Behavioural response of the house mosquitoes Culex quinquefasciatus and Culex pipiens molestus to avian odours and its reliance on carbon dioxide.

How Culex (Diptera: Culicidae) mosquitoes select and discriminate between potential avian hosts is critical for understanding the epidemiology of West Nile virus. Therefore, the present authors studied the behavioural responses of Culex quinquefasciatus (Say) and Culex pipiens molestus (Forsskål) to headspace volatiles of three avian species [chicken and pigeon (sexes analysed separately), and magpie], presented either alone or in combination with 600 p.p.m. carbon dioxide (CO2 ). The attraction of Cx. quinquefasciatus to the headspace volatiles of both sexes of chicken, and of female pigeon, in combination with CO2 was significantly higher than that achieved by the CO2 and solvent control. Although Cx. p. molestus was attracted to headspace volatiles of chickens and magpies, it was repelled by those of female pigeons when combined with CO2 . An increased effect between the avian volatiles and CO2 was observed for Cx. quinquefasciatus, whereas the addition of CO2 had no effect on the attraction of Cx. p. molestus females. The results of this study demonstrate that Cx. quinquefasciatus and Cx. p. molestus are attracted to the odour of potential avian hosts. Future studies aimed at identifying the bioactive volatile compounds in the headspace of chickens may contribute to the potential development of effective surveillance and control tools against Culex species.

Variation in adult longevity of Culex pipiens f. pipiens, vector of the West Nile Virus.

The common house mosquito, Culex pipiens (Diptera: Culicidae), which is considered the primary bridge vector of West Nile Virus (WNV) to humans, is a wide spread insect pest with medical importance and consists of two distinct bioforms, Cx. pipiens f. pipiens and Cx. pipiens f. molestus. Here, we consider the adult lifespan of male and female Cx. pipiens f. pipiens under controlled conditions at five constant temperature regimes (15, 20, 25, 27.5, and 30 °C). Our results show that adult longevity was affected by temperature, as it significantly decreased with increase in temperature. At the highest tested temperature, mean adult longevity did not exceed 12 days for both sexes and thus makes impossible the risk of WNV transmission. On the other hand at the lowest temperature, longevity was extremely high with some individuals surviving up to 129 and 132 days, males and females, respectively, and thus enable them to function as potential vectors of WNV for a prolonged period of time. As far as sex is concerned, adult females displayed a 1.2-1.4-fold longer longevity compared to the male ones. However, this difference was significant only at the lowest and highest tested temperature regime. This information is useful in determining the critical temperatures which may affect the distribution of Cx. pipiens and consequently the risk of WNV transmission. Moreover, the effect of environmental temperature should be considered when evaluating the abundance of these species.

Resurgence of the cotton bollworm Helicoverpa armigera in northern Greece associated with insecticide resistance.

Helicoverpa armigera has been controlled effectively with chemical insecticides in the major cotton crop production areas of northern Greece for many years. However, a resurgence of the pest was observed in 2010, which significantly affected crop production. During a 4-year survey (2007-2010), we examined the insecticide resistance status of H. armigera populations from two major and representative cotton production areas in northern Greece against seven insecticides (chlorpyrifos, diazinon, methomyl, alpha-cypermethrin, cypermethrin, gamma-cyhalothrin and endosulfan). Full dose-response bioassays on third instar larvae were performed by topical application. Lethal doses at 50% were estimated by probit analysis and resistance factors (RF) were calculated, compared to a susceptible laboratory reference strain. Resistance levels were relatively moderate until 2009, with resistance ratios below 10-fold for organophosphates and carbamates and up to 16-fold for the pyrethroid alpha-cypermethrin. However, resistance rose to 46- and 81-fold for chlorpyrifos and alpha-cypermethrin, respectively in 2010, when the resurgence of the pest was observed. None of the known pyrethroid resistance mutations were found in the pyrethroid-resistant insects. The possible association between resistance and H. armigera resurgence in Greece is discussed.

Effects of constant and changing temperature conditions on diapause induction in Helicoverpa armigera (Lepidoptera: Noctuidae).

