Joint action of the entomopathogenic fungus Isaria fumosorosea and four chemical insecticides against the whitefly Bemisia tabaci

The strain IfB01 of Isaria fumosorosea (Paecilomyces fumosoroseus) is a promising myco-insecticide candidate for control of Bemisia tabaci. Joint action of strain IfB01 and four chemical insecticides [spirotetramat (Spi), acetamiprid (Ace), imidacloprid (Imi) and thiamethoxam (Thi)] against the whitefly B. tabaci was evaluated using the cooperative virulence index (c.f.). Substantial synergistic actions were found in the mixtures of IfB01 conidia with Spi, Imi and Thi 2–4 days after treatment. The greatest synergism was recorded in the mixture of IfB01 (2.5 × 10⁶ conidia/L) and Imi (12.5 mg/L), which had a c.f. value of 184 at day 3 after treatment. Furthermore, the shorter LT₅₀ values recorded in mixtures indicated that effectiveness improved when the conidia were applied with the chemical insecticides. However, no substantial synergisms were recorded with Ace and from day 5 onwards.     

烟粉虱的飞行行为与害虫综合治理策略

烟粉虱具有较强的飞行潜力,飞行高度可以超过150 m,在田间的扩散距离最远可以超过150 km,但在食源丰富地区,较少作远距离的扩散,绝大部分飞行高度在距地面0.5 m左右.烟粉虱具有搜索飞行和迁飞飞行特性,它是寻找适宜寄主和扩大生境的重要方式.烟粉虱不具备“卵子发生与飞行共轭”的典型特征.可见光、温湿度、寄主质量和风等是影响烟粉虱飞行行为的重要生态因子.本文对烟粉虱的飞行能力、飞行生理学和影响飞行的生态因子进行了综述,对非露地越冬区利用烟粉虱的飞行特性实施IPM策略进行了探讨.     

Compatibility of the entomopathogenic fungus Lecanicillium muscarium and insecticides for eradication of sweetpotato whitefly, Bemisia tabaci

The compatibility of the entomopathogenic fungus Lecanicillium muscarium and chemical insecticides used to control the second instar stages of the sweetpotato whitefly, Bemisia tabaci, was investigated. The effect on spore germination of direct exposure for 24 h to the insecticides imidacloprid, buprofezin, teflubenzuron and nicotine was determined. Only exposure to buprofezin was followed by acceptable spore germination. However, all chemicals significantly reduced spore germination when compared to a water control. Infectivity of L. muscarium in the presence of dry residues of buprofezin, teflubenzuron and nicotine (imidacloprid is a systemic pesticide) on foliage were also investigated. No significant detrimental effects on the level of control of B. tabaci was recorded when compared with fungi applied to residue free foliage on either tomato or verbena plants. Fungi in combination with imidacloprid gave higher B. tabaci mortality on verbena foliage compared to either teflubenzuron or nicotine and fungi combinations. Use of these chemical insecticides with L. muscarium in integrated control programmes for B. tabaci is discussed.     

Machine vision algorithm for whiteflies (Bemisia tabaci Genn.) scouting under greenhouse environment

One of the main problems in greenhouse crop production is the presence of pests. Detection and classification of insects are priorities in integrated pest management (IPM). This document describes a machine vision system able to detect whiteflies (Bemisia tabaci Genn.) in a greenhouse by sensing their presence using hunting traps. The extracted features corresponding to the eccentricity and area of the whiteflies projections allow to establish differences among pests and other insects on both the trap surfaces and dust generated artefacts. Because of whiteflies geometrical characteristics, it was possible to design an efficient (related to manual counting) machine vision algorithm to scout and count units of this pest within a greenhouse environment. These algorithm results show high correlation indexes for both, sticky screens (R² = 0.97) and plant leaf situations (R² = 1.0). The machine vision algorithm reduces the scouting time and the associated human error for IPM‐related activities.     

