Marco Carrieri

Population structure of Aedes albopictus (Skuse): the mosquito which is colonizing Mediterranean countries

Multilocus electrophoresis analysis has been used to study the genetic structure of 18 populations of Aedes albopictus newly introduced to Italy, in comparison with two populations in the United States, four in Japan, and four in Indonesia. Allozyme analysis revealed that 15 out of the 18 studied loci were polymorphic among the 28 populations. No significant deviations from Hardy–Weinberg equilibrium were found at polymorphic loci. High genetic affinity was observed between the Italian populations and those from the United States and Japan. The analysis of variance in allele frequencies showed that variance among subpopulations accounted for most of the total variance, suggesting that isolation of the Italian populations is not related to distance. Analysis of linkage disequilibrium using Ohta’s method shows that the variance in the frequency of allele combinations could be explained by the action of the genetic drift which accompanies the establishment of new populations. The colonization process of Ae. albopictus in Italy is following a trend similar to that previously observed in the U.S.A., probably because both infestations derive from several successive introductions, each with large numbers of individuals.

On the Competition Occurring Between Aedes albopictus and Culex pipiens (Diptera: Culicidae) in Italy

Abstract Aedes albopictus (Skuse) was detected for the first time in Italy in 1990. Thanks to its high biological adaptability, it spread rapidly to colonize various urban areas in central and northern Italy. The purpose of this study was to examine the ecological niche occupied by Ae. albopictus and the role of competition with Culex pipiens L., which is a widespread indigenous species, in urban areas. The larval development cycle and competition for food between the two species was also studied under laboratory conditions. The study showed that both species can colonize any type of artificial water container. Ae. albopictus mainly occupies saucers under flower pots and other, usually smaller-sized, containers, whereas Cx. pipiens tends to develop more in large tanks (100–200 liters) used in residential gardens and other larger-sized containers. It would seem that the two species interact mainly in medium-sized containers (10–50 liters) such as manholes and tires. Laboratory studies showed that, at 25°C, the food-biomass conversion coefficient is significantly higher in Ae. albopictus than in Cx. pipiens, indicating that the new established species is more efficient in transforming food into biomass. As a consequence, it has a greater capacity to exploit food resources and therefore grow more rapidly than Cx. pipiens. In larval association where food was scarce, it was noted that competition between the two species takes place and is influenced by temperature: at 25 ± 2°C, Ae. albopictus prevails but is much less likely to do so at 20°C.

Mosquito, bird and human surveillance of West Nile and Usutu viruses in Emilia-Romagna Region (Italy) in 2010.

Background In 2008, after the first West Nile virus (WNV) detection in the Emilia-Romagna region, a surveillance system, including mosquito- and bird-based surveillance, was established to evaluate the virus presence. Surveillance was improved in following years by extending the monitoring to larger areas and increasing the numbers of mosquitoes and birds tested. Methodology/Principal Findings A network of mosquito traps, evenly distributed and regularly activated, was set up within the surveyed area. A total of 438,558 mosquitoes, grouped in 3,111 pools and 1,276 birds (1,130 actively sampled and 146 from passive surveillance), were tested by biomolecular analysis. The survey detected WNV in 3 Culex pipiens pools while Usutu virus (USUV) was found in 89 Cx. pipiens pools and in 2 Aedes albopictus pools. Two birds were WNV-positive and 12 were USUV-positive. Furthermore, 30 human cases of acute meningoencephalitis, possibly caused by WNV or USUV, were evaluated for both viruses and 1,053 blood bags were tested for WNV, without any positive result. Conclusions/Significance Despite not finding symptomatic human WNV infections during 2010, the persistence of the virus, probably due to overwintering, was confirmed through viral circulation in mosquitoes and birds, as well as for USUV. In 2010, circulation of the two viruses was lower and more delayed than in 2009, but this decrease was not explained by the relative abundance of Cx. pipiens mosquito, which was greater in 2010. The USUV detection in mosquito species confirms the role of Cx. pipiens as the main vector and the possible involvement of Ae. albopictus in the virus cycle. The effects of meteorological conditions on the presence of USUV-positive mosquito pools were considered finding an association with drought conditions and a wide temperature range. The output produced by the surveillance system demonstrated its usefulness and reliability in terms of planning public health policies.

Dispersal and survival of Aedes albopictus (Diptera: Culicidae) males in Italian urban areas and significance for sterile insect technique application.

ABSTRACT The dispersal and survival of laboratory-reared Aedes albopictus Skuse males were investigated during the summer of 2007 in three Northern Italy urban localities by mark-release-recapture techniques. Two marking methods were compared: one group of males was dusted with fluorescent pigments on the body (FP), and the other group was obtained from a strain whose natural infection of Wolbachia had been removed (WB0). FP- and WB0-marked males were released as adults and pupae, respectively, in one fixed station at each locality. Recaptures were performed by skilled technicians, within a radius of 350 m from the release site, on days 4, 5, and 7 after the release, and the males were collected while flying around the technicians body or in swarms. Recapture rates ranged from 0.63 to 4.72% for FP males and from 2.39 to 11.05% for WB0 males. The mean distance traveled for WBO males was significantly higher than for FP males; no difference was observed between the dispersal distance measured for the males recaptured on human host versus males recaptured while swarming. No further increase of the dispersal occurred during the postrelease period investigated (from day 4 to day 7 after release). The mean survival rate at the release was 0.51 for FP-marked males and 0.81 for WBO males. The data obtained are discussed for their significance in planning sterile insect technique programs against Ae. albopictus.

