From Bio-Prospecting to Field Assessment: The Case of Carvacrol Rich Essential Oil as a Potent Mosquito Larvicidal and Repellent Agent

Contemporary legislation tends to increase limitation on the use of all synthetic pesticides, promoting bio-pesticides as a safer alternative. Bio-prospecting efforts for bio-pesticides provide results, which rarely reach the industry. Present essay elaborates on our efforts to chart the path from the laboratory bench to field assessment. Eight Mediterranean wild gathered foods provided the essential oils that were assessed as mosquito control agents against the Asian tiger mosquito (Aedes albopictus). Three Lamiaceae essential oils, derived from Satureja thymbra, Origanum onites, and Thymbra spicata presented carvacrol as principal component. All exhibited DEET-like repellent performance and total larvae mortality defining the carvacrol rich essential oil (CREO) as a promising mosquito control agent. A commercial variety of Origanum vulgare ssp. hirtum, was selected as CREO source and subjected to dose-response and eco-toxicity studies. We have found significant larvicidal (LC90 of 58,747 mg/L), and repellent (0.2 μL/cm2) properties, but also severe toxicity (LC90 of 12,806 mg/L) against Macrocyclops albidus. This last figure was the limit for the larvicidal field assessment; while for the repellent evaluation was used double the minimum indication (0.4 μL/cm2). CREO was tested per se as larvicidal agent, and emulsified for both repellent and larvicidal field activity. The emulsified CREO's spatial repellent assessment showed maximum efficacy of 86% in day 1 that gradually declined in the following 2 days (81%, 69%). Both emulsified and crude CREO proved to be efficient larvicidal agents, with crude CREO (3 weeks) overrunning slightly the emulsified (2 weeks) in terms of endurance. Conclusively, CREO in its emulsified form may be considered as a promising mosquito larvicidal and repellent agent, applicable in both precautionary and emergency response measures

AIMSurv: First pan-European harmonized surveillance of Aedes invasive mosquito species of relevance for human vector-borne diseases

Human and animal vector-borne diseases, particularly mosquito-borne diseases, are emerging or re-emerging worldwide. Six Aedes invasive mosquito (AIM) species were introduced to Europe since the 1970s: Aedes aegypti, Ae. albopictus, Ae. japonicus, Ae. koreicus, Ae. atropalpus and Ae. triseriatus. Here, we report the results of AIMSurv2020, the first pan-European surveillance effort for AIMs. Implemented by 42 volunteer teams from 24 countries. And presented in the form of a dataset named “AIMSurv Aedes Invasive Mosquito species harmonized surveillance in Europe. AIM-COST Action. Project ID: CA17108”. AIMSurv2020 harmonizes field surveillance methodologies for sampling different AIMs life stages, frequency and minimum length of sampling period, and data reporting. Data include minimum requirements for sample types and recommended requirements for those teams with more resources. Data are published as a Darwin Core archive in the Global Biodiversity Information Facility- Spain, comprising a core file with 19,130 records (EventID) and an occurrences file with 19,743 records (OccurrenceID). AIM species recorded in AIMSurv2020 were Ae. albopictus, Ae. japonicus and Ae. koreicus, as well as native mosquito species

Reducing Aedes albopictus breeding sites through education: A study in urban area.

Aedes albopictus tends to proliferate in small, often man-made bodies of water, largely present in urban private areas. For this reason, education and community participation are considered crucial for source reduction and mosquito control. In the current study, we identify mosquito breeding habitat and evaluate the effectiveness of resident education. Since 2010 several outbreaks of West Nile virus infection occurred in Greece however urban population has no previous experience with mosquito-borne disease related to Aedes species, such as Dengue, Zika and Chikungunya. After the introduction of Ae. albopictus in Greece, urban areas have been considered to be at risk of epidemic arboviral outbreaks and identifying effective control strategies is imperative. Our study examines the relationship between mosquito breeding sources and socioeconomic or demographic characteristics of different households in a Greek municipality and evaluates efficacy of resident education. The results revealed that only a minority of residents knew where mosquitoes breed (18.6%) and only 46% felt that residents had any responsibility for managing breeding habitat. Our findings strongly suggest that only the presence of scientific staff inspecting possible habitats in their properties, could be enough to stimulate practices towards source reduction. However, educational interventions alone with printed education material cannot enhance significant community participation and source reduction

Curcumin Derivatives as Potential Mosquito Larvicidal Agents against Two Mosquito Vectors, Culex pipiens and Aedes albopictus

