How host larval age, and nutrition and density of the parasitoid Dinarmus basalis (Hymenoptera: Pteromalidae) influence control of Acanthoscelides obtectus (Coleoptera: Bruchidae)

Choice of the targeted host developmental stage, regulation of parasitoid numbers released and introduction of food supplements are operational factors with a potential to influence the level of biological control. In a closed laboratory storage system maintained over two generations of the host, the impact of these three parameters on the control potential of the parasitoid Dinarmus basalis Rondani was investigated for high populations of larvae of Acanthoscelides obtectus (Say) feeding inside dry common bean seeds Phaseolus vulgaris. The beans were already infested with immature bruchids at the beginning of the storage period to simulate harvest conditions, characterized in a previous study. Treatments resulted in a reduction of 48-75% of the bruchid population within 16 weeks of storage. The best timing of parasitoid release was at the simulated harvest, as later releases reduced the bruchid population only by about half this percentage. Host feeding is postulated to be the key factor involved in the observed difference. The effect of increasing the number of parasitoids strongly depended on host age and food supplement. Addition of vials with honey had no direct effect on the bruchid population or on the parasitoid progeny. The ecological significance of these findings and implications for biological control are discussed.     

Oviposition deterrents of Callosobruchus maculatus (Coleoptera: Bruchidae)

Abstract Many bruchids distribute their eggs uniformly among the host seeds available to them and tend to avoid adding eggs to seeds bearing any eggs if pristine seeds remain. Several authors have postulated that the adult cowpea seed beetles, Callosobruchus maculatus (F.) (Coleoptera: Bruchidae), produce pheromones inhibiting oviposition by females exposed to them. It is shown that both sexes produce compounds which can be washed from glass surfaces in a variety of organic solvents which do not deter oviposition as such, but seeds coated with such washings are avoided by ovipositing females when they are selecting seeds for egg-laying and uncoated seeds are available. Some components, at least, of the products persist for at least 30 days, longer than the normal development time of insects in the seeds. The compounds affect the distribution of eggs among the available seeds. At least three different biotypes of the species produce, recognize, and respond in the same way to each other's products. It is emphasized that the eggs themselves may not be the site of the product produced by females but that the physical presence of eggs cannot be overlooked in any discussion of the possible role of pheromones in oviposition and egg distribution by bruchids.     

Comparative seed resistance in eighty cowpea accessions to the seed bruchid,Callosobruchus maculatus (Fabricius) (Coleoptera: Bruchidae)

Eighty cowpea accessions comprising both improved and local were assessed for their resistance to Callosobruchus maculatus (F.) attack under laboratory conditions (29 ± 3?°C and 65 ± 5% R.H.). Sixty-nine accessions were derived from the International Institute for Tropical Agriculture breeding programmes, four were local cultivars, while seven were improved cultivars from national research institutes. Results showed that there were significant differences among the accessions in terms of number of initial bruchid eggs laid on them, adult emergence, number of seeds damaged and bruchid-induced seed weight loss at three months after artificial infestation with C. maculatus. On IT04K-334-2, IT04K-343-1, IT06K-141, IT99K-216-48-1, IT99K-494-6 and IT99K-529-2, fewer initial eggs were laid and no adult emergence, seed damage or weight loss and were considered highly resistant to C. maculatus. On the other hand, IT03K-337, IT04K-217-51, IT04K-332-1, IT04K-339-1, IT06K-188-49, IT81D-1137, IT845-2231-15, IT93K-452-1, IT04K-154-1, IT96K-610, IT97K-1069-6, IT97K-390-2, Ife Bpc, IT99K-573-2-1, Ife Modupe, IT845-2246, Drum, Ife Brown and Oloyin were the most susceptible with the highest damage indices values (p?<?0.05). The range of each of the resistance indices measured in the susceptible varieties was: number of initial eggs laid (61–100), adult emergence (40.0–62.10%), number of seeds damaged (53.0–85.5) and seed weight loss (58.0–66.25%).     

