Efficacy of Silwet L-77 against several arthropod pests of table grape

Silwet L-77, an organosilicone surfactant, was applied to several arthropod pests of California table grapes. Eggs of grape mealybug, Pseudococcus maritimus (Ehrhorn), and omnivorous leafroller, Platynota stultana Walsingham, were tolerant to 0.1, 0.25, and 0.5% treatment solutions; however, eggs of Pacific spider mite, Tetranychus pacificus McGregor, were highly susceptible with mortality >99.4% (0.1% Silwet L-77). Mortality of immature and adult stages of cotton aphid (Aphis gossypii Glover), Western flower thrips (Frankliniella occidentalis Pergande), and Pacific spider mite (Tetranychus pacificus McGregor) was > or = 93.8, > or = 98.5, and > or = 99.4% for 0.1, 0.25, and 0.5% Silwet L-77, respectively. Grape mealybug crawlers had 100% mortality when treated with 0.5 and 1.0% Silwet L-77 solutions; however, mortality was only 6.7% when 0.1% Silwet L-77 was applied. 'Thompson Seedless' table grapes were not damaged when treated with up to 1% Silwet L-77; however, grapes treated with the 0.5 and 1.0% solutions appeared wet after removal from cold storage because of the effect of the surfactant spreading the water condensation. Grapes dried with the normal bloom on the berries when they reached room temperature.     

Adjuvants enhance the biological control potential of an isolate of Colletotrichum gloeosporioides for biological control of sicklepod (Senna obtusifolia)

In greenhouse experiments, unrefined corn oil, Silwet L-77, and an invert emulsion were tested as adjuvants with the mycoherbicidal fungus Colletotrichum gloeosporioides, a weakly virulent pathogen of sicklepod (Senna obtusifolia). A 1:1 (v/v) fungus/corn oil tank mixture containing 0.2% (v/v) Silwet L-77 surfactant reduced the dew period requirements for maximum weed infection and mortality from 16 to 8 h, and delayed the need for free moisture for greater than 48 h. This formulation also resulted in the ability of the pathogen to infect and kill weeds in larger (>5 leaf) growth stages. The invert emulsion resulted in similar effects upon these parameters. These results suggest that invert emulsions, unrefined corn oil and Silwet L-77 surfactant greatly improve the bioherbicidal potential of this pathogen for control of sicklepod, a serious weed pest in the southeastern US.     

Potential of a combination of entomopathogenic fungal strains and a non-ionic surfactant to control the fall armyworm (Spodoptera frugiperda)

The fall armyworm, Spodoptera frugiperda, is an emerging invasive pest in Taiwan that feeds on a wide range of crops and causes serious damage. Herein, an entomopathogenic fungal library (EFLib) was constructed to identify potential microbes to control FAW. Twenty-eight indigenous entomopathogenic fungi (EPF) were isolated and investigated for their potential pathogenicity, with Metarhizium pinghaense (Mp-NCHU-124) and Beauveria bassiana (Bb-NCHU-157) exerting dose-dependent effects on the 4th instar FAW larvae. The non-ionic surfactant Silwet L-77 rapidly killed FAW larvae after spraying at a concentration of 300 mg/kg and the toxic effect of Silwet L-77 on FAW larvae was dose-dependent. When the EPF isolates (10⁶ conidia/mL) were applied to FAW larvae in combination with the non-ionic surfactant Silwet L-77 (30–90 mg/kg), the mortality rate dramatically increased and the LT₅₀ reduced, with increased fungal mycosis (Mp-NCHU-124: 38% to 72% and Bb-NCHU-157: 20 to 62%), indicating the high compatibility of EPF with the non-ionic surfactant. Thus, the Silwet L-77+EPF combined formulation has potential for practical field application for FAW pest control and sustainable agriculture in the future.     

