Legume type and temperature effects on the toxicity of insecticide to the genus Callosobruchus (Coleoptera: Bruchidae)

Three bruchid pest species, Callosobruchus maculatus, Callosobruchus chinensis and Callosobruchus rhodesianus, were studied for their response to insecticide toxicity taking into account the separate and interactive effects of temperature and pre-adult food. The food types used were cowpea (Vigna unguiculata) and mungbean (Vigna radiata). Callosobruchus maculatus was the most tolerant to malathion and the least affected by temperature change while C. rhodesianus was the least tolerant. Over a 4 °C range (23°, 25°, 27 °C), there was generally a significant impact of temperature on the tolerance of the three species to the insecticide. The food type on which the insects developed influenced considerably the degree of insecticide tolerance. Callosobruchus maculatus and C. chinensis populations reared on mungbean had higher tolerance to malathion than their counterparts reared on cowpea, but the opposite was observed in C. rhodesianus populations. The food influence in this study suggested an ancestral cause or fitness cost depending on the species. The interaction of food-by-temperature had no significant effect on malathion toxicity to this genus. Correlation analysis showed C. chinensis to be relatively less sensitive to insecticide concentration over the range studied compared with the other two species.     

Deterrent and insecticidal properties of bean seed (Phaseolus vulgaris L.) whole meal or protein extract incorporated into the diet of Callosobruchus maculatus (F.) (Coleoptera: Bruchidae)

Callosobruchus maculatus, a pest that causes serious damage to chickpea Cicer arietinum, cannot develop in the seeds of Phaseolus or Vigna spp. which contain lectins. The insecticidal activity towards C. maculatus in these seeds is attributed both to lectins with specific affinity to N-acetylglucosamine, the major component of insect chitin, and to alpha-amylase inhibitors (lectin-like proteins). The insecticidal properties of bean meal or bean protein extracts from different sources towards different pest species are variable and need to be experimentally evaluated. The main objective of this study was to determine through a feeding trial on artificial chickpea seed the potential of bean seed meal from a wild bean Vigna caracalla, four varieties of Phaseolus vulgaris, and of a protein extract of P. vulgaris seed, to alter different life history traits of C. maculatus. The chickpea weevil was set up on artificial chickpea seeds containing different amounts of bean meal to observe the effects on female oviposition, percentage of development to adulthood and juvenile development time. These traits were combined in a composite index measuring the alteration of the multiplication rate of C. maculatus fed on artificial seed. The activity of lectin-like extracts was observed on chickpea artificial seed spiked with bean seed extract. Incorporation of bean flour at a rate of 10 and 20% in chickpea artificial seed significantly decreased C. maculatus female fecundity, percentage of adult emergence, and greatly increased the development time. Feeding trials with protein extracts of P. vulgaris reduced fecundity and survival of C. maculatus. Incorporation of 10% V. caracalla bean seed meal in chickpea artificial seed, reduced the multiplication potential of C. maculatus by over 90% showing that bean seed lectin extracts are worthy of further investigation for post-harvest infestation control.     

Insecticidal activities of constituents identified in the essential oil from leaves of Chamaecyparis obtusa against Callosobruchus chinensis (L.) and Sitophilus oryzae (L.)

Analysis by gas chromatography-mass spectrometry led to the identification of 20 volatiles from the steam distilled oil of the leaves from Chamaecyparis obtusa Siebold et Zuccarni. Seven constituents of the oil were tested for contact and fumigant activity against adults of Callosobruchus chinensis (L.) and Sitophilus oryzae (L.). Responses varied with compound and dose rather than increasing exposure time beyond 1 or 2 days. In the impregnated-paper test with the constituents [bornyl acetate, (+)-limonene, myrcene, α-phellandrene, α-pinene, sabinene and terpinolene], at 0.1 mg/cm², bornyl acetate, α-phellandrene and terpinolene caused 97%, 97% and 87% mortality, respectively, against C. chinensis adults 1 day after treatment. At 0.05 mg/cm², moderate activity was achieved with bornyl acetate (56% mortality), α-phellandrene (75%) and terpinolene (55%). Against S. oryzae adults, at 0.26 mg/cm², terpinolene caused 93% mortality 2 days after treatment, whereas 80% mortality 4 days after treatment was observed with bornyl acetate and α-phellandrene. The toxicity of these compounds was significantly decreased at 0.18 mg/cm². In a fumigation test with S. oryzae adults, bornyl acetate and terpinolene were much more effective in sealed containers than in open ones, indicating that the insecticidal activity of these compounds was largely attributable to fumigant action. These naturally occurring materials derived from leaves of Chamaecyparis could be useful for managing populations of C. chinensis and S. oryzae.     

