Subchronic (90-day) toxicity assessment of Somacy-FP100, a lipopolysaccharide-containing fermented wheat flour extract from Pantoea agglomerans.

Pantoea agglomerans is a Gram-negative bacterium that is ubiquitous in the environment, colonizing animals, humans, and numerous plants, including cotton and wheat. A lipopolysaccharide-containing fermented wheat flour extract from P. agglomerans (Somacy-FP100) is proposed for use as a food ingredient for individuals seeking foods for healthy aging. Previously published genotoxicity studies with Somacy-FP100 reported its lack of genotoxicity in vitro, but a subchronic toxicity study has not yet been performed. Therefore, to demonstrate the safety of Somacy-FP100 for use as a food ingredient, a 90-day oral (gavage) toxicity study in rats was conducted. Male and female Han Wistar rats were administered vehicle (control) or Somacy-FP100 at 500, 1500, or 4500 mg/kg body weight/day at a dose volume of 10 mL/kg body weight, for at least 90 days. No test article-related adverse clinical signs or effects on body weight, food consumption, or clinical pathology were observed, and there were no macroscopic or microscopic findings related to the test article. Therefore, 4500 mg/kg body weight/day (the highest dose tested and highest feasible dose) was established as the no-observed-adverse-effect level. This absence of subchronic toxicity, in addition to the previously reported lack of genotoxicity, demonstrates the safety of Somacy-FP100 for use as a food ingredient.

Comparing six mathematical link function models of the antifeedant activity of lesser grain borer exposed to sub-lethal concentrations of some extracts from calotropis procera.

In the present study, Probit, Cauchy Fractional and three types of Log methods, i.e., Logit, Log-log, and Complementary log-log were employed to model the feeding deterrence of the lesser grain borer, Rhyzopertha dominica (F) (Coleoptera: Bostrichidae), when fed latex protein, crude flavonoid fraction, 3-O-rutinosides of quercetin, kaempferol and isorhamnetin, isolated from Calotropis procera (Ait.) (Gentianales: Asclepiadaceae). A nutritional study with treated flour discs at sub-lethal concentrations indicated that the tested natural products negatively affected the feeding behavior of the lesser grain borer, causing high feeding deterrent indices. Our results assure that Probit, Logit and Clog-log model the feeding deterrent indices with high goodness of fit. The models aim to support the management of the test insect when fed grains treated with sub-lethal doses of the tested phytochemicals in order to develop a viable, precise and long-term strategy to minimize the excessive reliance on the chemical pesticides currently in use.

Biological activity of Dittrichia viscosa (L.) Greuter extracts against adult Sitophilus granarius (L.) (Coleoptera, Curculionidae) and identification of active compounds.

Dittrichia viscosa (L.) Greuter, a perennial weed of the Mediterranean area, was reported to be source of active substances. Here, by means of both ingestion and contact assays, the biological activity of three different extracts (n-hexane, methanol, and distilled water) of D. viscosa aerial part has been evaluated against Sitophilus granarius (L.) adults, an important pest of stored grains. Ingestion assays showed negligible mortality and food deterrence for all the extracts, whereas only a slight reduction of some nutritional parameters (relative growth rate, relative consumption rate, food efficiency conversion) was recorded for water extract. High contact toxicity was found only for the n-hexane extract (24 h median lethal dose LD50 = 53.20 µg/adult). This extract was further subfractioned by silica gel column chromatography and then by thin layer chromatography. Further contact toxicity bioassays highlighted two active subfractions which were analyzed by GC-MS. This revealed the occurrence, in both subfractions, of two major peaks that were identified as α- and γ- costic acid isomers. Moreover, D. viscosa active subfractions, did not cause acetylcholinesterase (AChE) inhibition; therefore, in the light of progressive limitation of compounds acting by this mechanism of action, D. viscosa represents a promising eco-sustainable source of natural products for pest control.