Evaluation of three techniques used to determine surfactant phytotoxicity

Three techniques were used to measure and compare the phytotoxicities of Triton X-45 and Agral 90 (two organic surfactants) with two organosilicone surfactants (Silwet L-77 and L-7607), and to compare these four non-ionic compounds with a cationic tallow amine surfactant (Hyspray). Total ion efflux and ethylene response methods were used in vivo and in vitro, while a betacyanin efflux method was an in vitro system only. The first two methods, using intact leaves, were considered to be more closely related to normal spraying conditions than the betacyanin efflux test which used explant material. However, the use of intact leaves was thought to bias the results in favour of the leaf penetration properties of the surfactants rather than their phytotoxicities. The in vitro pigment efflux method provided a simple way of ranking the surfactants in order of their potential phytotoxicities, especially with regard to effects on membrane permeability. This ranking, in increasing order of toxicity, was: Silwet L-7607, Silwet L-77, Agral 90, Triton X-45 and Hyspray.     

Toxicological studies of the carbamates methomyl and bendiocarb in the bulb mite Rhizoglyphus echinopus (Acari: Acaridae)

Methomyl was 15 and 31.3 times more toxic than bendiocarb to bulb mites at the LC50 and LC90 values respectively. However, methomyl (pI50 3.0) was at least 126 times less active than bendiocarb (pI50 5.1) as an inhibitor of bulb mite cholinesterase in vitro. The disparity between the high toxicity of methomyl and its extremely low activity as an inhibitor of mite cholinesterase in vitro indicated that another mechanism was likely involved in its toxic action. Pharmacokinetic studies of methomyl and bendiocarb showed that penetration and metabolism were rapid and that there were no substantial differences in the internal levels of the respective parent carbamates during the 24 h test period. However, volatile radioactive material(s), some of which was carbon dioxide, was produced in appreciably greater amounts from methomyl than from bendiocarb. We speculate that the production of volatiles, such as carbon dioxide, acetonitrile and/or methylamine, may contribute to the toxicity of methomyl to bulb mites. © Rapid Science Ltd. 1998     

Limonene, a citrus extract, for control of mealybugs and scale insects

In a series of bioassays with mealybugs, aqueous solutions of 1% limonene were tested that used from 0.50 to 1.50% all purpose spray adjuvant (APSA)-80 as an emulsifier/surfactant. The two ingredients were added to water or to 0.1% Silwet L-77, an agricultural surfactant. Using 1% limonene, 0.75% APSA-80 and 0.1% Silwet L-77, a semitransparent mixture (primarily a microemulsion) was obtained that was safe for most plants and provided good control of mealybugs when sprayed or used in 1-min dips. Used at half strength, this mixture controlled > or =99% of whiteflies, whereas the full-strength mixture controlled from 69 to 100% of mealybugs and scales, including > or =93% control of root mealybugs. In side-by-side greenhouse tests, this mixture was superior to a 2% solution of insecticidal soap or a 2% solution of horticultural spray oil. Mortality of green scales on potted gardenia plants averaged 95, 89, and 88% on plants sprayed with limonene, insecticidal soap, or horticultural oil, respectively. In a related test, these same sprays killed 44.1, 22.7, or 12.5% of third and fourth instar clustering mealybugs, respectively. Limonene has promise as a safe, natural pesticide for insect pests on tolerant plants. Although 1% limonene solutions damaged certain species of ferns, gingers and delicate flowers, they caused no damage to ornamentals with thick, waxy leaves, such as palms, cycads, and orchids.     

