First evidence of pyrethroid resistance in Italian populations of West Nile virus vector Culex pipiens.

Culex pipiens (Linnaeus), one of the most abundant mosquito species in Europe, plays a crucial role in the endemic transmission of West Nile virus and caused the large outbreak with >1600 human cases in 2018. Although evidence of resistance to pyrethroids has been reported for Cx. pipiens populations from Spain and Greece, resistance monitoring has been largely neglected in Italy. Herein, we investigate susceptibility of Italian Cx. pipiens populations to the pyrethroids permethrin and deltamethrin. Results from WHO-tube-bioassays revealed mortalities ranging from 14-54%, indicating high levels of resistance, in four out of 10 populations exposed to permethrin (0.75%) and of 63% in one of three populations exposed to deltamethrin (0.05%). Reduced susceptibility (mortality<98%) was detected in almost all other populations. A clear association is shown between the resistant phenotype and the presence of kdr-alleles in position 1014 of the VSSC, strongly suggesting its role in reducing susceptibility. The study provides the first evidence of pyrethroid-resistance in Italian Cx. pipiens populations and reports levels of resistance paralleled in the European region only in Turkey. This highlights the urgent need to implement insecticide-resistance management plans to restore the efficacy of the nowadays only chemical weapon available to control arbovirus transmission in Europe.

A bacterium against the tiger: further evidence of the potential of noninundative releases of males with manipulated Wolbachia infection in reducing fertility of Aedes albopictus field populations in Italy.

BACKGROUND: Incompatible insect technique (IIT) is a population suppression approach based on the release of males with manipulated Wolbachia infection inducing egg inviability in wild females. We here present results of multiple field releases of incompatible ARwP males carried out in 2019 in a 2.7-ha green area within urban Rome (Italy) to assess the effect on Aedes albopictus egg viability. Data are compared with results obtained in 2018, when the approach was tested for the first time in Europe. RESULTS: An average of 4674 ARwP males were released weekly for 7 weeks, resulting in a mean ARwP:wild male ratio of 1.1:1 (versus 0.7:1 in 2018). Egg-viability dynamics in ovitraps significantly varied between treated and control sites, with an estimated overall reduction of 35% (versus 15% in 2018). The estimated proportion of females classified as mated with ARwP males was 41.8% and the viability rate of eggs laid by these females (9.5%) was on average significantly lower than that of females only mated with wild males (87.8%); however, high variability in fertility was observed. Values of ARwP male competitiveness were 0.36 and 0.73 based on the overall viability rate of eggs in ovitraps and on female fertility, respectively; thus, well above the conventional 0.2 threshold for an effective suppressive impact in the field. CONCLUSIONS: Results further support the potential of IIT as a tool to contribute to Ae. albopictus control in the urban context, stressing the need for larger field trials to evaluate the cost-efficacy of the approach in temperate regions. © 2023 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Is Côte D'Ivoire a new high hybridization zone for the two major malaria vectors, Anopheles coluzzii and An. gambiae (Diptera, Culicidae)?

Anopheles gambiae and An. coluzzii are very closely related and recently differentiated species representing the main malaria vectors in the Afrotropical region and responsible of up to >3 infective bites/person/night in Côte D'Ivoire, where prevention and control has stagnated in recent years. The aim of the present study was to genetically and ecologically characterize An. gambiae and An. coluzzii populations from two villages of Côte D'Ivoire, lying in the coastal forest belt and 250 km inland in the Guinean savannah mosaic belt, respectively. Results reveal high frequencies of both species in both study sites and high frequencies of hybrids (4-33%) along the whole year of sampling. Consistently with observations for the well-known high hybridization zone at the far-west of the species range, hybrid frequencies were higher in the coastal village and highest when the two species occurred at more balanced frequencies, supporting the "frequency-dependent hybridization" ecological speciation theory. Pilot genotyping revealed signatures of genomic admixture in both chromosome-X and -3. Coupled with previous reports of hybrids in the region, the results point to the coastal region of Côte D'Ivoire as a possible regions of high hybridization. Preliminary characterization of parameters relevant for malaria transmission and control (e.g. possibly higher sporozoite rates and indoor biting preferences in hybrids than in the parental species) highlight the possible relevance of the breakdown of reproductive barriers between An. gambiae and An. coluzzii not only in the field of ecological evolution, but also in malaria epidemiology and control.

