[The biological control of insects of medical importance].
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Fungal strains belonging to the genera Fusarium, Paecilomyces and Trichoderma were tested in vitro in order to study their effects against Schizaphis graminum, one of the major pests of cereal crops around the world. Biological assays were performed using a solid formulation that was obtained from fungal cultures grown on rice and then finely ground (<0.2 mm). The occurrence of toxic secondary metabolites (fumonisin B1 and beauvericin) produced by these fungi was also investigated. In each experiment, three groups of aphids: 15-hour old larvae, 5-day old nymphs with wing buds and wingless morphs were treated with a suspension of a fungal formulation. Some strains belonging to the genera Fusarium and Trichoderma significantly controlled the specimens of the three groups of S. graminum. The F proliferatum strain ITEM 1407, producing a high level of fumonisin B1 in the culture (1250 microg/g), and F larvarum strain ITEM 2139 had high insecticidal activity (>60%) within 10 minutes after application. As F. larvarum ITEM 2139 did not produce metabolites toxic to mammals, it might be a good candidate as a biocontrol agent of S. graminum in the field.
The granulosis viruses and nuclear polyhedrosis viruses are being considered for use as biological insecticides for control of their insect hosts. Many of these insect species, which include some of the most serious pests of agriculture and forests, have become difficult to control because they have developed resistance to chemical insecticides. Several laboratory and field studies have demonstrated that the baculoviruses (GV and NPV) are promising alternatives to chemicals for the control of economically important insects. These viruses are highly virulent, selective, and stable, and the impact on the environment following their application is minimal. A decision concerning the application of baculoviruses to stored grain and field crops must be based upon a prudent consideration of the benefits to be obtained and the potential risks of their use. Such decisions should be made only after consideration of the physical, chemical, and biological properties of these viruses. In addition, methods must be developed for the unequivocal identification of these viruses, and their effects on nontarget species at the cellular and molecular levels must be investigated. This can best be accomplished if a sufficient body of knowledge regarding the molecular properties of these viruses and their infection process is accumulated by an extensive quantitative approach. Much of this knowledge is lacking because, prior to their consideration for use as insecticides, the baculoviruses appeared to have little medical or economic importance. As a result, interest in studying them was limited. It has become obvious that the molecular properties of these viruses must be investigated if full advantage is to be taken of using them as insect control agents, and if present and future problems concerning their use as insecticides are to be handled properly. Fundamental research on the biochemical and biophysical properties of baculoviruses has concentrated mainly on a variety of nuclear polyhedrosis viruses (Harrap, 1972a,b; Harrap et al., 1977; Summers and Smith, 1975a,b; Arif and Brown, 1975). Much of this progress can be attributed to tissue culture-host cell systems available for the NPVs. The in vitro host system(s) has allowed insect virologists to make phenomenal strides in understanding the cellular and molecular events of virus infection, and, in addition, to enter the era of biochemical sophistication in which animal virology is found at present.(ABSTRACT TRUNCATED AT 400 WORDS)
The potency of novaluron on laboratory susceptible and field strains of S. littoralis resembles that of chlorfluazuron and both compounds are about 20-fold more potent than teflubenzuron. No appreciable resistance to novaluron or chlorfluazuron was observed in a field strain of Spodoptera littoralis collected from cucumber field in the central part of Israel. On the other hand, the field strain showed a mild resistance of about 4-fold to teflubenzuron as compared to the laboratory susceptible strain. A very resistant colony of Bemisia tabaci to pyriproxyfen (1,200- to 2,000-fold) showed no appreciable cross-resistance to novaluron. Two field colonies of B. tabaci pressurized with acetamiprid or thiamethoxam for 22 generations, resulting in a 30- to 50-fold resistance to acetamiprid and thiamethoxam, has no cross-resistance to novaluron. The above results are of special interest, indicating a possible alternation between novaluron, pyriproxyfen, and neonicotinoids in insecticide-resistance management programs aiming at preventing resistance development to these novel groups of insecticides against important pests such as whitefly and lepidopteran species.