Toxicity of the entomopathogenic bacteria Photorhabdus and Xenorhabdus to the mushroom mite (Luciaphorus sp.; Acari: Pygmephoridae)

The mushroom mite, Luciaphorus sp. is a serious pest of tropical mushrooms. We determined the pathogenicity and toxicity of species and strains of the entomopathogenic bacteria, Photorhabdus and Xenorhabdus to the mite. As these bacteria are known to produce antifungal substances, we first determined the effect of 21 species and strains of the bacteria on the mycelial growth of the mushroom, Lentinus squarrosulus. We then determined the toxicity of the eight species and strains of bacteria that did not show any effect on mushroom growth against both the female and male mites. All eight species and strains of the bacteria were toxic to the female mite resulting in significant mite mortality within 24-48 h. Cell-free supernatants from all the eight bacterial species and strains were also toxic to the female mite inflicting significant mortality within 24-48 h. The supernatants of two strains, GPS12 and GPS11, of Photorhabdus luminescens ssp. laumondii were significantly more toxic than the other species and strains to the female mite, resulting in 90-95% mite mortality within 48 h. Both the concentration and age of the bacteria had significant effect on the toxicity of the supernatants to the female mite. None of the bacteria showed toxicity to the male mite which has undeveloped mouthparts. These results indicate that P. luminescens ssp. laumondii and its byproducts are directly toxic to the female mite, suggesting the potential of developing a novel biological approach for the control of this mushroom pest. This is the first report on the miticidal activity of the entomopathogenic bacteria, Photorhabdus and Xenorhabdus.     

Miticide bioassays with spider mites (Acari: Tetranychidae): effect of test method,exposure period and mortality criterion on the precision of response estimates

Six different bioassay methods were evaluated using propargite (Omite 30% wettable powder (WP) and fenbutatin oxide (Torque 50% (WP) and 55% suspension concentrate (SC)) with twospotted spider mite, Tetranychus uriticae Koch (TSM) and European red mite, Panonychus ulmi Koch (ERM) to document their utility and precision for estimating median lethal concentrations (LC). For each method, two post-treatment exposure periods and mortality criteria were used. Post-treatment exposure period and mortality criterion had a significant influence on the precision of LC₅₀ estimates for all tested miticides with all bioassays methods. Twenty four hour (h) post-treatment exposure was found to be the most suitable for the slide dip and Petri dish methods while 48h was the most appropriate for leaf disc methods. Scoring moribund mites as dead was the most satisfactory criterion for ensuring that biossays were as simple and precise as possible. The Petri dish residue-Potter tower method (PDR-PT) estimated the responses of TSM and ERM to propargite with high precision. The same method was not as precise for fenbutatin oxide formulations. Because significant mite run-off occurred with the leaf disc methods, their precision was not fully established. The slide dip method gave less precise estimates of LC₅₀ values for propargite (WP) and fenbutatin oxide (WP), while the same method gave more precise LC₅₀ estimates for fenbutatin oxide (SC) than the PDR-PT method. The toxicity of candidate miticides was found to be method-and species-dependent.     

Residual bioassay to assess the toxicity of Acaricides against Aceria guerreronis (Acari: Eriophyidae) under laboratory conditions

Aceria guerreronis Keifer (Acari: Eriophyidae) is considered a major pest of the coconut (Cocos nucifera L.), and the use of pesticides is the current method to control it. However, no standard toxicological tests exist to select and assess the efficiency of molecules against the coconut mite. The aim of this study was to develop a methodology that allows for the evaluation of the relative toxicity of acaricides to A. guerreronis through rapid laboratory procedures. We confined A. guerreronis on arenas made out of coconut leaflets and tested two application methods: immersing the leaf fragments in acaricides and spraying acaricides on the leaf fragments under a Potter spray tower. In the latter application method, we sprayed leaf fragments both populated with and devoid of mites. We evaluated the comparative toxicity of two populations (Itamaracá and Petrolina, Pernambuco, Brazil) by spraying on leaflets without mites and submitted the mortality data to probit analysis after 24 h of exposure. No difference was observed in the LC50, regardless of whether the leaflets were immersed or sprayed with acaricide (abamectin, chlorfenapyr or fenpyroximate). The toxicity of chlorfenapyr and fenpyroximate did not differ, irrespective of whether it was applied directly to the leaflet or to the mite; however, the toxicity of abamectin was higher when applied directly to the mite. Chlorpyrifos and abamectin toxicities were lower for the Petrolina population than for the Itamaracá population. Immersing and spraying coconut leaflets can be used to assess the mortality of A. guerreronis under laboratory conditions.     