Toxicological effect of underutilized plant, Cleistanthus collinus leaf extracts against two major stored grain pests, the rice weevil, Sitophilus oryzae and red flour beetle, Tribolium castaneum.

Toxicity and repellency activity of Cleistanthus collinus (Roxb.) (CC) leaf extracts were evaluated against rice weevil, Sitophilus oryzae (L.) and red flour beetle, Tribolium castaneum (Herbst) under laboratory condition. Five concentration(s) (1%, 1.5%, 2%, 2.5% and 4%) with two controls (acetone and water) treatments along with deltamethrin were used for direct and contact residual toxicity. The insect that survived after CC exposure were transferred to an untreated feeding substrate and the population buildup of subsequent two generation were recorded after 30 (F1) and 60 days (F2). In the contact residual toxicity, highest CC concentration (4%) produced 75% mortality in S. oryzae and 62.5% mortality in T. castaneum during 7 days of exposure, whereas in direct toxicity the mortality were 81% and 58% respectively, for S. oryzae and T. castaneum. The long term effect of CC was apparent in both the insect species, where F2 populations were significantly decreased in the CC treatments. CC treatment at 4% produced similar adult mortality in comparison to deltamethrin at 1%. In addition, repellent activity of CC extracts was observed against both S. oryzae and T. castaneum. This is the first step towards assessing the scientific basis for the understanding the effectiveness of CC extracts against stored grain pests and it could be a viable eco-friendly option for stored grain insect pest management.

Typical Monoterpenes as Insecticides and Repellents against Stored Grain Pests.

Five monoterpenes naturally occurring in essential oils were tested for their insecticidal and repellent activities against the bruchid beetle Callosobruchus maculatus and the maize weevil Sitophilus zeamais. The monoterpenes were highly efficient as inducers of mortality or repellency against both insect species. They were more efficient in their fumigant activity against C. maculatus than against S. zeamais, while this profile of action was inverted when considering the repellent activities. Eugenol was one the most effective fumigants against both insects and one the most effective repellent against C. maculatus, while citronellal and geranial were one the most effective repellents against S. zeamais. Functional and positional isomerism of the monoterpenes pairs appears to exert little or no influence on theirs effects, especially in case of repellency. The validation of the insecticidal/repellent efficacy of isolated monoterpenes may permit a more advantageous, rapid, economic and optimized approach to the identification of promising oils for commercial formulations when combined with ethnobotanical strategies.

Acaricidal and Insecticidal Activities of Essential Oils against a Stored-Food Mite and Stored-Grain Insects.

Twenty plant-derived oils were evaluated for their acaricidal and insecticidal activities against Sitotroga cerealella, Sitophilus oryzae, Sitophilus zeamais, and Tyrophagus putrescentiae adults, by using the fumigant and filter paper diffusion methods. Responses varied with bioassay systems, insect or mite species, plant oils, and exposure time. Based on the 50% lethal dose (LD50) values against S. oryzae and S. zeamais in the fumigant bioassay, Anethum graveolens oil (4.12 and 1.12 µg/cm(3), respectively) induced the highest mortality, followed by Achillea millefolium (21.92 and 14.91 µg/cm(3)) and Eucalyptus dives (28.02 and 24.02 µg/cm(3)) oils, respectively. The most toxic oil based on the 50% lethal concentration values against T. putrescentiae was E. dives (3.13 µg/cm(3)), followed by Melaleuca leucadendron (3.93 µg/cm(3)) and Leptospermum pertersonii (4.41 µg/cm(3)). Neroli birgard oil (1.70 µg/cm(3)) was the most toxic based on the LD50 values against S. cerealella, followed by Citrus aurantium (1.80 µg/cm(3)) and Artemisia vulgaris (1.81 µg/cm(3)). The insecticidal and acaricidal activities of the plant oils in the filter paper diffusion bioassay were similar to those in the fumigant bioassay. In comparison, A. millefolium, A. graveolens, and E. dives oils were more effective against S. oryzae and S. zeamais in the fumigant bioassay than in the contact bioassay. These results indicate that the insecticidal activity of the three plant oils against S. oryzae and S. zeamais may be due to their fumigant action. Acaricidal activities of the A. millefolium, A. graveolens, and E. dives oils against T. putrescentiae were 2.62, 1.11, and 122 times higher than that of benzyl benzoate in the contact bioassay. These results indicate that A. millefolium, A. graveolens, and E. dives oils have potential for development as agents to control stored-grain insects and mites.