Characterization of Callosobruchus chinensis (L.) resistance in Vigna umbellata (Thunb.) Ohwi & Ohashi

Resistance to azuki bean weevil, Callosobruchus chinensis, was studied in a series of field and laboratory experiments in two accessions of rice bean (Vigna umbellata (Thunb.) Ohwi & Ohashi), one accession of black gram (V. mungo (L.) Hepper), and one accession of mungbean, (V. radiata (L.) Wilczek). Weevil damage to immature pods of the rice bean accessions, ‘Menaga’ and ‘Miyazaki’, was significantly less than to the susceptible mungbean, VC1973A. In mature pods, the pest damage to the pod wall of Menaga was significantly higher than to VC1973A, whereas the damage to Miyazaki was similar to VC1973A. Seeds within the pods of both rice bean accessions were resistant no matter when the pods were harvested. When the insects were exposed directly on dry seeds, both rice bean accessions and a black gram accession VM2164 were resistant to them. In artificial seeds made by mixing flour of the individual resistant Vigna accessions with VC1973A and subsequently exposed to bruchid oviposition, the higher the quantity of resistant Vigna flour the lower the number of bruchids that emerged from such seeds. No bruchids emerged from artificial seeds containing crude starch fraction from the three resistant Vigna accessions when such seeds were exposed to bruchid infestation, whereas many insects emerged from the seeds containing starch of VC1973A or flour of VC1973A alone. In artificial seeds made by mixing crude protein fractions of the three resistant Vigna accessions with flour of VC1973A, as the concentration of protein increased the number of C. chinensis adults that emerged decreased. Fractionation of crude proteins into acetone-precipitable proteins and peptide and amino acid portions resulted in the loss of antibiosis effect. Artificial seeds made from purified starch-polysaccharides fraction, however, exhibited antibiosis effects if prepared from the rice bean seed of Menaga and Miyazaki but not if made from the black gram seed, VM2164.     

Comparative study of field and laboratory evaluations of the ethnobotanical Cassia sophera L. (Leguminosae) for bioactivity against the storage pests Callosobruchus maculatus (F.) (Coleoptera: Bruchidae) and Sitophilus oryzae (L.) (Coleoptera: Curculionidae)

The powdered leaves of Cassia sophera along with hot- and cold-water leaf extracts of this plant were tested in laboratory experiments in the UK and in field trials in Tamale, Northern Ghana, using traditional storage containers, to determine their inhibitory and toxic effects against Sitophilus oryzae and Callosobruchus maculatus infestation of stored rice and cowpea, respectively. Laboratory and field experiments with cowpea showed that the use of C. sophera hot-water extracts was more effective at reducing C. maculatus infestation and adult emergence on cowpea than the traditional leaf-powder application (1% and 5% w/w) or the use of a cold-water extract of C. sophera. Hot-water extracts of C. sophera might be a more effective technique of applying the plant material on to stored cowpea than using powdered C. sophera leaves, the currently used application by small-scale farmers. In contrast, experiments with S. oryzae on rice showed that C. sophera leaf powder (5% w/w) effectively reduced adult emergence in the laboratory, but this could not be confirmed under field conditions. The hot and dry climatic conditions in the field might impart a natural protection against rice infestation by S. oryzae, making the use of protectants and pesticides less necessary for farmers. This was supported by the negligible rice grain damage after 6 months of field storage and by the failure of the S. oryzae population to establish itself under field conditions. The implications of using botanicals in pest control are discussed.     