喷雾:一种简便的拟南芥转化方法

目的：建立一种改良的且操作更为简便可行的拟南芥转化方法。方法：用含有OD600等于0．8的农杆菌,5%蔗糖和0. 2 ml/L表面活性剂Silwet L-77的MS液体培养基喷洒正在发育的花器官即可实现基因转化。植物生长到高约4cm时进行转化,此时植物具有大量的花和极少数的果荚,可以得到比较高的转化效率。喷洒溶液后将植物用保鲜膜等材料包裹起来以保持湿度,可有利于转化。结果：得到了365棵转化植株,转化效率和花器官浸泡法的基本一致。结论：这种方法可适用于多种生态型的拟南芥植物转化,并且方便大规模的拟南芥植物的转化,便于人们快速获得带有T-DNA标记的植株或基因遗传互补工作的顺利开展。     

真空渗透法转化油菜及转化种子的筛选

真空渗透遗传转化法首先在拟南芥(Arabidopsis thaliana)中获得成功, 是一种简便、快速且无需经过组织培养阶段即可获得大量转化植株的基因转化方法。该研究以不同遗传背景的3个甘蓝型油菜(Brassica napus)品种(系)陕3B、L45和Mg23为材料, 对真空渗透遗传转化方法中真空渗透时间和Silwet L-77浓度与遗传转化效果的关系进行了比较, 同时对转化种子的筛选方法进行了优化。结果表明, 卡那霉素(Km)对油菜种子的萌发影响不显著, 但对其生长发育有明显的抑制作用。不同油菜品种对卡那霉素的敏感性不同, 各自的致死浓度也不一样。在0%-0.05%的浓度范围内, 随着Silwet L-77浓度的增加, 在相同的真空渗透时间内, 3个油菜品种的转化率逐渐升高。当Silwet L-77浓度为0.05%时, 10分钟的真空渗透时间可获得最高转化效率, 此时陕3B、L45和Mg23的转化率分别达到1.97%、2.09%和2.30%。     

Nitric oxide level, protein oxidation and antioxidant enzymes in rats infected by Trypanosoma evansi

The acaricidal (miticidal) activity of 90% ethanolic extracts of leaves and stem bark of Swietenia mahogani and Swietenia macrophylla were tested against Varroa destructor mite. Four concentrations were used over two different time intervals under laboratory and field conditions. In general, it was noticed that the acaricidal effect based on mortality and LC(50) of all tested extracts against the Varroa mite was concentration and time dependant. The acaricidal action against Varroa mites was relatively the least for the S. macrophylla stem bark extract at 500 ppm concentration after 48 h while it reached 100% and 95% in case of S. mahogani bark and S. macrophylla leaves, respectively. The% infestation with Varroa in colonies treated with the different extracts at various time intervals showed that the rate of infestation decreased to 0.0% after 12 days from the beginning of treatments with 500 ppm of S. mahogani leaves extract compared to 0.79% decrease after treatment with Mitac, a reference drug (60 mg/colony). The rate of infestation in case of treatments with S. mahogani bark, S. macrophylla leaves and S. macrophylla bark was decreased to 0.11%, 2.41% and 1.08%, respectively. The highest reduction was observed with S. mahogani leaves extract followed by S. mahogani bark. All the tested extracts showed less or no effect on honey bees at the different concentrations and at different bioassay times. This study suggested that the use of natural plant extracts or their products as ecofriendly biodegradable agents could be of high value for the control of Varroa mite.     