Protocol for semi‐automatic identification of whiteflies Bemisia tabaci and Trialeurodes vaporariorum on yellow sticky traps

Yellow sticky traps (YSTs) are commonly used in greenhouse crops to monitor flying pest species. Whiteflies like Trialeurodes vaporariorum (Westwood) (Hemiptera: Aleyrodidae) and Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) are typically monitored using YSTs in tomato and sweet pepper crops. By counting the whiteflies on a YST, growers get an idea of the pests density in space and time in the greenhouse and can take pest control measurements accordingly. The downside is that manual counting of whiteflies on a YST is very time‐consuming and thus costly. A protocol to semi‐automate counting and identification of whiteflies on YSTs using image analysis software was developed to speed up the monitoring process. Bemisia tabaci is on average smaller than T. vaporariorum and by discriminating by size based on the amount of pixels in digital images, ratios of both species in a mixed population on YSTs could be estimated accurately. At low densities, the countings of different YSTs should be pooled till a 200 density threshold is reached in order to get accurate ratio estimates of both species. This study provides a protocol to reliably count and identify whiteflies semi‐automatically on standardized pictures. More research is required to develop alternative techniques to make standardized pictures in the field (e.g., with smartphone).     

Proteomics as a tool for tapping potential of entomopathogens as microbial insecticides

Biopesticides are collective pest control harnessing the knowledge of the target pest and its natural enemies that minimize the risks of synthetic pesticides. A subset of biopesticides; bioinsecticides, are specifically used in controlling insect pests. Entomopathogens (EPMs) are micro‐organisms sought after as subject for bioinsecticide development. However, lack of understanding of EPM mechanism of toxicity and pathogenicity slowed the progress of bioinsecticide development. Proteomics is a useful tool in elucidating the interaction of entomopathogenic fungi, entomopathogenic bacteria, and entomopathogenic virus with their target host. Collectively, proteomics shed light onto insect host response to EPM infection, mechanism of action of EPM’s toxic proteins and secondary metabolites besides characterizing secreted and membrane‐bound proteins of EPM that more precisely describe relevant proteins for host recognition and mediating pathogenesis. However, proteomics requires optimized protein extraction methods to maximize the number of proteins for analysis and availability of organism's genome for a more precise protein identification.     

Effects of three insecticides on adult bionomics of the parasitoid Encarsia formosa

Pesticides can negatively affect many life history traits of natural enemies. In this study, we studied the effects of three insecticides with different modes of action on the bionomics of Encarsia formosa, a parasitoid of the greenhouse whitefly, Trialeurodes vaporariorum (Westwood). Buprofezin (800 mg (a.i.)/L) and pyriproxyfen (50 mg (a.i.)/L) were selected among IGRs, while fenpropathrin (250 mg (a.i.)/L) was selected from the Pyrotheroids. Adults of E. formosa were treated via exposure to residues of insecticides on leaf discs. Our results revealed that buprofezin and pyriproxyfen did not affect longevity and fecundity, while fenpropathrin significantly reduced the longevity and fecundity of treated wasps. Results of logistic regression revealed that control, buprofezin- and pryproxyphen-treated E. formosa was a function of host density and followed a type II functional response. In contrast, E. formosa treated by fenpropathrin showed a type III functional response. Estimated attack rate for buprofezin and pyriproxyfen did not differ significantly from the control, whereas fenpropathrin-treated wasps showed a lower attack rate than the control. According to the obtained results, handling time of fenpropathrin-treated wasps was significantly higher (4.57 ± 0.5) than the control (2.83 ± 0.35). Our results showed that the maximum parasitism rate achieved by control wasps was 8.39, while the rate for buprofezin-, pyriproxyfen- and fenpropathrin-treated wasps was 6.99, 7.69 and 5.25, respectively. Overall, results suggest that buprofezin and pyriproxyfen can be used in an integrated pest management programme or biological control programme without destructive effects on the efficiency of this natural enemy in green houses.     

Further compatibility tests of the entomopathogenic fungus Lecanicillium muscarium with conventional insecticide products for control of sweetpotato whitefly, Bemisia tabaci on poinsettia plants

The effect on spore germination of the entomopathogenic fungus Lecanicillium muscarium following direct exposure for 24 h to the insecticides Majestik, Spray Oil, Agri-50E, Savona and Oberon for the control of both egg and second instar stages of the sweetpotato whitefly, Bemisia tabaci , was determined. Exposure to both Agri-50E and Oberon was followed by acceptable spore germination. Infectivity rates of L. muscarium on poinsettia foliage in the presence of dry residues of the insecticides were also investigated. No significant detrimental effects on the levels of control of B. tabaci were recorded compared with fungus applied to residue-free foliage. Sequential application of the chemicals Savona, Spray Oil and Majestik with the fungus all produced mortalities of second instar B. tabaci above 90%. Incorporation of these chemicals with L. muscarium into integrated control programs for B. tabaci is discussed.     

