Toxicity of Carum copticum essential oil‐loaded nanogel against Sitophilus granarius and Tribolium confusum

Myristic acid–chitosan (MA‐chitosan) nanogels were synthesized with the technique of self‐assembly and loaded with Carum copticum (L.) essential oil. Scanning electron microscopy (SEM) study indicated nanogels with smooth surfaces and near spherical appearance, and according to transmission electron microscopy (TEM) image, core–shell stricter of nanogels was reported. Subsequently, fumigant toxicity of C. copticum oil and oil‐loaded nanogels was assessed against Sitophilus granarius (L.) and Tribolium confusum Jacquelin du Val. Adults were exposed to different concentrations of the oil and oil‐loaded nanogels at 27 ± 1°C and 55 ± 5% r.h. in continuous darkness. The mortality was counted after 3, 6, 10, 24 and 48 h of exposure. Mortality increased with increasing concentration levels. Fumigant toxicity of oil‐loaded nanogels was significantly more than the oil even after 48 h. According to the findings, oil‐loaded nanogels were 8.9‐ and 3.7‐fold more toxic than the oil against S. granarius and T. confusum, respectively. The persistence of oil and oil‐loaded nanogels was also investigated over period of time. Oil‐loaded nanogels lost the insecticidal effectiveness after 21 days post‐application for S. granarius and 33 days in the case of T. confusum, while the efficacy of oil decreased in the early days of application. Chemical composition of C. copticum essential oil was measured, and six components were identified. Thymol, ρ‐cymene and γ‐terpinolene were the major components of the oil. The essential oil‐loaded nanogels may have potential to be used as an alternative to synthetic insecticides for stored products protection. However, more research is necessary to improve the findings.     

The antagonistic/synergistic effects of some medicinal plant essential oils,extracts and powders combined with Diatomaceous earth on red flour beetle,Tribolium castaneum Herbst (Coleoptera: Tenebrionidae)

Abstract

Red flour beetle, Tribolium castaneum Herbst (Coleoptera: Tenebrionidae) is a prominent pest of stored products particularly cereal flour. Since resistance of this pest to common chemical insecticides is well documented, we were examined the synergistic/antagonistic interaction between Satureja hortensis L., Trachyspermum ammi L., Ziziphora tenuior L., Cuminum cyminum L. and Foeniculum vulgare Miller essential oils, ethanolic extracts and powders with Diatomaceous earth (DE) against T. castaneum adults under laboratory conditions at 27 ± 1 °C, 65 ± 5% RH and continuous darkness. We assayed repellency of ethanolic extracts and essential oils of mentioned plants on the pest. Results showed that DE had high toxicity to the pest. Plant essential oils and ethanolic extracts (except ziziphora) synergized the performance of DE. Nevertheless, plant powders elicited antagonistic effects (except ziziphora that exhibited synergistic effect). The most repellent EO and extract was cumin which exhibited mean repellency value on adult insect equivalent to 92.58 and 51.47%, respectively.     

Synergistic activity of coriander oil and conventional antibiotics against Acinetobacter baumannii

In this study we investigated the existence of synergistic antibacterial effect between coriander (Coriandrum sativum L.) essential oil and six different antibacterial drugs (cefoperazone, chloramphenicol, ciprofloxacin, gentamicin, tetracycline and piperacillin). The antibacterial activity of coriander oil was assessed using microdilution susceptibility testing and synergistic interaction by checkerboard assays. The association of coriander essential oil with chloramphenicol, ciprofloxacin, gentamicin and tetracycline against Acinetobacter baumannii showed in vitro effectiveness, which is an indicator of a possible synergistic interaction against two reference strains of A. baumannii (LMG 1025 and LMG 1041) (FIC index from 0.047 to 0.375). However, when tested the involvement between coriander essential oil and piperacillin or cefoperazone, the isobolograms and FIC index showed an additive interaction. The in vitro interaction could improve the antimicrobial effectiveness of ciprofloxacin, gentamicin and tetracycline and may contribute to resensitize A. baumannii to the action of chloramphenicol.     

