Fast molecular assay to detect the rate of decay of <Emphasis Type="Italic">Bactrocera oleae</Emphasis> (Diptera: Tephritidae) DNA in <Emphasis Type="Italic">Pterostichus melas</Emphasis> (Coleoptera: Carabidae) gut contents

The molecular analysis of predation through specific DNA amplification has been utilized extensively over the last decade, and has been shown to be fast and effective. However, it is necessary to evaluate the prey detectability half-life if we are to correctly infer the relevance of particular predators to particular pests and to accurately model the effect of biocontrol. We present here the design and analysis of a set of primers to amplify olive fruit fly (Bactrocera oleae) DNA in predator gut contents, allowing fast evaluation of the digestion time. We modified the existing protocol by solubilizing the prey DNA directly from the gut, and we applied this modified protocol to demonstrate that Pterostichus melas, one of the most common carabids in olive groves in Italy, feeds on B. oleae pupae. After feeding carabids with a single pupa, traces of the pest were found to be detectable more than 20 h after ingestion. This method could also be applied to other predators to evaluate trophic interactions of the olive fruit fly. The relevance of generalist predation to the mortality of the pupal stage of B. oleae is of great economic interest since B. oleae causes serious damage during olive production, reducing the commercial value of olive oil and table olives.     

Olive fruit fly (Diptera: Tephritidae) ovipositional preference and larval performance in several commercially important olive varieties in California

The olive fruit fly, Bactrocera oleae (Gmelin) (Diptera: Tephritidae),is an invasive pest of olives (Olea spp.) in the United States. The objectives of this study were to determine whether B. oleae exhibits ovipositional preference under California field conditions similar to that demonstrated in European populations and whether the resulting larvae fare better in preferred varieties. Female B. oleae exhibited strong ovipositional preference for certain varieties of the domesticated olive, Olea europaea L, and the resulting larvae performed better by some measures in preferred varieties than in lesser preferred varieties. Ovipositional preference was observed in the field from 2003 to 2005, and laboratory assays were conducted to evaluate larval performance in 2005 and 2006. Among the olive varieties tested, Sevillano, Manzanillo, and Mission olives were the most heavily infested during three consecutive years. The larval performance measurements used were pupal yield, pupal weight, larval developmental time, and pupal emergence time. Ovipositional preference and pupal yield do not seem associated. There were significant differences in pupal emergence time, but these also measures did not reflect ovipositional preference. Two measures on performance did seem related to ovipositional preference; there were significant effects of variety on pupal weight and larval developmental time. Pupae developing in Manzanillo and Sevillano olives were heavier than those developing in less preferred varieties, and larval developmental time was significantly shorter in Sevillano olives relative to the other varieties. Oviposition preference and enhanced larval performance has implications for the pest status of this invasive insect in California.     

Evaluation of Fopius arisanus as a biological control agent for the olive fruit fly in California

1 The egg‐prepupal parasitoid Fopius arisanus (Hymenoptera: Braconidae) was evaluated in quarantine facilities as a potential biological control agent for the olive fruit fly Bactrocera oleae (Diptera: Tephritidae) in California, U.S.A. 2 Nontarget testing of two weed biological control agents confirmed that F. arisanus will not attack Tephritidae that feed in inflorescences or galls. It may, however, pose risks to native Tephritidae that feed in fruit. 3 Females preferentially oviposited in eggs, although first‐instar B. oleae were also attacked. Low lifetime reproductive potential and high rates of direct mortality inflicted on host eggs indicate that rearing on B. oleae may prove difficult. 4 In multiparasitized B. oleae, F. arisanus prevailed in competition against two species of larval–pupal parasitoids, Diachasmimorpha kraussii and Psyttalia concolor (both Hymenoptera: Braconidae). 5 The broad host‐range of F. arisanus with respect to fruit‐feeding Tephritidae may preclude its introduction to California, as may its low fecundity and its intrinsic competitive superiority over larva l–pupal parasitoids, which include specialists on B. oleae that are currently being introduced to California. High rates of direct mortality, however, point to potential uses in augmentative biological control. Whether or not F. arisanus is released in California, its biology as a parasitoid of B. oleae has been little studied to date and the results herein may be applied in other regions worldwide where B. oleae is a problem.     

