Behavioural responses of bethylid parasitoid species of the coffee Berry borer to chemicals cues from host and non-host dust/frass

The bethylid wasps Cephalonomia stephanoderis Betrem and Prorops nasuta Waterston are parasitoids of the coffee berry borer, Hypothenemus hampei (Ferrari), the most serious insect pest of coffee worldwide. Recent studies on the host location behaviour of these parasitoids have shown that females are attracted to volatile compounds released by immature stages and dust/frass of H. hampei. Also, these studies have reported that the locomotory behaviour of parasitoids is influenced by contact chemical cues from host dust/frass. In this study, we investigated the responses of females of both species to volatiles and contact cues from dust/frass from different sources. Volatiles from dust/frass from H. hampei-infested coffee and maize infested with Sitophilus sp were attractive to P. nasuta females. In contrast, C. stephanoderis females were only attracted to the volatiles from dust/frass from H. hampei-infested coffee. Volatiles from an artificial diet used for rearing H. hampei were not attractive to females of both species. The effect of methanolic extracts from dust/frass from different sources on the patch-searching time of parasitoids was analysed using the Ethovision software. P. nasuta females spent more time in the patches treated with extracts of dust/frass from H. hampei-infested berries, artificial diet, and from maize infested with Sitophilus sp than on patches treated with dry coffee extract or solvent control. C. stephanoderis females spent more time on patches treated with dust/frass extracts from H. hampei-infested berries and artificial diet than on patches treated with an extract of dust/frass from maize infested with Sitophilus sp, an extract of dry coffee, and methanol control.     

Coffee volatiles induced after mechanical injury and beetle herbivory attract the coffee berry borer and two of its parasitoids

The coffee berry borer (CBB), Hypothenemus hampei (Ferrari), is the most important insect pest of coffee worldwide. In this study, we used headspace solid-phase microextraction coupled with gas chromatography–mass spectrometry to sample and identify volatile compounds from Robusta coffee berries, Coffea canephora Pierre ex Froehner, infested with CBB and with mechanical damage. Furthermore, we evaluated the behavioral responses of the CBB and two of its parasitoids, Prorops nasuta Waterstone and Phymastichus coffea LaSalle, to three selected coffee volatile compounds in a Y-tube olfactometer. We found in the effluvia of red coffee berry compounds not previously reported for this coffee species. Our results show that Robusta coffee berries release induced volatiles either by insect herbivory or by mechanical damage. Small amount of butyl acetate, unknown compound 2, α-longipinene, longiborneol and longiborneol acetate are produced only in infested coffee berries fruits. Quantitatively, nine compounds account for the difference between healthy berries, infested, or mechanically damaged berries. Trans-ocimene, 4,8-dimethyl-3,7-nonadien-2-ol, α-copaene and kaurene increased amount levels in infested berries, while amount of methyl salicylate and linalool increased in mechanically damaged coffee berries. The olfactometric bioassays showed that CBB females and its two parasitoids were attracted to methyl salicylate. In addition, H. hampei and P. nasuta were attracted to linalool, and P. nasuta and P. coffea were attracted to trans-ocimene.     

Where to sample? Ecological implications of sampling strata in determining abundance and impact of natural enemies of the coffee berry borer,Hypothenemus hampei

Several parasitoids of African origin have been introduced to coffee producing areas of the Americas and Asia as biological control agents of the coffee berry borer (CBB) Hypothenemus hampei (Coleoptera: Curculionidae). These parasitoids have become established in the field but their effect on the CBB has been limited. A two-year field study in Western Kenya has found Prorops nasuta (Hymenoptera: Bethylidae) to be the predominant parasitoid emerging from CBB-infested coffee berries collected on coffee trees or from the ground. P. nasuta comprises more than 75% of the total natural enemies collected. The density of P. nasuta was 90% higher in the berries collected from the ground than from the trees. Its hyperparasitoid, Aphanogmus sp. (Hymenoptera: Ceraphronidae), also emerged from both type of berries. Across the two seasons, the average P. nasuta density per berry was 18–35 times higher than that of Aphanogmus sp. Throughout the two years sampled, significantly higher numbers of P. nasuta and Aphanogmus sp. occurred between February and March, which coincides with the beginning of the rainy season. Higher numbers of live CBB females were recorded in berries collected from the trees. Nevertheless, mortality of adult CBB was considerably higher from January to March and started to decrease from April onwards. The possibly negative effects of cultural control practices in Latin America which include the removal of berries fallen to the ground on biological control of CBB are discussed, and the use of screened collection devices for these berries which would permit the release of parasitoids but prevent escape of the pest is proposed.     