The effects of photoperiod and temperature on the induction and termination of facultative pupal diapause in Helicoverpa armigera (Lepidoptera: Noctuidae) were investigated under laboratory conditions. Exposing H. armigera larvae to both constant and fluctuating temperature regimes with a mean of 25°C and 20°C resulted in a type-III photoperiodic response curve of a short-long day insect. The long-day critical daylengths for diapause induction were ten hours and 12 hours at the constant temperatures of 25°C and 20°C, respectively. Higher incidences of diapause and higher values both for the longer and the shorter critical photoperiods for diapause induction were observed at fluctuating regimes compared with the corresponding constant ones. At alternating temperatures, the incidence of diapause ranged from 4.2% to 33.3% and was determined by the temperature amplitude of the thermoperiod and by the interaction of cryophase or thermophase with the photoperiod. Helicoverpa armigera larvae seem to respond to photoperiodic stimuli at temperatures >15°C and <30°C; all insects entered diapause at a constant temperature of 15°C, whereas none did so at a constant temperature of 30°C under all the photoperiodic regimes examined. Although chilling was not a prerequisite for diapause termination, exposure of diapausing pupae to chilling conditions significantly accelerated diapause development and the time of adult emergence. Therefore, temperature may be the primary factor controlling the termination of diapause in H. armigera.

Cold tolerance of field-collected and laboratory reared larvae of Sesamia nonagrioides (Lepidoptera: Noctuidae).

Sesamia nonagrioides Lefébvre (Lepidoptera: Noctuidae) is considered one of the most destructive pests of corn in the Mediterranean region. The purpose of the present study was to investigate some aspects of the cold tolerance of non-diapausing and diapausing laboratory reared larvae of S. nonagrioides, as well as of field-collected larvae, taking into consideration various parameters, such as supercooling ability, mean lethal temperature and accumulation of cryoprotectant substances, in relation to diapause. Our results provide evidence that S. nonagrioides has limited cold tolerance as it displays a low ability of supercooling. This is strongly supported by the fact that mortality of the individuals occurred after extended exposure to subzero temperatures, equivalent or slightly lower to their mean supercooling point. However, lethal temperatures of diapausing larvae were significantly lower in relation to that of non-diapausing larvae, indicating the existence of a direct link between diapause and cold tolerance. Regarding the role of cryoprotectant substances, accumulation of glycerol seems to be closely related to diapause, in contrast to accumulation of trehalose, which is more related to exposure to low temperatures slightly higher than 0 degree C. Finally, non-diapausing larvae of different instars displayed a similar ability of supercooling and tolerance to low temperatures as well as accumulation of cryoprotectant substances. The ecological significance of our findings on cold tolerance of this species is being discussed with particular reference to the microclimate observed in northern Greece.

Overwintering survival and spring emergence of Helicoverpa armigera (Lepidoptera: Noctuidae) in northern Greece.

Overwintering survival of Helicoverpa armigera (Hübner) was studied under field conditions during the winter of 2004-2005 and 2005-2006 to clarify whether a local population of this insect is established in northern Greece. Additionally, the postdiapause eclosion times of the overwintering generation was compared with adult male dynamics using pheromone traps. Our study supplies strong evidence that part, if not all, of the late-season generation of H. armigera overwinters as diapausing pupa in northern Greece, suggesting that a local population exists in this area. Pupae enter diapause during September and October in response to declining temperatures and photoperiod. A limited number of degree-days in autumn prevented part of the population from developing to the diapausing pupal stage. Larvae of H. armigera that were placed in field conditions late in October were not able to pupate and died. The termination of diapause and the eclosion of adults in the following spring were determined by the local soil temperatures. Diapause terminated between mid-April and early May, and adult emergence occurred within a 4- to 6-wk period from late April to early June. Females emerged slightly earlier than the males. The emergence of 10, 25, 50, 75, and 90% of adults required 153, 199, 252, 303, and 347 DD, respectively. Differences in timing and degree of overlap between adult emergence from the experimental population and pheromone trap catches suggests that the spring population of this species also includes immigrants.

Artificial diets for larvae of Anarsia lineatella Zeller (Lepidoptera: Gelechiidae).

