Are Aedes albopictus or other mosquito species from northern Italy competent to sustain new arboviral outbreaks?

The Asian tiger mosquito Aedes albopictus (Skuse) (Diptera: Culicidae), native to Southeast Asia, has extended its geographical distribution to invade new temperate and tropical regions. This species was introduced in 1990 to Italy and has since become the main pest in urban settings. It was incriminated as a principal vector in the first European outbreak of chikungunya virus (CHIKV) in the province of Ravenna (Italy) in 2007. This outbreak was associated with CHIKV E1-226V, efficiently transmitted by Ae. albopictus . The occurrence of this outbreak in a temperate country led us to estimate the potential of Ae. albopictus to transmit CHIKV and dengue virus (DENV), and to determine the susceptibility to CHIKV of other mosquito species collected in northern Italy. Experimental infections showed that Ae. albopictus exhibited high disseminated infection rates for CHIKV (75.0% in Alessandria; 90.3% in San Lazzaro) and low disseminated infection rates for DENV-2 (14.3% in San Lazzaro; 38.5% in Alessandria). Moreover, Ae. albopictus was able to attain a high level of viral replication, with CHIKV detectable in the salivary glands at day 2 after infection. In addition, the other three mosquito species, Anopheles maculipennis Meigen, Aedes vexans vexans (Meigen) and Culex pipiens L., showed variable susceptibilities to infection with CHIKV, of 0%, 7.7% and 0–33%, respectively. This information on vector competence is crucial in assessing the risk for an outbreak of CHIKV or DENV in Italy.     

Aedes albopictus in Italy and possible diffusion of the species into the Mediterranean area

Aedes albopictus (Skuse), the important Asian vector mosquito recently introduced in United States and Brazil, is reported from Genoa, North Italy. The infestation was discovered in a kindergarten pre-school center in September 1990 just after the summer holidays. Many discarded tires, well known to provide excellent breeding places for Ae albopictus, had been left in the school playground to be used as toys by the children. After sampling a few biting mosquito specimens for identification, the local health service carried out on September 18 an extensive indoor/outdoor treatment with pyrethroid insecticide. The extent of the infestation in the city of Genoa and in other areas of the Ligurian region has not been evaluated since the identification of the species was available in October, at the end of Ae albopictus reproductive period. A general survey in various Italian regions is being planned for the 1991 spring-summer period. The present record, together with the previous report of Ae albopictus in Albania, clearly supports the hypothesis of a spreading of the species in the Mediterranean area.     

Extension of the area of Aedes albopictus in Italy. 1st report of the species in central Italy]

A new infestation by Aedes albopictus was discovered in Civitavecchia, Central Italy, in October 1992. This follows previous records of the species in North Italy, where the first infestation was observed in 1990 in the city of Genoa. The present finding clearly indicates the capacity of Aedes albopictus to spread all over Italy and to become a permanent pest in Southern Europe. In spite of the expected embryonic diapause in October, larval breeding was observed in a fifteen-liter glass jar with a narrow neck, left outdoor.     

First finding of Dirofilaria repens in a natural population of Aedes albopictus

The invasive mosquito Aedes albopictus (Skuse) (Diptera: Culicidae) has become widespread in Italy during the past decade. Also Italy has foci of canine filariasis caused by Dirofilaria (Spirurida: Onchocercidae), due to subcutaneous D. repens Railliet & Henry as well as the dog heartworm D. immitis (Leidy) transmitted by various vector mosquitoes (Diptera: Culicidae). In 2002, at Fiumicino, west of Rome (Lazio Region), 17% of dogs were found to have D. repens microfilariae in peripheral blood. To evaluate the role of Ae. albopictus as a vector of Dirofilaria in this area, female mosquitoes were collected daily, June-October 2002, landing on dog or human bait in a rural house at Focene. Mosquitoes were maintained at 27 degrees C and 70% RH for 6 days, to allow development or purging of filaria larvae, then identified and frozen for subsequent molecular assay with filaria-specific ribosomal S2-S16 primers. To distinguish specimens harbouring infective L3 Dirofilaria larvae, DNA was extracted separately from the mosquito abdomen and head-thorax. Dirofilaria species were identified by sequencing, confirmed by polymerase chain reaction of positive specimens using primers specific for D. immitis and D. repens. Dirofilaria DNA was detected in 3/154 (2%) of Ae. albopictus females examined: D. repens DNA in head-thorax and abdomen of one collected 27th July; D. immitis in the abdomen of one collected 24th September; DNA of both D. immitis and D. repens in the head-thorax of one collected 11th October 2002. Thus Ae. albopictus is a potential vector of both Dirofilarias in Italy, representing risks for veterinary and human health.     