The integrated use of chemical insecticides and the entomopathogenic nematode,Steinernema carpocapsae (Nematoda: Steinernematidae), for the control of sweetpotato whitefly,Bemisia tabaci (Hemiptera: Aleyrodidae)

The integration of chemical insecticides and infective juveniles of the entomopathogenic nematode Steinernema carpocapsae (Wesier) (Nematoda: Steinernematidae), to control second instars of the sweetpotato whitefly, Bemisia tabaci Gennadius (Hemiptera: Aleyrodidae) was investigated. Using a sand bioassay, the effects of direct exposure of S. carpocapsae for 24 h to field rate dilutions of four insecticides (spiromesifen, thiacloprid, imidacloprid and pymetrozine) on infectivity to Galleria mellonella larvae were tested. Although all chemicals tested, except spiromesifen, produced acceptable nematode infectivity rates, they were all significantly less than the water control. The effect of insecticide treatment (dry residues of spiromesifen, thiacloprid and pymetrozine and soil drench of imidacloprid) on the efficacy of the nematode against B. tabaci was also investigated. Nematodes in combination with thiacloprid and spiromesifen gave higher B. tabaci mortality (86.5% and 94.3% respectively) compared to using nematodes alone (75.2%) on tomato plants. There was no significant difference in B. tabaci mortality when using the chemicals imidacloprid, pymetrozine and spiromesifen in conjunction with nematodes compared to using the chemicals alone. However, using thiacloprid in combination with the nematodes produced significantly higher B. tabaci mortality than using the chemical alone. The integration of S. carpocapsae and these chemical agents into current integrated pest management programmes for the control of B. tabaci is discussed.     

Indirect effect of elevated CO2 concentration on Bemisia tabaci MEAM1 feeding on Bt soybean plants

The development of herbivore insects is influenced by the quality of their host plants. Elevated CO₂ alters plant metabolism, which may change the nutritional quality of the plant, modifying the life history and feeding behaviour of herbivore insects. Understanding how insect pests respond to increasing CO₂ concentration is essential for predicting the impact of the pest on food security. In this study, we investigated the effects of elevated CO₂ (eCO₂) on the life history and feeding behaviour of the MEAM1 species of Bemisia tabaci on a Bt soybean cultivar. We found that eCO₂ increased the egg to adult development time and reduced the reproductive responses (fecundity and fertility) of B. tabaci. The whitefly B. tabaci that fed on the soybean plants grown under eCO₂ conditions was negatively influenced by several traits related to the host plant resistance, such as the time spent on phloem sap ingestion. Furthermore, we evaluated the changes in the C:N concentration and plant morphology of the Bt plants. The biomass (weight of leaves and stems) of the Bt soybean plants grown under eCO₂ conditions was significantly increased, and the elevated C:N ratio in the phenological stage V6 (i.e. when the plants had six trifoliate leaves developed) was the most pronounced difference in the Bt soybean plants subjected to eCO₂ treatment. Taken together, our results indicate that Bt plants cultivated under eCO₂ inhibit B. tabaci feeding, which can reduce whitefly infestations of the soybean fields.     

Establishment of a silverleaf whitefly, Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) population in western Sydney

A field survey in western Sydney found populations of Bemisia tabaci biotype B (silverleaf whitefly) that have the potential to damage greenhouse vegetable crops prevalent in the region. Adult whiteflies were collected from five farms at 2-week intervals and the proportion of silverleaf whitefly in the overall whitefly population was determined using polyacrylamide gel electrophoresis. High levels of silverleaf whitefly were detected at most sampling dates on two of the farms surveyed and a third farm exhibited lower but relatively continuous silverleaf whitefly abundance. The presence of permanent silverleaf whitefly populations must be considered by the greenhouse industry when formulating whitefly management plans for western Sydney.     

