Bioactivities of methyl allyl disulfide and diallyl trisulfide from essential oil of garlic to two species of stored-product pests, Sitophilus zeamais (Coleoptera: Curculionidae) and Tribolium castaneum (Coleoptera: Tenebrionidae)

Two of the major constituents of the essential oil of garlic, Allium sativum L., methyl allyl disulfide and diallyl trisulfide, were tested against Sitophilus zeamais Motschulsky and Tribolium castaneum (Herbst) for contact toxicity, fumigant toxicity, and antifeedant activity. The contact and fumigant toxicities of diallyl trisulfide were greater than that of methyl allyl disulfide to the adults of these two species of insects. These two compounds were also more toxic to T. castaneum adults than to S. zeamais adults. Older T. castaneum larvae were more susceptible to the contact toxicity of the two compound, whereas younger larvae were more susceptible to the fumigant toxicity of these compounds. Both compounds reduced egg hatching of T. castaneum and subsequent emergence of progeny. Diallyl trisulfide totally suppressed egg hatching at 0.32 mg/cm2, and larval and adult emergence at 0.08 mg/cm2. Methyl allyl disulfide significantly decreased the growth rate, food consumption, and food utilization of adults of both insect species, with feeding deterrence indices of 44% at 6.08 mg/g food for S. zeamais and 1.52 mg/g food for T. castaneum. However, it did not affect any nutritional indices of T. castaneum larvae. Diallyl trisulfide significantly reduced all of the nutritional indices in all of the insects tested. Feeding deterrence indices of 27 and 51% were obtained in S. zeamais adults and T. castaneum larvae, respectively, at the concentration of 2.98 mg/g food, whereas feeding deterrence of 85% was achieved in T. castaneum adults at a much lower concentration of 0.75 mg/g food. Hence, diallyl trisulfide is a more potent contact toxicant, fumigant and feeding deterrent than methyl allyl disulfide.     

Insecticidal activities of garlic substances against adults of grain moth,Sitotroga cerealella (Lepidoptera: Gelechiidae)

Abstract The angoumois grain moth, Sitotroga cerealella (Olivier), is one of the most serious stored grain pests around the world. In attempts to reduce the losses caused by the moth and to suppress its populations, the fumigant activities, behavioral influence and ovipositional inhibition of garlic (Allium sativum) essential oil and its two major components, diallyl disulfide and diallyl trisulfide, were investigated against the adult grain moth. Their effects on reduction in survival of first instar larvae to adult emergence were also evaluated. Results showed that these three materials (garlic essential oil, diallyl disulfide and diallyl trisulfide) had significant fumigant activity with 50% lethal concentration values at 1.33, 0.99, and 1.02 μL/L air space, respectively; meanwhile, the three materials possessed high behavioral deterrent activities against adults in the Y‐tube olfactometer. When applied to rice grains, these materials reduced adult longevity and inhibited oviposition, with ovipositional inhibition above 70% at a concentration of 1.5 μL/25 g in either no‐choice or two‐choice tests. In short, the study showed that both diallyl disulfide and diallyl trisulfide, like garlic essential oil, acted as fumigants, produced behavioral deterrence and inhibited oviposition against angoumois grain moth. Our work here indicates that diallyl disulfide and diallyl trisulfide may serve as potential alternatives for grain protectants since both of them can be prepared easily from readily available chemicals.     

Diallyl sulfide content and antimicrobial activity against food-borne pathogenic bacteria of chives (Allium schoenoprasum)

Chives, a member of the Alliaceae family, have been used in food and medicine in Thailand for a long time. Diallyl sulfides (diallyl monosulfide, dially disulfide, diallyl trisulfide, and diallyl tetrasulfide) are believed to be responsible for the antimicrobial activity of plants in this family. In this study, chive oil was examined for its diallyl sulfide content and its antimicrobial activity against some strains of food-borne pathogenic bacteria. Chive oil had a very low concentration of diallyl monosulfide in comparison with the other diallyl sulfides. They inhibited all pathogenic bacteria used in this study with a different degree of inhibition. Chive oil was also shown to be able to inhibit Escherichia coli O157:H7 in a food model. This study is the first report describing not only the diallyl disulfide content of chive oil, but also its antimicrobial activity against food-borne pathogens in both a test tube and food model.     