High Levels of Admixture in Anopheles gambiae Populations from Côte d'Ivoire Revealed by Multilocus Genotyping.

Anopheles coluzzii and An. gambiae-the two most recently radiated species of the An. gambiae complex and the major Afrotropical malaria vector species-are identified by markers in the X-centromeric IGS rDNA region. Putative IGS-hybrids are rarely found in the field, except in restricted areas where genomic studies have led to the hypothesis that the observed IGS-patterns are due to cryptic taxa rather than to hybridization between the two species. We investigated the genome-wide levels of admixture in two villages in Côte d'Ivoire where high levels of IGS-hybrids have been detected, confirming unparalleled high frequencies in the coastal village. Genotyping of 24 Ancestry Informative Markers (AIMs) along the three chromosomes produced discordant results between the IGS-marker and the multilocus genotype obtained for AIMs across the whole genome (29%) as well as AIMs on chromosome-X (considered to be fundamental for species reproductive isolation) only (21%). Results highlight a complicated pattern of admixture that deserves deeper genomic analyses to understand better possible underlying causes (from extensive processes of hybridization to the existence of different cryptic taxa), and stress the need of developing advanced diagnostics for An. coluzzii, An. gambiae and putative new taxa, instrumental for assessing taxon-specific epidemiological characters.

A bacterium against the tiger: preliminary evidence of fertility reduction after release of Aedes albopictus males with manipulated Wolbachia infection in an Italian urban area.

BACKGROUND: Novel tools are needed to reduce the nuisance and risk of exotic arbovirus transmission associated with the colonization of temperate regions by Aedes albopictus. The incompatible insect technique (IIT) is a population suppression approach based on cytoplasmic incompatibility between males with manipulated endosymbionts and wild females. Here, we present the results of the first field experiment in Europe to assess the capacity of an Ae. albopictus line (ARwP) deprived of its natural endosymbiont Wolbachia and transinfected with a Wolbachia strain from the mosquito Culex pipiens, to sterilize wild females. RESULTS: We released ∼ 4500 ARwP males weekly for 6 weeks in a green area within urban Rome (Italy) and carried out egg (N = 13 442), female (N = 128) and male (N = 352) collections. Egg (N = 13 783) and female (N = 48) collections were also carried out at two untreated control sites. The percentage of viable eggs during release was, on average, significantly lower in treated sites than in control sites, with the greatest difference (16%) seen after the fourth release. The ARwP to wild male ratio in the release spots between day 3 after the first ARwP male release and day 7 after the last release was, on average, 7:10. Released males survived up to 2 weeks. Approximately 30% of females collected in the release spots showed 100% sterility and 20% showed strongly reduced fertility compared with control sites. CONCLUSIONS: Results support the potential of IIT as a tool contributing to Ae. albopictus control in the urban context, and stress the need for larger field trials to evaluate the cost-efficacy of the approach in suppressing wild populations. © 2019 Society of Chemical Industry.

Complex interplay of evolutionary forces shaping population genomic structure of invasive Aedes albopictus in southern Europe.

BACKGROUND: In the last four decades, the Asian tiger mosquito, Aedes albopictus, vector of several human arboviruses, has spread from its native range in South-East Asia to all over the world, largely through the transportation of its eggs via the international trade in used tires. Albania was the first country invaded in Europe in 1979, followed by Italy in 1990 and other Mediterranean countries after 2000. METHODS/PRINCIPAL FINDINGS: We here inferred the invasion history and migration patterns of Ae. albopictus in Italy (today the most heavily-infested country in Europe), Greece and Albania, by analyzing a panel of >100,000 single nucleotide polymorphisms (SNPs) obtained by sequencing of double-digest Restriction site-Associated DNA (ddRADseq). The obtained dataset was combined with samples previously analyzed from both the native and invasive range worldwide to interpret the results using a broader spatial and historical context. The emerging evolutionary scenario complements the results of other studies in showing that the extraordinary worldwide expansion of Ae. albopictus has occurred thanks to multiple independent invasions by large numbers of colonists from multiple geographic locations in both native and previously invaded areas, consistently with the role of used tires shipments to move large numbers of eggs worldwide. By analyzing mosquitoes from nine sites across ~1,000-km transect in Italy, we were able to detect a complex interplay of drift, isolation by distance mediated divergence, and gene flow in shaping the species very recent invasion and range expansion, suggesting overall high connectivity, likely due to passive transportation of adults via ground transportation, as well as specific adaptations to local conditions. CONCLUSIONS/SIGNIFICANCE: Results contribute to characterize one of the most successful histories of animal invasion, and could be used as a baseline for future studies to track epidemiologically relevant characters (e.g. insecticide resistance).

