An extended multiplex PCR method was established to rapidly identify and classify Bacillus thuringiensis strains containing cry (crystal protein) genes toxic to species of Lepidoptera, Coleoptera, and Diptera. The technique enriches current strategies and simplifies the initial stages of large-scale screening of cry genes by pinpointing isolates that contain specific genes or unique combinations of interest with potential insecticidal activities, thus facilitating subsequent toxicity assays. Five pairs of universal primers were designed to probe the highly conserved sequences and classify most (34 of about 60) genes known in the following groups: 20 cry1, 3 cry2, 4 cry3, 2 cry4, 2 cry7, and 3 cry8 genes. The DNA of each positive strain was probed with a set of specific primers designed for 20 of these genes and for cry11A. Twenty-two distinct cry-type profiles were identified from 126 field-collected B. thuringiensis strains. Several of them were found to be different from all published profiles. Some of the field-collected strains, but none of the 16 standard strains, were positive for cry2Ac. Three standard and 38 field-collected strains were positive by universal primers but negative by specific primers for all five known genes of cry7 and cry8. These field-collected strains seem to contain a new gene or genes that seem promising for biological control of insects and management of resistance.
Susceptibility of Bacillus thuringiensis spores and toxins to the UV-B range (280--330 nm) of the solar spectrum reaching Earth's surface may be responsible for its inactivation and low persistence in nature. Spores of the mosquito larvicidal B. thuringiensis subsp. israelensis were significantly more resistant to UV-B than spores of the lepidopteran-active subsp. kurstaki. Spores of subsp. israelensis were as resistant to UV-B as spores of B. subtilis and more resistant than spores of the closely related B. cereus and another mosquito larvicidal species B. sphaericus. Sensitivity of B. thuringiensis subsp. israelensis spores to UV-B radiation depended upon their culture age; 24-h cultures, approaching maximal larvicidal activity, were still sensitive. Maximal resistance to UV-B was achieved only at 48 h.
Salinity varies considerably among temporary pools in the Dead Sea Basin, Israel. We experimentally assessed the effects of four salinity levels (0, 10, 20 and 30 g NaCl per liter) on the aquatic insect community in this basin in an artificial pool experiment. Each salinity level was randomly assigned to six pools (total ¼ 24 pools). Salinity did not affect total insect abundance but strongly affected abundance and distributions of different species, and consequently, community structure. Of 13 taxa colonizing the pools, 12 were Diptera including 10 mosquito species. Five taxa were sufficiently common to assess abundance in relation to salinity. Polypedilum nubiferum Skuse (Diptera: Chironomidae) was largely salinity intolerant being abundant only in the freshwater. Ephydra flavipes Macquart (Diptera: Ephydridae) was most abundant at the highest salinity level and was rare in freshwater. Ochlerotatus caspius Pallas (Diptera: Culicidae) abundance tended to be highest at 10 g/l and lowest at 30 g/l although the differences were not statistically significant. Anopheles multicolor Cambouliu (Diptera: Culicidae) was relatively euryhaline although numbers dropped significantly at the highest salinity. Cleon dipterum Linnaeus (Baetidae: Ephemeroptera) was also euryhaline and showed no significant differences in abundance across salinities. For the mosquito species, we also estimated survival to pupation. Survival to pupation was significantly lower for O. caspius in freshwater, but was not statistically significantly different across salinities for A. multicolor. Species diversity was highest at the two lowest salinities tested and then dropped with increasing salinity. Evenness was not significantly different across salinities. Community similarity generally decreased with increasing salinity differences though dissimilarity was greatest when comparing freshwater to other salinities. Thus, regional diversity is likely increased when there is a range of salinities among pools.
Oviposition habitat selection (OHS) is increasingly being recognized as playing a large role in explaining mosquito distributions and community assemblages. Most studies have assessed the role of single factors affecting OHS, while in nature, oviposition patterns are most likely explained by multiple, interacting biotic and abiotic factors. Determining how various factors interact to affect OHS is important for understanding metapopulation and metacommunity dynamics. We investigated the individual and interactive effects of three water salinities (0, 15 and 30 p.p.t. NaCl added) and the aquatic predator Anisops debilis Perplexa (Hemiptera: Notonectidae) on OHS and larval performance of the mosquitoes Ochlerotatus caspius Pallas and Culiseta longiareolata Macquart (Diptera: Culicidae) in outdoor-artificial-pool and laboratory experiments. C. longiareolata inhabited only freshwater pools, strongly avoided pools containing A. debilis, and larvae experienced lower survival in the presence of A. debilis. Salinity concentration interacted strongly with the predator in affecting OHS and larval survival of O. caspius; oviposition increased with increasing salinity in the absence of the predator and decreased with increasing salinity in the presence of the predator. O. caspius larval survival in predator-free pools was lowest in freshwater and highest at intermediate salinity. In predator pools, survival was highest at high salinity, where predation rate was shown to be lowest in the laboratory. Our results highlight that assessing the role of single factors in affecting mosquito distributions can be misleading. Instead, multiple factors may interact to affect oviposition patterns and larval performance.
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