Bioactivity and chemical constituents of the essential oil from Dendranthema indicum (L.) Des Moul. against two stored insects.

Dendranthema indicum (L.) Des Moul. is a perennial herb commonly used as a traditional Chinese medicine for a long time. In this work, we took Dendranthema indicum as a target plant and two stored insects which include Tribolium castaneum and Stegobium paniceum adults as target insects. Essential oil obtained from Dendranthema indicum was analyzed by gas chromatography-mass spectrometry (GC-MS). A total of 31 components representing 92.44% of the oil were identified and the main compounds were found to be chamazulene (15.93%), ß-caryophyllene (13.78%), germacrene D (9.11%), and b-cis-farnesene (6.59%). With a further isolation, three constituents were obtained from the essential oil and identified as chamazulene, ß-caryophyllene and eucalyptol. Significantly, in the progress of assay, it showed that the essential oil and chamazulene exhibited stronger insecticidal and repellent activities against Stegobium paniceum than Tribolium castaneum. The results indicate that additional to its traditional use as Chinese medicinal herb, the essential oil of Dendranthema indicum aerial parts and isolated compounds have potential to be developed into natural insecticides or repellents for control of insects in stored grains.

Can volatile organic metabolites be used to simultaneously assess microbial and mite contamination level in cereal grains and coffee beans?

A novel approach based on headspace solid-phase microextraction (HS-SPME) combined with comprehensive two-dimensional gas chromatography-time-of-flight mass spectrometry (GC×GC-ToFMS) was developed for the simultaneous screening of microbial and mite contamination level in cereals and coffee beans. The proposed approach emerges as a powerful tool for the rapid assessment of the microbial contamination level (ca. 70 min versus ca. 72 to 120 h for bacteria and fungi, respectively, using conventional plate counts), and mite contamination (ca. 70 min versus ca. 24 h). A full-factorial design was performed for optimization of the SPME experimental parameters. The methodology was applied to three types of rice (rough, brown, and white rice), oat, wheat, and green and roasted coffee beans. Simultaneously, microbiological analysis of the samples (total aerobic microorganisms, moulds, and yeasts) was performed by conventional plate counts. A set of 54 volatile markers was selected among all the compounds detected by GC×GC-ToFMS. Principal Component Analysis (PCA) was applied in order to establish a relationship between potential volatile markers and the level of microbial contamination. Methylbenzene, 3-octanone, 2-nonanone, 2-methyl-3-pentanol, 1-octen-3-ol, and 2-hexanone were associated to samples with higher microbial contamination level, especially in rough rice. Moreover, oat exhibited a high GC peak area of 2-hydroxy-6-methylbenzaldehyde, a sexual and alarm pheromone for adult mites, which in the other matrices appeared as a trace component. The number of mites detected in oat grains was correlated to the GC peak area of the pheromone. The HS-SPME/GC×GC-ToFMS methodology can be regarded as the basis for the development of a rapid and versatile method that can be applied in industry to the simultaneous assessment the level of microbiological contamination and for detection of mites in cereals grains and coffee beans.

Composition and toxic, repellent and feeding deterrent activity of essential oils against the stored-grain pests Tribolium castaneum (Coleoptera: Tenebrionidae) and Sitophilus oryzae (Coleoptera: Curculionidae).