Aedes albopictus (Diptera: Culicidae) in Rome: experimental study of relevant control strategy parameters

Since 1997, Aedes albopictus has colonised and then rapidly invaded the city of Rome (Italy) and its peripheral areas. Presently, the control of this mosquito in Italy relies mainly on larvicidal treatment of street storm sewer catch basins with the organophosphate temephos. We have therefore obtained baseline data on the susceptibility to temephos of the Roman Ae. albopictus population by laboratory bioassays on F1 fourth-instar larvae following standard WHO protocols. Estimated lethal concentrations were 0.014 mg/l (LC50) and 0.022 mg/l (LC90) indicating a lack of resistance to this compound. The persistence of temephos in sewer catch basins was evaluated by follow-up of catch basins treated with a dose of 1.5 mg of active ingredient. Mosquito larvae were recovered in 10% and 50% of the treated basins at 9 and 18 days posttreatment, respectively. In order to understand the relative contribution of this larval habitat to adult populations, we conducted a survey in the Zoo of Rome to estimate the larval density of mosquitoes breeding in sewer catch basins. A complete census of a 16.5 ha area mapped 243 catch basins, but only 25 (10.3%) contained water; of the latter 8 (32.0%) hosted mosquito larvae. All positive catch basins contained larvae of Culex pipiens, which were associated with Culiseta longiareolata and/or Ae. albopictus in 6 and 3 cases, respectively. A longitudinal survey of one catch basin over 4 months showed that the mean larval density of Ae. albopictus was markedly lower than that of Cx pipiens and Cs. Iongiareolata, ranging between 0 and 1.3 larvae/dip as compared to 0-33.2 and 0-22.7 larvae/dip, respectively. However, adult densities of Ae. albopictus in this area estimated during the same period with 20 ovitraps showed consistently high values (proportion of positive ovitraps around 100%). These preliminary observations suggest that whenever alternative larval biotopes other than sewer catch basins are widely available, they might be more productive and/or preferred substrates to catch basins for Ae. albopictus breeding.     

Aedes albopictus in Sardinia: reappearance or widespread colonization?

The Asian tiger mosquito Aedes albopictus (Skuse, 1894) was first discovered in the South of Sardinia in October 1994, in a tyre depot not far from Cagliari-Elmas airport. Insecticide treatment was thought to have successfully eradicated the mosquito, but in 1996 and 1997 new breeding sites were discovered, a few at some distance from the first. More recently two sites have been reported in the heart of the city of Cagliari. It is not known whether the mosquito has spread from the first breeding place discovered, where treatment may not have been definitive, or whether they have been newly introduced. The recent sighting of Ae. albopictus in Olbia in the Northeast of the island tends to suggest the latter. Cagliari and Olbia are actually Sardinia's two largest sea ports of entry.     

Interactions with native mosquito larvae regulate the production of Aedes albopictus from bromeliads in Florida

Abstract. 1. Immatures of the invasive container mosquito Aedes albopictus occur in water-holding tanks and axils of ornamental bromeliads in Florida, where this species established and became abundant in the late 1980s and early 1990s.
2. The numbers of A. albopictus in bromeliads in gardens vary geographically in Florida, being significantly higher in northern sites where two species of Wyeomyia mosquitoes, native specialists of this microhabitat, are absent.
3. Causes of the negative relationship between abundances of A. albopictus and Wyeomyia spp. were investigated experimentally using Billbergia pyramidalis , an introduced bromeliad popular in Florida gardens.
4. Egg laying by A. albopictus in B. pyramidalis confined in a large outdoor cage was unaffected by the presence or absence of Wyeomyia spp. fourth instars, indicating that immatures of the native bromeliad mosquito species do not deter oviposition by the invasive species in this microhabitat.
5. Growth and survivorship of A. albopictus first instars in tanks of B. pyramidalis were negatively affected by the presence of fourth instar, but not first instar, Wyeomyia spp. larvae at natural densities. Stimulative effects of leaf litter on A. albopictus growth and survivorship in this microhabitat were independent of the presence or absence of Wyeomyia spp. immatures.
6. These results demonstrate that interspecific larval interactions regulate the production and explain the geographic patterns in abundance of A. albopictus from ornamental bromeliads in Florida.     

Presence of Dirofilaria repens and an insect immunocyte (plasmatocyte) in a human subcutaneous nodule, induced by a mosquito bite.