Spiroacetals in the Colonization Behaviour of the Coffee Berry Borer: A ‘Push-Pull’ System

Coffee berries are known to release several volatile organic compounds, among which is the spiroacetal, conophthorin, an attractant for the coffee berry borer Hypothenemus hampei. Elucidating the effects of other spiroacetals released by coffee berries is critical to understanding their chemo-ecological roles in the host discrimination and colonization process of the coffee berry borer, and also for their potential use in the management of this pest. Here, we show that the coffee berry spiroacetals frontalin and 1,6-dioxaspiro [4.5] decane (referred thereafter as brocain), are also used as semiochemicals by the coffee berry borer for host colonization. Bioassays and chemical analyses showed that crowding coffee berry borers from 2 to 6 females per berry, reduced borer fecundity, which appeared to correlate with a decrease in the emission rates of conophthorin and frontalin over time. In contrast, the level of brocain did not vary significantly between borer- uninfested and infested berries. Brocain was attractive at lower doses, but repellent at higher doses while frontalin alone or in a blend was critical for avoidance. Field assays with a commercial attractant comprising a mixture of ethanol and methanol (1∶1), combined with frontalin, confirmed the repellent effect of this compound by disrupting capture rates of H. hampei females by 77% in a coffee plantation. Overall, our results suggest that the levels of frontalin and conophthorin released by coffee berries determine the host colonization behaviour of H. hampei, possibly through a ‘push-pull’ system, whereby frontalin acts as the ‘push’ (repellent) and conophthorin acting as the ‘pull’ (attractant). Furthermore, our results reveal the potential use of frontalin as a repellent for management of this coffee pest.     

Field superparasitism by Phymastichus coffea, a parasitoid of adult coffee berry borer, Hypothenemus hampei

Superparasitism by Phymastichus coffea LaSalle (Hymenoptera: Eulophidae), a parasitoid of adults of the coffee berry borer, Hypothenemus hampei (Ferrari) (Coleoptera: Curculionidae: Scolytinae), was recorded under field conditions in a coffee plantation in Colombia. Parasitoid adults were released 1, 5, and 9 days after artificial infestations of 90‐, 150‐, and 210‐day‐old coffee berries with H. hampei females. The position of the beetle inside the berry and the number of P. coffea larvae per female host were assessed 10 days after each parasitoid release. Under laboratory conditions, P. coffea usually lays two eggs per host, one female and one male. In our studies, we often recorded more than six P. coffea larvae in an individual host and mean numbers of larvae per host ranged from two to 4.45. Superparasitism by P. coffea under field conditions was influenced by the age of the coffee berries, which is the most important factor determining the speed of penetration by H. hampei, and therefore the time the beetles are exposed to a P. coffea attack. The number of parasitoid larvae in each H. hampei female gradually decreased with the age of the berry, and also linearly decreased with the time of parasitoid release. Age‐dependent effects of coffee berries that alter the ratio of available hosts to searching parasitoids by providing refuges to the herbivore, largely determine the extent of superparasitism of H. hampei by P. coffea under fields conditions in Colombia.     

Life history studies of <Emphasis Type="Italic">Prorops nasuta</Emphasis>, a parasitoid of the coffee berry borer