BACKGROUND: The composition and bioactivity of essential oils from Tagetes terniflora Kunth, Cymbopogon citratus Stapf. and Elyonurus muticus (Spreng) Kuntz were evaluated against stored-grain pests. RESULTS: Fumigant and contact toxicities were observed with T. terniflora on adults of both pests. In contact toxicity, this oil was less toxic to Tribolium castaneum (Herbst). Essential oils from C. citratus and E. muticus showed contact toxicity on S. oryzae. All essential oils produced: (a) repellency on larvae and adults of T. castaneum and adults of Sitophilus oryzae (L.); (b) post-ingestive toxicity on T. castaneum larvae and S. oryzae adults and alteration of nutritional index on T. castaneum and S. oryzae adults. Cymbopogon citratus reduced the relative growth rate and the efficiency of conversion of ingested food in T. castaneum larvae. Tagetes terniflora produced a feeding stimulant effect in T. castaneum adults. In addition, they had a feeding deterrent action against S. oryzae adults. The composition of essential oils from C. citratus and E. muticus varied only in one component. CONCLUSION: These results showed that the essential oils from T. terniflora, C. citratus and E. muticus should be studied further for their use in integrated pest management programmes for T. castaneum and S. oryzae control.

Toxicity of the essential oil of Illicium difengpi stem bark and its constituent compounds towards two grain storage insects.

During our screening program for new agrochemicals from Chinese medicinal herbs, the essential oil of Illicium difengpi stem bark was found to possess strong insecticidal activities against the maize weevil, Sitophilus zeamais (Motschulsky) (Coleoptera: Curculionidae) and red flour beetle, Tribolium castaneum Herbst (Coleoptera: Tenebrionidae). A total of 37 components of the essential oil of I. difengpi were identified. The main components of the essential oil were safrole (23.61%), linalool (12.93%), and germacrene D (5.35%). Bioactivities-directed chromatographic separation on repeated silica gel columns led to the isolation of two compounds: safrole and linalool. Safrole showed pronounced contact toxicity against both insect species and (LD50 = 8.54 for S. zeamais; 4.67 µg/adult for T. castaneum) and was more toxic than linalool (LD50 = 24.88 for S. zeamais; 8.12 µg/adult for T. castaneum). The essential oil acting against the two species of insects showed LD50 values of 13.83 and 6.33 µg/adult, respectively. Linalool also possessed strong fumigant toxicity against both insect species (LC50 = 10.02 for S. zeamais; 9.34 mg/L for T. castaneum) and was more toxic than safrole (LD50 = 32.96 and 38.25 mg/L), while the crude essential oil acting against the two species of insects showed LC50 values of 14.62 and 16.22 mg/L, respectively. These results suggest that the essential oil of I. difengpi stem bark and the two compounds may be used in grain storage to combat insect pests.

Biological activity of ethanolic extract fractions of Dracaena arborea against infestation of stored grains by two storage insect pests.

As part of on-going efforts to use eco-friendly alternatives to chemical pesticides, ethanolic extract of dried leaves of Dracaena arborea (Willd.) Link (Dragon tree; Dracaenaceae) dissolved in distilled water and partitioned between equal volumes of n-hexane, chloroform, ethyl acetate and butanol was assessed in the laboratory against infestation by Sitophillus zeamais Motsch. and Callosobruchus maculatus Walp. in stored maize and cowpea, respectively. One hundred grams each of maize grains and cowpea seeds were treated with 400 mg kg(-1) of each extract fraction to evaluate contact toxicity, damage assessment, effect on eggs and immature stages and progeny production in both insect species. Contact toxicity by topical application, toxicity upon filter paper application and repellency using area preference method were carried out on the two insect species. Results showed that the extract fraction caused significant (p < or = 0.05) mortality of both insect pests with a high residual contact activity against S. zeamais. Grain damage was significantly (p < or = 0.01) reduced, while progeny production and development of eggs within grains were inhibited. The extract fractions evoked a strong repellent action against S. zeamais but moderate action against C. maculatus. The full potentials of using extract fractions of D. arborea as grain protectant against infestation by insect pests is discussed.