Chemical irritants useful as repellents for brown treesnakes (Boiga irregularis) were identified. Exposure to various compounds produced a range of intensities for locomotory behavior in snakes. Essential oils comprised of 10 g liter-1 solutions of cedarwood, cinnamon, sage, juniper berry, lavender and rosemary each were potent snake irritants. Brown treesnakes exposed to a 2-s burst of aerosol of these oils exhibited prolonged, violent undirected locomotory behavior. In contrast, exposure to a 10 g liter-1 concentration of ginger oil aerosol caused snakes to locomote, but in a deliberate, directed manner. We also tested specific compounds, all derivative of food and flavor ingredients. 10 g liter-1 solutions delivered as aerosols of m-anisaldehyde, trans-anethole, cineole, cinnamaldehyde, citral, ethyl phenylacetate, eugenol, geranyl acetate or methyl salicylate all acted as potent irritants for brown treesnakes. The individual ingredients were classified using cluster analysis into groups that promoted different levels of response by snakes. This study is the first to systematically investigate the irritant potential of natural products for snakes. These data will be useful in the development of practical pest management tools for snakes.
Twenty-four serovars of Bacillus thuringiensis (Bt) were screened by polymerase chain reaction to detect the presence of vegetative insecticidal protein gene (vip)-like sequences by using vip3Aa1-specific primers. vip-like gene sequences were identified in eight serovars. These genes were cloned and sequenced. The deduced amino acid sequence of the vip3Aa14 gene from Bacillus thuringiensis tolworthi showed considerable differences as compared to those of Vips reported so far. The vip3Aa14 gene from Bt tolwarthi was expressed in Escherichia coli using expression vector pET29a. The expressed Vip3Aa14 protein was found in cytosolic supernatant as well as pellet fraction, but the protein was more abundant in the cytosolic supernatant fraction. Both full-length and truncated (devoid of signal sequence) Vips were highly toxic to the larvae of Spodoptera litura and Plutella xylostella. Truncation of Vip3Aa14 protein at N-terminus did not affect its insecticidal activity.
To conduct laboratory experiments aimed at quantifying secondary acquisition of fungal conidia by western flower thrips (Frankliniella occidentalis), an efficient assay technique using Beauveria bassiana as the model fungus was developed. Various application protocols were tested and it was determined that the percent mortality did not vary among protocols. Peak mortality of second-instar nymphs, under constant exposure to conidia, occurred 5 days post-inoculation. Second-instar thrips that were exposed to conidia within 24 h of the molt to second instar were more susceptible to Beauveria bassiana than thrips exposed after times greater than 24 h post-molt. Conidia efficacy, which was monitored at 24 h intervals, did not differ significantly within 72 h. A test of the final bioassay system was conducted in a series of assays aimed at determining the LD50 of B. bassiana technical powder against second-instar western flower thrips. It was determined that B. bassiana (strain GHA) is highly effective at very low doses (LD50 of 33-66 conidia/insect).
A native collection of Bacillus thuringiensis strains was screened, once a reliable bioassay technique to assess the toxicity against the coffee berry borer (CBB) first-instar larvae was developed. A first round of bioassays with 170 strains indicated that the great majority of them showed no or very little insecticidal activity and that very few showed significant levels of toxicity. Interestingly, only those strains that had previously been associated with mosquitocidal activity were also toxic to CBB. Qualitative bioassays (using one high dose) were carried out only with those native mosquitocidal strains, corroborating their significant toxicity towards the CBB first-instar larvae. Most of these strains belong to serovar israelensis. In a second approach, strains from the Institut Pasteur type collection, whose mosquitocidal activity had been previously demonstrated, were also subjected to bioassays. Only those strains that showed a comparable protein content in their parasporal crystals to the israelensis type strain also showed high levels of toxicity towards CBB. Finally, an accurate LC(50) was estimated, using purified parasporal crystals from B. thuringiensis serovar israelensis type strain, at 219.5 ng cm(-2) of diet. All the statistical requirements for a reliable estimator were fulfilled. This is the first report of B. thuringiensis serovar israelensis being active against a coleopteran species.