Phenotypic and genotypic pyrethroid resistance of Aedes albopictus, with focus on the 2017 chikungunya outbreak in Italy.

BACKGROUND: The highly invasive mosquito species Aedes albopictus has become a major health concern in temperate areas due to its role as vector of exotic arboviruses. Pyrethroid insecticides represent the main tools for limiting the circulation of such mosquito-borne viruses. The present work aim to extend previous reports on phenotypic pyrethroid-resistance in European Ae. albopictus, to identify its genetic basis and to monitor the geographical distribution of resistant genotypes, with a particular focus on sites experiencing the 2017 chikungunya outbreak in Italy. RESULTS: Bioassays, performed according to World Health Organization protocols, showed full susceptibility to deltamethrin (concentration = 0.05%) and varying levels of resistance to permethrin (0.75%) and/or α-cypermethrin (0.05%) across Italy, with highest levels in the core of the 2017 chikungunya outbreak. Partial genotyping of the VSSC gene revealed widespread distribution of V1016G mutation and confirmed its association with pyrethroid resistance. CONCLUSION: The results obtained show that the condition for the spread of pyrethroid resistance in Ae. albopictus in Europe exists under strong selective pressure due to intensive insecticide spraying to control exotic arbovirus outbreak or high levels of nuisance. The results draw attention to the need for an evidence-based implementation of mosquito nuisance control, taking insecticide resistance management into consideration. © 2019 Society of Chemical Industry.

First evidence of resistance to pyrethroid insecticides in Italian Aedes albopictus populations 26 years after invasion.

BACKGROUND: Aedes albopictus has spread during the last few decades all over the world. This has increased significantly the risk of exotic arbovirus transmission (e.g. chikungunya, dengue, and Zika) also in temperate areas, as demonstrated by the Chikungunya 2007 and 2017 outbreaks in northeastern and central Italy. Insecticides are an important tool for limiting the circulation of these mosquito-borne viruses. The aim of the present study was to address the gap in current knowledge of pyrethroid insecticide resistance of European Ae. albopictus populations, focusing on populations from Italy, Albania and Greece. RESULTS: Bioassays for resistance to permethrin (0.75%), α-cypermethrin (0.05%) or deltamethrin (0.05%) were performed according to World Health Organization (WHO) protocols and showed reduced susceptibility (<90% mortality) of some Italian populations to permethrin and α-cypermethrin, but not to deltamethrin. CONCLUSION: This study reports the first evidence of resistance to pyrethroids in adult Italian Ae. albopictus populations. Results refer to the season preceding the Chikungunya 2017 outbreak in central Italy and highlight the need to increase efforts to monitor the spread of insecticide resistance and the need to develop strategies to limit the spread of insecticide resistance, particularly in areas where extensive treatments have been carried out to contain disease outbreaks. © 2017 Society of Chemical Industry.

Intraductal papillary mucinous tumors of the pancreas. Surgical treatment: at what point should we stop?

The intraoperative management of the margins of intraductal papillary mucinous tumors (IPMNs) undergoing pancreatic resection is crucial. The surgeon must discontinue the resection whenever the pancreatic margin is negative and, of course, when a total pancreatectomy is indicated.Nevertheless, a wide gray area exists. The real surgical problems are represented by i) IPMNs involving only a segment of the pancreatic gland, thus necessitating intraoperative histological examination of the resection and the decision as to "when to stop the resection"; ii) the intraoperative management of those margins which are neither clearly negative nor clearly positive; iii) the actual indications either for surgery or for follow-up in those patients affected by peripheral IPMNs. In the literature, negative resection margins have a range of between 49 to 81% with a pancreatic recurrence rate of from 0 to 25% in follow-ups ranging from 6 months to 11 years after the first operation. In general, in this disease which mainly involves the head and uncinate process of the gland, decisions are a "balance" between the patient and the disease. In fact, on the one hand, there is the usual elderly patient with possible comorbidities, symptoms and the presence of diabetes; on the other hand, the disease, which usually involves the head and the uncinate process of the gland, tends to grow along the duct and be potentially malignant if carcinoma is not already present.