Life history studies were conducted in the laboratory on the African parasitoid Prorops nasuta Waterston (Hymenoptera: Bethylidae), a parasitoid of the coffee berry borer, Hypothenemus hampei (Ferrari) (Coleoptera: Scolytidae). The female wasp enters an infested coffee berry, kills the adult borer and seals the entrance of the berry with the body of the borer, impeding the entry of other organisms into the berry. The preoviposition period ranges from 3 to 14 days (mean 5.42 ± 0.37 SE). During this time females feed on the immature stages and paralyse fully grown larvae and pupae of the CBB. P. nasuta is an idiobiont solitary ectoparasitoid. Eggs are laid externally on the last instar larvae and pupae. Mean development time (egg to adult) for males and females was 27.7 (±0.37 SE) and 30 (±0.12 SE) days, respectively. Median survival for wasps fed on final instar CBB larvae was 27.7 days, significantly longer than any other treatment, while for females without food it was 2.5 days. In culture, females produced an average of 4.3 (±0.39 SE) progeny during their lifetimes. Adults began emerging at 30.6 days (±0.28 SE) after cultures were started and peak production was reached at 36 days, declining thereafter. Males normally emerged from coffee beans 2–3 days before females. Males usually emerged from 07:00 to 09:00h and females from 10:00 to 14:00 h. The culture sex ratio (proportion of males) was 0.21. Virgin females produced only male offspring.     

松墨天牛成虫对幼虫虫粪挥发性物质的触角电位反应及林间驱避试验

为了研制松墨天牛Monochamus alternatus Hope驱避剂,本文利用触角电位技术和田间试验的方法研究了松墨天牛雌雄成虫对幼虫虫粪挥发性物质的触角电位反应及幼虫虫粪己烷提取物、合成化合物混合物的林间驱避作用。结果表明：α-蒎烯、β-蒎烯、3-蒈烯、柠檬烯和长叶烯均能引起松墨天牛雌雄成虫一定的触角电生理反应,而4-甲基-2,6-二叔丁基苯酚不能引起松墨天牛雌雄成虫的触角电生理反应。林间驱避试验结果说明,幼虫虫粪的己烷提取物处理过的树干上的刻槽数明显少于对照树干上的刻槽数,二者具有极显著差异;有合成化合物的混合物存在的诱捕器诱捕到的雌虫数（平均值3.80±1.02）与对照诱捕到的雌虫数（平均值16.80±1.16）差异极显著。结果显示α-蒎烯、β-蒎烯、3-蒈烯、柠檬烯、长叶烯和4-甲基-2,6-二叔丁基苯酚的混合物对松墨天牛雌虫具有驱避作用,有望开发出一类新的驱避剂。     

Fly parasitoid Megaselia opacicornis uses defensive secretions of the leaf beetle Chrysomela lapponica to locate its host

Larvae of the leaf beetle Chrysomela lapponica derive a defensive secretion from salicyl glucosides found in the host plant Salix borealis. This secretion protects beetle larvae from some natural enemies, but does not appear to repel parasitoids. We tested the hypothesis that the fly parasitoid Megaselia opacicornis (Diptera, Phoridae) uses the larval defensive secretion of Ch. lapponica in its search for prey. In the field, nearly 30 times more M. opacicornis individuals were caught on leaves coated with sticky resin next to a source of secretion than on control leaves. In the laboratory, M. opacicornis females laid six times more eggs next to a cotton ball soaked in secretion than next to one soaked in water. Fly females also lay more eggs on prey rich in larval secretion than on secretion-poor prey. In the field, removal of defensive secretion from beetle prepupae resulted in a 7.5-fold reduction of oviposition by fly females. Parasitoids were nearly twice as likely to lay eggs on prepupae, rich in secretion, as on pupae, which contain little secretion. Fly offspring reared from beetle prepupae reached a 21% larger body mass than those reared from pupae. Finally, M. opacicornis females avoided host prepupae already parasitized by the tachinid fly Cleonice nitidiuscula, which possess little secretion. These experiments indicate that host plant-derived defensive secretions are used by this parasitoid for host location. Adaptation of parasitoids to use defensive secretions of hosts may selectively favor an increase in diet breadth in specialist herbivores.     