It is well known that the nematode Dirofilaria repens is transmitted to humans by vector mosquito bite. Examination of a fine needle aspiration biopsy drawn from a month-old nodule on the chest of a woman, residing in Garlasco, province of Pavia, Northern Italy, revealed the presence of not only one immature female of D. repens, but also some scattered cells that we believe to be mosquito's blood cells, plasmatocytes (immunocytes). We presume that plasmatocytes were carried into the bite wound with the mosquito's hemolymph that had oozed from a rupture in its mouthparts during feeding. Because Aedes albopictus recently colonized certain areas in the above region, we suspect that the nodule resulted from the bite of this mosquito.     

Results of a survey to detect the mosquito Aedes albopictus in the French Riviera.

A programme of surveillance was initiated in 1992, in the French Riviera, to detect a possible introduction of Aedes albopictus from Italy and to prevent nuisances caused by mosquitoes in the touristic localities of the C?te d'Azur. In five years, numerous mosquito breeding places have been located. Nine species have been collected: Anopheles claviger, An. plumbeus, Aedes geniculatus, Ae. vittatus, Culex hortensis, Cx. impudicus, Cx. pipiens, Culiseta fumipennis, Cs. longiareolata but no Ae. albopictus was found. Nuisances were mainly due to hypogean populations of Cx. pipiens. Breeding places in urban sites have been controlled or suppressed. The discovery of an important larval population of An. plumbeus in an urban area might further stress the importance of this species already suspected to transmit indigenous malaria in cities.     

The first report of Aedes (Stegomyia) albopictus in Haiti

Aedes albopictus was found in six of the 10 departments of Haiti and in 14 of the 35 communes surveyed. The survey found the larvae of Ae. albopictus in 13 different types of containers. Used tires and tins were by far the most common breeding sites used by this mosquito species. At the breeding sites, Ae. albopictus was associated with other mosquito species, such as Aedes aegypti, Culex nigripalpus and Aedes mediovittatus. The highest proportion of association was with Ae. aegypti. This study represents the first report of Ae. albopictus in Haiti.     

Critical review of the vector status of Aedes albopictus

The mosquito Aedes (Stegomyia) albopictus (Skuse) (Diptera: Culicidae), originally indigenous to South-east Asia, islands of the Western Pacific and Indian Ocean, has spread during recent decades to Africa, the mid-east, Europe and the Americas (north and south) after extending its range eastwards across Pacific islands during the early 20th century. The majority of introductions are apparently due to transportation of dormant eggs in tyres. Among public health authorities in the newly infested countries and those threatened with the introduction, there has been much concern that Ae. albopictus would lead to serious outbreaks of arbovirus diseases (Ae. albopictus is a competent vector for at least 22 arboviruses), notably dengue (all four serotypes) more commonly transmitted by Aedes (Stegomyia) aegypti (L.). Results of many laboratory studies have shown that many arboviruses are readily transmitted by Ae. albopictus to laboratory animals and birds, and have frequently been isolated from wild-caught mosquitoes of this species, particularly in the Americas. As Ae. albopictus continues to spread, displacing Ae. aegypti in some areas, and is anthropophilic throughout its range, it is important to review the literature and attempt to predict whether the medical risks are as great as have been expressed in scientific journals and the popular press. Examination of the extensive literature indicates that Ae. albopictus probably serves as a maintenance vector of dengue in rural areas of dengue-endemic countries of South-east Asia and Pacific islands. Also Ae. albopictus transmits dog heartworm Dirofilaria immitis (Leidy) (Spirurida: Onchocercidae) in South-east Asia, south-eastern U.S.A. and both D. immitis and Dirofilaria repens (Raillet & Henry) in Italy. Despite the frequent isolation of dengue viruses from wild-caught mosquitoes, there is no evidence that Ae. albopictus is an important urban vector of dengue, except in a limited number of countries where Ae. aegypti is absent, i.e. parts of China, the Seychelles, historically in Japan and most recently in Hawaii. Further research is needed on the dynamics of the interaction between Ae. albopictus and other Stegomyia species. Surveillance must also be maintained on the vectorial role of Ae. albopictus in countries endemic for dengue and other arboviruses (e.g. Chikungunya, EEE, Ross River, WNV, LaCrosse and other California group viruses), for which it would be competent and ecologically suited to serve as a bridge vector.     