Maintenance of an insect colony under laboratory conditions is prerequisite for its further study. However, numerous artificial diet formulas, such as dietary replacements or supplements, influence species growth and survivorship and display difficulties in utilization in laboratory settings. In this work, successful rearing in the laboratory is reported for the peach twig borer A. lineatella on artificial diet. The diet contains dry pindo beans (380 g), brewer's yeast (64 g) agar 31 g, 1360 ml distilled water and preservatives. It is a modification of an artificial rearing medium proposed for the development of the Tortricid Cydia pomonella. Larval survivorship, when developed on the above diet, is significantly higher (-90%) when compared to peach fruits (-60%) and to other diets that were initially tested (5-35%). Diet had no effect on larval developmental time when compared to fruits (27.1 +/- 0.4 and 27.2 +/- 0.5 days, respectively, at 25 degrees C and 65 +/- 5% RH). Light presence of 16:8h L:D did not appear to be a critical factor for a successful rearing of A. lineatella larvae in the laboratory. Type of diet had a significant effect on male (d.f. = 4,103, F = 18.562, P < 0.05) and female (d.f. = 4,91, F = 14.990, P < 0.05) pupal weights. Pupal weights, when they developed as larvae on the proposed 'pindo-bean' diet, ranged from 7.7 +/- 0.3-8.2 +/- 0.2 mg. First-instar larvae exhibited lower survivorship during development, regardless of rearing medium. Sex ratio, for individual larval rearing, was in all cases close to 1:1 regardless of tested rearing medium. More than five generations of A. lineatella were reared under constant conditions without observable adverse effects on development.

Temperature-dependent bionomics and modeling of Anarsia lineatella (Lepidoptera: Gelechiidae) in the laboratory.

Laboratory studies were conducted to assess the effect of constant temperatures (15, 20, 25, 30, and 35 degrees C) on life history traits of peach twig borer, Anarsia lineatella Zeller (Lepidoptera: Gelechiidae). Developmental rate, survival, longevity, and fecundity were studied in environmental chambers from individuals reared on peach, Prunus persica L., twigs until adulthood. Temperature-dependent relationships of various developmental events were characterized, and applied models were evaluated. Total developmental time ranged from 20.4 d (30 degrees C) to 124 d (15 degrees C). Lower developmental thresholds for egg-to-adult development assessed to 11.2 or 11.8 degrees C, according to a linear weighted regression or the reduced major axis method, whereas development required on average 400 degree days (DD) or 424.4, respectively. Survival was substantially reduced at lower (15 degrees C) and higher (35 degrees C) temperatures. First-instar larvae exhibited higher sensitivities during development in all treatments. Mean longevity ranged from 12.1 d (35 degrees C) to 45.8 d (15 degrees C) and from 10.4 d (15 degrees C) to 50.3 d (35 degrees C) for females and males, respectively. A significantly higher number of eggs was laid at the moderate temperatures (126.9 at 25 degrees C), whereas at the extremes, females laid the fewest (40.4 and 26.3 at 15 and 35 degrees C, respectively). A new model, based on a beta type distribution function, fitted on the temperature-dependant developmental data to predict population dynamics. Relative accuracy of the above-mentioned formula was validated using root mean square error (RMSE), index of agreement (d) and the mean square error quotient (E1,2) with respect to a proved model.

Development, survivorship, and reproduction of Helicoverpa armigera (Lepidoptera: Noctuidae) under constant and alternating temperatures.