Effect of host quality of Callosobruchus maculatus (Coleoptera: Bruchidae) on performance of the egg parasitoid Uscana lariophaga (Hymenoptera: Trichogrammatidae)

Development and reproductive success of the solitary egg parasitoid Uscana lariophaga Steffan were examined after development in eggs of the bruchid storage pest Callosobruchus maculatus Fabricius reared at either low or high densities on cowpea seeds and laid at day 1 and 4 of maternal life. Both bruchid larval competition and maternal age negatively affected egg size, but the latter more than the former. Uscana lariophaga reared in small hosts developed slower, were smaller and produced fewer eggs compared to parasitoids reared in large hosts. Fecundity of the parasitoid was heavily influenced by host egg size. This was reflected in the values for the intrinsic rate of increase of U. lariophaga, which differed for wasps that developed in host eggs laid by bruchid females of different age. Wasps allocated marginally more female offspring to larger hosts.     

Impact of the egg parasitoid Uscana lariophaga and the larval-pupal parasitoid Dinarmus basalis on Callosobruchus maculatus populations and cowpea losses

In West Africa, Uscana lariophaga Steffan (Hymenoptera: Trichogrammatidae) and Dinarmus basalis (Rondani) (Hymenoptera: Pteromalidae) parasitize the eggs and larvae, respectively, of Callosobruchus maculatus Fab. (Coleoptera: Bruchidae), an important pest of stored cowpea. The impact of the parasitoids on pest populations was evaluated in clay pots similar to the ones used in traditional storage in Niger. At the beginning of the storage period, cowpeas were infested with one density of C. maculatus and with either one or both of the parasitoid species. If the parasitoids we inoculated as single species in stores, both D. basalis and U. lariophaga significantly suppressed the bruchid population, but the former did so more effectively than the latter. A combination of D. basalis and U. lariophaga resulted in the same suppression of bruchid populations as when D. basalis was the only parasitoid. Fifteen weeks after storage, the parasitoids reduced the number of grains damaged significantly by 38–56%. The effect of inoculating single or multiple parasitoid species on C. maculatus populations in an augmentation strategy is discussed.     

Polymorphism of competition type and its genetics in Callosobruchus maculatus (Coleoptera: Bruchidae)

Callosobruchus maculatus has both contest and scramble competition strategies. The currently existing theoretical models using game theory suggest that the contest strategy should be selected for. However, most geographic strains of C. maculatus show scramble competition. We experimentally crossed the representative strains of contest and scramble. We expressed the degree of contest competition by a continuous value named the C-value, which ranges from zero (pure scramble) to unity (pure contest). The competition types expressed in the C-value were genetically additive. Their larval developmental rates were negatively correlated with C-values. Multiple-generation experiments of the mixed strains confirmed that there were no overwhelming advantages of contest over scramble type. Most of the mixed strains remained in the intermediate states. We discuss the results in terms of the resource size necessary for developmental success and developmental speed. Received: April 19, 2001 / Accepted: November 2, 2001     

Sperm Storage and Use After Multiple Mating in Dinarmus basalis (Hymenoptera: Pteromalidae)

Multiple mating and its effects on the sex ratio in Dinarmus basalis (Hymenoptera: Chalcidoidea, Pteromalidae), an ectoparasitoid of Callosobruchus maculatus (Coleoptera: Bruchidae), were investigated under controlled conditions. Once-mated females suffer a sperm depletion about 21 days after mating and thereafter are constrained to produce only haploid males. On the other hand, three-times-mated females store more sperm in their spermathecae (335 vs 147) and produce daughters during the major part of their reproductive life. Consequently, once-mated females showed a male-biased life time sex ratio (mean = 0.31) as opposed to three-times-mated females (mean = 0.63). Females can copulate only at the beginning of their reproductive life, and multiple mating must occur before egg-laying activity. This behavioral strategy could be an avoidance of consanguinity in a promiscuous environment. This ectoparasitoid species reveals a reproductive strategy which promotes polyandry and a very high sperm efficiency.     