Reproduction and feeding behavior of Myzus persicae on four cereals.

Green peach aphid, Myzus persicae (Sulzer), does not overwinter outdoors in Minnesota; it arrives each spring on low-level jet streams from the south. After arrival, anholocylic reproduction occurs on numerous herbaceous species, including many common weeds, before movement to potato, Solanum tuberosum L. In investigating aphid feeding behavior on barrier crops, we observed winter wheat, Triticum aestivum L., colonized by green peach aphid. The Northern Great Plains grows 94,000 ha of potatoes and 20.5 million ha of small grain cereals each year, the latter potentially providing an early emerging and widely distributed green peach aphid host to influence early season potato colonization. Life tables statistics indicated green peach aphid had its highest reproductive potential among cereals on winter wheat, with rye (Secale cereale L.) > barley (Hordeum vulgare L.) > oats (Avena sativa L.). Green peach aphid was found to colonize barley, rye, and winter wheat, but not oats. Mean generation time, net reproductive rate, doubling time, and finite rate of increase were significantly different between host plants. Electrical penetration graph technique indicated mean nonpenetration duration by green peach aphid was significantly different among plant species, and significantly longer on winter wheat than on the other cereals. Mean xylem phase duration was not significantly different among plant species but sieve element salivation was of longest duration on potato. Phloem sap ingestion (E2) was also significantly different among plant species with longest E2 duration on winter wheat. This study demonstrates that this aphid can effectively use key cereals at the vegetative stage.

Effect of volatile constituents from Securidaca longepedunculata on insect pests of stored grain.

Securidaca longepedunculata Fers (Polygalaceae) is commonly used as a traditional medicine in many parts of Africa as well as against a number of invertebrate pests, including insects infesting stored grain. The present study showed that S. longepedunculata root powder, its methanol extract, and the main volatile component, methyl salicylate, exhibit repellent and toxic properties to Sitophilus zeamais adults. Adult S. zeamais that were given a choice between untreated maize and maize treated with root powder, extract, or synthetic methyl salicylate in a four-way choice olfactometer significantly preferred the control maize. Methyl salicylate vapor also had a dose-dependant fumigant effect against S. zeamais, Rhyzopertha dominica, and Prostephanus truncates, with a LD100 achieved with a 60microl dose in a 1-l container against all three insect species after 24 hr of exposure. Probit analyses estimated LD50 values between 34 and 36 microl (95% CI) for all insect species. Furthermore, prolonged exposure for 6 days showed that lower amounts (30 microl) of methyl salicylate vapor were able to induce 100% adult mortality of the three insect species. The implications are discussed in the context of improving stored product pest control by small-scale subsistence farmers in Africa.

Potential use of essential oils from Cameroon applied as fumigant or contact insecticides against Sitophilus zeamais Motsch. (Coleoptera: Curculionidae).

Essential oils from seven plants species currently found in Cameroon were extracted by steam distillation and tested for their insecticidal activities against Sitophilus zeamats Motsch. Responses varied with the test applied and the plant species. For the contact toxicity, the acetone was used in order to dilute the pure essential oil. Formulations of 1% of essential oils of Xylopia aethiopica and Ocimum gratissimum were the most toxic and led to 96 and 98% of mortality respectively after 24 h. There was no death in control (0% of mortality). Fumigation test were done by applying 300 microl of pure essential oils in the bottom of closed 800 ml glass containers. In that case, the essential oil of Hyptis spicigera was the most toxic (fumigant) after 48 hours of fumigation followed by Annona seregalensis and Xylopia aethiopica 96 and 95% of mortality respectively. Some of the essential oils of plant species tested are promising for pest control in farmer granaries. However, further investigations are to be done on formulation and side effects.

Effect of plant a-amylase inhibitors on sunn pest, Eurygaster integriceps Puton (Hemiptera: Scutelleridae), alpha-amylase activity.