Effect of Ifchit1 gene of Isaria fumosorosea on mortality,oviposition and oxidase activities of Bemisia tabaci

Isaria fumosorosea is one of important entomopathogenic fungi showed a good potential in controlling Bemisia tabaci. The effects of I. fumosorosea ⊿Ifchit1 mutant (Ifchit1 gene deletion mutant) on the mortality, oviposition, and host immunological response of B. tabaci, on Brassica campestris L. plant, were evaluated under laboratory conditions. The wild-type fungal strain infection significantly increased insect mortality and reduced the oviposition effeciency of B. tabaci, whereas the ⊿Ifchit1 mutant was much less effective, resulting in higher survival and ovipositing of B. tabaci. The activities of four insect enzymes were examined during a time course of fungal infection. Insect phenoloxidase, perioxidase, and catalase activities were decreased in whiteflies treated with the wild type and mutant I. fumosorosea strain at 12–36?h post treatment. However, these enzyme activities increased in fungal-treated whiteflies as compared to controls between 36 and 60?h post-infection, reaching peak values. Superoxide dismutase activity in fungal-treated whiteflies was higher than that in controls during the entire experimental time course examined. The overall enzyme activity profiles in ⊿Ifchit1 mutant-treated whiteflies were significantly different from wild-type strain treatments. Our results showed that loss of the Ifchit1 gene in I. fumosorosea affects whitefly mortality, ovipositioning and various antioxidant enzyme activities, providing new insights into the role of chitinases in I. fumosorosea-insect host–pathogen interactions.     

Common pitfalls when testing additivity of treatment mixtures with chi‐square analyses

Studying interactions of multiple pesticides applied simultaneously in a mixture is a common task in phytopathology. Statistical methods are employed to test whether the treatment components influence each other's efficacy in a promotive or inhibitory way (synergistic or antagonistic interaction) or rather act independent of one another (additivity). The trouble is that widely used procedures based on chi‐square tests are often seriously flawed, either because people apply them in a preposterous way or because the method simply does not fit the problem at hand. Browsing recent volumes of entomological journals, we found that numerous researchers have (in all likelihood unwittingly) analysed their data as if they had had a sample size of 100 or, equally bad, a sample size of one! We show how to avoid such poor practices and further argue that chi‐square testing is, even if applied correctly (meaning that no technical errors are made), a limited purpose tool for assessing treatment interactions.     

Genetics and evolution of resistance to insecticides

The evolution of resistance to insecticides has become a serious problem world-wide. It is important to identify patterns of insecticide use whereby insecticides can be used in integrated pest management programmes to help control insect numbers, but in such a manner that the evolution of resistance to insecticides will be retarded. The principal mechanisms of insecticide action and of resistance to these are reviewed. Some generalizations that can be made about the evolution of resistance are examined. In general, to control resistance it appears better to use an intense dose of non-persistent pesticides over a circumscribed area. Some features of the problem where population genetics and evolutionary theory might contribute to controlling resistance are discussed.     

Delayed development of the whitefly (Bemisia tabaci) and increased parasitism by Encarsia bimaculata in response to sublethal doses of piperonyl butoxide

Abstract Effects of sublethal piperonyl butoxide (PB) on parasitization of Bemisia tabaci (Gennadius ) (Hemiptera: Aleyrodidae) by Encarsia bimaculata Heraty et Polaszek (Hymenoptera: Aphelinidae) were evaluated both in cage and greenhouse experiments. When first, second and third instar B. tabaci nymphs were treated with PB, all but the first instar were significantly prolonged. Data indicated that sublethal PB could improve E. bimaculata parasitism rates without influencing parasitoid eclosion rates. Prolonged development increased rates of parasitism by E. bimaculata, from 17.6% to 24.7% in cages, presumably by increasing the duration of host exposure. Sublethal PB combined with E. bimaculata as an integrated approach to control B. tabaci was evaluated using life table parameters under greenhouse conditions. Indices of population trend (I) calculated from life tables were estimated at 4.6 for B. tabaci exposed to PB and parasitoids compared to 14.1 with parasitoids alone and 23.5 in untreated controls. The results showed that after PB was sprayed and parasitoids introduced, development of B. tabaci was delayed and the peak of each stage was postponed. The older nymphal stage had highest mortality, primarily due to mortality caused by parasitism by E. bimaculata.     