Relation of fruit color,elongation, hardness,and volume to the infestation of olive cultivars by the olive fruit fly,Bactrocera oleae

The susceptibility of olive cultivars to the olive fruit fly, Bactrocera oleae (Rossi) (Diptera: Tephritidae), has seldom been studied. This article examines factors associated with olive fruit fly infestation of 16 commonly planted Sicilian olive cultivars. Total infestation data were simultaneously correlated with categorical and quantitative factors using ordinal logistic regression. When all factors were included in the analysis, year, sampling date, cultivar, and fruit color were highly significant, but the quantitative factors fruit volume, fruit elongation, and fruit hardness were not. When the analysis was repeated excluding cultivar, all quantitative factors were significant, and elongation and volume were highly significant. Spherical, large, and hard fruit seemed to be preferred by B. oleae over fruit that are elongate, small, and soft. Therefore, fruit color, elongation, volume, and hardness provide useful information regarding the susceptibility of cultivars. In both organic and conventional olive cultivation, information about olive cultivar susceptibility to olive fruit fly will help orchard managers to produce quality oil and table olives while reducing treatments for olive fruit fly control.     

Diachasmimorpha longicaudata and D. kraussii (Hymenoptera: Braconidae), potential parasitoids of the olive fruit fly

The olive fruit fly, Bactrocera oleae (Tephritidae), is a significant threat to California's olive industry. As part of a classical biological control program started in 2002, the parasitoids Diachasmimorpha kraussii and D. longicaudata (Hymenoptera: Braconidae) were imported to California from laboratory colonies in Hawaii. Studies on their biology and behavior as parasitoids of the olive fruit fly were conducted in quarantine. Both species tend to oviposit into 2nd and young 3rd instars, with the offspring completing development in the flies’ puparia. Most eggs are deposited in the first two weeks of adult life. Observed lifetime fecundity was low, possibly as a consequence of the relatively poor quality of the harvested olives used as a host substrate. Both pre-imaginal development and adult longevity were limited at constant temperatures above 30°C, which may indicate that these species will have difficulty establishing in the warmest regions of California.     

Psyttalia ponerophaga (Hymenoptera: Braconidae) as a potential biological control agent of olive fruit fly Bactrocera oleae (Diptera: Tephritidae) in California

The olive fruit fly, Bactrocera oleae (Rossi), is a newly invasive, significant threat to California's olive industry. As part of a classical biological control programme, Psyttalia ponerophaga (Silvestri) was imported to California from Pakistan and evaluated in quarantine. Biological parameters that would improve rearing and field-release protocols and permit comparisons to other olive fruit fly biological control agents were measured. Potential barriers to the successful establishment of P. ponerophaga, including the geographic origins of parasitoid and pest populations and constraints imposed by fruit size, were also evaluated as part of this investigation. Under insectary conditions, all larval stages except neonates were acceptable hosts. Provided a choice of host ages, the parasitoids' host-searching and oviposition preferences were a positive function of host age, with most offspring reared from hosts attacked as third instars. Immature developmental time was a negative function of tested temperatures, ranging from 25.5 to 12.4 days at 22 and 30 degrees C, respectively. Evaluation of adult longevity, at constant temperatures ranging from 15 to 34 degrees C, showed that P. ponerophaga had a broad tolerance of temperature, living from 3 to 34 days at 34 and 15 degrees C, respectively. Lifetime fecundity was 18.7 +/- 2.8 adult offspring per female, with most eggs deposited within 12 days after adult eclosion. Olive size affected parasitoid performance, with lower parasitism levels on hosts feeding in larger olives. The implications of these findings are discussed with respect to field manipulation and selection of parasitoid species for olive fruit fly biological control in California and worldwide.     