Toxicity, feeding deterrence, and effect of activity of 1,8-cineole from Artemisia annua on progeny production of Tribolium castanaeum (Coleoptera: Tenebrionidae)

1,8-Cineole isolated from Artemisia annua was tested against Tribolium castaneum (Herbst) for contact toxicity, fumigant toxicity, and antifeedant activity. The adults of T. castaneum were more susceptible than larvae to both contact and fumigant toxicity of 1,8-cineole, and LD50 and LC50 values of 108.4 microg/mg body weight of adult insect and 1.52 mg/liter air were found, respectively. Furthermore, 14-d-old T. castaneum larvae were more tolerant than 16- and 18-d-old larvae and adults to the contact toxicity of 1,8-cineole, but the 16- and 18-d-old larvae have similar susceptibility. In contrast, all the larvae (14-18 d old) of T. castaneum were much more tolerant than the adults to the fumigant action, but larvae of different ages had similar susceptibility. The compound 1,8-ciineole applied to filter paper at a concentration of 3.22-16.10 mg/cm2 significantly (P < 0.05) reduced the hatching of T. castaneum eggs and the subsequent survival rate of the larvae. Adult emergence was also reduced by 1,8-cineole. Feeding deterrence of 81.9% was achieved in T. castateum adults by using a concentration of 121.9 mg/g food, whereas larvae showed 68.8% at the same concentration.     

Toxic and deterrent effects of two volatile oils against cowpea weevil,Callosobruchus chinensis (Coleoptera: Bruchidae)

The fumigant effect of Mentha piperita and Ocimum basilicum oils and their mixture against adults and eggs of Callosobruchus chinensis (L.) was evaluated. Bioassay experiments showed that Ocimum basilicum oil was significantly effective against adult and egg stage which was the least effective. However, in the fumigant toxicity experiments against adult stage, Ocimum basilicum oil at a dose of 1.0 μl/38.5 ml air caused 100% mortality (LC₅₀ = 1.88), the mixture of both oils at a dose of 6.0 μl/38.5 ml air caused 100% mortality (LC₅₀ = 10.3) and Mentha piperita oil at a dose of 80.0 μl/38.5 ml air caused 80.0% mortality (LC₅₀ = 41.224) during a one day exposure period. Regarding the oviposition deterrent activity, Ocimum basilicum oil achieved 100% oviposition deterrent (at a dose of 0.5 μl/38.5 ml air) followed by the mixture of Mentha piperita and Ocimum basilicum oils, which achieved 71.22% oviposition deterrent (at a dose of 1.0 μl/38.5 ml air) and Mentha piperita oil, which achieved 39.6% oviposition deterrent (at a dose of 5.0 μl/38.5 ml air). The essential oils and their mixtures studied here determined a significant decrease in the number of eggs hatched and in the emergence of adults. The eggs failed to hatch on using Ocimum basilicum oil at a dose of 0.6 μl/38.5 ml air. However, the number of eggs hatched decreased to 7.4 on using a mixture of oils at a dose of 2.0 μl/38.5 ml air and 14.0 with Mentha piperita oil at a dose of 80.0 μl/38.5 ml air compared with 25.0 eggs hatched in the control experiments. On the other hand, the percentage reduction in emerging adults were 100% for Ocimum basilicum oil, 90.9% for a mixture of the two oils and 72.7% for Mentha piperita oil. Results showed that Ocimum basilicum oil and Ocimum basilicum oils plus Mentha piperita oils in blend are potential alternatives to synthetic fumigants in the treatment of durable agricultural products. Successful adoption of plant oils in the protection of food commodities promises an eco-friendly option compatible with international biosafety regulations.     

Synthesis of radioiodine labeled dibenzyl disulfide for evaluation of tumor cell uptake

Benzyl 4-halobenzyl and ally benzyl disulfide were synthesized as diallyl disulfide analogues and their tumor growth inhibitory effects on the cancer cells (SNU C5 and MCF-7) were comparable to that of diallyl disulfide, indicating that the disulfide functional group was responsible for the tumor growth inhibitory effects. Cu(I)-assisted radioiodination of benzyl 4-bromobenzyl disulfide gave benzyl 4-[123I/125I]iodobenzyl disulfide in 30-40% radiochemical yield. The radiolabeled disulfide was taken up by the cancer cells in a time-dependent manner, and the uptake was inhibited by the pretreatment of S-methyl methanethiosulfonate (MMTS), phorone and diallyl disulfide. This study suggested that the radiolabeled dibenzyl disulfide was taken up by the cancer cells via thiol-disulfide exchange and retained inside the cells.     

