the course of the study. The new genetic material with the improved grain quality indicators is recommended for use in wheat selection.     

Effect of entomopathogenic fungus <Emphasis Type="Italic">Metarhizium robertsii</Emphasis> on non-target organisms,water bugs (Heteroptera: Corixidae,Naucoridae, Notonectidae)

The influence of the fungus Metarhizium robertsii Bischoff, Rehner and Humber on the mortality of four water bug species, Cymatia coleoptrata (Fabricius), Sigara assimilis (Fieber), Ilyocoris cimicoides cimicoides (Linnaeus), and Notonecta reuteri Hungerford, and bloodsucking mosquito Anopheles messeae Falleroni, was investigated under various concentrations of conidia and different treatment types. We found that the mortality of adults of the water bug species was similar or higher than that of A. messeae, with C. coleoptrata and S. assimilis being more susceptible to M. robertsii than N. reuteri, I. c. cimicoides, and the mosquito A. messeae. Treatment with dry conidia at concentrations of 5 × 10⁴ and 5 × 10⁵ conidia/ml caused higher mortality of the water bug species than did treatment at the same concentrations with conidia in an aqueous suspension. In contrast, higher concentrations (5 × 10⁶ conidia/ml) led to higher mortality after treatment with the aqueous suspension, relative to treatment with dry conidia. Our studies showed that water bugs exhibited the classical development of a mycosis with hemocoel colonization, mummification, and conidia formation on cadavers directly on the surface of the water. Possible changes in invertebrate communities in aquatic ecosystems after treatment with Metarhizium are discussed.     

New Weevils (Coleoptera,Curculionoidea) from the Eocene of the Green River,United States: Part 1

New genera, Pseudochirotenon gen. nov. (with the type species P. eocaenicus sp. nov.), Archaeoheilus gen. nov. (type species A. scudderi sp. nov.), Primocentron gen. nov. (type species P. wickhami sp. nov.), and Pseudophaops gen. nov. (type species Otiorhynchus perditus Scudder, 1876), and new species, Pseudochirotenon eocaenicus sp. nov., Perapion rasnitsyni sp. nov., Archaeoheilus scudderi sp. nov., A. ovalis sp. nov., Primocentron wickhami sp. nov., and Eudiagogus vossi sp. nov., from the Early–Middle Eocene of the Green River are described. New combinations of names (Apionion evestigatum (Scudder, 1893), comb. nov., Archaeoheilus packardii (Scudder, 1893), comb. nov., A. provectus (Scudder, 1876), comb. nov., A. deleticius (Scudder, 1893), comb. nov., A. lacoei (Scudder, 1893), comb. nov., Pseudophaops perditus (Scudder, 1876), comb. nov.) are established. The first fossil records of the tribe Ecelonerini from the New World and genus Perapion from the Green River Formation are provided.     

Analysis of 5S rDNA changes in synthetic allopolyploids <Emphasis Type="Italic">Triticum</Emphasis> × <Emphasis Type="Italic">Aegilops</Emphasis>

Changes of 5S rDNA at the early stage of allopolyploidization were investigated in three synthetic allopolyploids: Aegilops sharonensis × Ae. umbellulata (2n = 28), Triticum urartu × Ae. tauschii (2n = 28), and T. dicoccoides × Ae. tauschii (2n = 42). Fluorescent in situ hybridization (FISH) revealed quantitative changes affecting separate loci of one of the parental genomes in S₃ plants of each hybrid combination. Southern hybridization with genomic DNA of the allopolyploid T. urartu × Ae. tauschii (TMU38 × TQ27) revealed a lower intensity of signals from Ae. tauschii fragments compared with those derived from T. urartu. This confirmed the signal reduction revealed for chromosome 1D of this hybrid by FISH. Neither Southern hybridization nor PCR testing of 5–15 plants of the S₂-S₃ generations revealed an appearance of new 5S rDNA fragments or a complete disappearance of parental fragments from the allopolyploids under study. No changes were found by aligning nine 5S rDNA sequences of the allopolyploid TMU38 × TQ27 with corresponding sequences of the parental species. The similarity between one of the synthetic allopolyploids examined and a natural allopolyploid with the same genome composition points to an early formation of the 5S rDNA organization unique for each allopolyploid.     

