Daniele Arnoldi

Comparison of social networks derived from ecological data: implications for inferring infectious disease dynamics.

1. Social network analyses tend to focus on human interactions. However, there is a burgeoning interest in applying graph theory to ecological data from animal populations. Here we show how radio-tracking and capture-mark-recapture data collated from wild rodent populations can be used to generate contact networks. 2. Both radio-tracking and capture-mark-recapture were undertaken simultaneously. Contact networks were derived and the following statistics estimated: mean-contact rate, edge distribution, connectance and centrality. 3. Capture-mark-recapture networks produced more informative and complete networks when the rodent density was high and radio-tracking produced more informative networks when the density was low. Different data collection methods provide more data when certain ecological characteristics of the population prevail. 4. Both sets of data produced networks with comparable edge (contact) distributions that were best described by a negative binomial distribution. Connectance and closeness were statistically different between the two data sets. Only betweenness was comparable. The differences between the networks have important consequences for the transmission of infectious diseases. Care should be taken when extrapolating social networks to transmission networks for inferring disease dynamics.

Climatic factors driving invasion of the tiger mosquito (Aedes albopictus) into new areas of Trentino, northern Italy.

Background The tiger mosquito (Aedes albopictus), vector of several emerging diseases, is expanding into more northerly latitudes as well as into higher altitudes in northern Italy. Changes in the pattern of distribution of the tiger mosquito may affect the potential spread of infectious diseases transmitted by this species in Europe. Therefore, predicting suitable areas of future establishment and spread is essential for planning early prevention and control strategies. Methodology/Principal Findings To identify the areas currently most suitable for the occurrence of the tiger mosquito in the Province of Trento, we combined field entomological observations with analyses of satellite temperature data (MODIS Land Surface Temperature: LST) and human population data. We determine threshold conditions for the survival of overwintering eggs and for adult survival using both January mean temperatures and annual mean temperatures. We show that the 0°C LST threshold for January mean temperatures and the 11°C threshold for annual mean temperatures provide the best predictors for identifying the areas that could potentially support populations of this mosquito. In fact, human population density and distance to human settlements appear to be less important variables affecting mosquito distribution in this area. Finally, we evaluated the future establishment and spread of this species in relation to predicted climate warming by considering the A2 scenario for 2050 statistically downscaled at regional level in which winter and annual temperatures increase by 1.5 and 1°C, respectively. Conclusions/Significance MODIS satellite LST data are useful for accurately predicting potential areas of tiger mosquito distribution and for revealing the range limits of this species in mountainous areas, predictions which could be extended to an European scale. We show that the observed trend of increasing temperatures due to climate change could facilitate further invasion of Ae. albopictus into new areas.

Effects of temperature and rainfall on the activity and dynamics of host-seeking Aedes albopictus females in northern Italy.

The Asian tiger mosquito, Aedes albopictus, has colonized nearly all the regions of Italy as well as other areas of Europe. During the summer of 2007 the tiger mosquito was responsible for an outbreak of Chikungunya in Italy, when this virus was brought in by a tourist of Indian origin returning from an endemic area. To increase the knowledge of tiger mosquito population dynamics, a survey was carried out from April to November 2008 in the municipalities of Arco and Riva del Garda (northern Italy) through a Biogents Sentinel™ (BG)-trap sampling. In particular, the aim of the study was to evaluate the influence of temperature and rainfall on the activity and dynamics of A. albopictus host-seeking females. The seasonal emergence of host-seeking females was strongly influenced by the minimum temperature, and a lower threshold of 13°C was identified. In addition, the threshold for the end of adult activity was found at a minimum temperature of 9°C. Host-seeking female abundance was positively affected by the accumulated temperatures over the period 3 and 4 weeks before the sampling week, possibly as a consequence of the positive effect of accumulated temperatures on larval density. Instead, accumulated precipitation over 1-4 weeks before sampling was negatively correlated with host-seeking female abundance. Finally, the activity of host-seeking females, estimated by the weekly increment in female abundance, was positively affected by the total abundance of females and by mean weekly temperatures. Our study provides useful information for predicting the dynamics of host-seeking Ae. albopictus females in northern Italy and for designing control strategies for preventing arbovirus outbreaks in areas colonized by Ae. albopictus.