Control of the rootknot nematode meloidogyne javanica by Bacillus cereus

Exposure of Meloidogyne javanica second‐stage juveniles to the bacterium Bacillus cereus in soil inhibited the penetration of the juvenile nematodes into tomato roots. Culture filtrate of the bacterium grown on nutrient broth and tryptic soy broth revealed nematocidal activity on M. javanica juveniles and eggs. Loss of the nematocidal activity of the media by lowering pH, boiling or dialysis raised the possibility that the active ingredient in the culture filtrate was ammonia, released during the breakdown process of peptides in the media by bacterial activity. Free ammonia (NH₃) concentrations in the nutrient broth and tryptic soy broth culture filtrates measured after 48 h were 140 and 190 µg ml^?1 respectively. Exposure of second‐stage juveniles to 9.3 µg ml^?1 ammonia for 40 h in vitro was lethal to 95% of the nematode population. In a nitrate medium, nitrite accumulated up to 250 µg ml^?1 during the growth of the bacterium, and its culture filtrate revealed nematocidal activity. The nematocidal activity of the bacterium increased when the bacterium was applied with various proteinaceous supplements to soil. Soil treated with the bacteria and peptone showed an earlier nematocidal activity than either the bacteria or peptone applied alone, and also had a higher level of ammonia than the individual treatments. However, the level of ammonia was lower than the lethal level for second‐stage juveniles recorded in vitro. The nematocidal activity exhibited by the bacterium‐proteinaceous amendment combination is not fully understood; the ammonia released during protein degradation by the bacterium may contribute significantly to the recorded nematocidal activity.     

双斑恩蚜小蜂寄生对烟粉虱体内保幼激素和20-羟基蜕皮酮含量的影响

用反相高效液相色谱法（RP-HPLC）测定了烟粉虱Bemisia tabaci体内保幼激素和20-羟基蜕皮酮的滴度。结果显示,被双斑恩蚜小蜂Encarsia bimaculata寄生的烟粉虱高龄若虫体内的保幼激素滴度显著高于（P<0.05）对照即未被寄生的高龄若虫,20-羟基蜕皮酮（20-E）滴度显著低于（P<0.05）对照,与未被寄生的低龄若虫滴度差异不显著,说明双斑恩蚜小蜂寄生后可以将烟粉虱高龄若虫的激素含量保持在低龄若虫的水平,从而调节烟粉虱高龄若虫的生长发育。烟粉虱伪蛹和成虫与对照体内保幼激素含量,烟粉虱伪蛹与对照之间20-E含量差异均不显著（P>0.05）。     

Compatibility and interaction between Bacillus thuringiensis and certain insecticides: perspective in management of Tuta absoluta (Lepidoptera: Gelechiidae)

Bacillus thuringiensis var. kurstaki (Berliner) (Bt) has been suggested as a biological control agent for Tuta absoluta (Meyrick). The objective of this study was to determine the interaction between abamectin, azadirachtin, indoxacarb, chlorantraniliprole, dichlorvos and metaflumizone with Bt. Effect of recommended doses of the chemical insecticides on colonisation of Bt was also investigated in culture medium. Except for metaflumizone, none of the chemicals tested reduced the colonisation of Bt compared with control. Interaction between Bt and the chemical insecticides on 2nd-instar larvae was also assessed. In interaction tests, Bt was applied at LC₅₀ level, 0, 12, 24 or 36 h after treating the larvae with LC₁₀ or LC₂₅ of the chemical insecticides. An antagonistic effect was observed in all treatments where Bt was applied immediately after the chemical insecticide. Also, antagonism was observed when treatment with Bt was done 12 h after azadirachtin and metaflumizone applications. Applying Bt 12 and 24 h after treatment with LC₂₅ of chlorantraniliprole, dichlorvos and abamectin resulted in synergism. But, synergism with LC₁₀ of dichlorvos and abamectin was observed only after 12 h. Additive effect was observed in the rest of the time and concentration combinations. Based on the results obtained, simultaneous use of the chemical insecticides tested and Bt is not recommended for T. absoluta control; and an appropriate time interval should be taken into consideration accordingly.     