Chikungunya epidemic outbreak in Emilia-Romagna (Italy) during summer 2007

During summer 2007, an outbreak due to the local transmission of CHIKV by Aedes albopictus mosquitoes occurred moreover in Italy, Emilia-Romagna Region, in the areas of Ravenna, Forli-Cesena, Rimini and Bologna cities. The original outbreak developed in Castiglione di Cervia and Castiglione di Ravenna, two small villages divided by a river. The first case was recorded on August 9th the epidemic outbreak then spread out, thus giving rise to smaller secondary outbreaks and further sporadic cases in the same area, for a total of 337 suspected cases, 217 of which confirmed by blood analysis. CHIKV has been isolated and characterized on both blood and mosquito samples.     

Molecular and functional characterization of the odorant receptor2 (OR2) in the tiger mosquito Aedes albopictus

In mosquitoes, the olfactory system plays a crucial role in many types of behavior, including nectar feeding, host preference selection and oviposition. Aedes albopictus, known also as the tiger mosquito, is an anthropophilic species, which in the last few years, due to its strong ecological plasticity, has spread throughout the world. Although long considered only a secondary vector of viruses, the potential of its vector capacity may constitute a threat to public health. Based on the idea that an improved understanding of the olfactory system of mosquitoes may assist in the development of control methods that interfere with their behavior, we have undertaken a study aimed at characterizing the A. albopictus Odorant Receptors. Here we report the identification, cloning and functional characterization of the AalOR2 ortholog, that represents the first candidate member of the odorant receptor (OR) family of proteins from A. albopictus. AalOR2 is expressed in the larval heads and antennae of adults. Our data indicate that A. albopictus OR2 (AalOR2) shares a high degree of identity with other mosquito OR2 orthologs characterized to date, confirming that OR2 is one of the most conserved mosquito ORs. Our data indicate that AalOR2 is narrowly tuned to indole, and inhibited by (-)-menthone. In agreement with this results, these two compounds elicit two opposite effects on the olfactory-based behavior of A. albopictus larvae, as determined through a larval behavioral assay. In summary, this work has led to the cloning and de-orphaning of the first Odorant Receptor in the tiger mosquito A. albopictus. In future control strategies this receptor may be used as a potential molecular target.     

Detection of and monitoring for Aedes albopictus (Diptera: Culicidae) in suburban and sylvatic habitats in north central Florida using four sampling techniques

A sampling study using a BG-Sentinel trap baited with CO(2), a gravid trap baited with an oak-pine infusion, a human subject, and a vegetative aspirator was conducted to compare their reliability at detecting Aedes albopictus Skuse in suburban and sylvatic habitats. We collected 73,849 mosquitoes, representing 29 species from 11 genera over a 20-wk period. The BG-Sentinel trap accounted for over 85% of all Ae. albopictus captured and was significantly more effective at detecting the presence of Ae. albopictus compared with the other three techniques. Landing counts provided the fewest mosquito species (n = 10), yet provided a quick and effective weekly assessment of the major biting species and were the most effective method for sampling Ae. albopictus within a 10-min period. Fewer Ae. albopictus were sampled from sylvatic habitats compared with suburban ones. Sampling criteria advantageous for surveying Ae. albopictus and other mosquito species are discussed.     

Larvicidal activity of tectoquinone isolated from red heartwood-type Cryptomeria japonica against two mosquito species

Mosquito larvicidal activities of methanolic extracts from different plant parts of red heartwood-type Cryptomeria japonica D. Don against the fourth-instar larvae of Aedes aegypti and Aedes albopictus were examined. Results of mosquito larvicidal tests demonstrated that the n-hexane fraction of C. japonica sapwood methanolic extract had an excellent inhibitory effect against the larvae of A. aegypti and A. albopictus and its LC50 values were 2.4 and 3.3 microg/ml, respectively, in 24h. Following the bioactivity-guided fractionation procedure, the active constituent isolated from C. japonica sapwood was characterized as tectoquinone by spectroscopic analyses. The LC50 values of tectoquinone against A. aegypti and A. albopictus in 24h were 3.3 and 5.4 microg/ml, respectively. In addition, comparisons of mosquito larvicidal activity of anthraquinone congeners demonstrated that anthraquinone skeleton with a methyl group at C-2 position, such as tectoquinone, exhibited the strongest mosquito larvicidal activity. Results of this study show that the methanolic extract of C. japonica sapwood may be considered as a potent source and tectoquinone as a new natural mosquito larvicidal agent.     