EcoPred: an educational individual based model to explain biological control,a case study within an arable land

ABSTRACT

Individual based models (IBMs) are up-to-date tools both in research and educational areas. Here we introduce an IBM built on NetLogo platform that simulates a top-down trophic cascade controlled by the pressure exerted by two model predators (web-building spiders and ground runner spiders) on a model pest (the olive fruit fly) within a hypothetical agricultural landscape (the olive crop). EcoPred is an IBM that aims to be an educational tool that can help teachers to explain concepts related to ecology in a modern, enjoyable and comprehensive way. EcoPred reflects the changes on a fly population within a simulated olive crop according to (1) the mortality rate caused by the predation of two spider species and energy loss, (2) the energy gain by feeding on flowers and (3) the reproduction rate in olive trees. The model was tested with 26 students achieving very good results in terms of acceptance and interest on the learning method. EcoPred can be used for educational purposes with 16 year old students and older to explain ecological concepts such as trophic level, species interactions, limiting factor and biological control in an interactive way simultaneously introducing students to biology oriented programming languages.     

Psyttalia cf. concolor (Hymenoptera: Braconidae) for biological control of olive fruit fly (Diptera: Tephritidae) in California

The larval parasitoid, Psyttalia cf. concolor (Szépligeti), reared on Mediterranean fruit fly, Ceratitis capitata (Weidemann), by the USDA-APHIS-PPQ, Guatemala City, Guatemala, was imported into California for biological control of olive fruit fly, Bactrocera oleae (Gmelin), in olives, Olea europaea L. Mean percentage parasitism of olive fruit fly third instars infesting fruit in field cages ranged from 7.0 in Grapevine to 59.7 in Santa Barbara and in free releases ranged from 0 in Grapevine to 10.6 in Santa Barbara after 4- to 6-d exposures. In the laboratory, more parasitoids developed to adults in olive fruit fly larvae that were 11-13 d old than in larvae 8-10 d old. Adult parasitoids lived significantly longer when provided with water than adults without water in environmental chambers at 5 degrees C, 85% RH; 15 degrees C, 65% RH; 25 degrees C, 25% RH; and 35 degrees C, 25% RH. Adult parasitoids lived for 48 d with honey for food and water and 32 d with food and sugar solution at 15 degrees C and 65% RH. Survival of adult parasitoids without food and water in greenhouse tests was approximately 4 d in a simulated coastal climate and 1 d in a simulated inland valley climate and was significantly increased by providing food and water. The parasitoid did not develop in the beneficial seedhead fly, Chaetorellia succinea (Costa), in yellow star thistle. The rate of parasitism of walnut husk fly, Rhagoletis completa Cresson, larvae in green walnut husks was 28.4% in laboratory no-choice tests. In choice tests, the rate of parasitism of walnut husk fly versus olive fruit fly larvae in olives was 11.5 and 24.2%, respectively.     

Olive Fruit Fly (Bactrocera oleae) Population Dynamics in the Eastern Mediterranean: Influence of Exogenous Uncertainty on a Monophagous Frugivorous Insect

Despite of the economic importance of the olive fly (Bactrocera oleae) and the large amount of biological and ecological studies on the insect, the factors driving its population dynamics (i.e., population persistence and regulation) had not been analytically investigated until the present study. Specifically, our study investigated the autoregressive process of the olive fly populations, and the joint role of intrinsic and extrinsic factors molding the population dynamics of the insect. Accounting for endogenous dynamics and the influences of exogenous factors such as olive grove temperature, the North Atlantic Oscillation and the presence of potential host fruit, we modeled olive fly populations in five locations in the Eastern Mediterranean region. Our models indicate that the rate of population change is mainly shaped by first and higher order non-monotonic, endogenous dynamics (i.e., density-dependent population feedback). The olive grove temperature was the main exogenous driver, while the North Atlantic Oscillation and fruit availability acted as significant exogenous factors in one of the five populations. Seasonal influences were also relevant for three of the populations. In spite of exogenous effects, the rate of population change was fairly stable along time. We propose that a special reproductive mechanism, such as reproductive quiescence, allows populations of monophagous fruit flies such as the olive fly to remain stable. Further, we discuss how weather factors could impinge constraints on the population dynamics at the local level. Particularly, local temperature dynamics could provide forecasting cues for management guidelines. Jointly, our results advocate for establishing monitoring programs and for a major focus of research on the relationship between life history traits and populations dynamics.     