Development ofTrichogramma australicum (Hym.: Trichogrammatidae) at low and high population density in artificial diet

We compared the development ofTrichogramma australicum Girault in artificial-diet filled artificial “eggs” containing 10–15 (low density) or 35–50 (high density) immature parasitoids. The size of low- and high-densityT. australicum eggs at 1, 15, 30 and 36 h was not significantly different. Also, incubation time of low- and high-density eggs in artificial diet was similar. Appearance of urate bodies in larvae developing low and high density occurred at the same time (100–108 h). Size of immatureT. australicum in low density significantly increased after the urate bodies had appeared. Black deposits appeared on the surface ofT. australicum prepupae soon after all diet had been consumed (at 108–120 and 144–168 h in low and high density, respectively). We concluded that the appearance of the black deposits was an indication of the onset of the prepupal stage. The pupal size was significantly larger at low density than high density. Mortality of prepupae and pupae was significantly higher at low density than at high density. Proportion of larvae at high density developed to abnormal and normal wasps. Wasps did not emerge from larvae reared at low density.     

Pre- and post-imaginal experience in a specialist parasitoidDiadromus pulchellus (Hym.: Ichneumonidae)

The locomotory behaviour of femaleDiadromus pulchellus Wesm. (Ichneumonidae), a specialist endoparasitoid ofAcrolepiopsis assectella Zell. chrysalids, was analyzed in an olfactometer. Three parameters were measured: distance walked, number of turns and female distribution. The volatiles emitted by the larval frass ofA. assectella which feeds exclusively onAllium plants modified all three parameters. When females were placed in contact with frass and/or with the host before the olfactometer test, the number of turns was lower in comparison to that of naive females tested without prior contact with the host and/or its frass. This indicated that the trajectory of experienced females towards the odour source was more direct. Associative learning is a possible mechanism behind this change. No evidence of pre-imaginal experience was found because parasitoids emerging from the chrysalids of a non-host moth,P. interpunctella, responded to the frass volatiles of the usual host,A. assectella, in the same way as parasitoids reared on this latter host.     

Comparative Development and Performance of Artificially Reared versus Host-RearedDiapetimorpha introita(Cresson) (Hymenoptera: Ichneumonidae) Wasps

Diapetimorpha introita(Cresson) (Hymenoptera: Ichneumonidae), a native ectoparasitoid ofSpodopteraspp. pupae, was reared in the laboratory on an artificial diet devoid of any insect host components. Diet-reared wasps demonstrated a propensity to search for and parasitize natural hosts in a field cage trial. Longevity of the diet-reared wasps was comparable with the longevity of wasps reared on host pupae. Survival rate ofD. introitawas 61.3% when reared on diet and 76.3% when reared on host pupae. Wasps reared on the artificial diet had longer developmental times, reduced fecundity, and reduced adult weights compared to wasps reared on host pupae. These studies suggest that future research efforts should focus on increasing fecundity and weight of diet-reared wasps and decreasing the mortality and developmental time of wasps reared on the artificial diet. The ability to rearD. introitaon an inexpensive, artificial diet significantly enhances the potential of mass rearing this parasitoid for inundative releases againstSpodopteraspp.     

Interactions among bethylid parasitoid species attacking the coffee berry borer,Hypothenemus hampei (Coleoptera: Scolytidae)

The question of whether biological control is most likely achieved by deploying single or multiple species of biological control agents is much debated. While utilizing several natural enemies may enhance control, there is also the potential for disruptive inter-specific interactions. Such interactions may be studied in the laboratory by focusing on the details of the interactions themselves and attempting to infer population level consequences from their sum, or by focusing more directly on the overall effects on natural enemy populations: we term these approaches ‘reductionist’ and ‘holistic.’ Here we conduct a holistic laboratory study on interactions between three species of parasitoid wasps that are parasitoids of the coffee berry borer, Hypothenemus hampei (Ferrari) (Coleoptera: Scolytidae): Cephalonomia stephanoderis Betrem, C. hyalinipennis Ashmead and Prorops nasuta Waterston (all Hymenoptera: Bethylidae). We find evidence for both intra- and inter-specific resource competition. Interactions between C. stephanoderis and P. nasuta, both indigenous to Africa, appear to be approximately symmetrical, while C. hyalinipennis, naturally found in the coffee plantations of Chiapas, Mexico, may exert a disruptive influence. C. hyalinipennis also has a low population growth rate. We now consider it to be a detrimental invader of the Mexican coffee agro-ecosystem that should not be encouraged by augmentative release or introduced into other regions. Overall, the most successful species, in terms of both emergence and female production, was P. nasuta. We compare these results with those from prior reductionist and holistic studies, and with observations on patterns of establishment of these bethylid species in the field. Given that it is increasingly clear that disruptive inter-specific interactions are generally common when multiple species are deployed in biological control, screening of potential agents should consider such interactions alongside the more ‘traditional’ focus on host specificity.     