Biological activity of tree marigold, Tithonia diversifolia, on cowpea seed bruchid, Callosobruchus maculatus (Coleoptera: Bruchidae)

The powder and ethanol extract of Tithonia diversifolia leaves were tested for their efficacy at five different concentrations (0.0%, 0.5%, 1.0%, 1.5% and 2.0%) on bruchid mortality, oviposition and adult emergence of cowpea seed beetle Callosobruchus maculatus at ambient tropical storage conditions of temperature and relative humidity. Results showed that the leaf extract had a higher bioactivity on oviposition, adult emergence and mortality of C. maculatus. The mean number of eggs laid on seeds treated with extract was reduced from 20.7 in the solvent-treated to 4.7 at 2%Tithonia extract concentration while adult emergence reduced from a mean of 92.2 in the solvent-treated seeds to 72.2 at 2.0% extract treatment. Mortality was 100% at higher concentrations of 3%, 4% and 5% within 24 h of extract application but at lower concentrations mortality was 73.3% and 93.3% at 1% and 2%, respectively after 24 h. Fourty-eight hours after application, 100% mortality of adult C. maculatus was obtained at all concentrations. The mean number of eggs laid was reduced from 41.3 in the untreated to 17.3 at 2.0% powder concentration while adult emergence dropped from 98.5% in the control to 74.2% at the highest powder concentration. The powder was effective at higher concentrations and longer exposure time. At 3%, 4% and 5%, 63–75% mortalities occurred within 24 h of application while at 48 h, mortality was 76–98% at 3–5% powder application rates. The results of this study revealed T. diversifolia as a potential candidate for bioinsecticide preparations because of antiovipositional, ovicidal and knockdown properties of its products, which have some volatile components.     

Insecticidal and biological efficacy of two vegetable oils against Callosobruchus maculatus (Fabricius) (Coleoptera: Bruchidae) under laboratory conditions

Bioefficacy of two vegetable oils, namely ethyl oleate and Acorus calamus, were conducted against the pulse beetle Callosobruchus maculatus (Fabr.) under a constant temperature of 29 ± 1 °C and a relative humidity of 65 ± 5% R.H. Four concentrations (0.125, 0.25, 0.5 and 1.0%) of each oil were used against the adult and tested their latent effect on the immature stages of the beetle. The results obtained show insecticidal effect of both ethyl oleate and A. calamus oil; however, A. calamus oil proved to be more toxic to the adults at all the concentrations tested in comparison to the ethyl oleate oil. As the concentration of the oils decreased, the percentage mortality decreased. The obtained results concentrated on biological efficacy of the oils tested against the adults. Both oils affected the longevities of both male and female C. maculatus adults in comparison to the control. The oviposition periods of the adult female were affected significantly in comparison to the control. A. calamus oil prohibited oviposition completely at high concentrations of 0.5 and 1.0% and caused complete sterility to the females. Lower concentrations decreased the oviposition period greatly and reduced the number of eggs laid in comparison to the control. Ethyl oleate had an inconsiderable latent effect on the incubation period as well as the percentage of hatching of the resulting eggs. A. calamus oil decreased the percentage of egg hatchability with serious increase in the percent sterility. The two tested oils had slight extension in either larval or pupal stage of the resulting progeny of C. maculatus. Both oils show oviposition deterrency towards C. maculatus adults. The oviposition deterrent index was 100% when mung bean seeds were treated with A. calamus oil. The bruchid females laid all eggs on control seeds and no eggs were laid on treated seeds.     

Sperm production and mating potential of males after a cold shock on pupae of the parasitoid wasp Dinarmus basalis (Hymenoptera: Pteromalidae)

For ectothermic species, temperature is a key environmental factor influencing several aspects of their physiology and ecology, acting particularly on reproduction. To measure the consequences of a severe thermal stress during development on male reproduction, a cold shock (1h at -18 degrees C) was tested on Dinarmus basalis pupae. D. basalis (Hymenoptera: Pteromalidae) is a parasitoid wasp in which sperm management in both male and female is of prime importance. After a cold shock, developmental success was reduced, with a quarter of cold-shocked males not emerging correctly. The stress effects were estimated at the level of sperm stock in seminal vesicles of males at different ages and on the ability of 2-day-old males to access females in single and multiple mating and in male-male competition. Cold-shocked males had a reduced sperm stock compared to control males and this difference persisted with age. The rate of sperm production was similar in both groups. The consequences of a cold shock on male reproductive ability were perceptible in multiple mating and male-male competition but not in single mating. Cold-shocked males were at a disadvantage, inseminating fewer females and copulating less frequently. Finally, male pupae of D. basalis were able to withstand severe temperature stresses and their reproductive functions were partially preserved.     