Olive fruit fly (Diptera: Tephritidae) populations in relation to region, trap type, season, and availability of fruit

Olive fruit fly, Bactrocera oleae (Gmelin), was monitored with adult captures by season and trap type, and was related to fruit volume and nonharvested fruit to elucidate the occurrence of the newly introduced pest in California. The highest numbers of adults captured in ChamP traps in olive trees, Olea europaea, were in October in an inland valley location, and in September in a coastal location. Comparisons of trap types showed that the number of olive fruit fly adults captured in Pherocon AM traps in a commercial orchard was significantly greater than in ChamP traps. A significantly greater number of females were captured in Pherocon AM traps with bait packets and pheromone lures than traps with pheromone lures alone, while a significantly greater number of adults and males were captured in traps with pheromone lures alone. Significantly more adults were captured in ChamP traps with bait packets and pheromone lures versus traps with bait packets alone. Fruit volume increased by four times from mid-June to mid-November. Olive fruit fly was found to oviposit on small olive fruit <1 cm3 shortly after fruit set, the maximum number of ovipositional sites per fruit occurred in October, and the greatest number of pupae and adults were reared from fruit collected in September and October. The highest numbers of pupae were collected from nonharvested fruit in March when high numbers of adults were captured in the same orchard.     

Insecticidal activity of strains of Bacillus thuringiensis on larvae and adults of Bactrocera oleae Gmelin (Dipt. Tephritidae).

The olive fly, Bactrocera oleae, is the key pest on olives in the Mediterranean area. The pest can destroy, in some cases, up to 70% of the olive production. Its control relies mainly on chemical treatments, sometimes applied by aircraft over vast areas, with their subsequent ecological and toxicological side effects. Bacillus thuringiensis is a spore-forming soil bacterium which produces a protein crystal toxic to some insects, including the orders of Lepidoptera, Diptera, and Coleoptera and other invertebrates. The aim of this study was to search for isolates toxic to B. oleae. Several hundred B. thuringiensis isolates were obtained from olive groves and olive presses in different areas of Greece, Sardinia (Italy), and Spain and from cooperating scientists throughout the world. Some isolates were found toxic only to adults or larvae and some to both stages of the olive fly. In addition, the most toxic isolates were assayed on Opius concolor Szepl. (Hym. Braconidae), the most important parasitoid of the olive fruit fly. Only 3 isolates out of 14 gave significant mortality against this parasitoid. Several of the most toxic crystalliferous isolates may contain novel toxins since they gave no PCR products when probed with primers specified for 39 known toxin genes.     

Evaluation de deux méthodes de lutte intégrée contre les ravageurs en vergers d'agrumes

Abstract:  Evaluation of two IPM methods to control main pests in citrus orchards This study aims to the consolidation of the acquired knowledge regarding integrated pest management (IPM) against the main pests in citrus orchards. We provide evidence that the principles of this method already tested on small scales, are quite applicable on larger ones. In addition to this aspect, the purpose of our work has been the evaluation of two IPM methods: the first one, has been tested in Morocco for 3 years in areas not exceeding 1.2 ha; the second one originating from Australia, is presently recommended by the Plant Protection Authorities in Morocco. The results obtained confirm the complementarity between these two methods which are not mutually exclusive, despite some advantages recorded in favour of this method or the other. Our results show that the Moroccan method is more efficient in monitoring aphids and citrus leaf miner (CLM) population as it leads to the same results while reducing by half the number of the samples inspected. In contrast, mites monitoring is more convenient with the Australian method, allowing a good evaluation of the numbers of phytoseiid predators. Moreover, this method enabled us to reduce the quantity of pesticides used against Mediterranean fruit fly through treatment by lured spots reducing pulp volume by more than 92.8% and costs by 20%. These two methods gave the same results for monitoring population dynamics of California red scale and Mediterranean fruit fly but, fruit infestation at harvest was higher with the Australian method than the method developed locally (70.95% against 74.75 for California red scale and 2.35 against 3.35 for Mediterranean fruit fly).     