Plant-insect interaction is a dynamic system, subjected to continual variation and change. In order to reduce insect attack, plants developed different defence mechanisms including chemical and physical barriers such as the induction of defensive proteins, volatiles that attract predators of the insect herbivores and secondary metabolites. Proteinaceous inhibitors of alpha-amylase and proteases are widely distributed in cereals, legumes and some other plants. Because of the possible importance of these inhibitors in plant physiology and animal nutrition, extensive research has been conducted on their properties and biological effects. Sunn pest like other insect pests of wheat lives on a polysaccharide-rich diet and depends to a large extent on effectiveness of their alpha-amylases for survival, a-amylase (1-4-alpha-D-glucan glucanohydrolase) hydrolyses starch, and related polysaccharides by randomly cleaving internal alpha-1,4-glucosidic linkages and has a major role in the utilization of polysaccharides. The enzyme inhibitors act on key insect gut digestive hydrolyses, alpha-amylase. Several kinds of a-amylase inhibitors present in seeds and vegetative organs of plant, act to regulate number of phytophagous insects. Therefore, the aim of the current study is to study cereal proteinaceous inhibitors of insect digestive enzymes and their potential use as resistance factors against Sunn pest. The proteinaceous inhibitors from different cereal species including barley (Hordeum vulgare L.) and wheat (Triticum aestivum L.) were extracted and tested in in vitro condition against Sunn pest alpha-amylase. Extraction was made with NaCl (0.15 M) at room temperature and further purification was done by ammonium sulphate precipitation. It was found that fractions obtained from barley had more inhibitory effect on amylase activity of Sunn pest than fractions obtained from wheat. Knowledge gained through these studies can be used to select resistant plant against insect pest.

Laboratory evaluation of Melia azedarach L. and Eucalyptus globulus Labill. Extracts in order to control Ocneridia volxemi Bolivar (Orthoptera, Pamphaginae) hoppers.

This study was carried out to evaluate the effect of Melia azedarach L. (Meliaceae) dry fruit extracts (oil and water) and Eucalyptus globulus Labill. (Myrtaceae) dry leaves extracts (water) on Oneridia volxemi hoppers (fifth-instar) feeding on cereals leaves (Triticum durum) under laboratory conditions. Larva feeding reduced and mortality was significatly (p < 0.05) on cereals leaves sprayed with three concentrations 0, 5, 1 and 2% of oil extracted from Melia azederach. In addition, O. volxemi feeding and mortality was significantly (p < 0.05) on cereals leaves sprayed with two concentrations 50 g/l and 80 g/l of water extracts from Melia azedarach dry fruits soaked for 24 hours. the water extract solution of 80 g/l significantly reduced feeding than the other concentrations 50 and 25 g/l. This study showed also the water extract solution from Eucalyptus globulus dry leaves reduced also feeding and induced the mortality of O. volxemi but not better than Melia azedarach dry fruit extracts water at the same concentration (80 g/l). The aim of this study is in an integrated management program for control of O.

Identification and in situ and in vitro characterization of secreted proteins produced by plant-parasitic nematodes.

Secretions of plant-parasitic nematodes which are released into plant tissue may play critical roles in plant-nematode interactions. The identification and characterization of these molecules are of fundamental importance and may help to facilitate the development of novel strategies to interfere with nematode infection of plants and thereby decrease nematode-induced damage to crops. An antibody-based approach was used to isolate molecules present on the nematode surface and in nematode secretions. Monoclonal antibodies (MAbs) were produced to secretions and to whole Heterodera avenae 2nd-stage juveniles; several of these MAbs recognized molecules present in nematode secretions produced in vitro. Three of these molecules have been partly characterized in H. avenae, Globodera rostochiensis, G. pallida and Meloidogyne incognita. A MAb reacting with the surfaces of these nematodes recognized antigens of different molecular weight in each of the species tested. This difference in antigenicity might be related to specific functions in these nematodes. Preliminary results show that this antibody also localized the antigen in root cells surrounding the feeding site induced by M. incognita in Arabidopsis thaliana.