茚虫威和六伏隆对小菜蛾卵、幼虫和成虫的药效（英文）

化学防治是世界各地防治小菜蛾Plutella xylostella (L.)的主要方法。选择对小菜蛾各发育阶段高效的杀虫剂有助于很好地控制这一害虫。本研究用浸叶法测定了茚虫威和六伏隆对小菜蛾卵、幼虫和成虫的毒性。茚虫威和六伏隆对小菜蛾卵的LC₅₀值分别为201.40 和 37.32 mg/L。茚虫威和六伏隆对3龄幼虫也有很好的毒性（茚虫威的LC₅₀ 为 4.82 mg/L; 六伏隆的LC₅₀ 为1.48 mg/L)。对成虫的毒性结果表明, 这两种杀虫剂对成虫均没有理想的毒性（茚虫威的LC₅₀ 为845.20 mg/L; 六伏隆的LC₅₀ 为3 438 mg/L)。根据计算的LC₅₀值, 六伏隆对小菜蛾卵和幼虫的毒性比茚虫威高, 但是茚虫威对成虫的毒性更高。     

Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana

The Agrobacterium vacuum infiltration method has made it possible to transform Arabidopsis thaliana without plant tissue culture or regeneration. In the present study, this method was evaluated and a substantially modified transformation method was developed. The labor-intensive vacuum infiltration process was eliminated in favor of simple dipping of developing floral tissues into a solution containing Agrobacterium tumefaciens, 5% sucrose and 500 microliters per litre of surfactant Silwet L-77. Sucrose and surfactant were critical to the success of the floral dip method. Plants inoculated when numerous immature floral buds and few siliques were present produced transformed progeny at the highest rate. Plant tissue culture media, the hormone benzylamino purine and pH adjustment were unnecessary, and Agrobacterium could be applied to plants at a range of cell densities. Repeated application of Agrobacterium improved transformation rates and overall yield of transformants approximately twofold. Covering plants for 1 day to retain humidity after inoculation also raised transformation rates twofold. Multiple ecotypes were transformable by this method. The modified method should facilitate high-throughput transformation of Arabidopsis for efforts such as T-DNA gene tagging, positional cloning, or attempts at targeted gene replacement.     

Extending the host range of the bioherbicidal fungus Colletotrichum gloeosporioides f. sp. aeschynomene

Spore formulations of the bioherbicidal fungus Colletotrichum gloeosporioides f. sp. aeschynomene (ATCC No. 20358) (CGA) were evaluated for control of three weed species: northern jointvetch (Aeschynomene virginica), Indian jointvetch (A. indica), and hemp sesbania (Sesbania exaltata) in greenhouse experiments. Mortality, dry weight reduction, and plant height reduction of A. virginica seedlings ranged from 98% to 100%, 15 days after inoculation with CGA in water, in an invert emulsion, or in Silwet L-77 surfactant. However, CGA in water caused no effects of these parameters on S. exaltata, and only slight effects on A. indica. A. indica and S. exaltata were also severely injured (mortality, dry weight and plant height reduction, 98–100%) by CGA in the invert emulsion or in Silwet L-77. The CGA in Silwet formulation incited severe disease development more rapidly than the invert emulsion or water formulations of CGA in all species. These results suggest that the host range of CGA can be expanded though formulation modification to enable this bioherbicide to control multiple weeds, thus making this product more economically acceptable.     

Evaluation of phloxine B as a possible control agent against citrus thrips (Thysanoptera: Thripidae)

