Bettina Maccagnani

γ Ray Dosimetry and Mating Capacity Studies in the Laboratory on Aedes albopictus Males

ABSTRACT In Italy, Aedes albopictus Skuse is currently recognized as the most dangerous mosquito, and as currently applied conventional control methods gave unsatisfactory results, we are developing alternative strategies such as the sterile insect technique. To find the optimal sterilizing dose, male pupae were exposed to different doses of &ggr; rays in the range 20–80 Gy, generated by a Cesium-137 source. The effects of male pupal age at irradiation and &ggr; ray dose on adult male emergence, sterility level, longevity, and mating capacity were evaluated, and dose-response curves of residual fertility were calculated. Radiation tests were also performed on female pupae to observe their reproductive capacity in case of accidental release. Results confirmed that the age at which the male pupa is irradiated is an important factor that affects the longevity of the adult, whereas the effect of age on the induced sterility level is less pronounced. When male pupae older than 30 h were irradiated, the longevity of the adults was not affected by doses up to 40 Gy. The 40-Gy dose appeared sufficient to induce high level of sterility (>99%) at any male pupal age for all the strains tested. The duration of coupling and the number of mated females per male appeared to be affected by the radiation received by male pupae only at doses higher than 40 Gy. The female pupae were more sensitive to radiation than male pupae, with strong reduction in fecundity and fertility at 20 Gy and complete suppression of oviposition at higher doses.

Aedes albopictus (Diptera: Culicidae) Population Size Survey in the 2007 Chikungunya Outbreak Area in Italy. I. Characterization of Breeding Sites and Evaluation of Sampling Methodologies

ABSTRACT The population of Aedes albopictus (Skuse) (Diptera: Culicidae) was measured in four towns in northern Italy by means of Stegomyia indices (house index: percentage of houses with at least one active breeding site; container index: percentage of containers with larvae; and Breteau index: number of active breeding sites per premises), pupae per premises index (PPI), pupae per hectare index (PHI), and ovitrap monitoring. A detailed investigation on types and number of productive or potential larval habitats in the inspected premises was performed. Catch basins were the most productive breeding sites in both private and public areas. The influence of the premises maintenance and premises shading indices (PMI and PSI) on the population indices and the correlations between the latter and egg density were assessed. The highest PPI was found in badly maintained premises, most times combined with shady coverage. Stegomyia indices and PPI were well correlated with each other, but not with the PHI, which also included the pupae collected in public areas. We obtained a highly positive correlation (R = 0.86) between the PHI and weekly mean egg density estimated 7–14 d after inspection of the premises. The number of females per hectare, calculated using the Focks model, was correlated with the mean egg density of the week after sampling (R = 0.79), and we propose that ovitrap monitoring can be used to evaluate the efficiency of the control activities or to determine a mean egg density threshold for epidemic risk evaluation.

Larvicidal activity and influence of Bacillus thuringiensis var. israelensis on Aedes albopictus oviposition in ovitraps during a two-week check interval protocol.

Abstract Toxicity persistence of Bacillus thuringiensis var. israelensis (Bti) was evaluated in laboratory and field trials to develop a new protocol for Aedes albopictus monitoring. In the laboratory, the residual toxicity of 5 Bti formulations was compared using the percentage mortality at days 4, 8, 12, and 16 of Culex pipiens larvae exposed for 24 h to Bti treatments. The results showed a good performance of all tested formulations (>97% mortality at day 14, for all the formulations), but only Vectobac™ 12AS at the concentration of 1 ml/liter showed an efficacy of 100% for 2 wk. A field study was designed to test the effect of Bti on the ovitrap check interval or influence of ovipositional response of gravid Ae. albopictus females. Three different ovitrap treatments were used: ovitraps with tap water checked weekly; ovitraps with tap water checked every 2 wk; ovitraps with Bti (Vectobac 12AS, dose of 1 ml/liter) checked every 2 wk. Our study demonstrated that in the ovitrap, the toxic action of a 1% solution of Bti was maintained for at least 14 days with mortality of 100% and that rainfall did not seem to negatively influence the residual action of Bti. Therefore the probability that the larvae may complete the developmental cycle in ovitraps with Bti seems to be very low. The oviposition activity index showed that Bti enhances the oviposition rate of Ae. albopictus by 17.4%.

Seasonal pattern of daily activity of Aedes caspius, Aedes detritus, Culex modestus, and Culex pipiens in the Po Delta of northern Italy and significance for vector-borne disease risk assessment

ABSTRACT: The increasing concern about vector-borne diseases such as West Nile disease in northern Italy motivated our analysis of data on the mosquito fauna and the seasonal and daily flight patterns collected in 1998 in the Po Valley. Collections were performed once a week from May to November, with human landing collections and CO2 traps. Culex pipiens was present from July to October and showed a clearly unimodal nocturnal flight habit. Culex modestus appeared in July–August and showed a bimodal flight pattern, (main peak during the evening and a minor one in the morning). Aedes caspius was present from May to November (highest densities in July–August) and showed a bimodal flight pattern with a major crepuscular peak and a minor dawn peak in the morning. Aedes detritus was the most abundant species in May, with a crepuscular sharply bimodal flight pattern, particularly according to human landing collections. Sunset and sunrise time, in combination with the solar phase (that determines daylight duration and its trend of changing) were the main factors affecting flight behavior. Temperature, relative humidity, and wind speed differently affected the flight behavior of mosquito females according to the species.

