Isolation,morphological and molecular characterization of Bacillus thuringiensis strains against Hypothenemus hampei Ferrari (Coleoptera: Curculionidae: Scolytinae)

The coffee berry borer Hypothenemus hampei Ferrari, 1876 (Coleoptera: Curculionidae: Scolytinae) is considered the most serious pest of the coffee crop and is controlled primarily with the use of chemical insecticides. An alternative to this control method is the use of the entomopathogenic bacterium, Bacillus thuringiensis Berliner, 1911. Therefore, the objective of this work was to select strains of B. thuringiensis virulent against H. hampei and characterize them by morphological and molecular methods to identify possible genes for the production of genetically modified plants. To achieve this objective, 34 strains of B. thuringiensis underwent a selective bioassay to evaluate their toxicity to H. hampei first-instar larvae. Among the strains tested, 11 and the standard B. thuringiensis subspecies israelensis (IPS-82) caused mortality above 90%. Then, the median lethal concentration (LC₅₀) was estimated for these strains followed by characterization using morphological, biochemical and molecular methods. The lowest LC₅₀ was obtained for strain BR58, although this concentration did not differ significantly from that of the standard strain IPS-82 or from that of strains BR137, BR80 and BR67. The molecular characterization detected cry4A, cry4B, cry10, cry11 and cyt1 genes in 10 of the most virulent strains (BR58, BR137, BR80, BR81, BR147, BR135, BR146, BR138, BR139, BR140). Strain BR67 differed completely from the others and amplified only the cry3 gene. This strain was more virulent than BR135, BR146, BR138, BR139 and BR140, but it did not differ from BR58, BR137, BR80, BR81 and BR147. The protein profile revealed proteins of 28, 65, 70 and 130 kDa, and the morphological analysis identified spherical crystalline inclusions in all strains. The results showed that the 11 strains studied have potential for use as a gene source for insertion into coffee plants for the control H. hampei, especially the cry3, cry4A, cry4B, cry10, cry11 and cyt1 genes, that were repeated in the most virulent isolates.     

Reproductive compatibility between mite populations previously identified as Euseius concordis (Acari: Phytoseiidae)

The objective of the present research is to study the reproductive compatibility between populations of predatory mites previously identified as Euseius concordis (Chant) based on morphological characteristics. Colonies of these mite populations were established in the lab with specimens collected from different localities and host plants. Reproductive compatibility was evaluated through crosses and backcrosses within and between populations and the subsequent observation of females' oviposition, over a period of 10 days. The levels of oviposition obtained in the crosses between individuals from the same population were higher than those obtained in the crosses between individuals from different populations. Results indicate the occurrence of post-mating reproductive incompatibility between the mite population from Petrolina and the other populations studied. Crosses and backcrosses between populations involving female mites from Petrolina did not produce offspring, although endospermatophores were present inside the spermathecas of those females. Oviposition was reduced, and only sons were obtained, in crosses between populations with males from Petrolina. Crosses of females from Pontes e Lacerda and males from Jaguariúna and vice versa produced only male progeny. Our results established that the populations originating from Arroio do Meio, Pontes e Lacerda, Jaguariúna and Viçosa, are reproductively compatible. However, the latter populations and the population from Petrolina are genetically isolated. Based on these results we suggest that more cytological and genetic studies are needed to establish if this reproductive isolation represents a species barrier.     

咖啡天牛的生存对策,防治策略及其应用

通过分析咖啡旋皮天牛Ａｃａｌｏｌｅｐｔａ ｃｅｒｖｉｎｕｓ 和咖啡灭字虎天牛Ｘｙｌｏｔｒｅｃｈｕｓ ｑｕａｄｒｉｐｅｓ Ｃｈｅｖｒ;的生态对策,种间竞争策略,并结合有害生物综合管理理论,咖啡林生态系统的特性,咖啡天牛的发生特点,提出了咖啡天牛的防治策略。     

The biology of Phymastichus coffea LaSalle (Hymenoptera: Eulophidae) under field conditions