Chemical constituents and medicinal properties of Allium species

Allium species, belonging to Alliaceae family, are among the oldest cultivated vegetables used as food. Garlic, onions, leeks and chives, which belong to this family, have been reported to have medicinal properties. The Allium species constituents have been shown to have antibacterial and antioxidant activities, and, in addition, other biological properties. These activities are related to their rich organosulfur compounds. These organosulfur compounds are believed to prevent the development of cancer, cardiovascular, neurological, diabetes, liver diseases as well as allergy and arthritis. There have also been reports on toxicities of these compounds. The major active compounds of Allium species includes, diallyl disulfide, diallyl trisulfide, diallyl sulfide, dipropyl disulfide, dipropyl trisulfide, 1-propenylpropyl disulfide, allyl methyl disulfide and dimethyl disulfide. The aim of this review is to focus on a variety of experimental and clinical reports on the effectiveness, toxicities and possible mechanisms of actions of the active compounds of garlic, onions, leek and chives.

     

Food mixing by generalist grasshoppers: Plant secondary compounds structure the pattern of feeding

Plant secondary compounds can prevent feeding by phytophagous insects or, if ingested, can be poisonous to them. Less attention has been paid to the additional effects they have on feeding behavior when they are only weakly deterrent or not deterrent at all. Experiments were carried out on the generalist grass-hopperSchistocerca americana. Individuals were presented either with two cakes of high-quality artificial food with a single deterrent compound added or with two cakes, each with a different added deterrent compound. The deterrents consisted of single plant secondary compounds that were either marginally or strongly deterrent. There were profound differences in feeding behavior between those individuals given identical and those given different cake types, including longer feeding bouts on single cakes when a choice of different cake types was available. The behavioral effects demonstrate that the presence of secondary compounds in one food can influence the patterns of feeding on other available foods and suggest that such chemicals could impact foraging activities in a complex manner.     

Bioactivities of the leaf essential oil of Curcuma longa (var. ch-66) on three species of stored-product beetles (Coleoptera)

Essential oil extracted from the leaves of turmeric, Curcuma longa L., was investigated for contact and fumigant toxicity and its effect on progeny production in three stored-product beetles, Rhyzopertha dominica F. (lesser grain borer), Sitophilus oryzae L. (rice weevil), and Tribolium castaneum Herbst (red flour beetle). Oviposition-deterrent and ovicidal actions of C. longa leaf oil were also evaluated against T. castaneum. The oil was insecticidal in both contact and fumigant toxicity assays. The adults of R. dominica were highly susceptible to contact action of C. longa leaf oil, with LD50 value of 36.71 microg/mg weight of insect, whereas in the fumigant assay, adults of S. oryzae were highly susceptible with LC50 value of 11.36 mg/liter air. Further, in T. castaneum, the C. longa oil reduced oviposition and egg hatching by 72 and 80%, respectively at the concentration of 5.2 mg/cm2. At the concentration of 40.5 mg/g food, the oil totally suppressed progeny production of all the three test insects. Nutritional indices indicate >81% antifeedant action of the oil against R. dominica, S. oryzae and T castaneum at the highest concentration tested.     

Impact of Spodoptera littoralis (Boisd.) and Agrotis ipsilon (Hufn.) larval frass on oviposition of conspecific insects

Ethanolic extracts of larval frass of the cotton leaf worm and the black cutworm were prepared and tested to deter the eggs lay of the adults of the same insects. Two different types of food were used for larval feeding. Extracts fractions were identified using gas chromatography–mass spectrometry analysis. High concentrations were more deterrent to oviposition than low. Extracted frass of fed larvae on semi-artificial diet was more effective than others fed on natural host. Sensitivity of the black cutworm adult females to the cotton leaf worm frass extract was clearly observed at high tested concentration of L_1–3 and L₄ frass extracts which resulted completely oviposition deterrent. Several fatty acids were identified qualitatively and quantitatively in frass extracts of different larval instars of both target insects. Type and quantity of fatty acid depends mainly on larval food source and larval instar, except palmitic acid which recorded at all larval instars and food sources. Oleic acid and ethyl 9-hexadecaenoic acid were found when semi-artificial diet used as a food source while myrisitic acid was observed only in extracted frass of fed larvae on castor oil leaves.     