Fumigant toxicity of Carum copticum L. (Apiaceae) essential oil on Trialeurodes vaporariorum (Westwood) (Hemiptera: Aleyrodidae) adults under greenhouse conditions

Greenhouse whitefly Trialeurodes vaporariorum (Westwood) (Hemiptera: Aleyrodidae) is one of the most injurious pests of greenhouse crops and ornamental plants in worldwide, both outdoor and indoor. This insect, feeding on plant sap, producing honeydew and transmitting plant viruses, causes quantitative and qualitative damages in plants. For controlling this pest in greenhouse, plant essential oils are mentioned instead of chemical insecticidal. So, in this research, fumigant toxicity of Carum copticum L. (Fam.: Apiaceae) plant oil on mentioned adult pest was surveyed. Dry ground seeds were subjected to hydrodistillation using a modified Clevenger-type apparatus and the resulting oil contained Thymol (50.07%), Gama-Terpinene (23.99%), P-Cymene (22.9%), Myrcene (0.51%) and 1,8-Cineole (0.5%). All bioassay tests were conducted at 27?±?2?°C, 65?±?5 RH and a photoperiod of 16:8?h (light: dark). This research was performed with a completely random design with six treatments (five different concentrations of essential oils plus control). Each concentration includes of three replicates and each replicate consisted of 20 adult of pests. The results showed that aforementioned essential oil shows significant mortality of adults 24?h after exposure. The value LC₅₀ of mentioned plant oil on T. vaporariorum was 1.03?μl?L/L air. And mortality percentage shows higher sensitivity of T. vaporariorum against application of essential oil. The value LT₅₀ estimated for T. vaporariorum in concentration of 1.03?μl?L/L air was 7.18?h. Fumigant toxicity this essential oil has had an ordered relationship with the concentration and time exposure. The results of this research showed that mentioned plant oil had appropriate insecticidal effects on these greenhouse pests. The findings showed that C. copticum L. oil has high impact on the above-mentioned pests and it is suggested because of its high potential in fumigant toxicity and its use in integrated pest management programmes in greenhouses is mentioned.     

Larvicidal,ovicidal activities and histopathological alterations induced by Carum copticum (Apiaceae) extract against Culex pipiens (Diptera: Culicidae)

An experiment was carried out, firstly, to determine the possible toxicity of Carum copticum (Apiaceae) extract against Culex pipiens (Diptera: Culicidae), and, secondly, to study the histopathological alterations in the midgut of Cx. pipiens as a result of treatment with C. copticum extract. Larvicidal and ovicidal activities of C. copticum extract against the larvae of Cx. pipiens was determined according to World health organization (WHO). The inhibition effect of C. copticum was assessed by determining the mortality of the treated larvae and eggs. The histopathological effect of the C. copticum extracts on midgut epithelium of the larvae was examined under both light and transmission electron microscopy. The crude extract of C. copticum exerted 100% mortality for Cx. pipiens after 24 h at 200 μm/ml, and zero hatchability (100% mortality) at 150 μm/ml for Cx. pipiens. The histopathological study showed that larvae treated with C. copticum extract had cytopathological alterations of the midgut epithelium. The study provided information on various effects of C. copticum extract against Cx. pipiens.     

Bioactive extracts of Carum copticum L. enhances efficacy of ciprofloxacin against MDR enteric bacteria

The widespread occurrence of extended spectrum β-lactamases (ESβLs) producing enteric bacteria and their co-resistance with flouroquinolones has impaired the current antimicrobial therapy. This has prompted the search for new alternatives through synergistic approaches with herbal extracts. In this study Carum copticum (seeds) was extracted first in methanol and then subsequently extracted in different organic solvents. MIC of plant extracts, ciprofloxacin and thymol was determined by broth micro-dilution method using TTC. Synergism between plant extracts and ciprofloxacin was assayed by the checkerboard method. Chemical constituents of active extracts were analyzed by GC-MS. Methanolic, hexane and ether extract of Carum copticum exhibited significant antibacterial activity with MIC values ranged from 0.25 mg/ml to 2.0 mg/ml. Synergy analysis between Carum copticum extracts and ciprofloxacin combinations revealed FIC index in the range of 0.093–0.25. About 81% ciprofloxacin resistant ESβL producing enteric bacteria were re-sensitized in the presence of 15.6–250 μg/ml of methanolic extract of Carum copticum. Moreover, ciprofloxacin showed 8 to 64 folds reduction in MIC in presence of 250 and 500 μg/ml of hexane extract. Whereas, 4–32 folds reduction in MIC of ciprofloxacin was achieved in the presence of 31.25 and 62.5 μg/ml of ether extract, indicating synergistic enhancement of drug activity. The chemical analysis of hexane and ether extracts by GC-MS revealed the common occurrence of one or more phenolic hydroxyl at different locations on benzene ring. This study demonstrated the potential use of herbal extract of Carum copticum in combination therapy against ESβL producing bacteria.     