The coffee berry borer (CBB) Hypothenemus hampei (Ferrari) (Coleoptera: Curculionidae: Scolytinae) was accidentally introduced into México in 1978, and rapidly became the main pest of coffee. As an exotic pest, its management has been mainly based on biological control methods through the introduction of parasitoids from Africa. In this context, at the beginning of the present decade, the parasitoid Phymastichus coffea LaSalle (Hymenoptera: Eulophidae) was imported to Mexico. Since then, several studies have been carried out as part of the post introduction evaluation of this parasitoid. In this paper, information concerning the parasitism and life-cycle of P. coffea in coffee farms is presented with the objective of providing information that elucidates its role as a biological control agent. P. coffea showed highly significant preferences for allocation of two eggs per host, usually one female and one male. Both offspring are able to develop and reach the adult stage successfully. Lifespan of adults is 2–3 days only. The degree of parasitism by P. coffea was more than 95% at the three altitudes tested, when releases consisted of a ratio of 10 CBB:1 parasitoid. The median survivorship of CBB parasitized by this wasp was 13, 15 and 19 days at the low, medium and high altitude coffee zones, respectively. The parasitism by P. coffea was higher when parasitoid releases were carried out simultaneously with the CBB, and decreased with the time between host and parasitoid releases. We showed that using P. coffea at a density of 1 parasitoid per 10 hosts resulted in a 3- to 5.6-fold decrease in CBB damage to the coffee seeds when compared to the control. The importance and value of these results are discussed in terms of the use of P. coffea as a biological control agent of the CBB in Latin America.     

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.     

Biological control through provision of additional food to predators: a theoretical study

In this paper we analyze a variation of a standard predator-prey model with type II functional response which represents predator-prey dynamics in the presence of some additional food to the predator. The aim is to study the consequences of providing additional food on the system dynamics. We conclude that handling times for the available foods play a key role in determining the eventual state of the ecosystem. It is interesting to observe that by varying the quality and quantity of additional food we can not only control and limit the prey, but also limit and eradicate the predators. In the context of biological pest control, the results caution the manager on the choice of quality and quantity of the additional food used for this purpose. An arbitrary choice may have opposite effects leading to increase in pest concentration and eradication of the predator. This study offers insight into the possible management strategies that involve manipulation of quality and supply level of additional food to predators, for the benefit of biological control. The theoretical conclusions agree with results of some practical biological control experiments.     

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.     

Evaluation of pest control efficiencies for different banker plant systems with a simple predator–prey model

The banker plant system has been introduced for the biological control of various pest species in Japanese greenhouses. With the banker plant system, non-crop plants infested with a host insect (a non-commercial crop pest) are placed in the greenhouse to provide alternative resources for the parasitoids or predators. We want to evaluate the effectiveness for controlling pests on the crop in a quantitative way by immigrating predators from the banker plant. Therefore, we developed a simple model for the interaction of the pest and predator in the crop and included the banker plant only as a source for predators. For three different pest-predator systems we parameterised the model and used these models to predict under what conditions biological control in a banker plant system is successful. We defined successful as keeping the pest below the economic injury level of the crop estimated from damage analysis. Because the crop is mostly grown during a period that lasts less than a year our analysis should not only focus on the equilibrium dynamics. In contrast, it should also focus on the transient dynamics. Our main analytical result, from the equilibrium analysis, is that for successful control the maximum lifetime consumption of immigrating predators should exceed the daily prey growth at half the value of the maximum consumption rate. For practical purpose this translates into the fact that the immigration of predators at a low initial pest density is crucial for successful control.     

咖啡化学成分及其生物活性研究进展

咖啡为茜草科(Rubiaceae)咖啡属(Coffea)植物,位居世界三大饮品之首,具有降低血糖、保护肝脏和神经保护等作用.咖啡化学成分类别较多,包括生物碱、酚酸类、黄酮类、萜类等.咖啡中的化学成分是发挥其生物学功能和形成特色风味的基础,对其化学成分来源和生物活性进行综述,为进一步发展咖啡产业提供依据和理论支撑.     

Pyrethroid-Acarophenax lacunatus interaction in suppressing the beetle Rhyzopertha dominica on stored wheat

Any biocontrol agent used against stored grain insects will have to interact with insecticides used as grain protectants due to their heavy use especially in tropical areas. Therefore, the interaction between the pyrethroid insecticides bifenthrin and deltamethrin and Acarophenax lacunatus (Cross & Krantz) (Prostigmata: Acarophenacidae), an egg parasite of the stored cereal pest Rhyzopertha dominica(F.) (Coleoptera: Bostrichidae), was assessed in a range of mite densities and insecticide doses for each compound. Despite the mite species presence on all insecticide doses, the lowest instantaneous rate of increase of A.lacunatus was recorded with the highest doses of insecticides. The presence of the biological control agent at all doses of both insecticides suggests its tolerance to these compounds. A. lacunatusshowed higher parasitism rate with bifenthrin than with deltamethrin indicating a higher selectivity of the first compound to the biocontrol agent. This revised version was published online in August 2006 with corrections to the Cover Date.     