Updating the geographical distribution and frequency of Aedes albopictus in Brazil with remarks regarding its range in the Americas

The geographical distribution of Aedes albopictus in Brazil was updated according to the data recorded across the country over the last eight years. Countrywide house indexes (HI) for Ae. albopictus in urban and suburban areas were described for the first time using a sample of Brazilian municipalities. This mosquito is currently present in at least 59% of the Brazilian municipalities and in 24 of the 27 federal units (i.e., 26 states and the Federal District). In 34 Brazilian municipalities, the HI values for Ae. albopictus were higher than those recorded for Ae. aegypti, reaching figures as high as HI = 7.72 in the Southeast Region. Remarks regarding the current range of this mosquito species in the Americas are also presented. Nineteen American countries are currently infested and few mainland American countries have not confirmed the occurrence of Ae. albopictus. The large distribution and high frequency of Ae. albopictus in the Americas may become a critical factor in the spread of arboviruses like chikungunya in the new world.     

Accidental importation of the mosquito Aedes albopictus into the Netherlands: a survey of mosquito distribution and the presence of dengue virus

Abstract.  In the summer of 2005, the Asian tiger mosquito, Aedes albopictus (Skuse) (Diptera: Culicidae) was found for the first time in the Netherlands. It was collected on the premises of several horticultural companies that import the ornamental plant Dracaena sanderiana (Sparagalus: Dracaenaceae [Agavaceae]), known as Lucky bamboo, from southern China, an area endemic for this mosquito species and for arboviruses transmitted by this vector. Here we report the results of a 1-year survey of the distribution and vector status of Ae. albopictus in Lucky bamboo nurseries in the Netherlands (July 2006–June 2007). As it had been established previously that the presence of this species was linked to the import of Lucky bamboo, the survey was conducted only on sites owned by relevant import companies. In total, 569 adult Ae. albopictus were collected with mosquito traps from 15 of the 17 (88%) glasshouses used by Lucky bamboo importers, none of which were found to be infected with dengue virus. On two occasions there was evidence that Ae. albopictus had escaped from the glasshouses, but, overall, there was no evidence that a population had become established in the greenhouses or elsewhere.     

Differential Behavioral Responses to Water-Borne Cues to Predation in Two Container-Dwelling Mosquitoes

Larvae of the mosquito Toxorhynchites rutilus (Coquillett) prey upon other container-dwelling insects, including larvae of Aedes albopictus (Skuse), which is native to Asia but was introduced into the United States, and on the native tree hole mosquito Ochlerotatus triseriatus (Say). Previous work has established that O. triseriatus adopts low-risk behaviors in the presence of predation risk from T. rutilus. It is unknown whether introduced A. albopictus show a similar response to this predator. Behavior of fourth instars of A. albopictus or O. triseriatus was recorded in water that had held either A. albopictus or O. triseriatus larvae alone (control) and in water that had held T. rutilus larvae feeding on either A. albopictus or O. triseriatus (predation). Activity and position of larvae were recorded in 30-min instantaneous scan censuses. In response to water-borne cues to predation, O. triseriatus adopted low-risk behaviors (more resting, less feeding and movement), but A. albopictus did not change its behavior. We also tested the species specificity of the cues by recording the behavior of A. albopictus in water prepared using O. triseriatus and vice versa. O. triseriatus adopted low-risk behaviors even in predation water prepared by feeding T. rutilus with A. albopictus, but A. albopictus did not alter its behavior significantly between predation and control treatments prepared using O. triseriatus. Thus, A. albopictus does not seem to respond behaviorally to cues produced by this predator and may be more vulnerable to predation than is O. triseriatus.     

Variations in insecticidal activity and chemical compositions of leaf essential oils from Cryptomeria japonica at different ages

The larvicidal effects of the essential oils extracted from the leaves of Cryptomeria japonica at different ages (58, 42, and 26 years old) against 2 mosquito species, Aedes aegypti and Aedes albopictus, were studied. The analysis of major constituents of these essential oils was also investigated. Results obtained from the larvicidal tests, using essential oil from the leaves of 58-year-old C. japonica was found to be most effective against both A. aegypti and A. albopictus larvae, indicating tree age has significant influence on mosquito larvicidal activity. In addition, the eleven pure constituents from C. japonica leaf essential oil were also tested individually against the two mosquito larvae. Among them, alpha-terpinene, gamma-terpinene, p-cymene, 3-carene, terpinolene, and beta-myrcene shows strong larvicidal effect against the two mosquito larvae. Among these pure constituents, 3-carene exhibits the best larvicidal effect against A. aegypti and terpinolene shows an excellent inhibitory action against A. albopictus larvae. The results of this study show that the leaf essential oil and its effective constituents might be considered as a potent source for the production of fine natural larvicides.