We evaluated the toxicity of phloxine B photoactive dye combined with a cane molasses bait against citrus thrips, Scirtothrips citri (Moulton). Laboratory bioassays conducted under artificial light showed that thrips mortality followed a log-dose probit-response model with an estimated LC50 of 0.0079% dye. Diluted cane molasses plus 0.01% phloxine B then was used as a standard for comparison of eight additional baits, including three formulations of concentrated citrus peel liquor (CCPL1-3). Citrus thrips mortality ranked highest to lowest with CCPL1 and CCPL3 > CCPL2, Mo-Bait and cane molasses > concentrated beet molasses, concentrated cane molasses, hemicellulose extract, and whey. Several commercial surfactants were tested to see if their addition to the standard increased efficacy. Hyper-Active, Kinetic, and Tween 60 at 1% and Cohort, Hyper-Active, Kinetic, and Silwet at 0.25% when added to the standard, reduced citrus thrips mortality, whereas Tween 60 and Dyne-Amic at 0.25% had no effect. Cane molasses with one or 5% phloxine B dye and CCPL1 with 1% dye were sprayed on citrus trees and allowed to weather in the field. Laboratory bioassays conducted after leaves had weathered for up to 8 d indicated that bait-dye toxicity was persistent. Possible use of the bait-dye mixture in commercial control of citrus thrips is discussed.     

Toxicity of neem (Azadirachta indica A. Juss) formulations for twospotted spider mite and Euseius alatus de leon and Phytoseiulus macropilis (Banks) (Acari: Phytoseiidae)

The toxicity of selected commercial formulations of neem on Tetranychus urticae Koch (Acari: Tetranychidae) and two predatory mites Euseius alatus De Leon and Phytoseiulus macropilis (Banks) was studied. Topical toxicity was tested with the commercial formulations (Natuneem, Neemseto and Callneem) and extract of neem's seeds at concentration 1%, compared to the standard acaricide abamectin at concentration of 0.3 ml/L and the control treatment (distilled water). Based on the best performance against T. urticae through topical contact, the formulation Neemseto was selected to be evaluated using different concentrations against eggs, and residual and repellent effects on adults of the mites. Egg treatment consisted of dipping eggs into Neemseto dilutions and control treatment for five seconds. In addition, residual and repellent effects of Neemseto for adult mites consisted of using leaf discs dipped into the dilutions for five seconds. The toxicity of Neemseto on eggs and adults was greater for T. urticae compared to the toxicity observed for the predatory mites. Neemseto was repellent for T. urticae and E. alatus when tested at the concentrations of 0.25, 0.50 and 1.0%, and did not affect P. macropilis. Neemseto using all concentrations, while for the predatory mites significant reduction of mite fecundity was only observed at the largest concentrations reduced the fecundity of T. urticae significantly. So Neemseto, among tested neem formulations, performed better against the twospotted spider mite and exhibited relatively low impact against the predatory mites studied.     

Acaricidal activity of Juglans regia leaf extracts on Tetranychus viennensis and Tetranychus cinnabarinus (Acari: Tetranychidae)

Leaf extracts of the walnut, Juglans regia L., were evaluated under laboratory conditions to determine their acaricidal activity on the mites Tetranychus cinnabarinus (Boisduval) and Tetranychus viennensis Zacher (Acari: Tetranychidae). Extracts had both contact and systemic toxicity to these mites. The four solvents tested for preparing crude extracts were petroleum ether, chloroform, ethyl acetate, and methanol. Methanol was the most efficient solvent, with an extraction rate from 17.06 + 0.80 to 20.27 +/- 0.28%. Petroleum ether was the least effective solvent, with extraction rates from 2.30 +/- 0.13 to 2.71 +/- 0.13%. However, the crude extracts with petroleum ether resulted in the highest mite mortality (79.04 +/- 0.52%) in a slide dip bioassay. Mites mortalities from the concentrated extracts prepared by chloroform, ethyl acetate, methanol, or distilled water were significantly lower than petroleum ether. The mean lethal concentrations (LC50) of the extracts from petroleum ether, chloroform, ethyl acetate, methanol, and distilled water to the two mite species were 0.73 +/- 0.04, 1.66 +/- 0.28, 4.96 +/- 0.35, 7.45 +/- 0.67, and 9.91 +/- 0.32 mg/ml, respectively. After liquid chromatography and thin-layer chromatography, the concentrated extracts of petroleum ether were separated into eight fractions and tested for acaricidal activity. Fraction 6 produced significantly higher mite mortality rates than the other groups, killing approximately 90% of both species.     