Interspecific Relationships Between the Solitary Ectoparasitoid, Eupelmus vuilleti (Crw.) (Eupelmidae), and Its Sympatric Species, Dinarmus basalis (Rond.) (Pteromalidae), in the Presence of Their Host, Callosobruchus maculatus Pic (Coleoptera Bruchidae)

The eupelmid Eupelmus vuilleti CRW. and the pteromalid Dinarmus basalis Rond. are sympatric in West Africa. The reproduction of E. vuilleti and D. basalis females was analyzed in 10, 20, and 40 host patches when present alone or together. In 10- and 20-hosts patches, the presence of E. vuilleti affected the reproduction and offspring survival chances of D. basalis. In 40-hosts patch, only the offspring survival of D. basalis was affected by the presence of E. vuilleti. In contrast, the presence of D. basalis did not influence the reproduction of E. vuilleti and the survival chances of its offspring at all host densities tested. Multiparasitism did not occur at random. In a choice test, E. vuilleti showed a preference for hosts previously parasitized by D. basalis on healthy hosts. This attraction is mediated by chemical markers deposited by D. basalis females on the surface of the seed. At high host density, the likelihood of encountering a seed containing a host previously parasitized by D. basalis was lower for E. vuilleti, affecting moderately D. basalis reproductive success.     

Characterization of resistance to three bruchid species (Callosobruchus spp., Coleoptera,Bruchidae) in cultivated rice bean (Vigna umbellata)

Resistance of wild and cultivated rice bean (Vigna umbellata [Thunberg] Ohwi and Ohashi) to three bruchid species, Callosobruchus chinensis L., Callosobruchus maculatus F., and Callosobruchus analis F., was evaluated. All but three accessions of cultivated, and all wild rice bean accessions tested, exhibited complete resistance to all three bruchid species. Rice bean seeds with seed coat removed also showed complete resistance to the three bruchid species. Results indicate that physical attributes and/or chemical(s) present in the seed coat of rice bean are not the main factors responsible for resistance. Feeding tests were performed by using artificial beans prepared with varying proportions of rice bean (resistant) and azuki bean (susceptible) flour. Number of bruchid adults that emerged decreased, and larval developmental period (days) was extended, when artificial beans with an increasing proportion of rice bean flour were used. These tests revealed that a chemical compound(s) contained in the cotyledon of rice bean has an inhibitory effect on the growth of these bruchid species. The results also indicate that the chemical(s) in rice bean cotyledon is most effective against C. maculatus.     

Host selection by virgin and inseminated females of the parasitic wasp, Dinarmus basalis (Pteromalidae, Hymenoptera)

1. Fitness is related to reproduction and survival. There apparently exists a negative correlation between the numbers of male and female offspring. There also exists a trade-off between survival and reproduction. This paper investigates optimal decisions with the reproduction and survival trade-off in host selection by wasps.
2. Whereas inseminated female wasps could manipulate the sex of their offspring, virgin females produced only male offspring. I surveyed behavioural differences and the consequences of oviposition by inseminated and virgin females of a solitary parasitic wasp in host choice situations.
3. Two host types were available at the same time to both inseminated and virgin female wasps: one (a 17-day-old host in one bean) presenting difficulties for the laying of eggs, but more benefits for the offspring and the other (five 12- or 13-day-old hosts in one bean) easier for the female wasp for laying of eggs but less beneficial for the offspring.
4. Inseminated female wasps chose more 17-day-old hosts than 12-day-old hosts, but more 13-day-old hosts than 17-day-old hosts in each pair-wise choice. Virgin females chose the smaller hosts in both situations.
5. Virgin females, having greater longevity than inseminated females, laid larger numbers of eggs than the inseminated females during their lifetime by adopting an energy-saving host choice that had little effect on male offspring fitness.     

Talisia esculenta lectin and larval development of Callosobruchus maculatus and Zabrotes subfasciatus (Coleoptera: Bruchidae)

Bruchid larvae cause major losses in grain legume crops throughout the world. Some bruchid species, such as the cowpea weevil and the Mexican bean weevil, are pests that damage stored seeds. Plant lectins have been implicated as antibiosis factors against insects, particularly the cowpea weevil, Callosobruchus maculatus. Talisia esculenta lectin (TEL) was tested for anti-insect activity against C. maculatus and Zabrotes subfasciatus larvae. TEL produced ca. 90% mortality to these bruchids when incorporated in an artificial diet at a level of 2% (w/w). The LD(50) and ED(50) for TEL was ca. 1% (w/w) for both insects. TEL was not digested by midgut preparations of C. maculatus and Z. subfasciatus. The transformation of the genes coding for this lectin could be useful in the development of insect resistance in important agricultural crops.     