Microbial control of cotton pests: use of the naturally occurring entomopathogenic fungus Aspergillus sp. (BC 639) in the management of Bemisia tabaci (Genn.) (Hemiptera: Aleyrodidae) and beneficial insects on transgenic cotton crops

The commercial adoption of transgenic Bacillus thuringiensis (Bt) cotton (Bollgard II®) reduced the use of insecticides to control Helicoverpa spp. However, the ineffectiveness of the Bt toxin against sucking pests such as silverleaf whiteflies (Bemisia tabaci) resulted in a marked increase in B. tabaci populations and in the use of insecticides to control this pest. The effect of the entomopathogenic fungus Aspergillus sp. BC 639 on B. tabaci and beneficial insects (predominantly predatory insects) was studied in commercial cotton field trials. The results showed that oil-based extracts of the entomopathogenic fungus BC 639 control the number of B. tabaci adults and nymphs in commercial transgenic cotton crops. The BC 639 fungus caused 60.0%, 67.2%, and 68.8% mortality in adults, and 54.6%, 62.3%, and 51.7% in nymphs at 7, 14, and 21 days after treatment, respectively, relative to the unsprayed controls. The effect of BC 639 at concentrations of 125, 250, and 500?ml/ha on low-density B. tabaci (～10 nymphs/leaf) did not differ significantly from that of the commercial insecticide (pyriproxifen). However, at higher densities (>50 nymphs per leaf), low concentrations of BC 639 (125 and 250?ml/ha) were not as effective as 500?ml/ha BC 639 in successfully controlling the pest. A simple graphic analysis suggested that the more B. tabaci nymphs per leaf, the fewer adults per leaf, and that once the number of nymphs increased to ～70 per leaf, a negative feedback regulatory effect reduced the survivorship of the nymphs and adults and/or caused the emigration of the adults from the contaminated leaves in search of new resources. Therefore, the ability of BC 639 to control B. tabaci adults and nymphs with minimal effects on predatory insects indicates its potential utility in supplementing integrated pest management programmes for cotton crops.     

The effect of simulated seasonal temperatures on Metarhizium brunneum-associated mortality in Agriotes spp. click beetles,and a degree-day infection model

Entomopathogens tend to have a slow speed of kill when used for targeting agricultural insect pests. Relating temperature as a driver of this speed is important to predict pest mortality, and extending this to a degree-day infection model has rarely been studied. Many species of wireworms (Coleoptera: Elateridae), the larvae of click beetles, are subterranean and generalist agricultural pests that can be difficult to control with pesticides. Targeting adult beetles, however, may be an effective method to reduce larval recruitment. Metarhizium brunneum Petch (Hypocreales), an entomopathogenic fungus, kills click beetles but the mortality rate and speed of kill are expected to vary according to temperature. Using a thermal gradient plate to simulate daily oscillating temperatures in Agassiz, British Columbia, Canada, for April, May, and June, the effectiveness of M. brunneum strains LRC112 and F52 in causing mortality to Agriotes obscurus (L.) and Agriotes lineatus (L.) click beetles was studied in the laboratory. Mortality was fastest in beetles exposed to June temperatures and slowest in those exposed to April temperatures, with differences among beetle species × M. brunneum strain combinations. Higher temperatures resulted in more rapid mycelial outgrowth and conidiation in beetle cadavers, with only A. obscurus infected with M. brunneum LRC112 attaining near 100% conidiation. The number of degree days required to kill 50% of the beetles (LDD₅₀) was least for A. obscurus infected with M. brunneum LRC112 (176) followed by A. obscurus × M. brunneum F52 (212), A. lineatus × M. brunneum LRC112 (215), and A. lineatus × M. brunneum F52 (292). Hypothetical calculations showed that M. brunneum exposure earlier in the season resulted in a longer time to kill 50% of the beetles (LT₅₀) but the earliest LT₅₀ calendar date. Later M. brunneum exposure dates resulted in lower LT₅₀'s, but later LT₅₀ dates. This conceptual work demonstrates that daily temperature oscillations, seasonality, and degree days must be considered to predict the efficacy and speed of kill of different fungal entomopathogen strains when targeting different click beetle species.