Developmental changes in urea concentrations in the haemolymph of Daizo (T), an original strain of the silkworm,Bombyx mori reared on an artificial diet and on fresh mulberry leaves

Urea concentrations in the haemolymph of Daizo (T), an original strain of the silkworm, Bombyx mori, reared on an artificial diet and on fresh mulberry leaves were determined by a urease-indophenol method during the larval-pupal-adult development. Urea concentrations in the fourth and fifth instar larvae reared on an artificial diet (diet I) were between 0.10 and 0.15 mg urea N/ml haemolymph, and increased during the larval-pupal transformation to reach 0.33 mg/ml at the larval-pupal ecdysis. A further increase was observed during pupal-adult development and finally reached 0.48 mg/ml at day 7 pharate adult. In the fourth and fifth instar larvae reared on fresh mulberry leaves, the concentrations were low (0.05 mg/ml). From the larval-pupal ecdysis until day 8 pharate adult, further low urea concentrations (0.04 mg/ml) were observed. By starvation from 72 hr of the fifth instar larvae reared on another artificial diet (diet II), the elevation of urea concentrations (between 1.4- and 3.0-fold against the controls) was observed from just after starvation until day 1 spinning. From day 5 pupae, both the starved and the control insects showed a marked elevation of urea concentrations in the haemolymph, which was never observed on animals reared on diet I.     

Development ofBrachymeria ovata [hymenoptera: chalcididae] in three noctuid hosts reared on artificial diet and insect-resistant and susceptible soybean genotypes

The development of the pupal parasitoidBrachymeria ovata (Say) was studied in 3 lepidopterous hosts reared on artificial diet and insect-susceptible and insectresistant soybena genotypes.Pseudoplusia includens (Walker),Anticarsia gemmatalis Hubner andHeliothis zea (Boddie) pupae from larvae reared on soybean leaves were less suitable forB. ovata development than pupae from larvae reared on artificial diet.B. ovata emergence rates, size and sometimes development period were adversely affected on plant-reared hosts. Dissection of hosts 6 days after parasitisation byB. ovata showed a higher proportion of diet-reared host pupae contained large parasitoid larvae than plant-reared hosts. Suitability differences were also detected among hosts reared on different soybean genotypes but these differences did not parallel closely the response of the lepidopterous hosts to soybean genotypes.      

Behavior and Activity Pattern of Cephalonomia stephanoderis (Hymenoptera: Bethylidae) Attacking the Coffee Berry Borer,Hypothenemus hampei (Coleoptera: Scolytidae)

We describe behavioral sequences and daily activities of pre-ovipositing and ovipositing females of Cephalonomia stephanoderis (Hymenoptera: Bethylidae), an ectoparasitoid of the coffee berry borer, Hypothenemus hampei (Coleoptera: Scolytidae). Noticeable behavioral differences among preovipositing and ovipositing females include host examination, host stinging—probing, host feeding, and the oviposition per se. The female of C. stephanoderis feeds primarily on host eggs, but pupae are also exploited, mainly by pre-ovipositing females. After the onset of the oviposition period, C. stephanoderis examines the pupae repeatedly, stings them at frequent intervals, and spends more time feeding than during the pre-oviposition period. Host paralysis is linked both to host feeding and oviposition. It induces irreversible developmental arrest, which presumably allows preservation of the host until subsequent utilization, and contributes to successful offspring development, particularly by reducing host motility. Oviposition consists in a host selection process, a prolonged period of preparation of the potential host, and the egg-laying phase itself. Under our experimental conditions, pre-ovipositing and ovipositing females are active 17% and 36% of the day, respectively. Host handling time averages 6% and 23% in pre-ovipositing and ovipositing females, respectively. All coffee berry borer developmental stages are exploited by C. stephanoderis females, either for host feeding and/or oviposition activities. Such flexible behavior is advantageous given that host availability is limited inside the coffee berries.     