Bioactivities of l-carvone, d-carvone, and dihydrocarvone toward three stored product beetles

The rice weevil, Sitophilus oryzae L., adults were highly susceptible by contact to l-carvone, d-carvone, and dihydrocarvone when compared with the lesser grain borer, Rhyzopertha dominica F., adults and red flour beetle, Tribolium castaneum (Herbst.). Adults of R. dominica were more susceptible than the other species to fumigant vapors of l-carvone, d-carvone, and dihydrocarvone. The three larval stages (14-, 16-, and 18-d-old) of T. castaneum progressively became more susceptible with age toward contact toxicity of three test compounds but in fumigant toxicity, 16-d-old larvae of T. castaneum were more susceptible to the three compounds. Comparison of contact and fumigant toxicity of the test compounds indicates that l-carvone and d-carvone possess 24 times more fumigant toxicity toward adults of R. dominica than its contact toxicity. Overall order of toxicity was l-carvone > d-carvone > dihydrocarvone. Egg hatching and subsequent larval and adult survival of T. castaneum were significantly reduced when the eggs of T. castaneum were treated with l-carvone, d-carvone, and dihydrocarvone. l-Carvone completely suppressed egg hatching at the concentration of 7.72 mg/cm2. Data on feeding-deterrent indices indicate the high potency of l-carvone as feeding-deterrent in order of S. oryzae adults > T. castaneum adults > R. dominica adults > T. castaneum larvae.     

Thiol‐disulfide organization in alliin lyase (alliinase) from garlic (Allium sativum)

Alliinase, an enzyme found in garlic, catalyzes the synthesis of the well-known chemically and therapeutically active compound allicin (diallyl thiosulfinate). The enzyme is a homodimeric glycoprotein that belongs to the fold-type I family of pyridoxal-5′-phosphate-dependent enzymes. There are 10 cysteine residues per alliinase monomer, eight of which form four disulfide bridges and two are free thiols. Cys368 and Cys376 form a S—S bridge located near the C-terminal and plays an important role in maintaining both the rigidity of the catalytic domain and the substrate-cofactor relative orientation. We demonstrated here that the chemical modification of allinase with the colored —SH reagent N-(4-dimethylamino-3,5-dinitrophenyl) maleimide yielded chromophore-bearing peptides and showed that the Cys220 and Cys350 thiol groups are accesible in solution. Moreover, electron paramagnetic resonance kinetic measurements using disulfide containing a stable nitroxyl biradical showed that the accessibilities of the two —SH groups in Cys220 and Cys350 differ. Neither enzyme activity nor protein structure (measured by circular dichroism) were affected by the chemical modification of the free thiols, indicating that alliinase activity does not require free —SH groups. This allowed the oriented conjugation of alliinase, via the —SH groups, with low- or high-molecular-weight molecules as we showed here. Modification of the alliinase thiols with biotin and their subsequent binding to immobilized streptavidin enabled the efficient enzymatic production of allicin.     

Food discrimination and induction of preference for artificial diets in the tobacco hornworm, Manduca sexta

