Mediterranean fruit fly population phenological patterns are strongly affected by elevation and host presence.

The Mediterranean fruit fly (medfly) (Ceratitis capitata, Diptera: Tephritidae), is an extremely polyphagous pest that threatens the fruit production and trading industry worldwide. Monitoring C. capitata populations and analysing its dynamics and phenology is considered of outmost importance for designing and implementing sound management approaches. The aim of this study was to investigate the factors regulating the population dynamics of the C. capitata in a coastal and semi-mountainous area. We focused on effects of topography (e.g. elevation), host presence and seasonal patterns of ripening on the phenological patterns considering data collected in 2008. The experimental area is characterized by mixed fruit orchards, and Mediterranean climate with mild winters. Two trap types were used for population monitoring. The female targeted McPhail type and the male targeted Jackson type. Traps were placed in farms located at different elevations and landscape morphology (coastal and semi-mountainous areas). The main crops included citrus, apples, peaches, plums, pears, figs, quinces and apricots. Adult captures were first recorded in May, peaked in mid-summer and mid-autumn and almost ceased at the end of the season (January 2008). Captures in the coastal areas preceded that of highlands by 15 days. Most of the adults detected during the fruit ripening of late stone fruit cultivars (first peak) and citrus (second peak). The probability of capturing the first adults preceded almost three weeks the peak of adult captures either considering the elevation or host focus analyses. The results provide valuable information on the seasonal population trend of C. capitata in mixed fruit Mediterranean orchards and can support the set-up of IPM systems in areas with various landscapes and different hosts throughout the fruit growing season.

Electronic recording of lifetime locomotory activity patterns of adult medflies.

Age-specific and diurnal patterns of locomotory activity, can be considered as biomarkers of aging in model organisms and vary across the lifetime of individuals. Τhe Mediterranean fruit fly (medfly), Ceratitis capitata, is a commonly used model-species in studies regarding demography and aging. In the present study, we introduce a modification of the automated locomotory activity electronic device LAM25system (Locomotory Activity Monitor)-Trikinetics, commonly used in short time studies, to record the daily locomotory activity patterns of adult medflies throughout the life. Additionally, fecundity rates and survival of adult medflies were recorded. Male and female medflies were kept in the system tubes and had access to an agar-based gel diet, which provided water and nutrients. The locomotory activity was recorded at every minute by three monitors in the electronic device. The locomotory activity of females was higher than that of males across the different ages. For both sexes locomotory rates were high during the first 20 days of the adult life and decreased in older ages. The activity of males was high in the morning and late afternoon hours, while that of females was constantly high throughout the photophase. Negligible locomotory activity was recorded for both sexes during the nighttime. Males outlived females. Fecundity of females was higher in younger ages. Our results support the adoption of LAM25system in studies addressing aging of insects using medfly as a model organism.

Adaptation of an Invasive Pest to Novel Environments: Life History Traits of Drosophila suzukii in Coastal and Mainland Areas of Greece during Overwintering.

Drosophila suzukii is a polyphagous pest of small and soft fruit, originating from Asia, which has spread and established in Europe and the USA. Adults exhibit seasonal phenotypes, i.e., summer morphs (SM) and winter morphs (WM) to cope with fluctuating environmental conditions. WM have a darker cuticle and larger wings compared to SM, while WM females experience reproductive dormancy. We studied the life history traits (lifespan, female reproductive status and number of produced offspring) of WM and SM that were exposed to winter field conditions of a coastal and a mainland agricultural area, with mild and cold winter climates, respectively. Mated adults of each phenotype were individually placed in vials bearing nutritional/oviposition substrate, and transferred to the field from November 2019 to May 2020, when the death of the last individual was recorded. Almost all SM females (90%) and no WM female carried mature ovarioles before being transferred to the field. WM exhibited a longer lifespan than SM adjusting for location and sex. Differences in survival between the two phenotypes were more pronounced for adults kept in the mainland area. The majority of SM females produced offspring during overwintering in the mild coastal area, but only a few SM were reproductively active in the cold mainland area. Some WM females produced progeny during overwintering in the mild conditions of the coastal area, but all WM females were in reproductive arrest in the mainland area. Overwintering females in the coastal area had a shorter lifespan and produced more progeny than those kept in the mainland area. High survival rates of WM provide indications of the successful performance of this phenotype in the adverse conditions of the cold climates. Additionally, the continuous reproductive activity of SM females and the onset of progeny production by WM females during overwintering in the coastal area indicate that the insect remains reproductively active throughout the year in areas with mild climatic conditions. Our findings support the successful adaptation of D. suzukii in both areas tested and can be used for the development of area-specific population models, based on the prevailing climatic conditions.