Microsatellite analysis revealed a different approach of control of olive fly population (Bactrocera oleae) in Slovenia

The olive fruit fly (Bactrocera oleae Gmelin) is the most important olive pest in the north‐eastern Adriatic coast region. Despite the importance of olive production in the region, and the significance of the olive fruit fly, no information with respect to genetic diversity, population structure or dispersion patterns of this pest is available. The aims of this study were to investigate the genetic structure of the olive fly population in the Slovenian Istria region using microsatellite markers to determine olive fruit fly migration between locations and to establish an appropriate and effective strategy for controlling the pest population. Analysis was performed on a sample of 117 flies, collected from attacked olive fruits at three different locations. Olive fruit flies were genotyped using eight microsatellite loci. Sixty‐six alleles were identified over all microsatellite loci with an average of 8.25 alleles per locus. The population structure was determined with methods based on Bayesian principles using the BAPS 6.0 and STRUCTURE 2.3 programs. Genetic analysis confirmed unlimited migration and random mating between individuals of different microlocations, which suggests time‐coordinated first treatment in the region would be the best solution.     

Prevalence of Candidatus Erwinia dacicola in wild and laboratory olive fruit fly populations and across developmental stages

The microbiome of the olive fruit fly, Bactrocera oleae (Gmelin), a worldwide pest of olives (Olea europaea L.), has been examined for >100 yr as part of efforts to identify bacteria that are plant pathogens vectored by the fly or are beneficial endosymbionts essential for the fly's survival and thus targets for possible biological control. Because tephritid fruit flies feed on free-living bacteria in their environment, distinguishing between the transient, acquired bacteria of their diet and persistent, resident bacteria that are vertically transmitted endosymbionts is difficult. Several culture-dependent and -independent studies have identified a diversity of species in the olive fruit fly microbiome, but they have not distinguished the roles of the microbes. Candidatus Erwinia dacicola, has been proposed to be a coevolved endosymbiont of the olive fruit fly; however, this was based on limited samples from two Italian populations. Our study shows that C. Erwinia dacicola was present in all New and Old World populations and in the majority of individuals of all life stages sampled in 2 yr. Olive fruit flies reared on olives in the laboratory had frequencies of C. Erwinia dacicola similar to that of wild populations; however, flies reared on artificial diets containing antibiotics in the laboratory rarely had the endosymbiont. The relative abundance of C. Erwinia dacicola varied across development stages, being most abundant in ovipositing females and larvae. This uniform presence of C. Erwini dacicola suggests that it is a persistent, resident endosymbiont of the olive fruit fly.     

Pupal development of Ceratitis capitata (Diptera: Tephritidae) and Diachasmimorpha longicaudata (Hymenoptera: Braconidae) at different moisture values in four soil types

This study aimed to evaluate adult emergence and duration of the pupal stage of the Mediterranean fruit fly, Ceratitis capitata (Wiedemann), and emergence of the fruit fly parasitoid, Diachasmimorpha longicaudata (Ashmead), under different moisture conditions in four soil types, using soil water matric potential. Pupal stage duration in C. capitata was influenced differently for males and females. In females, only soil type affected pupal stage duration, which was longer in a clay soil. In males, pupal stage duration was individually influenced by moisture and soil type, with a reduction in pupal stage duration in a heavy clay soil and in a sandy clay, with longer duration in the clay soil. As matric potential decreased, duration of the pupal stage of C. capitata males increased, regardless of soil type. C. capitata emergence was affected by moisture, regardless of soil type, and was higher in drier soils. The emergence of D. longicaudata adults was individually influenced by soil type and moisture factors, and the number of emerged D. longicaudata adults was three times higher in sandy loam and lower in a heavy clay soil. Always, the number of emerged adults was higher at higher moisture conditions. C. capitata and D. longicaudata pupal development was affected by moisture and soil type, which may facilitate pest sampling and allow release areas for the parasitoid to be defined under field conditions.     

Evaluation of Beauveria bassiana and B. brongniartii strains and four wild-type fungal species against adults of Bactrocera oleae and Ceratitis capitata

The virulence of two isolates of the hyphomycete fungi, Beauveria bassianaand B. brongniartii, and additional fungal species isolated from diseased Bactrocera oleae pupae and Sesamia nonagrioideslarvae were assessed against adults of the olive fruit fly B. oleae and the Mediterranean fruit fly Ceratitis capitata (Diptera: Tephritidae). Contact and oral bioassays revealed that moderate to high mortality rates for the olive fruit fly occurred when the adults were exposed to conidia of Mucor hiemalis, Penicillium aurantiogriseum, P. chrysogenum and B. bassianaisolates. A strain of M. hiemalis isolated from S. nonagrioides larvae was the most toxic resulting in 85.2% mortality to the olive fruit fly adults. B. brongniartiiand B. bassiana were the most pathogenic to the C. capitataadults causing 97.4 and 85.6% mortality. Metabolites collected from the M. hiemalis and P. chrysogenum isolates were toxic to adults of both species.     

