Random mutations induced by a sub-sterilizing dose of gamma ray on Aedes albopictus male pupae and transmission to progeny.

The application of the Sterile Insect Technique (SIT) to mosquito control is based on the systematic release of large numbers of adult males that have been previously sterilized by irradiation. Ionizing radiation doses inducing full sterility also cause somatic damages that reduce the capacity of the treated males to compete with wild males. The optimal dose inducing high levels of male sterility and minimal impact on competitiveness can be assessed by establishing a dose-response curve. Sub-sterile males are, to a variable degree, still fertile and might be able to transmit to the progeny and following generation(s) sub-lethal random mutations resulting from irradiation. To investigate this, we treated Ae. albopictus male pupae with a sub-sterilizing (2-4 % of egg hatching) dose of gamma rays and explored expressed mutated genes in treated males and their progeny using RNA-seq. Single nucleotide polymorphisms (SNPs) were called using two independent pipelines. Only SNPs common to both pipelines (less than 5 % of the total SNPs predicted) were considered reliable and were annotated to genes. Over 600 genes with mutations likely induced by irradiation were found in the treated Ae. albopictus males. A part of the genes found mutated in irradiated males were also found in (and therefore probably passed on to) males of the F1 and F2 progeny, indicating that genetic variations induced by irradiation may be transmitted along generations. The mutated genes in irradiated males did not seem to significantly affect biological processes, except in one case (i.e., oxidative phosphorylation). Only in four cases (i.e., oxidative phosphorylation, UDP-glucose metabolic process, proton transmembrane transport and riboflavin metabolism) we found biological processes to be significantly affected by mutated genes that were likely transmitted to the male progeny. Our results suggest that random mutations induced by a sub-sterilizing dose of gamma ray in Ae. albopictus male pupae and transmitted to the male progeny of the irradiated mosquitoes do not affect biological processes potentially harmful, from a public-health point of view.

Optical recognition of the eggs of four Aedine mosquito species (Aedes albopictus, Aedes geniculatus, Aedes japonicus, and Aedes koreicus).

The continuous expansion of exotic Aedine mosquito species potential vectors of pathogens into new areas is a public health concern. In continental Europe, the surveillance of these mosquitoes is hindered by the simultaneous presence of three main invasive species (i.e., Aedes albopictus, Ae. japonicus, and Ae. koreicus). Standard low-cost surveillance methods (i.e., the deployment of oviposition traps and count of eggs under stereoscopic microscope) fail to distinguish the eggs of the different species. Identification of eggs by molecular methods is costly and time consuming and prevents measuring the density of invasive species and detecting early new invaders. Here we tested whether certain species could be identified by the patterns on the exochorionic membrane of their eggs. In a first step, we examined Aedine eggs of the three mentioned invasive and one indigenous (i.e., Ae. geniculatus) species with a high-resolution stereomicroscope and we identified each egg by MALDI-TOF mass spectrometry. In a second step, we submitted images of the eggs to 60 entomology experts and non-experts and tested their ability to distinguish among the species after an initial short training. The results obtained were consistent. Participants did not encounter difficulties in determining Ae. albopictus and Ae. geniculatus, while they had more difficulties in distinguishing Ae. japonicus from Ae. koreicus. In general, the quality of the exochorion seemed to play a more important role than the expertise level of the rater. The feasibility to differentiate Ae. albopictus from the other two invasive species is a significant achievement, as this is currently the most problematic species at the level of public health in Europe. Due to the presence of multiple invasive species that might prevent the correct quantification of mosquito population densities using standard surveillance methods and due to Ae. aegypti threat, it is recommended to optically determine also other species.

Estimating the Impact of Consecutive Blood Meals on Vector Competence of Aedes albopictus for Chikungunya Virus.