Plant Essential Oils Synergize and Antagonize Toxicity of Different Conventional Insecticides against Myzus persicae (Hemiptera: Aphididae)

Plant-derived products can play an important role in pest management programs. Essential oils from Lavandula angustifolia (lavender) and Thymus vulgaris (thyme) and their main constituents, linalool and thymol, respectively, were evaluated for insecticidal activity and synergistic action in combination with insecticides against green peach aphid, Myzus persicae (Sulzer) (Hemiptera: Aphididae). The essential oils and their main constituents exerted similar insecticidal activity when aphids were exposed by direct sprays, but were non-toxic by exposure to treated leaf discs. In synergism experiments, the toxicity of imidacloprid was synergized 16- to 20-fold by L. angustifolia and T. vulgaris essential oils, but far less synergism occurred with linalool and thymol, indicating that secondary constituents of the oils were probably responsible for the observed synergism. In contrast to results with imidacloprid, the insecticidal activity of spirotetramat was antagonized by L. angustifolia and T. vulgaris essential oils, and linalool and thymol. Our results demonstrate the potential of plant essential oils as synergists of insecticides, but show that antagonistic action against certain insecticides may occur.     

Sensitive voltammetric determination of thymol in essential oil of Carum copticum seeds using boron-doped diamond electrode

Essential oil of Carum copticum seeds, obtained from a local shop, was extracted and content of thymol was analyzed using square-wave voltammetry at boron-doped diamond electrode. The effect of various parameters, such as pH of supporting electrolyte and square-wave voltammetric parameters (modulation amplitude and frequency), was examined. In Britton–Robinson buffer solution (pH 4), thymol provided a single and oval-shaped irreversible oxidation peak at +1.13 V versus silver/silver chloride potassium electrode (3 M). Under optimal experimental conditions, a plot of peak height against concentration of thymol was found to be linear over the range of 4 to 100 μM consisting of two linear ranges: from 4 to 20 μM (R² = 0.9964) and from 20 to 100 μM (R² = 0.9993). The effect of potential interferences such as p-cymene and γ-terpinene (major components in essential oil of C. copticum seeds) was evaluated. Thus, the proposed method displays a sufficient selectivity toward thymol with a detection limit of 3.9 μM, and it was successfully applied for the determination of thymol in essential oil of C. copticum seeds. The Prussian blue method was used for validation of the proposed electroanalytical method.     

Fumigant Toxicity of Plant Essential Oils to Plutella xylostella (Lepidoptera: Yponomeutidae) and Cotesia glomerata (Hymenoptera: Braconidae)

The fumigant toxicity of 66 plant essential oils to Plutella xylostella (L.) larvae and Cotesia glomerata (L.) adults was examined using a vapor-phase toxicity bioassay and compared with that of dichlorvos. Responses varied according to oil and insect species used. Based on 24 h LD₅₀ values, pennyroyal oil [10.77 mg/filter paper (4.25 cm diameter)] was the most toxic fumigant, followed by rosemary and sage (Dalmatin) oils (15.15 mg/paper). Potent fumigant toxicity was also produced from armoise, buchu leaf, cedarleaf, coriander, eucalyptus, howood, lavender, myrtle, niaouli, peppermint, and rosewood oils (LD₅₀, 21.29–27.31 mg/paper). All essential oils were less effective than dichlorvos (LD₅₀, 0.52 mg/paper). Against adult C. glomerata, dichlorvos (LD₅₀, 0.03 mg/paper) was the most toxic fumigant, whereas the LD₅₀ values of the 14 essential oils ranged from 1.59 to 8.51 mg/paper. Based on selective toxicity ratio (STR, P. xylostella LD₅₀/C. glomerata LD₅₀), the 14 essential oils (STR, 2.5–14.5) are more selective than dichlorvos (STR, 17.3). The essential oils tested merit further study as potential fumigants for the control of P. xylostella in greenhouses because of their selective toxicity to adult C. glomerata and their much greater activity as a fumigant.     