Population Dynamics of Drosophila suzukii in Coastal and Mainland Sweet Cherry Orchards of Greece.

Despite the recent invasion and wide spread of Drosophila suzukii Matsumura (Diptera: Drosophilidae) in Europe, little is known regarding its population trends in coastal areas of the southern Mediterranean countries. Using adult trapping and fruit sampling, we studied the population dynamics of D. suzukii in coastal and mainland (semi-highland) cherry orchards of Greece, from 2018 to 2020. Adults were captured in traps baited with apple cider vinegar, placed in conventional and unmanaged sweet-cherry orchards, and in neighbouring wild growing hosts. Sampling of sweet-cherry fruit to assess infestation levels was conducted from early and late-ripening cherry cultivars in both areas. Adults were captured throughout the year in the coastal area with two peaks registered in spring and late-autumn. Captures were nearly zero during the hot summer months. Flight activity exhibited only one peak in autumn at the mainland area, and ceased during winter and spring. Captures in wild hosts were lower during the sweet-cherry ripening period than later in the season. Higher sweet-cherry infestation levels were recorded in the coastal than in the mainland area and in unmanaged than in commercial orchards. Both early and late-ripening cultivars were highly susceptible to D. suzukii infestation in the coastal area. Infestation rates were higher in late-ripening cultivars than in early-ripening ones in the mainland area. We conclude that D. suzukii has well adapted to the Mediterranean climate of Greece, and is able to progressively exploit several crops and wild hosts of mainland and coastal areas.

Oviposition-deterrent effect of linalool - a compound of citrus essential oils - on female Mediterranean fruit flies, Ceratitis capitata (Diptera: Tephritidae).

BACKGROUND: Linalool is a natural scent, found in essential oils (EOs) of several plants. It is widely used as a fragrant, also possessing toxic, oviposition-deterrent and repellent properties against many insect species. Ceratitis capitata (medfly) is one of the most notorious insect pests for international fresh fruit production. Despite the available information on the repellent effects of EOs on herbivorous insects, possible oviposition-deterrent effects of linalool against C. capitata remain unknown. OBJECTIVES AND RESULTS: We tested whether different linalool concentrations, application methods on fruit hosts and time since application elicited oviposition-deterrence on medfly, from bitter oranges, apples and nectarines, under constant laboratory conditions. We also tested possible repellent properties of linalool on adult medflies in field conditions. Females drilled fewer oviposition stings and deposited fewer eggs per fruit in all linalool-treated hosts. Spraying and topical-droplet application were the most efficient methods on bitter oranges, although spraying was the least effective method in apples. Bitter oranges and nectarines that were offered to females immediately post-exposure to linalool received more oviposition stings and eggs than those offered 3 days post-exposure to linalool solutions. Conversely, apples tested 3 days following linalool application received significantly more oviposition stings and eggs than those tested immediately and 1 day post-exposure. More adults in total, and more females, were captured in traps located on untreated-control than on linalool-treated trees, especially in citrus hosts. CONCLUSION: Linalool has a potential future use in environmentally friendly control strategies against C. capitata. © 2020 Society of Chemical Industry.

Effect of host fruit, temperature and Wolbachia infection on survival and development of Ceratitis capitata immature stages.

The Mediterranean fruit fly, Ceratitis capitata (Diptera: Tephritidae), holds an impressive record of successful invasions promoted by the growth and development of international fruit trade. Hence, survival of immatures within infested fruit that are subjected to various conditions during transportation seems to be a crucial feature that promotes invasion success. Wolbachia pipientis is a common endosymbiont of insects and other arthropods generating several biological effects on its hosts. Existing information report the influence of Wolbachia on the fitness traits of insect host species, including the Mediterranean fruit fly. However, little is known regarding effects of Wolbachia infection on immature development in different host fruits and temperatures. This study was conducted to determine the development and survival of immature stages of four different Mediterranean fruit fly populations, either infected or uninfected with Wolbachia, in two hosts (apples, bitter oranges) under three constant temperatures (15, 25 and 30°C), constant relative humidity (45-55 ± 5%), and a photoperiod of 14L:10D. Our findings demonstrate both differential response of two fruit fly lines to Wolbachia infection and differential effects of the two Wolbachia strains on the same Mediterranean fruit fly line. Larva-to-pupa and larva-to-adult survival followed similar patterns and varied a lot among the four medfly populations, the two host fruits and the different temperatures. Pupation rates and larval developmental time were higher for larvae implanted in apples compared to bitter oranges. The survival rates of wildish medflies were higher than those of the laboratory adapted ones, particularly in bitter oranges. The Wolbachia infected medflies, expressed lower survival rates and higher developmental times, especially the wCer4 infected line. High temperatures constrained immature development and were lethal for the Wolbachia infected wCer4 medfly line. Lower temperatures inferred longer developmental times to immature stages of all medfly populations tested, in both host fruits. Implications on the ecology and survival of the fly in nature are discussed.