The continuous expansion of Aedes albopictus in Europe and the increases in autochthonous arboviruses transmissions in the region urge a better understanding of the virus transmission dynamic. Recent work described enhanced chikungunya virus (CHIKV) dissemination in Aedes aegypti mosquitoes exposed to a virus-free blood meal three days after their infection with CHIKV. Our study investigated the impact of a second blood meal on the vector competence of Ae. albopictus from southern Switzerland infected with CHIKV. Seven-day-old Ae. albopictus females were exposed to CHIKV-spiked blood and incubated at constant (27 °C) and fluctuating (14-28 °C) temperatures. Four days post-infection (dpi), some of these females were re-fed with a non-infectious blood meal. Virus infectivity, dissemination, transmission rate, and efficiency were investigated at seven and ten dpi. No enhanced dissemination rate was observed among females fed a second time; however, re-fed females have shown higher transmission efficiency than those fed only once after seven days post-infection and incubated under a fluctuating temperature regime. Vector competence for CHIKV was confirmed in Ae. albopictus from southern Switzerland. We did not observe an increase in dissemination rates among mosquitoes fed a second time (second blood meal), regardless of the temperature regime.

The effects of microclimatic winter conditions in urban areas on the risk of establishment for Aedes albopictus.

The tiger mosquito, Aedes albopictus, has adjusted well to urban environments by adopting artificial water containers as oviposition sites. Its spread in temperate regions is favoured by the deposition of cold-tolerant diapausing eggs that survive winter temperatures to a certain degree. The probability of establishment in new geographical areas is estimated using predictive models usually based on meteorological data measured at coarse resolution. Here, we investigated if we could obtain more precise and realistic risk scenarios for the spread of Ae. albopictus when considering the winter microclimatic conditions of catch basins, one of the major sites of oviposition and egg overwintering in temperate urban areas. We monitored winter microclimatic conditions of catch basins in four Swiss cities and developed a regression model to predict the average microclimatic temperatures of catch basins, based on available meteorological parameters, accounting for the observed differences between cities. We then used the microclimatic model to correct the predictions of our previously developed risk model for the prediction of Ae. albopictus establishment. Comparison of the predictive model's results based on local climate data and microclimate data indicated that the risk of establishment for Ae. albopictus in temperate urban areas increases when microhabitat temperatures are considered.

Risk-Based Mapping Tools for Surveillance and Control of the Invasive Mosquito Aedes albopictus in Switzerland.

BACKGROUND: In Switzerland, Aedes albopictus is well established in Ticino, south of the Alps, where surveillance and control are implemented. The mosquito has also been observed in Swiss cities north of the Alps. Decision-making tools are urgently needed by the local authorities in order to optimize surveillance and control. METHODS: A regularized logistic regression was used to link the long-term dataset of Ae. albopictus occurrence in Ticino with socioenvironmental predictors. The probability of establishment of Ae. albopictus was extrapolated to Switzerland and more finely to the cities of Basel and Zurich. RESULTS: The model performed well, with an AUC of 0.86. Ten socio-environmental predictors were selected as informative, including the road-based distance in minutes of travel by car from the nearest cell established in the previous year. The risk maps showed high suitability for Ae. albopictus establishment in the Central Plateau, the area of Basel, and the lower Rhone Valley in the Canton of Valais. CONCLUSIONS: The areas identified as suitable for Ae. albopictus establishment are consistent with the actual current findings of tiger mosquito. Our approach provides a useful tool to prompt authorities' intervention in the areas where there is higher risk of introduction and establishment of Ae. albopictus.

Effectiveness of integrated Aedes albopictus management in southern Switzerland.