Insecticidal Activity and Chemical Composition of Artemisia sieben Besser Essential Oil from Karaj,Iran

Atremisia sieberi Besser is a widely distributed plant that grows in many areas of Iran and has strong insecticidal activity against stored product pests, so an experiment was conducted to investigate fumigant toxicity of the A. sieberi oil collected from Karaj region of Iran. The oil was applied against one to seven day old adults of three major stored product insects including: Callosobruchus maculatus (Fab.), Sitophilus oryzae (L.), and Tribollium castaneum (Herbst). The potency of fumigant toxicity of A. sieberi on C. maculatus was higher (LC₅₀: 1.64 μL per L) than S. oryzae (LC₅₀: 4.41 μL per L) and T. castaneum (LC50: 20.31 μ.L per L). The relationships between the time exposure and oil concentration on mortality show that the mortality was increased as oil concentration and exposure time was increased. The concentration of 185 μL per L and exposure time of 24h was enough to obtain 100% kill of the insects. It was also found that the regions where A. sieberi grows affect essential oil components of the plant and can play an important role in properties of fumigant toxicity.     

The in vitro antimicrobial activity of Cymbopogon essential oil (lemon grass) and its interaction with silver ions

BackgroundIt is well known that Cymbopogon (lemon grass) essential oil exhibits antimicrobial activity while the efficacy of silver ions as a disinfectant is equally well reported.HypothesisThe antimicrobial activity of CEO and Ag⁺ and their synergistic combinations will be useful in improving the current treatment strategies for various infections.Study designIn the present study, we determined the chemical composition and in vitro antimicrobial activity of six different Cymbopogon essential oils (CEO's) alone and in combination with silver ions (Ag⁺) against two Gram-positive (Staphylococcus aureus and Enterococcus faecalis), two Gram-negative (Escherichia coli and Moraxella catarrhalis) and two yeast species (Candida albicans and Candida tropicalis). The nature of potential interactions was determined by fractional inhibitory concentration indices (FICIs) for CEO's and Ag⁺ calculated from microdilution assays and time–kill curves.ResultsGas chromatography–mass spectrometry results confirmed the presence of nerol, geranial and geraniol as major volatile compounds. Minimum inhibitory concentration (MIC) values confirmed that all the tested pathogens are variably susceptible to both CEO's as well as Ag⁺. The MIC of CEO's and Ag⁺ against all the tested pathogens ranged from 0.032 mg/ml to 1 mg/ml and 0.004 and 0.064 mg/ml respectively, whereas when assayed in combination the FICI values were drastically reduced to range between 0.258 and 2.186, indicating synergy, additive and indifferent interactions. The most prominent interaction was observed between Cymbopogon flexuosus essential oil and Ag⁺ against C. albicans with ∑FIC = 0.254. The synergistic interactions were further confirmed through the construction of isobolograms and time–kill plots. Transmission electron microscopy showed disturbance in the cell envelope upon the concomitant treatment of CEO's and Ag⁺, which ultimately leads to cell death.ConclusionResults suggest that CEO's and Ag⁺ when used in combination offers an opportunity to the formulation scientist to produce novel combinations acting synergistically in the continued quest to control important infectious pathogens.     

Insecticidal activity and chemical composition of the essential oil of Artemisia vestita from China against Sitophilus zeamais

In our screening program for new agrochemicals from local wild plants, essential oil of Artemisia vestita Wall (Asteraceae) was found to possess strong insecticidal activity against maize weevil, Sitophilus zeamais Motsch. Essential oil of aerial parts of A. vestita was obtained from hydrodistillation and was investigated by GC and GC–MS. The main components of essential oil were grandisol (40.29%), 1,8-cineol (14.88%) and camphor (11.37%). The essential oil of A. vestita possessed strong fumigant toxicity against S. zeamais adults with a LC₅₀ value of 13.42 mg/L air. The essential oil of A. vestita also showed contact toxicity against S. zeamais adults with a LD₅₀ value of 50.62 mg/adult.     

Rice weevil response to basil oil fumigation

Basil oil, Ocimum basilicum L., is a volatile plant essential oil that is known to have insecticidal activity against stored product pests such as rice weevil, Sitophilus oryzae (L.). Basil oil was diluted in acetone and applied to a sponge held inside a tea strainer for fumigations in containers with and without rice. Basil oil fumigation (3 ml, 10% concentration) caused high mortality in adult rice weevils when weevils were exposed in air in a sealed 1 L plastic container. However, when basil oil was placed in packaged rice, weevil mortality was low and reproduction was not affected. Effectiveness of plant essential oil fumigation should be conducted under realistic conditions to avoid experimental artifacts and misleading results.     