Laboratory studies were conducted to assess the effect of temperature on the survival, development, fecundity, and longevity of Helicoverpa armigera (Hübner) at 11 constant temperatures ranging from 12.5 to 40 degrees C, as well as at five alternating temperature regimes (25-10, 30-15, 32.5-17.5, 35-20, and 35-27.5 degrees C) and under a photoperiod of 16:8 (L:D) h. H. armigera reared at constant temperatures did not develop from egg to adult (emergence) outside the temperature range of 17.5-32.5 degrees C. The alternating conditions expanded this range from 10 to 35 degrees C. The lowest developmental thresholds of the immature stages were estimated by a linear model and ranged from 10.17 (pupal stage) to 11.95 degrees C (egg stage) at constant temperature regimes and from 1.1 to 5.5 degrees C, respectively at alternating temperatures. The values of developmental thresholds estimated using the nonlinear (Lactin-2) model were lower than those estimated by the linear model for constant and alternating temperature regimes except for larval and pupal stages at constant temperatures. Mean adult longevity fluctuated from 34.4 d at 15 degrees C to 7.6 d at 35 degrees C. Females reared under all alternating temperature regimes laid more eggs than females reared at any, except the 25 degrees C, constant temperature treatment. The intrinsic rate of increase was highest at 27.5 degrees C, at both the constant and the corresponding alternating temperature regimes (0.147 and 0.139, respectively). Extreme temperatures had a negative effect on life table parameters.

Frequency of resistance to Bacillus thuringiensis toxin CrylAb in Greek and Spanish population of Sesamia nonagrioides (Lepidoptera: Noctuidae).

The high-dose/refuge strategy is considered as the main strategy for delaying resistance in target pests to genetically modified crops that produce insecticidal proteins derived from Bacillus thuringiensis Berliner. This strategy is based on a key assumption that resistance alleles are initially rare (<10(-3)). To test this assumption, we used an F2 screen on natural populations of Sesamia nonagrioides Lefebvre (Lepidoptera: Noctuidae) from Greece and Spain. In total, 75 lines from Greece and 85 lines from Spain were screened for survival of F2 larvae on Cry1Ab corn, Zea mays L., leaves. No major resistance alleles were found. The frequency of resistance alleles in the Greek population was <9.7 x 10(-3) with 95% probability, which was very similar to that of the Spanish population (<8.6 x 10(-3) with 95% probability), and the expected frequencies were 3.2 x 10(-3) (0-0.0097) and 2.9 x 10(-3) (0-0.0086) in Greece and Spain (pooled 1.5 x 10(-3)). The experiment-wise detection probability of resistance was 94.0 and 97.5% for the Greek and the Spanish population, respectively. Evidence of alleles conferring partial resistance to Cry1Ab was found only for the Greek population. The frequency of alleles for partial resistance was estimated as 6.5 x 10(-3) with a 95% credibility interval between 8 x 10(-4) and 17.8 x 10(-3) and a detection probability of 94%. Our results suggest that the frequency of alleles conferring resistance to CrylAb, regarding the population of S. nonagrioides, may be rare enough so that the high-dose/refuge strategy could be applied with success for resistance management.

Flight patterns of Anarsia lineatella (Lep., Gelechiidae) in relation to degree--days heat accumulation in Northern Greece.

The flight patterns of the peach twig borer Anarsia lineatella (Zeller) (Lepidoptera: Gelechiidae) were studied in two important and representative regions of peach production in Northern Greece. In order to determine the seasonal occurrence of the species, adult male moth flight was monitored from April till October (2005-2006) using 8 pheromone traps. The observation areas were located at the regions of Veria (lat. 40.32 degrees North) and Velvendos (lat. 40.16 degrees North). The first flight was observed from early May to early June in Veria and from late May to mid June in Velvendos. The 2nd and 3rd flights were observed from late July to early August and from late August to late September in both regions. The degree-days required for the first generation (431 Veria, 371 Velvendos) were fewer than for the 2nd (661 Veria, 504 Velvendos) and the 3rd generation (675 Veria, 506 Velvendos). A lower threshold temperature of 100 C was used in calculating daily day-degrees from March 1st. All the above data are valuable for the development of prediction models and for decision making in order to achieve better timing of treatments. Considering the increasing interest of biorational insecticides where precise timing of treatments is extremely important, degree-day models could be a useful tool for improving their efficacy in IPM.

Developing optimum sample size and multistage sampling plans for Lobesia botrana (Lepidoptera: Tortricidae) larval infestation and injury in northern Greece.