BACKGROUND: The exotic invasive tiger mosquito, Aedes albopictus, appeared in southern Switzerland in 2003. The spread of the mosquito has been surveyed constantly since then, and an integrated vector management (IVM) has been implemented to control its numbers. The control measures focus on the aquatic phase of the mosquito with removal of breeding sites and applications of larvicides in public areas whereas private areas are reached through extensive public information campaigns. Here, we evaluated the efficacy of the IVM. METHODS: Since all the municipalities with Ae. albopictus in southern Switzerland are currently implementing the IVM, Italian municipalities just across the Swiss-Italian border, where Ae. albopictus is present but no coordinated intervention programme is in place, served as control. Ovitraps and adult female traps were used to measure mosquito abundance in 2019. Generalised mixed-effects models were used to model the numbers of Ae. albopictus eggs and adult females collected. These numbers of Ae. albopictus eggs were compared to the numbers of eggs collected in 2012 and 2013 in a previous assessment of the IVM, using a hurdle model. RESULTS: Mean numbers of Ae. albopictus eggs and adult females in 2019 were consistently higher in the municipalities not following an IVM programme. In these municipalities, there were about four times (3.8) more eggs than in the municipalities implementing an IVM programme. Also, the numbers of eggs and adult females increased steadily from the beginning of the Ae. albopictus reproductive season, reaching a peak in August. In contrast, the increase in numbers of Ae. albopictus was much more contained in the municipalities implementing an IVM programme, without reaching an evident peak. Comparison with data from 2012 and 2013 indicates that the gap between intervention and non-intervention areas may have almost doubled in the past 6 years. CONCLUSIONS: The results of the survey strongly support the efficacy of the IVM programme implemented in southern Switzerland compared to municipalities without defined control measures. With the constant implementation of an IVM, it appears possible to contain the numbers of Ae. albopictus at a manageable level, reducing the nuisance for the human population and the risk of arbovirus epidemics.

Emerging Aedes-borne infections in southern Switzerland: Preparedness planning for surveillance and intervention.

BACKGROUND: The Preparedness Plan for Surveillance and Interventions on Emerging Vector-Borne Diseases (VBDs) in Southern Switzerland outlines the strategy for preventing and managing potential outbreaks, as well as the surveillance and control activities with a specific focus on Aedes-borne diseases transmitted by Aedes albopictus mosquitoes. The objective of the plan is to provide Public Health Authorities with a framework of preventive and control measures according to the situation and level of epidemic risks. MATERIAL AND METHODS: The plan is divided into various phases representing the different steps for all potential situations, ranging from no vectors and no transmission risk to epidemic levels with multiple autochthonous/local cases of hospitalization (and deaths) until the end of the epidemic. An algorithm presents how decisions are taken to move from one phase of the plan to another, with detailed activities for different partners and strategies for each specific phase. RESULTS: The different phases of the plan include activities on disease surveillance and clinical case management, on vector surveillance and control, communication and coordination of activities. The plan is divided into five phases of activities and decision levels. From phase 0 (no cases) to phase 1 (low number of local cases, less than 5), phase 2 (small outbreak with more than 5 local cases), phase 3 (epidemic) and phase 4 (return to no more cases). CONCLUSION: The plan has been approved by the cantonal authorities and will be submitted to federal authorities. The required implementation tests will begin shortly.

Effect of Powdered Activated Carbon as Advanced Step in Wastewater Treatments on Antibiotic Resistant Microorganisms.

BACKGROUND: Conventional wastewater treatment plants discharge significant amounts of antibiotic resistant bacteria and antibiotic resistance genes into natural water bodies contributing to the spread of antibiotic resistance. Some advanced wastewater treatment technologies have been shown to effectively decrease the number of bacteria. Nevertheless, there is still a lack of knowledge about the effectiveness of these treatments on antibiotic resistant bacteria and antibiotic resistant genes. To the best of our knowledge, no specific studies have considered how powdered activated carbon (PAC) treatments can act on antibiotic resistant bacteria, although it is essential to assess the impact of this wastewater treatment on the spread of antibiotic resistant bacteria. METHODS: To address this gap, we evaluated the fate and the distribution of fluorescent-tagged antibiotic/ antimycotic resistant microorganisms in a laboratory-scale model simulating a process configuration involving powdered activated carbon as advanced wastewater treatment. Furthermore, we studied the possible increase of naturally existing antibiotic resistant bacteria during the treatment implementing PAC recycling. RESULTS: The analysis of fluorescent-tagged microorganisms demonstrated the efficacy of the PAC adsorption treatment in reducing the load of both susceptible and resistant fluorescent microorganisms in the treated water, reaching a removal efficiency of 99.70%. Moreover, PAC recycling did not increase the resistance characteristics of cultivable bacteria neither in the sludge nor in the treated effluent. CONCLUSION: Results suggest that wastewater PAC treatment is a promising technology not only for the removal of micropollutants but also for its effect in decreasing antibiotic resistant bacteria release.