Blood Meal Analysis, Flavivirus Screening, and Influence of Meteorological Variables on the Dynamics of Potential Mosquito Vectors of West Nile Virus in Northern Italy

ABSTRACT: An extended area of northern Italy has experienced several West Nile virus (WNV) outbreaks and the emergence of Usutu virus (USUV) during previous years. Our aim was to study some of the factors that could explain disease patterns in the Trentino region, where circulation was detected in human sera and sentinel chickens, but no human or equine cases were reported. We collected Culex species (Diptera: Culicidae) in peridomestic environments. The collected specimens were analyzed for feeding behavior, the influence of temperature and rainfall on the abundance of mosquitoes, and the occurrence of flaviruses. Analysis of blood meals showed that Culex pipiens fed mainly on blackbirds (Turdus merula) and house sparrows (Passer domesticus), while Culex hortensis fed strictly on lizards. The abundance of Cx. pipiens females correlated positively with mean temperature and negatively with rainfall (one to four weeks before capture). This negative relationship could be due to the direct effect of the flushing of habitats together with an indirect effect of oviposition repellency. The mean weekly temperature influenced the abundance of Cx. hortensis. No flaviviruses were detected in the analyzed Culex mosquitoes. These data suggest a silent cycle at low enzootic transmission levels in the area. Furthermore, we present the first contribution to understanding the transmission role of Cx. pipiens mosquitoes in Italy by identifying vertebrate hosts to species level.

Detection of a new insect flavivirus and isolation of Aedes flavivirus in Northern Italy

BackgroundDuring recent years, numerous novel ‘insect flaviviruses’ have been discovered in natural mosquito populations. In a previous study we described the presence of flavivirus DNA sequences integrated in Aedes albopictus (Asian tiger mosquito) populations from Northern Italy in 2007.MethodsDuring 2008 we collected and tested Aedes females for flavivirus presence and developed phylogenetic analysis, virus isolation, electron microscopy studies and RNAse treatments.ResultsWe detected a high prevalence of flavivirus in Ae. albopictus (77.5%). The phylogenetic analysis identified the insect flavivirus sequences as Aedes flavivirus (AEFV) recently described in Japan, and that may have been introduced in Italy travelling with the tiger mosquito. Some of these pools grew in C6/36 cells, producing cytopathic effects, and the RNase treatment results showed the presence of the detected sequences in RNA forms. Furthermore, we detected a new insect flavivirus in one pool of Aedes cinereus/geminus mosquitoes. Phylogenetic analysis of this virus shows that it forms a distinct cluster within the clade of insect flavivirus.ConclusionsThis is the first study to report a high prevalence, to describe the seasonal activity and an isolation of the insect flavivirus Aedes flavivirus in Europe. Moreover we describe the detection of a new insect flavivirus detected from Ae. cinereus mosquitoes from Italy. These flavivirus may be common, ubiquitous and diverse in nature and we discuss the implications of the insect flavivirus group in virus evolution and transmission.

Avian malaria parasites in the last supper: identifying encounters between parasites and the invasive Asian mosquito tiger and native mosquito species in Italy

BackgroundThe invasive Asian tiger mosquito Aedes albopictus has dramatically expanded its distribution range, being catalogued as one of the world’s 100 worst invasive alien species. As vectors of pathogens, Ae. albopictus may create novel epidemiological scenarios in the invaded areas.MethodsHere, the frequency of encounters of Ae. albopictus with the avian malaria parasite Plasmodium and the related Haemoproteus was studied in an area with established populations in northeastern Italy and compared with those from four native mosquito species, Anopheles maculipennis s.l., Culex hortensis, Culex pipiens, and Ochlerotatus caspius. The abdomens of mosquitoes with a recent blood meal were used to identify both the blood meal source and the parasites harboured.ResultsAedes albopictus had a clear antropophilic behaviour while An. maculipennis and Oc. caspius fed mainly on non-human mammals. Birds were the most common hosts of Cx. pipiens and reptiles of Cx. hortensis. Parasites were isolated from three mosquito species, with Cx. pipiens (30%) showing the highest parasite prevalence followed by Cx. hortensis (9%) and Ae. albopictus (5%).ConclusionsThese results are the first identifying the avian malaria parasites harboured by mosquitoes in Italy and represent the first evidence supporting that, although Ae. albopictus could be involved in the transmission of avian malaria parasites, the risk of avian malaria parasite spread by this invasive mosquito in Europe would be minimal.

Current distribution of the invasive mosquito species, Aedes koreicus (Hulecoeteomyia koreica) in northern Italy

BackgroundThe invasive species Aedes (Finlaya) koreicus was first identified in north-eastern Italy in 2011, during the ongoing surveillance activity of Aedes albopictus. Following this finding, a more intensive monitoring was carried out to assess the distribution of the species and to collect biological data. Herein, we report the new records obtained by four years of surveillance.FindingsThe presence of Ae. koreicus was checked using ovitraps, adults traps and by larval collections in all possible breeding sites from May 2011 to July 2015. The monitoring started in the site of the first detection (Province of Belluno) and was then extended in the neighbouring Provinces belonging to four Regions. Aedes koreicus was found in 73 municipalities out of 155 monitored (47.1 %), including 23 municipalities (14.8 %) previously not infested. The area of first detection of Ae. koreicus (Province of Belluno) was also the most infested (68 %). However the mosquito has also been found to the west (Province of Trento) and to the south and south-west (Provinces of Vicenza and Treviso) of the initially infested area.ConclusionsThe spread of Ae. koreicus is directed towards south and west from the original infested area, likely due to the dense road connections and the habitat suitability of the new areas. According to these records, northern Italy has a high probability to be invaded by Ae. koreicus in the next decade. These data can be useful to validate predictive models of potential distribution and dispersal of this species in Italy or in Europe.