Analysis of the insecticidal activity of methylisothiocyanate on Callosobruchus maculatus (F.) (Coleoptera: Bruchidae) and its parasitoid Dinarmus basalis (Rondani) (Hymenoptera: Pteromalidae)

The bruchid Callosobruchus maculatus causes major losses during the storage of the seeds of Vigna unguiculata in West Africa. An endemic parasitoid, the pteromalid Dinarmus basalis is present in the stores and can reduce the increase in bruchid populations. African farmers often place in the stores the leaves of a shrub, Boscia senegalensis Lam.(Capparaceae), which release methylisothiocyanate (MITC). This compound is toxic to adult bruchids and could reduce seed losses but its influence on the bruchid natural enemy D. basalis was hitherto unknown. The susceptibility of C. maculatus and D. basalis to MITC was assessed under laboratory conditions. The susceptibility of adults of both species towards MITC were similar. After a 24 h exposure, the LC₅₀ was respectively 0.48 and 0.54 mg/l for C. maculatus and D. basalis. The ovicidal activity of this compound was high; the LC₅₀ of C. maculatus eggs was 0.04 mg/l after a 24 h exposure to MITC. High MITC concentrations only slightly affected the survival of C. maculatus during its post-embryonic development in the seeds of V. unguiculata. Gas chromatographic analysis demonstrated that 25-35% of the MITC present in the atmosphere of the experimental jars was absorbed by the seeds but concentrations inside the cotyledons were too low to influence the survival of the C. maculatus larvae. The D. basalis larvae developing at the expense of their host inside the larval galleries, were more affected by the treatment. The introduction of B. senegalensis releasing MITC in the storage systems could reduce the density of the parasitoid population and so increase the seed losses by permitting the development of the bruchid population.     

Automatic, rapid screening of seed resistance in cowpea, Vigna unguiculata (L.) Walpers, to the seed beetle Callosobruchus maculatus (F.) (Coleoptera: Bruchidae) using acoustic monitoring

The biomonitor technique was investigated as a rapid and automatic method for measuring the resistance of cowpea varieties to the seed beetle Callosobruchus maculatus. This technique measures the activity of internally feeding insect larvae by counting ultrasonic emissions produced as they feed. Activity throughout the development of C. maculatus larvae in known susceptible and resistant cowpea varieties was recorded. This showed details of the development of each larval instar, and showed clear differences between the resistant and susceptible cowpeas. A rapid method for comparing cowpeas was proposed in which the activity of larvae was recorded for 24 h starting 14 days after oviposition. Using this method, significant differences in activity were apparent between one susceptible and two resistant cowpea varieties. Further comparisons using a randomised block experimental design also showed a clear difference in activity between one susceptible and one known resistant cowpea variety. This experimental protocol took 21 days. It is suggested that the biomonitor offers a promising method for screening seeds for resistance to insect pests with a reduction in the time and effort required over conventional bioassay methods.     

Inhibition of Callosobruchus maculatus (F.) (Coleoptera: Bruchidae) development with essential oil extracted from Cymbopogon schoenanthus L. Spreng. (Poaceae), and the wasp Dinarmus basalis (Rondani) (Hymenoptera: Pteromalidae)

The control of the development of Callosobruchus maculatus was studied using a method that combined exposure to essential oil extracted from Cymbopogon schoenanthus and the introduction of a pteromalid natural enemy of the bruchid, Dinarmus basalis. The effect of the essential oil used was evaluated on all developmental stages of C. maculatus and on adults of D. basalis. At the highest concentration tested (33.3 μl/l) all adults of C. maculatus were killed within 24 h of exposure to the oil and the development of newly laid eggs and neonate larvae was also inhibited. However, the oil had variable efficacy against the bruchid instars developing inside the seeds: 5-day-old larvae (63% LI and 37% LII) of C. maculatus developing inside the seeds proving to be highly susceptible while 15-day-old insects (84% of pupae and 16% of larvae) were tolerant. Under the same conditions (33.3 μl/l), adults of D. basalis were very susceptible to oil vapours and to the residual activity of the oil after 3 or 6 days. However, the introduction of 10 pairs of adult D. basalis into a jar containing 100 hosts aged 10 days, 3 or 6 days before the oil application, gave respectively an emergence of 26 or 18 adults of the parasitoid compared to 28 in the control and there was no adult emergence of the host. The possibility of an integrated pest management strategy by using allelochemicals such as essential oils and indigenous natural enemies to control C. maculatus development in cowpea stocks is discussed.     