Macrophytes as dispersal vectors of zooplankton resting stages in a subtropical riverine floodplain

The resting stages of freshwater zooplankton constitute a special mechanism for passive dispersal, often displaying a variety of adaptations so as to ease transport. In floodplain systems, macrophytes are one of the most representative biotic groups showing interactions with the zooplankton community. The annual fluctuations in the hydrometric level of the Paraná River favour the displacement of this aquatic vegetation in floodplain environments. This paper hypothesizes that the roots and submerged portions of different macrophytes contain zooplankton resting stages which are able to hatch when environmental conditions are favourable. In turn, this contributes to the dispersal of zooplankton by plants when they are displaced by the flood pulse. Six macrophyte species were sampled (Eichhornia crassipes, Azolla filiculoides, Limnobium spongia, Pistia stratiotes, Eichhornia azurea and Nymphoides indica) from lakes within the Paraná River floodplain. Roots and submerged portions of vegetation were stored (90 days) at 4 °C then incubated at 25 °C for 90 days. Hatchling emergence was recorded at 2-day intervals during this period. In total, 70 zooplankton taxa were recorded in all macrophyte samples; rotifers were the most representative group (69%) followed by cladocerans (28%) and copepods (3%). The roots and submerged parts of aquatic vegetation house viable zooplankton resting stages. This phenomenon allows the dispersal of resting stages and therefore colonization of new habitats during the displacement of macrophyte species.     

Genetic mapping of a novel recessive allele for non-glaucousness in wild diploid wheat <Emphasis Type="Italic">Aegilops tauschii</Emphasis>: implications for the evolution of common wheat

Cuticular wax on the aerial surface of plants has a protective function against many environmental stresses. The bluish–whitish appearance of wheat leaves and stems is called glaucousness. Most modern cultivars of polyploid wheat species exhibit the glaucous phenotype, while in a wild wheat progenitor, Ae. tauschii, both glaucous and non-glaucous accessions exist. Iw2, a wax inhibitor locus on the short arm of chromosome 2D, is the main contributor to this phenotypic variation in Ae. tauschii, and the glaucous/non-glaucous phenotype of Ae. tauschii is usually inherited by synthetic hexaploid wheat. However, a few synthetic lines show the glaucous phenotype although the parental Ae. tauschii accessions are non-glaucous. Molecular marker genotypes indicate that the exceptional non-glaucous Ae. tauschii accessions share the same genotype in the Iw2 chromosomal region as glaucous accessions, suggesting that these accessions have a different causal locus for their phenotype. This locus was assigned to the long arm of chromosome 3D using an F₂ mapping population and designated W4, a novel glaucous locus in Ae. tauschii. The dominant W4 allele confers glaucousness, consistent with phenotypic observation of Ae. tauschii accessions and the derived synthetic lines. These results implied that glaucous accessions of Ae. tauschii with the W2W2iw2iw2W4W4 genotype could have been the D-genome donor of common wheat.     

Incidence of Keratinophilic Fungi from the Selected Soils of Kaziranga National Park,Assam (India)

Seventy-eight soil samples were collected from the various locations in the vicinity of Kaziranga National Park (Assam), India, during April to October 2009 and screened for the presence of keratinophilic fungi using the hair baiting techniques for isolation. Thirty-nine isolates were recovered and identified by recognition of their macro- and micromorphological features. Their identification was also confirmed by the BLAST search of sequences of the ITS1-5.8S-ITS2 rDNA region against the NCBI/GenBank data and compared with deposited sequences for identification purpose. Eleven species related to seven genera were recorded viz. Aphanoascus durus (1.28%), Arthroderma tuberculatum (3.84%), Arthroderma corniculatum (1.28%), Chrysosporium indicum (16.66%), C. tropicum (3.84%), Ctenomyces serratus (5.12%), Keratinophyton punsolae (1.28%), Microsporum appendiculatum (1.28%), Microsporum gypseum complex (11.53%), Trichophyton mentagrophytes (11.28%) and T. terrestre (2.56%).     

Laboratory and simulated-field bioassays for assessing mixed cultures of <Emphasis Type="Italic">Lysinibacillus sphaericus</Emphasis> against <Emphasis Type="Italic">Aedes aegypti</Emphasis> (Diptera: Culicidae) larvae resistant to temephos

Aedes aegypti (L.) is the main vector of tropical diseases such as dengue, chikungunya and Zika. Due to the overuse of insecticides, Ae. aegypti resistant populations have increased. Biological control with Lysinibacillus sphaericus (Ahmed) has been used against Culex sp. and Anopheles sp. Although Ae. aegypti is refractory to the binary toxin of L. sphaericus spores, vegetative cells have been shown to be effective against Ae. aegypti larvae. In this work, the effect of L. sphaericus vegetative cells on Ae. aegypti temephos-resistant larvae was assessed under lab and simulated field conditions. L. sphaericus caused about 90% mortality of insecticide-resistant Ae. aegypti larvae under simulated field conditions. Likewise, Ae. aegypti larvae were more sensitive to mixed cultures of L. sphaericus than to individual strains; then, the most effective mixed culture exhibited an LC₅₀ of 1.21 × 10⁵ CFU/mL with Rockefeller larvae and 8.04 × 10⁴ CFU/mL with field-collected larvae. Additionally, we found that mixed cultures composed of two L. sphaericus strains were more effective than a culture formed by the three strains. Our results suggest that mixed cultures comprising L. sphaericus vegetative cells could be useful for controlling temephos-resistant populations of Ae. aegypti, as evidenced by the effectiveness demonstrated under laboratory and simulated field conditions.