Two types of Barbarea vulgaris var. arcuata, the G-type and the P-type, differed in resistance to larvae of the diamondback moth (DBM) Platella xylostella. Rosette plants of the G-type were fully resistant to the DBM when grown in a greenhouse or collected in the summer season, but leaves collected during the late fall were less resistant, as previously found for flea beetle resistance. The P-type was always susceptible. Extracts of resistant leaflets inhibited larval growth in a bioassay, and a growth-inhibiting fraction was isolated by activity-guided fractionation. A triterpenoid saponin (1) was isolated from this fraction and identified as 3-O-beta-cellobiosyloleanolic acid from spectroscopic data and analysis of hydrolysis products. The decrease in resistance of the G-type in the fall was correlated with a decrease in the level of 1, from 0.6-0.9 to < 0.2 micromol/g dry wt. Compound 1 was not detected in the susceptible P-type. We conclude that 1 is correlated with the variable resistance of B. vulgaris foliage to the DBM.
Entomopathogenic nematodes are widely used as biological insecticides for agricultural and horticultural pests. In the vast majority of cases, commercial products contain nematodes partially dehydrated on to inert solid carriers. Nematode survival in these products is generally poor, they are difficult to handle and are not suitable for use with all nematode species. We have developed a non-viscous, non-adhesive and non-toxic liquid formulation for nematode storage and transport based on neutral density colloidal silica suspensions. Survival and virulence of nematodes stored in this formulation without aeration was superior to nematodes stored in aerated quarter strength Ringer's solution.
Teretrius nigrescens is considered to be a specialized predator of Prostephanus truncatus, a serious pest of stored maize and dried cassava roots. Using a bait-bag behavioral bioassay, this investigation found T nigrescens to be strongly arrested by dust and frass produced by P. truncatus on maize, whereas responses to maize flour were weak. Attempts to increase the arrestiveness of flour by altering its physical properties (coarseness or particle size range) were unsuccessful. The arrestive property of dust/frass did not degrade with long-term storage nor did it volatilize or degrade with oven baking up to 150 degrees C. However, extraction with methanol and reapplication was successful in transferring the arrestive property from the dust/frass onto maize flour. The prey dust/frass also induced more oviposition than did maize flour, as judged by production of F1 offspring. The results provide strong evidence for the existence of a high boiling point compound(s) in P. truncatus dust/frass that acts as a contact kairomone for T nigrescens. This, or another compound(s) also seems to act as an oviposition stimulant for female T. nigrescens.
Lagenidium giganteum was cultivated on solid media in the absence of free water and evaluated for efficacy against second-instar Aedes aegypti mosquito larvae in 100-ml bioassays. Bioassay variables included level of media addition, CO(2) evolution rate (CER) and cell density. Logistic regression was performed on bioassay infection observations to determine if the tested variables were correlated to the probability of attaining at least 80% infection. Both CER (p=0.003) and number of cells (p=0.017) were significantly correlated, while level of media addition was not (p=0.42). Although media addition did not correlate with efficacy, media levels greater than 2 g/l reduced water clarity and infection. Media from cultures younger than 3 days performed poorly under all conditions tested.
Strategies for delaying pest resistance to genetically modified crops that produce Bacillus thuringiensis (Bt) toxins are based primarily on theoretical models. One key assumption of such models is that genes conferring resistance are rare. Previous estimates for lepidopteran pests targeted by Bt crops seem to meet this assumption. We report here that the estimated frequency of a recessive allele conferring resistance to Bt toxin Cry1Ac was 0.16 (95% confidence interval = 0.05-0.26) in strains of pink bollworm (Pectinophora gossypiella) derived from 10 Arizona cotton fields during 1997. Unexpectedly, the estimated resistance allele frequency did not increase from 1997 to 1999 and Bt cotton remained extremely effective against pink bollworm. These results demonstrate that the assumptions and predictions of resistance management models must be reexamined.