Effect of chemical complexity of essential oils on feeding deterrence in larvae of the cabbage looper

The relationship between chemical composition and feeding deterrent activity of seven essential oils is explored, including those from Syzygium aromaticum (L.) Merr. & Perry (Myrtaceae), Cinnamomum zeylanicum Blume (Lauraceae), Lavendula latifolia (L.) Medicus, Lavendula angustifolia L., Mentha crispa L., Mentha arvensis L. and Mentha piperita L. (Lamiaceae) against an important agricultural pest, the cabbage looper (Trichoplusia ni Hübner, Noctuidae), using a leaf‐disc choice bioassay. Comparison of the deterrent activity of ‘full mixtures' with respective artificial blends missing individual constituents demonstrates that, for most oils, minor constituents in a mixture can be as important as major constituents with respect to the overall feeding deterrent effect. There is a lack of correlation between the feeding deterrent effect of an individual constituent of an essential oil and its contribution to the overall activity of the ‘full mixture’ in some cases. The effect of removing an individual constituent from the mixture of an essential oil depends on the unique properties of the mixture, which in turn may reflect the interaction of its constituents. Understanding the role and contribution of each constituent to the overall activity of the oil can facilitate the creation of artificial blends that optimize their efficacy against different pests.     

Control of stored grain pest,Callosobruchus maculatus (F.) (Coleoptera: Bruchidae), using the essential oils isolated from Zingiber officinale (L.) and Mentha pulegium (L.) in laboratory condition

Fumigant activity of essential oil vapours distilled from Zingiber officinale (L.) and Mentha pulegium (L.) was tested against eggs, larvae and adults of Callosobruchus maculatus (F.). Fumigant toxicity was assessed at 27?±?1?°C and 60?±?5% RH, in dark condition. The influence of different concentrations of the essential oil vapours on egg hatchability, larval and adult mortality was significant. Data probit analysis showed that lethal concentration of the essential oil to kill 50% of the population (LC₅₀) for egg, larvae and adult was found to be 1.151, 2.336 and 2.183?μl/l air of Z. officinale, followed by 0.072, 0.113 and 0.093?μl/l air essential oil of M. pulegium, respectively. Between these essential oils, Z. officinale is almost more toxic than M. pulegium on all growth stages of C. maculatus. The present study suggests that essential oils from these medicinal plants may be potential grain protectants as botanical alternative fumigants and could be used in the management of various life stages of C. maculatus.     

Spermidine promotes stress resistance in Drosophila melanogaster through autophagy-dependent and -independent pathways

The naturally occurring polyamine spermidine (Spd) has recently been shown to promote longevity across species in an autophagy-dependent manner. Here, we demonstrate that Spd improves both survival and locomotor activity of the fruit fly Drosophila melanogaster upon exposure to the superoxide generator and neurotoxic agent paraquat. Although survival to a high paraquat concentration (20 mM) was specifically increased in female flies only, locomotor activity and survival could be rescued in both male and female animals when exposed to lower paraquat levels (5 mM). These effects are dependent on the autophagic machinery, as Spd failed to confer resistance to paraquat-induced toxicity and locomotor impairment in flies deleted for the essential autophagic regulator ATG7 (autophagy-related gene 7). Spd treatment did also protect against mild doses of another oxidative stressor, hydrogen peroxide, but in this case in an autophagy-independent manner. Altogether, this study establishes that the protective effects of Spd can be exerted through different pathways that depending on the oxidative stress scenario do or do not involve autophagy.     

Chemical constituents and fumigant toxicity of essential oil from Carum copticum against two stored product beetles

Plant secondary metabolites play an important role in plant-insect interactions and therefore such compounds may have insecticidal or antifeedant activity against insects. Carum copticum C. B. Clarke （Apiaceae） is one of these plants that have medicinal effects on humans. The chemical composition of the essential oil from dry seeds of C. copticum was studied by gas chromatography （GC） and gas chromatography mass spectrometry （GC-MS）. Thymol （41.34%）,α-terpinolene （17.46%） and ρ-cymene （11.76%） were found to be the major constituents of the oil. In fumigant toxicity tests with the essential oil against adults of Sitophilus oryzae （L.） and Tribolium castaneum （Herbst） at 27 ±1℃ and 60% ± 5% RH, it was observed that S. oryzae （LC50 = 0.91 μL/L） were significantly susceptible than T. castaneum （LC50 = 33.14 μL/L）. The mortalities of the insect species reached 100% at concentrations higher than 185.2 μL/L and 12-h exposure time. The findings indicate the strong insecticidal activity of C. copticum oil and its potential role as a fumigant for storedproduct insects.     