ABSTRACT. Feeding discrimination of Manduca larvae reared on leaves and a variety of artificial media was tested against leaves, various media, solvent fractions of leaves and media, and media components, including pure chemicals. Field-collected animals showed the same preferences as did animals cultured in the laboratory for many generations. Rearing larvae on a leaf species, an artificial diet, or homogenized leaves added to artificial diet induced a preference for that food in subsequent choice tests. Extracts of these foods using organic solvents (particularly hexane) elicited feeding choices resembling those evoked by the foods themselves: water extracts were effective as stimulants or deterrents, but responses to them differed considerably from responses to the foods. Animals raised on a defined artificial diet showed an increased preference (or decreased deterrence) for the lipid component (linolenic acid) of that diet in behavioural tests. Thus induction of preference was shown to be influenced by a specific nutrient compound. Fifteen artificial diets were tested; three were successful, including a completely defined medium. Various components of the diets were tested for feeding preferences, both as omissions from the main diet and as pure compounds. Some were stimulatory, most were neutral or slightly deterrent; a few were strongly deterrent. With the exception of linolenic acid, preferences for pure compounds were not significantly correlated with the food on which the animals were reared. The data suggest that food discrimination depends on the perception of a complex chemical message comprised of both polar and non-polar compounds, with the latter being of somewhat greater importance. The results are consistent with the explanation of the induction of feeding preferences being due to a reduced responsiveness to deterrents, to an increased preference for feeding stimulants or to both.     

Effect of diallyl disulfide on physiological performance of Ephestia kuehniella Zeller (Lepidoptera: Pyralidae)

Ephestia kuehniella Zeller is a key pest by severe deficiencies on quantity and quality of stored products. Due to concerns made by synthetic insecticides, use of natural products (like botanical insecticides) is of great importance as an alternative pest management. In the current study, the effect of diallyl disulfide was determined on digestive enzymes and those involved in intermediary metabolism of E. kuehniella. Concentration of 0.31% was obtained as LC₅₀ of diallyl disulfide followed by bioassay experiments. Activities of α-amylase, proteases, alanine aminotransferase, alkaline phosphatase and lactate dehydrogenase significantly decreased in the treated larvae, while the adverse effects were observed in case of triacylglyceride-lipase, aspartate aminotransferase, γ-glutamyl transferase, aldolase and acid phosphatase. Results confirmed that diallyl disulfide interrupted larval digestion by lower activities of digestive enzymes indicating significant effect on intermediary metabolism.     

Suppressive effect of exposure to Chenopodium album var. centrorubrum on the feeding response of the tortoise beetle, Cassida nebulosa, to spinach

Depending upon preceding dietary exposure, the tortoise beetle, Cassida nebulosa L. (Coleoptera: Chrysomelidae), exhibits various feeding responses to spinach, Spinacia oleracea L. (Chenopodiaceae). Adult tortoise beetles reared as adults on the main host Chenopodium album var. centrorubrum Makino (CAC) (Chenopodiaceae) eat little spinach, whereas those reared on spinach accept this plant as food. In this study, we investigated the suppressive effect of exposure to CAC on the feeding response of tortoise beetles to spinach. To investigate the effect of chemical factors in CAC, we provided beetles with Gomphrena globosa L. (Amaranthaceae) as the base food and exposed them to a CAC extract, because bioassays have indicated that exposure to G. globosa did not affect the feeding response of beetles to spinach. Adults reared on G. globosa treated with CAC extract consumed significantly less than adults reared on untreated G. globosa . Thus, we determined that exposure to CAC components is responsible for the suppressive effect on the feeding response to spinach observed in the beetles. In a choice feeding test with membrane filter discs, we found that adults reared on G. globosa treated with CAC extract exhibited a greater feeding deterrent response to the spinach deterrent than adults reared on untreated G. globosa leaves. Therefore, the suppression of spinach consumption due to exposure to CAC is probably caused by the development of a response to the feeding deterrents present in spinach.     

Ovipositional deterrent on mature stage of sweet pepper, Capsicum annuum, against Liriomyza trifolii (Burgess)

Liriomyza trifolii (Burgess), the American serpentine leafminer fly, is a well-known serious pest in the world. This insect species attacks plants of more than 21 families including Solanaceae plants. A sweet pepper, Capsicum annuum (Solanaceae), on mature stage, however, shows resistance to this leafminer fly. This resistance is based on the ovipositional deterrent in the sweet pepper leaf against the fly species. Based on the bioassay guided fractionation, phytol [(2E)-3,7,11,15-tetramethyl-2-hexadecen-1-ol] was isolated and identified as an ovipositional deterrent against this insect species. The yield of this compound was 815 microg/g fresh leaf of C. annuum. This compound completely deterred the females from laying their eggs on host plant leaves treated at 35.2 microg/cm2.     