A laboratory assessment of the potential of different strains of the entomopathogenic fungi Beauveria bassiana (Balsamo) Vuillemin and Metarhizium anisopliae (Metschnikoff) to control Callosobruchus maculatus (F.) (Coleoptera: Bruchidae) in stored cowpea

Twelve indigenous and exotic isolates of Beauveria bassiana and Metarhizium anisopliae were evaluated for their virulence and their ability to suppress populations of Callosobruchus maculatus in stored cowpea. LT₅₀ values ranged from 3.11 to 6.12 days following immersion in aqueous suspensions containing 1×10⁸ conidia ml⁻¹. Indigenous isolates that had been recovered from C. maculatus were more virulent in laboratory bioassays than exotic isolates from other insects. The two isolates with the shortest LT₅₀ values were compared in dose-response assays by immersion and by exposure to cowpea grains treated with dry conidia. In both assays B. bassiana 0362 was consistently more virulent than M. anisopliae 0351. By immersion, LC₅₀ values on day 6 post-treatment were 9.10×10⁴ and 7.10×10⁵ conidia ml⁻¹ for B. bassiana 0362 and M. anisopliae 0351, respectively. Exposure to treated grains gave LC₅₀ values of 1.15×10⁷ and 4.44×10⁷ conidia g⁻¹ grain for B. bassiana 0362 and M. anisopliae 0351, respectively. In 1 kg batches of cowpea stocked with 50 adult C. maculatus, B. bassiana 0362 at both 1×10⁷ and 1×10⁸ conidia g⁻¹ grain led to significant adult mortality and reduced F₁ emergence relative to untreated populations. At 1×10⁸ conidia g⁻¹ the effect of the fungus persisted into the F₁ generation. The net reproductive rates, R₀, measured 26 days after insects were released were 5.16 and 7.32 for the high and low doses compared to 9.52 for the untreated control.     

Insecticidal activities of aromatic plant extracts and essential oils against Sitophilus oryzae and Callosobruchus chinensis

Methanol extracts from 30 aromatic medicinal plant species and five essential oils were tested for their insecticidal activities against adults of Sitophilus oryzae (L.) and Callosobruchus chinensis (L.), using direct contact application and fumigation methods. Responses varied with plant material, insect species, and exposure time. In a test with a filter paper diffusion method at 3.5 mg/cm², potent insecticidal activity against both species was produced by an extract from Cinnamomum cassia bark, cinnamon (C. cassia) oil, horseradish (Cocholeria aroracia) oil, and mustard (Brassica juncea) oil within 1 day after treatment. Over 90% mortality at 3 or 4 days after treatment was achieved using extracts of Acorus calamus var. angustatus rhizome, Acorus gramineus rhizome, Illicium verum fruit, and Foeniculum vulgare fruit. An extract from Cinnamomum sieboldii root bark gave 100% mortality at 2 days after treatment. At 0.7 mg/cm², extracts from C. cassia, C. sieboldii, and F. vulgare as well as cinnamon oil, horseradish oil and mustard oil were highly effective against both species. In a fumigation test with S. oryzae adults, the oils described were much more effective in closed containers than in open ones, indicating that the insecticidal activity of the oils was attributable to fumigant action. The plant extracts and essential oils described could be useful for managing field populations of S. oryzae and C. chinensis.     