冬青油与肉桂油对淡色库蚊的生物活性

为进一步明确冬青油（wintergreen oil）与肉桂油（cinnamon oil）对蚊虫的生物活性, 本研究采用浸液法、 “Y”型嗅觉仪法和密闭三角瓶熏蒸法, 分别测定了两种精油对淡色库蚊Culex pipiens pallens幼虫的毒杀作用以及对成虫的驱避和熏杀活性。结果显示： 肉桂油和冬青油对淡色库蚊幼虫均具有较强的毒杀作用, 处理24 h的LC₅₀分别为71.87 mg/L和 102.83 mg/L。在0.5 μL的供试剂量下, 冬青油与肉桂油在20 min内对淡色库蚊的驱避率均在80%以上, 显示了较好的驱蚊活性。冬青油对淡色库蚊具有明显的熏蒸击倒作用, 12 μL/L浓度下的KT₅₀为3.97 min; 而肉桂油则具有较好的熏蒸致死活性, 熏蒸5 h的LC₅₀ 为0.31 μL/L。冬青油与肉桂油按1∶1体积比混配后对淡色库蚊表现出较好的击倒和致死效果。冬青油和肉桂油既对淡色库蚊幼虫具有较强的毒杀作用, 又对淡色库蚊雌成虫表现出较好的驱避和熏杀活性, 具有开发成为植物源蚊虫防控剂的潜力。     

Pharmacological interactions of essential oil constituents on the in vitro growth of Plasmodium falciparum

The pharmacological properties of essential oils have been widely studied. However the interaction and contribution of the individual constituents assayed singularly and in combination remains relatively unexplored. To investigate these possible interactions, various essential oil constituents (EOC's) were combined and the interactions on their antimalarial and toxicity properties determined using the tritiated hypoxanthine incorporation and the tetrazolium-based cellular viability assays, respectively. In combination, two inactive EOC's (IC50 values greater than 1 mM), ρ-cymene and carvacrol interacted in a synergistic manner (∑ FIC = 0.02) against a chloroquine-resistant strain of Plasmodium falciparum. This interaction was comparable to that between the potent E- and Z-(±)-nerolidol (IC50 value: 0.9 ± 0.3 μM) and the standard antimalarial, quinine (IC50 value: 0.13 ± 0.04 μM) (∑ FIC = 0.01). However, this latter combination also potentiated the toxicity (∑ FIC = 0.001) of each molecule. A similar increase in toxicity was noted between ρ-cymene and γ-terpinene, as well as E- and Z-(±)-nerolidol and (−)-pulegone. These results show that the pharmacological activities of individual EOC's can vary greatly when used in combination, and show potential as adjuvants in the elimination of malaria infections.     

Insecticidal activity of mixtures of Bacillus thuringiensis crystals with plant oils of Sinapis alba and Azadirachta indica

The main purpose of this study was to determine the insecticidal activity of mixtures of Bacillus thuringiensis crystalline toxins and plant substances that could contribute to create a new bioinsecticide of high efficiency. We decided to determine the toxicity of crystals of B. thuringiensis MPU B9 isolate, azadirachtin (NeemAzal), mustard oil and their mixtures against lepidopteran pests: Cydia pomonella, Spodoptera exigua and Dendrolimus pini. The interaction of the components in mixtures against insects was evaluated and their utility in plant protection was assessed. The crystals and plant substances showed synergistic effect against all insect species. The occurrence of synergism between two components depended on the ratio of their concentration in mixtures. Slight amount of one of the components added to the mixture led to enhanced toxicity of the preparation. The supplementation of mustard oil with B. thuringiensis toxins from up to 0.3% of the mixture resulted in more than twofold higher observed mortality of L1 larvae of D. pini in comparison with expected mortality. The beneficial insecticidal effect of the mustard oil and crystals mixture was also noted when crystals comprised up to 1.3% and from 7 to 20% against L1 caterpillars of C. pomonella and L3 larvae of S. exigua, respectively. The insecticidal efficient effect was achieved when mixture of B. thuringiensis crystals and azadirachtin was used against the three species of lepidopteran pests. The combinations in which components interacted synergistically against caterpillars are potentially useful for making a new bioinsecticide for controlling lepidopteran pests in plant protection.