Laboratory bioassays were conducted to evaluate the effect of the diatomaceous earth (DE) formulation SilicoSec (Biofa GmbH, Münsingen, Germany), against two stored-product mite species, Tyrophagus putrescentiae (Shrank) and the predator Cheyletus malaccensis Oudemans. For this purpose, DE was applied in wheat, oat, rye, and maize, at the dose rates 0, 0.5, 1, and 2 g/kg grain. The mortality of the exposed mites was assessed after 24 h, 48 h, 7 d, and 14 d of exposure in the treated substrate. After this interval, the treated grains were checked for oviposition or progeny. The tests were conducted at 80% RH and at two temperatures, 20 and 25 degrees C. Generally, for both species, mortality was higher at 25 degrees C than at 20 degrees C. For T. putrescentiae, at both temperatures, the mortality in grains treated with the highest DE rate was 100% after only 24 h of exposure, with the exception of maize at 20 degrees C, where mortality was 91.7%. The mortality of C. malaccensis after 24 h of exposure to the treated grains, in the absence of prey, did not exceed 29% at any of the temperature- grain-dose combinations, whereas no mites were dead in rye and maize treated with 0.5 and 1 g of DE. Even after 14 d of exposure at the highest DE rate, mite mortality did not reach 100%. The presence of T. putrescentiae individuals as prey in the treated substrate enhanced C. malaccensis survival. Hence, after 14 d of exposure, the mortality of C. malaccensis, in wheat, oat, rye, and maize treated with the highest DE rate was 51.7, 59.7, 70, and 36.9, respectively. No progeny production was recorded in the treated substrate for T. putrescentiae; in contrast, oviposition and F1 progeny were recorded for C. malaccensis. Our results suggest that the use of C. malaccensis with low doses of DE may be an appealing integrated pest management (IPM) approach against T. putrescentiae, and probably against other stored-grain mite species.
The susceptibility of three strains of Aedes aegypti (L.) to four strains of Ascogregarina culicis were studied under laboratory conditions. The parasite was found to be pathogenic to the mosquito; although pathogenicity varied among geographical strains of A. culicis and susceptibility varied among geographical strains of Ae. aegypti. In addition to affecting the survival of the mosquito larvae, infection with the parasite was found to shorten larval development time. Selected strains of A. culicis may be useful as a biological control agent against Ae. aegypti.
The influence of environmental factors during isolation on the composition of potential biocontrol isolates is largely unknown. Bacterial isolates that efficiently suppressed wheat seedling blight caused by Fusarium culmorum were found by isolating psychrotrophic, root-associated bacteria and by screening them in a bioassay that mimicked field conditions. The impact of individual isolation factors on the disease-suppressive index (DSI) of almost 600 isolates was analyzed. The bacteria originated from 135 samples from 62 sites in Sweden and Switzerland. The isolation factors that increased the probability of finding isolates with high DSIs were sampling from arable land, Swiss origin of samples, and origination of isolates from plants belonging to the family Brassicaceae. The colony morphology of the isolates was characterized and compared to DSIs, which led to identification of a uniform morphological group containing 57 highly disease-suppressive isolates. Isolates in this group were identified as Pseudomonas sp.; they were fluorescent on King's medium B and had characteristic crystalline structures in their colonies. These isolates were morphologically similar to seven strains that had previously been selected for suppression of barley net blotch caused by Drechslera teres. Members of this morphological group grow at 1.5 degrees C and produce an antifungal polyketide (2,3-deepoxy-2,3-didehydrorhizoxin [DDR]). They have similar two-dimensional polyacrylamide gel electrophoresis protein profiles, phenotypic characteristics, and in vitro inhibition spectra of pathogens. In summary, in this paper we describe some isolation factors that are important for obtaining disease-suppressive bacteria in our system, and we describe a novel group of biocontrol pseudomonads.
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