Effect of relative humidity on the susceptibility of Callosobruchus maculatus (Fabricius) (Coleoptera: Bruchidae) to two modified atmospheres

It is known that modified atmospheres (MA) created in a storage environment, involving high carbon dioxide (CO₂) levels (hypercarbia) or low oxygen (O₂) levels (anoxia) are detrimental to bruchid pests of grain legumes. The possibility of enhancing MA action by increasing or decreasing the relative humidity (r.h.) conditions in storage at 25±2°C was investigated against Callosobruchus maculatus (F.), a major bruchid pest of stored cowpea seeds. Mortality of eggs and adults of the bruchid in 70% CO₂ in air and 1.0% O₂ in nitrogen (N₂) was higher at 10±3% and 34±2% than at 70±2% and 90±3% r.h. Mortality of larvae and pupae of the bruchid in the atmospheres was not affected by the r.h. in storage. Development period was longer for adults emerging from bruchid eggs exposed for 12 h to these atmospheres at the lower r.h. Freshly emerged adult bruchids from treated eggs also laid fewer eggs after 12 h of exposure to these atmospheres at the lower r.h.     

Low and high temperatures for the control of cowpea beetle, callosobruchus maculatus (F.) (coleoptera: Bruchidae) in chickpeas

Chickpea (Cicer arietinum L.; Leguminasae) is an important pulse crop grown, around the world. The whole grain of chickpea is damaged by the cowpea seed beetle, Callosobruchus maculatus (Coleoptera: Bruchidae), which is the most important field-carry-over storage pest of pulses. The management of this insect in storage using chemicals leads to insecticide residues in grains and insecticide resistance development in insects. Thermal disinfestation is one of the means of physical insect control. Eggs, larvae, pupae and adults were held at 42 or 0 °C for varying durations. Pupae and adults were equally heat tolerant. The lethal time to reduce survival by 50% (LT₅₀) at 42 °C for eggs, larvae, pupae and adults were 18, 57, 78 and 71 h, respectively. Pupa was the most cold-tolerant stage. The LT₅₀ at 0 °C for eggs, larvae, pupae and adults were 3, 8, 10 and 4 d, respectively. The LT₅₀ for pupae were 4907, 4262, 336, 36 and 13 min at the grain temperature of 42, 45, 50, 55 and 60 °C, respectively. The LT₅₀ of pupae at 0,−5,−10 and −15 °C were 274, 122, 7 and 2 h, respectively.     

Ecological factors associated with pest status in Callosobruchus (Coleoptera: Bruchidae): high host specificity of non-pests to Cajaninae (Fabaceae)

Larval host plants of six Callosobruchus species (Coleoptera: Bruchidae), including pest species, were discovered by collecting the seeds of wild (or inedible) and cultivated edible legumes in the field in Taiwan, Thailand, continental China, Myanmar, Nepal and the Philippines. A close relationship between Asian Callosobruchus species and the leguminous subtribe Cajaninae was revealed: Rhynchosia species were commonly used by three Callosobruchus species, wild and cultivated species of Cajanus and two Dunbaria species, respectively, by single species. Two Taiwanese species were confirmed to be conspecific with continental species. We further reviewed host legumes of 11 species of Callosobruchus including nine species of pests. There were significant positive correlations between geographic distribution range and host range at all three taxonomic levels of hosts. Principal component analysis on geographic distribution range, host range, altitude of distribution, utilizations of cultivated hosts, of Cajaninae and of Phaseolinae showed that the first axis (PC1) described 52% of total variance, which was related significantly with the frequencies of utilization of cultivated legumes (0.93), and of Cajaninae (Cajanus, Dunbaria and Rhynchosia) (−0.85). PC1 was also positively correlated with the frequency of utilization of Phaseolinae (Vigna and Lablab) (0.68), geographical range (0.67) and with host range (0.67) before Bonferroni corrections. Contrary to the polyphagy of the widely distributed pest Callosobruchus, non-pest species exhibited fidelity to single specific genera of wild or inedible legumes, and pests with limited distribution are specific to leguminous subtribes. Non-pests are characterized by tight association with Cajaninae. We concluded that specialization to non-economic Cajaninae prevents a species of Callosobruchus from becoming a pest of cultivated legumes.     