稻纵卷叶螟绒茧蜂对寄主的搜索行为

稻纵卷叶螟绒茧蜂是稻纵卷叶螟幼虫的重要天敌。以Y形嗅觉仪测定了稻纵卷叶螟2—3龄幼虫虫粪、4—5龄幼虫虫粪、健康水稻、稻纵卷叶螟幼虫为害后水稻在纵卷叶螟绒茧蜂对寄主的搜索过程中的作用。结果表明已交配的雌蜂显著趋向稻纵卷叶螟2—3龄幼虫虫粪和4—5龄幼虫虫粪,不趋向健康水稻、稻纵卷叶螟幼虫为害后的水稻。雄蜂、处女雌蜂不显著趋向健康水稻、稻纵卷叶螟幼虫为害后水稻、稻纵卷叶螟2—3龄幼虫虫粪、稻纵卷叶螟4—5龄幼虫虫粪。利用固相微萃取(SPME)及气相色谱/质谱(GC/MS)联用分析稻纵卷叶螟2—3龄幼虫虫粪的化学成分,并通过Y-形嗅觉仪测试已交配雌蜂对稻纵卷叶螟2—3龄幼虫虫粪化学成分标准品的嗅觉反应,结果表明已交配的雌蜂显著趋向1-十一烯和异缬草醛,稻纵卷叶螟幼虫虫粪中的1-十一烯和异缬草醛可能是稻纵卷叶螟绒茧蜂搜索寄主的利他素。     

A fungal metabolite masks the host plant odor for the pine weevil (Hylobius abietis)

The pine weevil is one of the most important pest insects of conifer reforestation areas in Europe. Female pine weevils cover their eggs with chewed bark and feces (frass) resulting in avoidance behavior of feeding conspecifics towards egg laying sites. It has been suggested that microorganisms present in the frass may be responsible for producing deterrent compounds for the pine weevil. The fungi Ophiostoma canum, O. pluriannulatum, and yeast Debaryomyces hansenii were isolated from aseptically collected pine-weevil frass. The isolated fungi were cultured on weevil frass broth and their volatiles were collected by SPME and identified by GC–MS. D. hansenii produced methyl salicylate (MeS) as a major compound, whereas, in addition, O. canum and O. pluriannulatum produced 6-protoilludene. In a multi-choice lab bioassay, MeS strongly reduced pine weevil's attraction to the Pinus sylvestris volatiles. Thus, a fungal metabolite was found that strongly affects the pine weevil host-odor search.     

Field Evaluation of Steinernema feltiae Against the Web-spinning Larch Sawfly Cephalcia lariciphila

Field trials were conducted in Rheola Forest, Wales, Great Britain, to determine the effectiveness of Steinernema feltiae UK strain in controlling the web-spinning larch sawfly Cephalcia lariciphila. Foliar sprays at the rate of 5,000-20,000 nematodes/100 cm branch resulted in 3.4-29.4% infection of sawfly larvae. Soil application of 200 nematodes/cm² resulted in 61% infection of sawfly prepupae and 17.3% of pupae. Prepupal infection ranged from 4.8 to 14.7% 1 year after nematode application. Soil applications of this nematode show that it has potential for biological control of sawfly prepupae.     

Altitude and coffee production systems influence extent of infestation and bean damage by the coffee berry borer

The coffee berry borer (CBB), Hypothenemus hampei Ferrari (Coleoptera: Curculionidae: Scolytinae), is one of the major insect pests of coffee worldwide. The present study was designed to assess the level of infestation of coffee berries at different developmental stages across different altitudes and coffee management systems. The experiment was carried out at three locations in southwestern Ethiopia under two coffee management systems and four coffee berry development stages with three replications. Results of the study showed significantly highest proportion of damaged berries (37.5%), number of holes per berry (10.88) and number of adult CBB per berry (7.55) on dried leftover berries at low-altitude study sites. On the other hand, the lowest mean percent damaged berries, number of holes per berry and number of adults were recorded at mid- and high-altitude study sites. The study also showed that, CBB caused significantly highest damage in plantation coffee management system than garden coffee. Results of this study highlight proper harvesting at red ripe stage in order to minimise incidence of CBB. It is also important to design integrated management strategies to mitigate CBB damage especially in lowland plantation coffee production systems.