Low-Density Releases of Neoseiulus Fallacis Provide for Rapid Dispersal and Control of Tetranychus Urticae (Acari: Phytoseiidae, Tetranychidae) on Apple Seedlings

Releases of Neoseiulus fallacis (Garman) at 1500--6000 per ha when prey were at 0.1-0.3 per leaf provided seasonal control of Tetranychus urticae Koch (all stages) at 1-2 per leaf in an apple seedling rootstock nursery. Predaceous mites (all stages) increased to 0.3-0.4 per leaf after releases and predator prey ratios of < or = 1:3-7 provided pest regulation thereafter. Such low-density releases were thought to be effective because multiple dispersal bouts allowed predators to locate widely distributed spider mites (on 2-6% of leaves). A random-diffusion model simulating predator dispersal (incorporating wind speed and direction parameters) adequately explained movement and pest control patterns. An upright, dense, uniform planting of apple seedlings was an effective producer and recipient for dispersing predators and these attributes seemed to explain why biological control was so effective. Low-density releases of N. fallacis for control of T. urticae are predicted to be less effective on other crops with less prominent profiles and soil coverage.     

Susceptibility of the predatory stinkbug <Emphasis Type="Italic">Picromerus bidens</Emphasis> to selected insecticides

The Palaearctic stinkbug Picromerus bidens L. (Heteroptera: Pentatomidae) has been considered as a potential biocontrol agent of several defoliator pests in various agricultural and forest ecosystems. It may therefore be a valuable alternative for the Nearctic pentatomid Podisus maculiventris (Say) (Heteroptera: Pentatomidae), the augmentative use of which has largely been abandoned in Europe given its possible environmental impact as an exotic polyphagous predator. However, no study has yet documented the impact of insecticides on P. bidens, which is essential to evaluate the possible combination of this predatory pentatomid with current insecticide applications in integrated pest management (IPM) programmes. This study reports on laboratory experiments investigating the susceptibility of P. bidens to five insecticides with different modes of action: the pyrethroid deltamethrin, the diacylhydrazine methoxyfenozide, the juvenile hormone mimic pyriproxyfen, the spinosyn spinosad, and the neonicotinoid imidacloprid, all of which are used to control defoliator pests. Fourth-instar nymphs and female adults of the predator were exposed to formulated materials of the insecticides by residual contact. Methoxyfenozide and spinosad did not cause significant mortality to 4th instars and female adults of P. bidens. In contrast, deltamethrin and imidacloprid were harmful to nymphs and adults of the predator, with LC₅₀ values ranging between 1.5 and 9.9 mg a.i./l. Pyriproxyfen was toxic to 4th instars with an LC₅₀ value of 13.9 mg a.i./l but did not affect female adults. Reproduction and longevity of P. bidens were not adversely affected when the predator was exposed to field concentrations of spinosad, methoxyfenozide, or pyriproxyfen, or sublethal concentrations (around LC₁₀) of deltamethrin and imidacloprid. The results from residual contact experiments suggest that methoxyfenozide, spinosad, and to a lesser extent, pyriproxyfen may be compatible with P. bidens in an IPM programme. Further experiments assessing food chain toxicity of these compounds should offer a more complete picture of their selectivity.     

Bacillus thuringiensis serovar israelensis is highly toxic to the coffee berry borer, Hypothenemus hampei Ferr. (Coleoptera: Scolytidae)

A native collection of Bacillus thuringiensis strains was screened, once a reliable bioassay technique to assess the toxicity against the coffee berry borer (CBB) first-instar larvae was developed. A first round of bioassays with 170 strains indicated that the great majority of them showed no or very little insecticidal activity and that very few showed significant levels of toxicity. Interestingly, only those strains that had previously been associated with mosquitocidal activity were also toxic to CBB. Qualitative bioassays (using one high dose) were carried out only with those native mosquitocidal strains, corroborating their significant toxicity towards the CBB first-instar larvae. Most of these strains belong to serovar israelensis. In a second approach, strains from the Institut Pasteur type collection, whose mosquitocidal activity had been previously demonstrated, were also subjected to bioassays. Only those strains that showed a comparable protein content in their parasporal crystals to the israelensis type strain also showed high levels of toxicity towards CBB. Finally, an accurate LC(50) was estimated, using purified parasporal crystals from B. thuringiensis serovar israelensis type strain, at 219.5 ng cm(-2) of diet. All the statistical requirements for a reliable estimator were fulfilled. This is the first report of B. thuringiensis serovar israelensis being active against a coleopteran species.