Wide detection of Aedes flavivirus in north-eastern Italy - a European hotspot of emerging mosquito- borne diseases

The pattern of flavivirus infection in mosquitoes belonging to the genera Aedes and Culex collected in two regions of north-eastern Italy (Trentino and Veneto) was assessed. Mosquitoes were collected during 2012 and screened for flaviviruses using a generic reverse transcription-nested-PCR targeted on a region of the non-structural NS5 gene. The phylogenetic analysis was performed on a fragment of ~1000 bp. Virus isolation was attempted in C6/36 insect cell lines and the infected cell cultures were studied by electron microscopy. We detected a wide distribution of Aedes flavivirus (AeFV) in Aedes albopictus, with higher infection prevalence in Trentino than in Veneto. In Culex pipiens collected in Veneto, we detected a new sequence of an insect-specific flavivirus and one of Usutu virus. Interestingly, we detected AeFV in C. pipiens, for the first time to our knowledge, in both regions. Viral isolation in cell culture was successful for AeFV. AeFV sequences found in Veneto showed a high percentage of similarity to those detected in Trentino and to those previously reported in other areas of northern Italy. Co-infections with different flaviviruses were not detected.

Ecological Distribution and CQ11 Genetic Structure of Culex pipiens Complex (Diptera: Culicidae) in Italy.

Mosquitoes in the Culex pipiens complex are considered to be involved in the transmission of a range of pathogens, including West Nile virus (WNV). Although its taxonomic status is still debated, the complex includes species, both globally distributed or with a more limited distribution, morphologically similar and characterised by different physiological and behavioural traits, which affect their ability as vectors. In many European countries, Cx. pipiens and its sibling species Culex torrentium occur in sympatry, exhibiting similar bionomic and morphological characters, but only Cx. pipiens appears to play a vector role in WNV transmission. This species consists of two biotypes, pipiens and molestus, which can interbreed when in sympatry, and their hybrids can act as WNV-bridge vectors, due to intermediate ecological features. Considering the yearly WNV outbreaks since 2008 and given the morphological difficulties in recognising species and biotypes, our aim was to molecularly identify and characterised Cx. pipiens and Cx. torrentium in Italy, using recently developed molecular assays. Culex torrentium was not detected; as in other European countries, the pipiens and molestus biotypes were widely found in sympatry with hybrids in most environments. The UPGMA cluster analysis applied to CQ11 genotypic frequencies mainly revealed two groups of Cx. pipiens populations that differed in ecological features. The high propensity of the molestus biotype to exist in hypogean environments, where the habitat’s physical characteristics hinder and preclude the gene flow, was shown. These results confirmed the CQ11 assay as a reliable diagnostic method, consistent with the ecological and physiological aspects of the populations analysed. Since the assessment of the actual role of three biotypes in the WNV circulation remains a crucial point to be elucidated, this extensive molecular screening of Cx. pipiens populations can provide new insights into the ecology of the species and may give useful indications to plan and implement WNV surveillance activities in Italy.

The effect of interspecific competition on the temporal dynamics of Aedes albopictus and Culex pipiens

BackgroundAedes albopictus and Culex pipiens larvae reared in the same breeding site compete for resources, with an asymmetrical outcome that disadvantages only the latter species. The impact of these interactions on the overall ecology of these two species has not yet been assessed in the natural environment. In the present study, the temporal patterns of adult female mosquitoes from both species were analysed in north-eastern Italy, and substantial temporal shifts between abundance curves of Cx. pipiens and Ae. albopictus were observed in several sites. To understand which factors can drive the observed temporal shifts, we developed a mechanistic model that takes explicitly into account the effect of temperature on the development and survival of all mosquito stages. We also included into the model the effect of asymmetric interspecific competition, by adding a mortality term for Cx. pipiens larvae proportional to the larval abundance of Ae. albopictus within the same breeding site. Model calibration was performed through a Markov Chain Monte Carlo approach using weekly capture data collected in our study sites during 2014 and 2015.ResultsIn almost half of observation sites, temporal shifts were due to competition, with an early decline of Cx. pipiens caused by the concurrent rise in abundance of its competitor, and this effect was enhanced by higher abundance of both species. We estimate that competition may reduce Cx. pipiens abundance in some sites by up to about 70%. However, in some cases temporal shifts can also be explained in the absence of competition between species resulting from a “temporal niche” effect, when the optimal fitness to environmental conditions for the two species are reached at different times of the year.ConclusionsOur findings demonstrate the importance of considering ecological interactions and, in particular, competition between mosquito species in temperate climates, with important implications for risk assessment of mosquito transmitted pathogens, as well as the implementation of effective control measures.