Effects of early-life protein starvation on longevity and sexual performance of male medfly.

Using a well-established model species for demographic, behavioural and aging research, the Mediterranean fruit fly (Ceratitis capitata), we explored whether nutritional stress early in adult life affects the sexual performance and survival in older ages. To do so we established two different protein starvation (PS) protocols that included the elimination of proteinaceous diet either before or after sexual maturity of male medflies. The frequency of sexual signalling and the age of death were daily recorded. Sexual signalling is directly related with male mating success in this model system. PS early in adult life results in high mortality rates (similar to sugar-only fed males), which are gradually restored in more advanced ages. Provision of a proteinaceous diet following early-life PS increases straightaway male sexual signalling to levels similar with those having continuous access to proteinaceous diet. Switching diet regimes from a protein-free to a protein-rich one progressively compensates mortality rates. Apparently, males prioritize sexual signalling over lifespan. PS after attaining sexual maturity significantly reduces both longevity and sexual performance. Access to protein only early in life is insufficient to support lifetime energy-consuming behaviours such as sexual signalling. Continuous access to a proteinaceous diet determines both lifetime sexual performance and longevity. Early in life PS males prioritize the allocation of nutritional elements, when available, in sexual activities over soma-maintenance.

Toxic and hormetic-like effects of three components of citrus essential oils on adult Mediterranean fruit flies (Ceratitis capitata).

Plant essential oils (EOs) and a wide range of their individual components are involved in a variety of biological interactions with insect pests including stimulatory, deterrent, toxic and even hormetic effects. Both the beneficial and toxic properties of citrus EOs on the Mediterranean fruit fly (medfly) have been experimentally evidenced over the last years. However, no information is available regarding the toxic or beneficial effects of the major components of citrus EOs via contact with the adults of the Mediterranean fruit fly. In the present study, we explored the toxicity of limonene, linalool and α-pinene (3 of the main compounds of citrus EOs) against adult medflies and identified the effects of sub-lethal doses of limonene on fitness traits in a relaxed [full diet (yeast and sugar)] and in a stressful (sugar only) feeding environment. Our results demonstrate that all three compounds inferred high toxicity to adult medflies regardless of the diet, with males being more sensitive than females. Sub-lethal doses of limonene (LD20) enhanced the lifespan of adult medflies when they were deprived of protein. Fecundity was positively affected when females were exposed to limonene sub-lethal doses. Therefore, limonene, a major constituent of citrus EOs, induces high mortality at increased doses and positive effects on life history traits of medfly adults through contact at low sub-lethal doses. A hormetic-like effect of limonene to adult medflies and its possible underlying mechanisms are discussed.

Description of Rhagoletis cerasi (Diptera: Tephritidae) pupal developmental stages: indications of prolonged diapause.

The European cherry fruit fly, Rhagoletis cerasi (L.) (Diptera: Tephritidae), is the key pest of sweet and sour cherries in many European countries and west Asia. It is a univoltine species of the west Palaearctic zone that undergoes obligatory pupal diapause. In this study, the development of R. cerasi pupae that were brought to an optimum temperature for postdiapause development following a long chilling period is described. The six most representative developmental stages within the puparium are illustrated, and the developmental progression among the stages after the end of the chilling period is quantified. Within 20 d postchilling, there was a gradual progress from stage I to pharate adult. However, ∼30% of the pupae remained at the transitional stage II, after 20 d at 25°C (optimum temperature for development). This suggests that a proportion of pupae remain at an intermediate developmental stage for an extended period of time that goes beyond 20 d postchilling. The pupal stage II might be related to diapause termination and responsiveness to environmental cues. It may also define the time before developmental progress to pharate adult. This finding agrees with previous studies proposing that a number of R. cerasi pupae undergo prolonged diapause, though the morphological characteristics of these pupae have never been described before.