Surveillance of the chikungunya vector Aedes albopictus (Skuse) in Emilia-Romagna (northern Italy): organizational and technical aspects of a large scale monitoring system.

ABSTRACT: The Chikungunya virus outbreak that occurred in 2007 in northern Italy (Emilia-Romagna region) prompted the development of a large scale monitoring system of the population density of Aedes albopictus (Skuse, 1894), comparable at the provincial and municipal levels. In 2007, egg density data presented an aggregated distribution (VMR>1) and Taylors power law was applied to calculate the minimum number of ovitraps needed to obtain the prefixed precision levels: D=0.2 in the areas where the Chikungunya epidemic occurred and D=0.3 in all the other urban areas >600 ha. The estimated minimum ovitrap number was then used to set up a monitoring network at the regional scale in season 2008 (May–October). In 242 municipalities 2,741 ovitraps were activated and the 2008 sampled data showed a similar aggregated distribution as in 2007. The adequacy of the monitoring design was evaluated by recalculating the Taylors coefficients and the minimum ovitrap number for each urban area >600 ha using the 2008 egg density data. The comparison between the two estimates showed that the minimum ovitrap number calculated in 2007 was underestimated by 2.7% in weeks 22–41 but was overestimated by 29.4% if referring to the period of highest population density (weeks 27–37). The low cost of the proposed monitoring system, based on the use of fortnightly checked ovitraps, could make it economically sustainable even in a non-epidemic season.

Tolerance Thresholds for Aedes albopictus and Aedes caspius in Italian Urban Areas

ABSTRACT With nuisance mosquito species, the goal of integrated pest management is to keep mosquito density below a tolerance level that is often set by economic, ecological, and political factors. This study compares actual human annoyance, as measured by a phone survey, with several measures of mosquito abundance, in order to determine a threshold that is both relevant and practical. The efficiency of CO2-baited traps, container index (CI), and oviposition traps for monitoring Aedes albopictus, and CO2-baited traps for monitoring Aedes caspius, was evaluated. CO2-baited traps were confirmed to be of low efficiency in Ae. albopictus collection, while correlation matrices showed a good relationship between CI and the number of eggs collected (R = 0.91), and between number of eggs and phone-survey nuisance level estimates (R = 0.88). Correlation between CI and phone-survey nuisance levels was slightly lower (R = 0.78). We found a close relationship between the nuisance level declared by residents and mosquito captures obtained with CO2-baited traps (Ae. caspius) and ovitraps (Ae. albopictus). An equation is presented to estimate annoyance according to dwelling characteristics and to the presence of children in the family.

Studies on Aedes albopictus Larval Mass-Rearing Optimization

ABSTRACT To set up a sterile male technique program to control Aedes albopictus (Skuse) in areas in northern Italy, a pilot mass-rearing facility is under development. For this purpose, experiments were carried out to find the optimal larval density for the optimization of the rearing parameters, i.e., to obtain the fastest larval development, the highest larval and pupal survival rate, and large-sized pupae. Several different larval densities, from 40 to 2,874 larvae per liter, were tested. For densities from 40 to 600 larvae per liter significant size differences were found among pupae obtained under different larval densities. The larvae raised at the lowest density tended to be smaller and to develop most slowly, i.e., longer pupation time. Also, increasing water volume and depth seemed to negatively affect the pupation success. Compared with the other larval densities tested, the larvae reared at a density of 2,874 larvae per liter developed slightly faster and showed higher survival rates, indicating this density as appropriate for the development of a mass rearing, at least using the current larval diet.

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%.

Sugar administration to newly emerged Aedes albopictus males increases their survival probability and mating performance

Aedes albopictus male survival in laboratory cages is no more than 4-5 days when kept without any access to sugar indicating their need to feed on a sugar source soon after emergence. We therefore developed a device to administer energetic substances to newly emerged males when released as pupae as part of a sterile insect technique (SIT) programme, made with a polyurethane sponge 4 cm thick and perforated with holes 2 cm in diameter. The sponge was imbibed with the required sugar solution and due to its high retention capacity the sugar solution was available for males to feed for at least 48 h. When evaluated in lab cages, comparing adults emerged from the device with sugar solution vs the device with water only (as negative control), about half of the males tested positive for fructose using the Van Handel anthrone test, compared to none of males in the control cage. We then tested the tool in semi-field and in field conditions with different sugar concentrations (10%, 15%, and 20%) and compared results to the controls fed with water only. Males were recaptured by a battery operated manual aspirator at 24 and 48 h after pupae release. Rather high share 10-25% of captured males tested positive for fructose in recollections in the vicinity of the control stations, while in the vicinity of the sugar stations around 40-55% of males were positive, though variability between replicates was large. The sugar positive males in the control test may have been released males that had access to natural sugar sources found close to the release station and/or wild males present in the environment. Only a slight increase in the proportion of positive males was obtained by increasing the sugar concentration in the feeding device from 10% to 20%. Surprisingly, modification of the device to add a black plastic inverted funnel above the container reduced rather than increased the proportion of fructose positive males collected around the station. No evidence of difference in the capacity of sterile (irradiated with 30 Gy) males to take a sugar meal relative to fertile males was observed in field comparison. A clear effect of temperature and relative humidity (RH) on the rate of sugar positive males was observed, with an increase of temperature and a decrease in RH strongly increasing the % of sugar positive males. In large enclosures we tested the effect of our sugar supplying tool on the mating competitiveness of sterile vs fertile males, which produced an evident favorable effect both on sterile and fertile males.

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.