Transinfection of the olive fruit fly Bactrocera oleae with Wolbachia: towards a symbiont‐based population control strategy

The olive fruit fly Bactrocera oleae is responsible for worldwide economic damage. In this report, we describe the first B. oleae lines transinfected with the Wolbachia strain wCer2, an endosymbiont of the cherry fruit fly Rhagoletis cerasi. Immunostaining followed by confocal microscopy, detects high numbers of Wolbachia in embryos as well as in ovarioles and sperm from individuals of both transinfected lines. wCer2 was uniformly distributed in B. oleae egg chambers and the cortex of preblastoderm embryos. Wolbachia is known to manipulate host reproduction with several strategies, one of which is cytoplasmic incompatibility (CI), resulting in embryonic mortality in incompatible crosses. Wolbachia was found to induce complete CI in the novel host, suggesting that symbiont‐based approaches can be used as novel environmentally friendly tools for the control of natural olive fruit fly populations.     

Delayed density-dependent parasitism of eggs and pupae as a contributor to the cyclic population dynamics of the autumnal moth

Many populations of forest Lepidoptera exhibit 10-year cycles in densities, with impressive outbreaks across large regions. Delayed density-dependent interactions with natural enemies are recognized as key factors driving these cyclic population dynamics, but emphasis has typically been on the larval stages. Eggs, pupae and adults also suffer mortality from predators, parasitoids and pathogens, but little is known about possible density relationships between mortality factors and these non-feeding life stages. In a long-term field study, we experimentally deployed autumnal moth (Epirrita autumnata) eggs and pupae to their natural enemies yearly throughout the 10-year population cycle in northern Norway. The abundance of another geometrid, the winter moth (Operophtera brumata), increased in the study area, permitting comparisons between the two moth species in predation and parasitism. Survival of autumnal moth eggs and pupae was related to the moth abundance in an inverse and delayed manner. Egg and pupal parasitoids dominated as density-dependent mortality factors and predicted the subsequent growth rate of the host population size. In contrast, effects of egg and pupal predators were weakly density dependent, and generally predation remained low. Parasitism rates did not differ between the autumnal and winter moth pupae, whereas predators preferred winter moth pupae over those of the autumnal moth. We conclude that parasitism of the autumnal moth by egg and pupal parasitoids can be related to the changes of the moth density in a delayed density-dependent manner. Furthermore, egg and pupal parasitoids cannot be overlooked as causal factors for the population cycles of forest Lepidoptera in general.     

Effect of exposure time on mass‐rearing production of the olive fruit fly parasitoid,Psyttalia lounsburyi (Hymenoptera: Braconidae)

Classical biological control programmes rely on mass production of high‐quality beneficial insects for subsequent releases into the field. Psyttalia lounsburyi (Silvestri) (Hymenoptera: Braconidae) is a koinobiont larval–pupal endoparasitoid of tephritid flies that is being reared to support a classical biological control programme for olive fruit fly in California. The mass‐rearing system for a P. lounsburyi colony, initiated with insects originally collected in Kenya, was evaluated with the goal of increasing production, while at the same time reducing time requirements for rearing in a quarantine facility. We tested the effect of exposure time of a factitious host Ceratitis capitata (Wiedemann) (Diptera: Tephritidae), on parasitization, adult production, superparasitism, and sex ratio of P. lounsburyi and survival of the host. Parasitization rates were highest (31%) at 3‐ and 4‐hr exposure times, while adult production (i.e., emergence of wasp progeny) was highest (16%) at the 2‐hr exposure time. Superparasitism over the course of the study was 1.5% and did not appear to be a factor affecting parasitoid production. The sex ratio of wasp progeny was male‐biased and did not vary significantly over different exposure times. The rate of stings on host larvae increased with exposure time and was consistent with decreases in pupal eclosion from larvae and emergence rate of adult flies. When compared to current rearing procedures, the 2‐hr exposure time resulted in an overall 2.8‐fold increase in P. lounsburyi production when standardized for time.