Interspecific Relations Between Two Sympatric Species of Hymenoptera, Dinarmus basalis (Rond) and Eupelmus vuilleti (Crw), Ectoparasitoids of the Bruchid Callosobruchus maculatus (F)

Dinarmus basalis (Rond) and Epelmus vuilleti (Crw) are two Hymenopteran species, which are solitary ectoparasitoids of bruchid larvae. In the presence of seeds of Vigna unguiculata (Walp) containing hosts parasitized by E. vuilleti, a high percentage of D. basalis females avoided multiparasitism whatever the age of the eggs or the larvae present on the host. The least avoidance was observed when the hosts were parasitized by E. vuilleti 30 min beforehand. This avoidance behavior is adaptive and is related to the low survival chances of the D. basalis larvae when they are in interspecific competition with E. vuilleti larvae. The analysis of the behavior of D. basalis demonstrated that the avoidance of multiparasitism could be due to the perception of two signals; an external signal deposited on the surface of the seeds during the E. vuilleti oviposition phase and an internal signal due to the presence of the eggs and larvae at the surface of the hosts. E. vuilleti females did not avoid multiparasitism and multiparasitized the hosts bearing D. basalis eggs or larvae. The behavior of E. vuilleti females was not disturbed by the presence of its competitor. Under these conditions of interspecific competition, the survival chances of E. vuilleti larvae were very high whatever the age of its competitor D. basalis. The two species of parasitoids could move in a column containing healthy seeds of V. unguiculata and patches with seeds containing parasitized or unparasitized larvae. The distribution of D. basalis females introduced into these columns depended on the host quality. They avoided the patches containing the hosts parasitized E. vuilleti and were found in the patches with healthy hosts. The behavior of E. vuilleti females was very different; the distribution of the females and the parasitism and multiparasitism rates were not affected by the quality of the hosts present in the patches. The adaptive significance of the behaviors of these two species was analyzed in relation to the survival chances of their offspring.     

Location and ultrastructure of sex pheromone glands in female Callosobruchus maculatus (Fabricius) (Coleoptera : Bruchidae)

The ultrastructure of the female sex pheromone glands in Callosobruchus maculatus (Coleoptera : Bruchidae) were localized using a masking technique, combined with eiectro-antennography and by a comparison of the glandular cells of sexually active (flightless) females and non-sexually active (flight-form) females. Each unicellular gland is an invagination of the integumental membrane capped by a single secretory cell. These glands are situated on the fine intersegmental membrane, which joins the pygidium to the ovipositor. The secretory cells of the glands of active females are characterized by well-developed microvilli, with many elongated mitochondria among the latter. The high metabolic activity of these cells is revealed by the presence of heterogeneous secretion vesicles, some of which contain abundant crystallized material. Deep basal invaginations indicate the uptake of substances from the haemolymph. The receptor canal is a network of fine cuticular filaments which have the same structure regardless of the female's sexual status. Cells from the glands of non-sexually active females are underdeveloped and show no invaginations of the basal membrane and very few microvilli. The localization of these glands was made possible by the use of SEM, TEM and EAG as well as by masking the suspected zones and by comparing females in different physiological states: flightless females, which were sexually active and producing pheromones; and flight-form females, non-sexually active and producing no sex pheromones. Only by adopting such a stringent method was it possible to confirm the function of the glands whose ultrastructure was studied.     

Mechanisms of the insecticidal action of TEL (Talisia esculenta lectin) against Callosobruchus maculatus (Coleoptera: Bruchidae)

Plant lectins have insecticidal activity that is probably mediated through their ability to bind carbohydrates. To examine the influence of sugars on the insecticidal activity of a lectin from Talisia esculenta seeds (TEL), the lectin was mixed with mannose, glucose, or mannose plus glucose. Mannose abolished the insecticidal activity. Affinity chromatography showed that TEL bound to midgut proteins of the insect Callosobruchus maculatus. Immunoblotting showed that TEL recognized some proteins, probably glycoproteins, present in the midgut membrane of this insect. The principal proteases responsible for digestive proteolysis in fourth instar larvae of C. maculatus were purified by chromatography on activated thiol-Sepharose. These purified proteases were unable to digest TEL after a 15-h incubation. These results suggest that the insecticidal activity of TEL involves a specific carbohydrate-lectin interaction with glycoconjugates on the surface of digestive tract epithelial cells, as well as binding to assimilatory glycoproteins present in midgut extracts and resistance to enzymatic digestion by cysteine proteinases.     