Weather Factors Influencing the Population Dynamics of Culex pipiens (Diptera: Culicidae) in the Po Plain Valley, Italy (1997–2011)

ABSTRACT The impact of weather variables on Culex pipiens L. (Diptera: Culicidae) population dynamics in the Po Valley, Northern Italy, a densely populated region containing the largest industrial and agricultural areas in Italy, was investigated. Monitoring of mosquitoes was carried out by using CO2-baited traps without light, collecting data weekly from 1700 to 0900 hours during the period May–September, from 1997 to 2011. Daily minimum, average, and maximum relative humidity; daily minimum, maximum, and average temperature; rainfall; and hydroclimatic balance (rainfall-potential evapotranspiration) were obtained from three weather stations within the surveillance zone. The average population dynamic trend over the 15-yr period showed a bell-shaped curve with a major peak in June and a secondary peak at the end of August in the rural areas, whereas bimodality was not evidenced in the urban areas. The correlation analyses showed that the mosquito seasonal population and the population in the period of maximum West Nile virus circulation (August–September) was mostly affected by the relative humidity registered from March to July, particularly in May, and, to a lower extent, also by hydroclimatic balance registered in April–July, and by the rainfall occurred in June–July. In addition, the rate of increase of the population during the spring months influenced the development of the mosquito population of the following months.

Macrocyclops albidus (Copepoda: cyclopidae) for the Biocontrol of Aedes albopictus and Culex pipiens in Italy

Abstract The aim of our study was to assess the potential of Macrocyclops albidus as a biological control agent against the 1st and 2nd instars of Culex pipiens and Aedes albopictus. Under laboratory conditions of prey saturation (50 1st instars/copepod), an average of 58.98% of Cx. pipiens and 54.99% of Ae. albopictus larvae were killed by 1 copepod in 24 h. Trials run in big drums containing 200 liters of water showed that the M. albidus population, inoculated in April, efficiently controlled the mosquito population for the entire season. The predator was particularly effective against Ae. albopictus, as only 2 larvae of this species were found in the treated drums, compared to 814 larvae in untreated control drums throughout the study period. No difference was observed in the control efficacy between the 2 initial densities of copepods used. The reduction in Ae. albopictus density in the drums with 100 and 500 M. albidus with respect to the control drums was 99.90 ± 0.35% and 100.0 ± 0.0%, respectively. For Cx. pipiens, the reduction in density was 88.69 ± 13.51% and 84.65% in drums inoculated with 100 and 500 copepods, respectively. Macrocyclops albidus populations survived through the winter and continued to keep the mosquito population under control during the 2008 season. The M. albidus population developed very well both in drums placed in sunny and shaded areas and proved to be tolerant to both high (summer) and low (winter) temperatures. Trials performed on M. albidus survival in catch basins showed that after a few weeks, the copepod population dramatically decreased and subsequently disappeared. The main problem for copepod survival in catch basins seemed to be the low oxygen tension and accumulation of toxic substances, rather than copepods being flushed out in heavy rainfall episodes. During the period when copepods were present, they maintained the mosquito population under control; their partial disappearance from the catch basins, however, would require more recolonization intervention to maintain mosquito control during the season.

Comparative study on the effectiveness of different mosquito traps in arbovirus surveillance with a focus on WNV detection.

The selection of the ideal trap for arbovirus surveillance is an issue of primary importance to increase the sensitivity of virus detection and the cost-effectiveness of the entomological surveillance. During the summer 2011, the effectiveness of five types of mosquito traps (CDC gravid trap, CO2(-)baited trap, BG-Sentinel™ and two experimental prototypes) to attract females potentially infected with West Nile virus were assessed. The study was carried out in three natural wetland sites located in the Emilia-Romagna Region (Northern Italy), using a Latin square scheme. Single night collections of adult females were performed and determination of species and physiological state (gravid, nulliparous or parous) was made upon return to the laboratory. The species most frequently collected in the gravid trap was Culex pipiens sl. L., being gravid females the large majority of the individuals. Species diversity was much higher in CO2(-)baited traps, which may therefore enable a more comprehensive description of the vector species composition and their role in arboviruses circulation. Our findings indicate that gravid traps can be a valid tool and should be integrated in the West Nile virus surveillance system in the Emilia-Romagna region, mainly based on collections made with CO2-baited traps.

Chikungunya and Dengue Risk Assessment in Greece

Objective: The aims of the study were: (1) To assess the mutated CHIKV E1-A226V and DENV II infection and dissemination rates of an Ae. albopictus population established in Athens (Greece) (2) To assess the risk of outbreaks in four Greek localities based on Ae. albopictus population density whose estimate was based on the number of eggs laid in ovitraps. Methods: Under laboratory conditions females were offered blood meal infected with the CHIKV titer of 1X106 TCID 50/mL and DENV II titer of 1.76 X106 TCID 50/mL; at day 11 after oral infection, females were sacrificed, legs were removed and processed for PCR analysis to assess the presence of viral replicates. In order to evaluate the risk of outbreak of CHIKV and DENV II, a pilot monitoring program was started in three Greek localities and in Chania (Crete), to estimate the adult female population density on the base of the number of eggs in the ovitraps. Results: We proved the vector competence of the Greek Ae. albopictus strain for E1-A226V mutated CHIKV and DENV II. Combining the data on the vector competence with those on the female population density, based on the egg density data, the estimated risk of outbreak was relatively low but not negligible. Conclusion: As the vector competence estimated under laboratory conditions was obtained by offering females moderately low initial virus titers, it can be expected a higher vector competence in the field. This consideration, together with a possible increase of the mosquito population due to the global warming effects, make the quantitative ovitrap-based monitoring a necessary and useful tool to estimate the risk of outbreaks.