The purpose of this research was to quantify the spatial pattern and develop a sampling program for larvae of Lobesia botrana Denis and Schiffermüller (Lepidoptera: Tortricidae), an important vineyard pest in northern Greece. Taylor's power law and Iwao's patchiness regression were used to model the relationship between the mean and the variance of larval counts. Analysis of covariance was carried out, separately for infestation and injury, with combined second and third generation data, for vine and half-vine sample units. Common regression coefficients were estimated to permit use of the sampling plan over a wide range of conditions. Optimum sample sizes for infestation and injury, at three levels of precision, were developed. An investigation of a multistage sampling plan with a nested analysis of variance showed that if the goal of sampling is focusing on larval infestation, three grape clusters should be sampled in a half-vine; if the goal of sampling is focusing on injury, then two grape clusters per half-vine are recommended.

Some attributes of cold hardiness of the gregarious ectoparasitoid Colpoclypeus florus (Hymenoptera: Eulophidae).

Cold hardiness of diapause and non-diapause larvae of the parasitoid Colpoclypeus florus Walker (Hymenoptera: Eulophidae) was examined in the laboratory. Mean supercooling point (SCP) for diapausing larvae was -26.7 degree C and for non-diapausing larvae immediately after their larval development, was -16.2 degree C. Mean SCP for non-diapause larvae at the prepupa stage was -19.1 degree C. A short period of acclimation (1 week at 5 degree C) had no influence on the mean SCPs of both diapause and non-diapause larvae. Pre-freeze mortality for diapause and non-diapause larvae was also studied. A constant exposure of diapause larvae to -6 degree C resulted in high mortality (70.7 percent) after a period of 40 days. In contrast, 6 days at -6 degree C were sufficient to cause the same level of mortality in non-diapause larvae. After exposure of 15 days at -9 degree C, mortality for non-diapause larvae was 70 percent, whereas after 20 days at the same temperature mortality of diapause larvae was 25 percent. The importance of these findings for the cryobiology of C. florus is discussed.

Biological control of Lobesia botrana (Lepidoptera: Tortricidae) larvae by using different formulations of Bacillus thuringiensis in 11 vine cultivars under field conditions.

This work describes the results of an experiment that was conducted in the vineyard of the American Farm School, Thermi, Thessaloniki Greece, during 2001. Its aim was to study the efficacy of two formulations of Bacillus thuringiensis Berliner (wettable powder and dust) to control the larvae of Lobesia botrana Denis & Schiffermueler (Lepidoptera: Tortricidae). The experimental results showed that the two formulations of B. thuringiensis are significantly more effective than the control, the dusting being more effective in most cultivars (Asyrtico, Sauvignon blanc, Debina, Athiri, Agiorgitico, Limnio, Syrah, and Cabernet sauvignon), and the spraying in a few cultivars (Xinomavro, Roditis, and Chardonnay). Dusting in the blue-black cultivars was found to be more effective than spraying, this not being the case in the yellow-green cultivars. This fact should be taken into consideration when cultivars of different color were planted in the same field. Single dusting proved to be better in cultivars with loose, average and dense berry cluster compactness, whereas double spraying gave better results in compact cultivars.

Temperature dependent development of the parasitoid Colpoclypeus florus (Hymenoptera: Eulophidae) in the laboratory.

Development, fecundity, and longevity of Colpoclypeus florus (Walker), a parasitoid of Adoxophyes orana (Fischer von Röslerstamm) (Lepidoptera: Tortricidae), were studied under laboratory conditions at different constant temperatures. Developmental time from egg to adult was inversely related to temperature and at 25 degrees C was 12.81 +/- 0.19 d. No adults emerged at 30 degrees C. The thermal units required for development from egg to adult were 232.56 +/- 19.5 degree-days. Female C. florus oviposited (mean +/- SEM) on average 30.7, 57.4, 46.6, and 34.1 eggs at 15, 17, 20, and 25 degrees C. respectively. Adult longevity was 11 +/- 1.25 d at 17, whereas it decreased to 4.5 +/- 0.4 at 25 degrees C. Honey significantly increased longevity for both males and females. The highest net reproductive rate was 30.739 at 17 degrees C, whereas the highest intrinsic rate of increase and the shortest doubling time occurred at 25 degrees C.