Laboratory evaluation of avermectin as a selective acaricide for use withMetaseiulus occidentalis (Nesbitt) (acarina: Phytoseiidae)

The suggestion that adding a light oil to avermectin B₁ would increase the toxicity of avermectin to spider mites and reduce its effect on predaceous mites was tested in laboratory trials withTetranychus urticae Koch andMetaseiulus occidentalis (Nesbitt) on almond and bean foliage. No differences were found in the toxicity of avermectin + oil vs. avermectin alone at the doses tested forT. urticae; all (0.025, 0.5, 1, and 5 ppm) were highly toxic. Mortality ofM. occidentalis females and larvae was not different on avermectin + oil vs. avermectin alone, but females produced more progeny on the avermectin + oil-treated foliage. At doses of 0.5 to 5 ppm, avermectin was sufficiently toxic to deplete predator populations in the field. Development of predator larvae on avermectin + oil and on avermectin alone was not different. Avermectin + oil on almond foliage aged outdoors was highly toxic after 96 h toT. urticae adults butM. occidentalis larvae survived well on residues by 96 h.M. occidentalis female survival and productivity were not different from the controls by 48 h. Hence a predator mite population might recover through larvae hatching onto residues. Avermectin + oil (3 ppm) residue on bean foliage held outdoors was still highly toxic toT. urticae after 33 days. In contrast,M. occidentalis females and larvae survived well on 48-to 96-hour-old residues. Neither predators nor spider mites placed on treated foliage (3 ppm) were able to reach untreated foliage in tests using bean plant seedlings with one leaf sprayed and one left unsprayed. Furthermore, whenM. occidentalis females were exposed to 3 ppm avermectin for 300 s or longer, mortality was significant and the fecundity of females that had been exposed for as few as 30 s was reduced significantly. Thus, while avermectin is significantly more toxic toT. urticae than toM. occidentalis, its value as a selective acaricide will depend upon learning to use it at rates that will allow the retention of sufficient prey so that surviving predators can persist. Based on these laboratory tests, such selective doses are likely to lie below 1 ppm and can best be determined in field trials.     

Local variation in susceptibility of Colorado potato beetle (Coleoptera: Chrysomelidae) to insecticide

The susceptibility status of Colorado potato beetle, Leptinotarsa decemlineata (Say), adults to phosalone was determined by dip and glass jar assay techniques. Bioassay results indicated a narrow variation in Colorado potato beetle insecticide susceptibility among sample sites. LC50 values were generally highest from specimens collected in field that received frequent phosalone applications for seven consecutive growing seasons. In five populations tested, LC50 values ranged from 503.72 to 827.95 ppm in dip test method. In glass jar technique, resistance ratio value of 1.72 for LC50 was obtained. A significant linear relationship between LC50 values of individual populations across test methods was detected. Both bioassay techniques were suitable for monitoring resistance to insecticide in Colorado potato beetle adult populations. Glass jar technique, however, exhibited less variability in LC50 estimates and showed a higher degree of sensitivity than the dip method. Filter paper and leaf disk techniques for larvae were two bioassay methods used to determine phosalone susceptibility in L. decemlineata populations. Both bioassay techniques exhibited a similar level of susceptibility of the larvae to phosalone; however, the fiducial limit values from filter paper method were narrow than the leaf disk assay technique. A significant direct relationship between LC50 values of individual population across test methods was observed. Differences in LC50 ranking among fields between adults and larvae indicated a differential susceptibility to insecticide between life stages. Low LC50 values obtained from Colorado potato beetle in sample sites indicated that phosalone resistance was not severe in these fields. The glass jar and filter paper testing methods are simple and sensitive test techniques for measuring susceptibility of Colorado potato beetle adults and larvae to phosalone, respectively.