A case for the standardization of protocols used in screening cowpea, Vigna unguiculata for resistance to Callosobruchus maculatus (Fabricius) (Coleoptera: Bruchidae)

Cowpea cultivars Ife Brown, Maiduguri-A, Maiduguri-B and TVu 2027, were screened for resistance to Callosobruchus maculatus (F.) attack using different seed and insect densities. The number of eggs laid on the seeds and most other demographic variables increased with increase in insect density (number of females per 40 seeds). Although the same trend was observed with increase in seed density (seeds per 8 females), this was less predictable. However, the number of eggs per seed decreased significantly as seed density increased, but increased with increase in female density. Adult emergence increased significantly with increase in both insect and seed density while the growth index did not vary. Significantly more eggs were laid on the seeds of TVu 2027 (the resistant control) than on all the other cultivars when 20 or 40 seeds were available to 8 females, or at the lowest insect density (2 females per 40 seeds). However, percentage adult emergence and growth were higher on Ife Brown (susceptible control) than on TVu 2027 and the two Maiduguri cultivars. Measurements of percentage adult emergence and growth index indicated clear cultivar differences at all seed and insect densities. Also, although the loss in weight of seed increased with increase in insect density and decreased significantly with increase in seed density, meaningful differences in resistance among test cultivars were not always apparent at all densities. Based on these findings, we propose using 40 seeds and 2 pairs of adult beetles in screening cowpea for resistance. In addition, we conclude that % adult emergence, growth index and % weight loss are the most reliable indicators for resistance of cowpea to damage by this insect. The need to standardize experimental procedures used for host plant resistance work on storage pests of cowpea is emphasized.     

Technique to isolate Callosobruchus chinensis (L.) (Coleoptera: Bruchidae) eggs

A simple technique to isolate the eggs of Callosobruchus chinensis from the surface on which they are laid is described. An aqueous extract of their natural food, green gram, Vigna radiata, is used to coat glass beads that attract adult females for oviposition. The thin layer of extract on the glass beads allows separation of the eggs from the bead surface after a 15–20 s immersion in water. When transplanted onto fresh grain treated with glue, the separated eggs exhibit normal development and up to 97% hatching.     

Repellent, toxic and reproduction inhibitory effects of essential oil vapours on Acanthoscelides obtectus (Say) (Coleoptera: Bruchidae)

Thirteen essential oils (Apium graveolens, Citrus sinensis, Eucalyptus globulus, Juniperus oxycedrus, Laurus nobilis, Lavandula hybrida, Mentha microphylla, Mentha viridis, Ocimum basilicum, Origanum vulgare, Pistacia terebinthus, Rosmarinus officinalis, and Thuja orientalis) were tested in their vapour form against Acanthoscelides obtectus. Choice and no-choice tests revealed that most of them have a repellent action, reduce fecundity, decrease egg hatchability, increase neonate larval mortality and adversely influence offspring emergence. Furthermore, some oils were strongly toxic to A. obtectus, males appearing more susceptible than females. Among the essential oils tested, the most toxic for males were those of M. microphylla and M. viridis whereas the most toxic against females were those of L. hybrida and R. officinalis.     

A new variant of antimetabolic protein,arcelin from an Indian bean, Lablab purpureus (Linn.) and its effect on the stored product pest, Callosobruchus maculatus

The anti-metabolic or insecticidal gene, arcelin (Arl) was isolated, cloned and sequenced using sequence specific degenerate primers from the seeds of Lablab purpureus collected from the Western Ghats, Tamil Nadu, India. The L. purpureus arcelin nucleotide sequence was homologous to Arl-3 and Arl-4 alleles from Phaseolus spp. The protein it encodes has 70% amino acid identity with the amino acid sequences of Arl-3I, Arl-3III, Arl-4 precursor, Arl-4 and Arl-4I. The partially purified arcelin from the seeds of L. purpureus using an artificial diet confirmed the complete retardation of development of the stored product pest Callosobruchus maculatus at 0.2% w/w arcelin-incorporated artificial seeds.     