Bacterial diversity in the water column of meromictic Lake Cadagno and evidence for seasonal dynamics.

The meromictic Lake Cadagno is characterized by a compact chemocline with high concentrations of anoxygenic phototrophic purple and green sulfur bacteria. However, a complete picture of the bacterial diversity, and in particular of effects of seasonality and compartmentalization is missing. To characterize bacterial communities and elucidate relationships between them and their surrounding environment high-throughput 16S rRNA gene pyrosequencing was conducted. Proteobacteria, Chlorobi, Verrucomicrobia, and Actinobacteria were the dominant groups in Lake Cadagno water column. Moreover, bacterial interaction within the chemocline and between oxic and anoxic lake compartments were investigated through fluorescence in situ hybridization (FISH) and flow cytometry (FCM). The different populations of purple sulfur bacteria (PSB) and green sulfur bacteria (GSB) in the chemocline indicate seasonal dynamics of phototrophic sulfur bacteria composition. Interestingly, an exceptional bloom of a cyanobacteria population in the oxic-anoxic transition zone affected the common spatial distribution of phototrophic sulfur bacteria with consequence on chemocline location and water column stability. Our study suggests that both bacterial interactions between different lake compartments and within the chemocline can be a dynamic process influencing the stratification structure of Lake Cadagno water column.

Multilocus sequence typing reveals clonality in Saprolegnia parasitica outbreaks.

The molecular epidemiology of fish pathogen Saprolegnia parasitica is still largely unknown. We developed a multilocus sequence typing scheme based on seven housekeeping genes to characterize 77 S. parasitica strains isolated from different fish host species at different times and from different geographic areas in Switzerland between 2015 and 2017. Ten different diploid sequence types (DSTs) were identified. The majority (52%) of outbreaks in Switzerland seemed to be caused by one genotype, namely DST3, which was recovered from farm-raised and wild-caught fish in all the geographic areas and river basins included in the study. DST3 was also recovered from the rivers Bienne (eastern France) and Doubs, where the episodes of massive mortality due to saprolegniosis started in 2009. Another genotype (DST7) showed, to a lesser extent, a distribution across different river basins, while eight DSTs were unique to a defined geographic area or river basin. The occurrence of sporadic DSTs indicates a certain degree of diversity within S. parasitica in the environment. The wide distribution of DST3 suggests that a clonal population may have spread in eastern France and Switzerland across geographic barriers, with strong implications for the management of both captive and wild fish populations.

Investigation of temperature conditions in Swiss urban and suburban microhabitats for the overwintering suitability of diapausing Aedes albopictus eggs.

BACKGROUND: In Switzerland, the invasive Asian tiger mosquito, Aedes albopictus, is firmly established in the Canton of Ticino, south of the Alps. According to a large-scale distribution model developed in 2013, suitable climatic conditions for the establishment of Ae. albopictus north of the Alps are found in Basel and Geneva while Zurich appears to be characterized by winters currently being too cold for survival of diapausing eggs. However, the spatial resolution of large-scale distribution models might not be sufficient to detect particular climatic conditions existing in urban settings, such as the presence of microclimatic temperatures, which may positively influence the probability of diapausing eggs to overwinter. In order to investigate this, microclimatic monitoring of potential diapausing sites (i.e. catch basins) and external controls was performed in January 2017 in Ticino and within the cities of Basel, Geneva and Zurich. RESULTS: Mean January temperatures in catch basins of Basel, Geneva and Zurich were always higher than the -1 °C temperature threshold previously set for survival probability of diapausing eggs, while mean January temperatures were below -1 °C in several catch basins south of the Alps, where Ae. albopictus eggs currently overwinter. The catch basin absolute January daily minimum temperatures both south and north of the Alps were in general higher than the external control temperatures. Absolute January daily minimum temperatures in catch basins in Basel, Geneva and Zurich were always above -10 °C, indicating that diapausing Ae. albopictus eggs could potentially survive winter nights in urban areas north of the Alps. CONCLUSIONS: The findings confirmed previous conclusions that urban catch basins can provide favourable conditions for overwintering of diapausing eggs compared to more cold-exposed sites. The results confirmed the presence of suitable winter conditions for the establishment of Ae. albopictus in the cities of Basel and Geneva. In addition, the microclimate-scale analysis added new information compared to the previous large-scale prevision model by showing that also the city of Zurich could provide winter conditions suitable for the establishment of Ae. albopictus. This illustrates the importance of the resolution of climate data in using models to predict Ae. albopictus distribution.