Condition-dependent effects of mating on longevity and fecundity of female Medflies: the interplay between nutrition and age of mating.

BACKGROUND: In various species mating exerts direct and indirect effects on female demographic traits ranging from life span shortening to behavioural shifts. A wealth of data regarding effects of nutrition on longevity and reproduction output also exists. Nonetheless, little is known regarding the interaction between the age of mating and nutrition on female fitness. METHODOLOGY: We studied, the effects of protein deprivation and age of mating on female fitness traits, using a wild population of the Mediterranean fruit fly (medfly). We tested the hypotheses that (a) protein availability increases female lifespan and fecundity, (b) female longevity and egg production are independent of mating and the age of mating, and (c) female mating behaviour is independent of their age and nutritional status. Thus, we recorded the mating success and the copulation characteristics, as well as the egg production and survival of females mated at young or at old age and fed a full or a protein-deprived diet. RESULTS: Mating boosts egg production and reduces longevity of protein-fed females. On the contrary, mating increases the longevity of protein-deprived females. Mortality responses (negative or positive) to mating are expressed after a long lag phase. Old females are more receptive and less selective than young females regardless of the food regime. CONCLUSIONS: Our findings suggest that condition (nutritional status and age) defines the positive or negative output of mating in female medflies. These results contribute towards understanding the effects of mating, aging, resource allocation and their interactions on survival and female reproduction.

Dual reproductive cost of aging in male Medflies: dramatic decrease in mating competitiveness and gradual reduction in mating performance.

Although age-based effects on the reproductive success of males have been reported in several animal taxa the cost of aging on male mating success in lekking species has not been fully explored. We used the Mediterranean fruit fly, a lekking species, to investigate possible cost of aging on male reproductive success. We performed no choice and choice mating tests to test the hypothesis that aging does not affect the mating performance (mating success in conditions lacking competition) or the mating competitiveness (mating success against younger rivals) of males. The mating probability of older males decreased significantly when competing with younger males. Aging gradually reduced the mating performance of males but older males were still accepted as mating partners in conditions lacking competition. Therefore, older males are capable of performing the complete repertoire of sexual performance but fail to be chosen by females in the presence of young rivals. Older males achieved shorter copulations than younger ones, and female readiness to mate was negatively affected by male age. Older and younger males transferred similar amount of spermatozoids to female spermathecae. Females stored spermatozoids asymmetrically in the two spermathecae regardless the age of their mating partner. Aging positively affected the amount of spermatozoids in testes of both mated and nonmated males. No significant differences were observed on the amount of spermatozoids between mated and nonmated males.

Physiological and biological patterns of a highland and a coastal population of the European cherry fruit fly during diapause.

Adult emergence of univoltine temperate insect species and its synchronization with specific host phenological stages is mainly regulated by obligatory pupal diapause. Although a few studies have investigated the factors affecting diapause intensity, little attention has been paid to the physiological alterations and metabolic regulation that take place during diapause. Here, we describe differences in diapause between a highland and a coastal Greek population of the European cherry fruit fly Rhagoletis cerasi, a major pest of sweet and sour cherries in many European countries. Pupae of both populations were exposed to the environmental conditions prevailing in the two areas and diapause termination was observed under laboratory conditions. The regulation of energetic metabolites during the long pupae stage was examined under both field and laboratory conditions. Differences in diapause intensity revealed that the two populations have adapted to the local geographical and climatic conditions and have different requirements for low temperatures to terminate diapause. The coastal population undergoes a shorter diapause and adults emerge more rapidly, especially in the highland area. The highland population failed to terminate diapause (<40% adult emergence) in the coastal area. Both populations draw on their major energetic reserves (lipids and protein) similarly during diapause. Nevertheless, regulation of carbohydrate and glycogen reserves seems to vary between the populations: major peaks of these stored nutrients occur on different dates in the two populations, suggesting a differential regulation. Differences in diapause intensity imply a genetic differentiation between the two populations. The importance of our findings in understanding the physiological patterns during obligatory diapause of a univoltine insect species, as well as the practical implications for the development of specific phenological models for the European cherry fruit fly are discussed.