Biological activity and persistence of four essential oils towards the main pest of stored cowpeas, Callosobruchus maculatus (F.) (Coleoptera: Bruchidae)

The use of essential oils extracted from native aromatic plants is perceived as a promising alternative to protect stored cowpeas in West Africa. However the optimal conditions for their efficiency remain to be determined. A study was therefore carried out to compare the biological activity and temperature-related persistence of four selected essential oils towards Callosobruchus maculatus, the main pest of stored cowpeas. Essential oil extracted from Ocimum americanum proved to be very toxic towards C. maculatus adults (LC₅₀ = 0.23 μl/l) while the oils from Hyptis suaveolens, Hyptis spicigera and Lippia multiflora exhibited higher LC₅₀ values (1.30 μl/l; 5.53 μl/l and 6.44 μl/l respectively). The persistence of the biological activity of the four oils was variable and that from O. americanum was most persistent. Fourteen days post-application, this oil was still as active on C. maculatus adults as it was immediately after its application. Exposure of this oil to a high temperature, close to temperatures occurring during storage in Burkina Faso in the dry season, however, resulted in a rapid decrease in its efficacy. Our results emphasize the need to take into account environmental factors such as temperature to optimize the use of promising essential oils for controlling stored-product pests in West Africa.     

Progress towards a lure to attract three stored product weevils, Sitophilus zeamais Motschulsky, S. oryzae (L.) and S. granarius (L.) (Coleoptera: Curculionidae)

The behavioural responses of Sitophilus granarius, S. oryzae and S. zeamais to synthetic 4S,5R-sitophinone alone and in combination with volatiles from kibbled carob have been investigated with a view to the development of a single lure to attract all three species. Sitophilus zeamais and S. oryzae were shown to respond to 4S,5R-sitophinone at amounts as low as 0.025 ng. Sitophilus granarius gave a significant response to 50 ng 4S,5R-sitophinone. Volatiles from kibbled carob were also shown to attract all three species. This is the first time that attraction of S. zeamais and S. oryzae to carob volatiles has been demonstrated. Fresh lures containing 4S,5R-sitophinone and carob volatiles attracted significantly more insects in pitfall bioassays for all three species than when either component was used alone. However, after 6 weeks a significant increase in response compared to the control was found only for S. oryzae and S. zeamais. The effect of insect age on response to the pheromone and carob volatiles was also studied for all three species. The response to carob volatiles decreased with increasing insect age for all three species. A significant response to 4S,5R-sitophinone was found only for 1-2-day-old adults of S. oryzae and S. zeamais but for all ages tested of S. granarius. The effect of the combination of pheromone and carob volatiles also decreased with increasing insect age.     

Polyphenolic compounds of Mediterranean Lamiaceae and investigation of orientational effects on Acanthoscelides obtectus (Say)

As a continuation of work on Mediterranean aromatic plants with insecticidal properties, the hydrodistillate fraction of five species of Lamiaceae showing insecticidal effects on Acanthoscelides obtectus (Say) (Coleoptera: Bruchidae) was analysed to determine if polyphenols were involved in the toxic effect. Chromatographic analyses indicated that the polyphenols were mainly cinnamic acids and flavones, the most abundant being rosmarinic acid, luteolin-7-glucoside and derivatives. A bioassay to assess the effect on the behaviour and survival of beetles was conducted with these main compounds compared to other polyphenolic acids and flavonoids (flavones, flavanones, flavonols and flavanols) aglycones and glycosides. The polyphenols tested were toxic to the beetles to different degrees. Caffeic and ferulic acids, vanillin and luteolin-7-glucoside induced a knockdown effect on the first day, gallic acid on the second day. Quercetin significantly decreased natural mobility from the first day, naringin, syringaldehyde, vanillic acid and gallic acid after the 4th day. On the 8th day all compounds caused significant mortality. Rosmarinic acid and luteolin-7-glucoside were the most active compounds. An attractive effect enhanced the toxicity.