Phylogenetic evidence that two distinct Trichuris genotypes infect both humans and non-human primates.

Although there has been extensive debate about whether Trichuris suis and Trichuris trichiura are separate species, only one species of the whipworm T. trichiura has been considered to infect humans and non-human primates. In order to investigate potential cross infection of Trichuris sp. between baboons and humans in the Cape Peninsula, South Africa, we sequenced the ITS1-5.8S-ITS2 region of adult Trichuris sp. worms isolated from five baboons from three different troops, namely the Cape Peninsula troop, Groot Olifantsbos troop and Da Gama Park troop. This region was also sequenced from T. trichiura isolated from a human patient from central Africa (Cameroon) for comparison. By combining this dataset with Genbank records for Trichuris isolated from other humans, non-human primates and pigs from several different countries in Europe, Asia, and Africa, we confirmed the identification of two distinct Trichuris genotypes that infect primates. Trichuris sp. isolated from the Peninsula baboons fell into two distinct clades that were found to also infect human patients from Cameroon, Uganda and Jamaica (named the CP-GOB clade) and China, Thailand, the Czech Republic, and Uganda (named the DG clade), respectively. The divergence of these Trichuris clades is ancient and precedes the diversification of T. suis which clustered closely to the CP-GOB clade. The identification of two distinct Trichuris genotypes infecting both humans and non-human primates is important for the ongoing treatment of Trichuris which is estimated to infect 600 million people worldwide. Currently baboons in the Cape Peninsula, which visit urban areas, provide a constant risk of infection to local communities. A reduction in spatial overlap between humans and baboons is thus an important measure to reduce both cross-transmission and zoonoses of helminthes in Southern Africa.

Rapid species specific identification and subtyping of Yersinia enterocolitica by MALDI-TOF mass spectrometry.

Yersinia enterocolitica are Gram-negative pathogens and known as important causes of foodborne infections. Rapid and reliable identification of strains of the species Y. enterocolitica within the genus Yersinia and the differentiation of the pathogenic from the non-pathogenic biotypes has become increasingly important. We evaluated here the application of matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) for rapid species identification and subtyping of Y. enterocolitica. To this end, we developed a reference MS database library including 19 Y. enterocolitica (non-pathogenic biotype 1A and pathogenic biotypes 2 and 4) as well as 24 non-Y. enterocolitica strains, belonging to eleven different other Yersinia spp. The strains provided reproducible and unique mass spectra profiles covering a wide molecular mass range (2000 to 30,000 Da). Species-specific and biotype-specific biomarker protein mass patterns were determined for Y. enterocolitica. The defined biomarker mass patterns (SARAMIS SuperSpectrum™) were validated using 117 strains from various Y. enterocolitica bioserotypes in a blind-test. All strains were correctly identified and for all strains the mass spectrometry-based identification scheme yielded identical results compared to a characterization by a combination of biotyping and serotyping. Our study demonstrates that MALDI-TOF-MS is a reliable and powerful tool for the rapid identification of Y. enterocolitica strains to the species level and allows subtyping of strains to the biotype level.