印楝素乳油对黄曲条跳甲种群控制作用评价

在生命表方法分析的基础上。建立了害虫累积为害量指标,用以描述和比较某一时间段内害虫动态持续为害力大小．应用该指标分析印楝素乳油对黄曲条跳甲(Phyllotreta striolata(F.))种群控制作用,结果表明,对成虫显著的忌避作用是其对种群控制的主要机理．印楝素0．50ml·L^-1对成虫的忌避率为83．75％,种群趋势指数I值降为0．3354,干扰作用控制指数为0．0863．采用田间试验获得的结果,避免了室内测定的空间限制,因而是评价异源植物次生化合物对害虫忌避作用的主要依据．该指标在以害虫控制为目标的动态研究中具有较好的应用前景．     

Insecticidal activity of Artemisia sieberi,Eucalyptus camaldulensis,Thymus persicus and Eruca sativa oils against German cockroach,Blattella germanica (L.)

The German cockroach, Blattella germanica (L.) (Blattodea: Blattellidae), has been recognized as a serious health problem because of its potential to harbor and transmit human disease-causing pathogens. The development of new and environmentally friendly control agents is essential as alternative tool to conventional chemical insecticides. The aim of the current study was to determine the fumigant and contact toxicity of essential oils (EOs), Artemisia sieberi Besser, Eucalyptus camaldulensis Dehn. and Thymus persicus (Ronniger ex Rech. f.) and cold press oil of Eruca sativa (Miller) (ESO) against first instar nymphs and adults. According to results, the LC₅₀ values of EOs were between 15.01 and 28.80 μl/l air for fumigant toxicity, 2202.8 and 5600.2 mg/l for dipping toxicity and 9484.2 and 34,421.1 mg/l for applied surface toxicity. Fumigant activity of the oils were significantly more toxic than other methods. In general, the toxicity of A. sieberi was higher than other EOs. Also, the first instar nymphs were more susceptible than adults. Additionally, the results showed that the EOs were significantly more toxic than ESO in both fumigant and dipping methods. In addition to contact effects, the fumigant activity of ESO was proved, although the toxicity was lower than the EOs. Hence, it is important to consider the production cost benefit of ESO in comparison with the EOs. So, the ESO could be considered as a good candidate in control of the cockroach but in enclosed spaces such as inaccessible cockroach hiding places, the EO of A. sieberi can be useful.     

Investigating antibacterial effects of garlic (Allium sativum) concentrate and garlic-derived organosulfur compounds on Campylobacter jejuni by using Fourier transform infrared spectroscopy, Raman spectroscopy, and electron microscopy

Fourier transform infrared (FT-IR) spectroscopy and Raman spectroscopy were used to study the cell injury and inactivation of Campylobacter jejuni from exposure to antioxidants from garlic. C. jejuni was treated with various concentrations of garlic concentrate and garlic-derived organosulfur compounds in growth media and saline at 4, 22, and 35°C. The antimicrobial activities of the diallyl sulfides increased with the number of sulfur atoms (diallyl sulfide < diallyl disulfide < diallyl trisulfide). FT-IR spectroscopy confirmed that organosulfur compounds are responsible for the substantial antimicrobial activity of garlic, much greater than those of garlic phenolic compounds, as indicated by changes in the spectral features of proteins, lipids, and polysaccharides in the bacterial cell membranes. Confocal Raman microscopy (532-nm-gold-particle substrate) and Raman mapping of a single bacterium confirmed the intracellular uptake of sulfur and phenolic components. Scanning electron microscopy (SEM) and transmission electron microscopy (TEM) were employed to verify cell damage. Principal-component analysis (PCA), discriminant function analysis (DFA), and soft independent modeling of class analogs (SIMCA) were performed, and results were cross validated to differentiate bacteria based upon the degree of cell injury. Partial least-squares regression (PLSR) was employed to quantify and predict actual numbers of healthy and injured bacterial cells remaining following treatment. PLSR-based loading plots were investigated to further verify the changes in the cell membrane of C. jejuni treated with organosulfur compounds. We demonstrated that bacterial injury and inactivation could be accurately investigated by complementary infrared and Raman spectroscopies using a chemical-based, "whole-organism fingerprint" with the aid of chemometrics and electron microscopy.