Consequences of inbreeding for the cowpea seed beetle, Callosobruchus maculatus (F.)(Coleoptera: Bruchidae)

Inbreeding is said to reduce vigour and fitness. It may also determine how a population responds to selection. Local populations of Callosobruchus maculatus , the cowpea seed beetle, are established annually from small numbers of founders and the species has been distributed to many parts of the world where isolated populations may have been founded by very small numbers of individuals. After more than 20 generations of inbreeding, inbred lines have been shown to diverge from a common ancestral stock in similar directions with respect of some variables such as developmental speed, but haphazardly in respect of other parameters such as male weight. The respective roles of drift and of selection as effective evolutionary forces in inbred lines are discussed in the light of these results. It is argued that some intraspecific differences in C. maculatus may be explained as a product of periodic inbreeding, but that the process does not impair the ability to adapt to local conditions so contributing to the status of the species as a pest of international importance.     

昆虫病原真菌与硅藻土混用对四纹豆象的毒力效果(英文)

在室内条件下检测了白僵菌Beauveria bassiana (Balsamo) Vuillemin 菌株IRAN-187C 和金龟子绿僵菌Metarhizium anisopliae (Metschnikow) Sorokin菌株A-396分别与硅藻土（DE）制剂InsectoSec混用对四纹豆象Callosobruchus maculatus成虫的毒力效果。结果表明：硅藻土与致死剂量和亚致死剂量的真菌分离物混用导致了四纹豆象成虫的高死亡率,且这些处理均缩短了真菌的致死中时(LT₅₀)。LC₅₀ 的白僵菌B. bassiana菌株IRAN-187C和 LC₅₀ 的DE混用对四纹豆象的LT₅₀ (122.2 h)最低,在95%置信限上与LC50 的金龟子绿僵菌M. anisopliae菌株A-396 与LC₅₀的DE混用得到的LT₅₀ (128.2 h) 相当,说明两种真菌分离物与DE混用对四纹豆象成虫都具有较好的毒力效果。用四纹豆象成虫检测3个温度（22, 27和30℃）和2个相对湿度（40%和55%）组合对DE活性的影响,结果显示30℃和40%相对湿度条件下得到的LT₅₀ (110.11 h)最低,说明高温低湿条件下DE活性较高。     

四纹豆象不同地理种群的遗传分化

【目的】通过对四纹豆象Callosobruchus maculatus Fabricius不同地理种群mtDNA-Cytb和COⅠ基因部分序列进行比较, 分析其不同地理种群间的遗传分化情况, 为揭示其与生物入侵的关系及入侵过程中种群系统发育地理格局与演变机制提供依据。【方法】用PCR产物直接测序法对分别来自中国海南、喀麦隆、韩国和泰国的四纹豆象4个地理种群的mtDNA-Cytb和COⅠ序列进行测序, 运用软件MEGA3.1对四纹豆象不同地理种群mtDNA-Cytb和COⅠ序列进行序列分析, 以绿豆象C. chinensis为外群构建了不同单倍型的分子系统树。【结果】34条420 bp Cytb序列中共检测到14个多态位点和5种单倍型, 33条822 bp COⅠ序列中检测到28个多态位点和9种单倍型, 其中4种单倍型为独享单倍型, 其余为全部或部分种群的共享单倍型。AMOVA分析结果显示, 四纹豆象4个地理种群间的遗传结构差异并不明显, 遗传差异主要发生在地理种群内。对4个地理种群进行了Fst值和基因流动统计, 结果表明4个地理种群间既存在着一定数量的基因交流, 也存在一定程度的遗传分化。【结论】根据单倍型分布格局初步推测, 中国不可能是四纹豆象的原产地, 而喀麦隆有可能是原产地之一, 并且喀麦隆种群与泰国种群之间的基因交流比较充分, 而中国种群与其他种群之间的遗传分化相对较大。