Vector-borne diseases have appeared or re-emerged in many Southern Europe countries making the transmission of infectious diseases by mosquitoes (vectors) one of the greatest worldwide health threats. Larvicides have been used extensively for the control of Aedes (Stegomyia) albopictus (Skuse, 1895) (Diptera: Culicidae) and Culex pipiens Linnaeus, 1758 (Diptera: Culicidae) mosquitoes in urban and semi-urban environments, causing the increasing resistance of mosquitoes to commercial insecticides. In this study, 27 curcuminoids and monocarbonyl curcumin derivatives were synthesised and evaluated as potential larvicidal agents against Cx. pipiens and Ae. albopictus. Most of the compounds were more effective against larvae of both mosquito species. Four of the tested compounds, curcumin, demethoxycurcumin, curcumin-BF2 complex and a monocarbonyl tetramethoxy curcumin derivative exhibited high activity against both species. In Cx. pipiens the recorded LC50 values were 6.0, 9.4, 5.0 and 32.5 ppm, respectively, whereas in Ae. albopictus they exhibited LC50 values of 9.2, 36.0, 5.5 and 23.6 ppm, respectively. No conclusive structure activity relationship was evident from the results and the variety of descriptors values generated in silico provided some insight to this end

Essential oil-in-water microemulsions for topical application: structural study, cytotoxic effect and insect repelling activity

International audienceEssential oils (EOs) extracted from aromatic plants have been well recognized for their insect repellent properties. Although EOs are effective when freshly applied, their protective effect usually dissipates quickly due to low chemical stability and rapid volatilization of their ingredients. To address this issue, encapsulation in protective carrier materials at the nanoscale is proposed

Efficacy Evaluation of Oregano Essential Oil Mixed with <i>Bacillus thuringiensis israelensis</i> and Diflubenzuron against <i>Culex pipiens</i> and <i>Aedes albopictus</i> in Road Drains of Italy

Mosquito management programs in the urban environment of Italian cities mainly rely on larval control with conventional insecticides, primarily targeting the road drains that constitute the principal mosquito breeding sites encountered in public. The repeated utilization of synthetic insecticides may have adverse effects on non-targets and lead to resistance development issues, while the performance of biopesticides encounters limitations in field use. Botanical insecticides as single larval control agents or in binary mixtures with conventional insecticides have been extensively studied in the laboratory as an effective and eco-friendly alternative mosquito control method with promising results. The study herein concerns the investigation, for the first time under realistic conditions in the field, of the joint action of a carvacrol-rich oregano Essential Oil (EO) with two conventional insecticides, namely, the insect growth regulator diflubenzuron and the bio-insecticide Bacillus thuringiensis israelensis (B.t.i.), in road drains in Crevalcore city, Italy, against Culex pipiens and Aedes albopictus. According to the obtained results, the application of both plain EO and its mixtures with diflubenzuron and B.t.i. exerted very high efficacy in terms of immature mosquito population reduction over a two-week period. Three weeks after treatment, the performance of the oil and its mixtures diminished but remained high, while the addition of diflubenzuron potentiated the persistent action of the oil against Cx. pipiens. These findings are indicative of the potential of mixing carvacrol-rich EO with diflubenzuron and B.t.i. as an efficient eco-friendly alternative to mono-insecticide applications in road drains against Cx. pipiens and Ae. albopictus larvae

Knowledge, Attitude, and Practices Survey in Greece before the Implementation of Sterile Insect Technique against <i>Aedes albopictus</i>

Community involvement in Aedes albopictus management can be very efficient and result in raising awareness among citizens. Toward this end, a door-to-door campaign can encourage active community participation in vector control. The current study describes the results of an intervention where a KAP (knowledge, attitude, practices) survey tool was paired with a door-to-door campaign and was implemented as an intervention method in Vravrona area (Attica, Greece) before the release of sterile males (sterile insect technique, SIT) against Aedes albopictus. The KAP tool was used to shed light on the knowledge, practices, and attitudes of local community members in order to better prepare and motivate participation in household mosquito control and to assess current understanding of SIT. Each household also received specific information about mosquito source habitat in their own yards at the time of the initial KAP survey. These household data were complemented by standardized mosquito trapping in the municipality. Our findings indicate that citizens’ attitude toward SIT ranged from indecisive to fully supportive, while 77.5% of the respondents agreed that the SIT has many advantages over chemical control methods. Furthermore, the results demonstrate that using the door-to-door campaign as an intervention and prerelease method before SIT can suppress the initial mosquito population and potentially improve its efficacy. Lastly, we show that the presence of local municipality officials during door-to-door visits was associated with increased willingness from the residents to participate in the intervention

Quality Control Methods for <i>Aedes albopictus</i> Sterile Male Transportation

Genetic based mosquito control methods have been gaining ground in recent years for their potential to achieve effective suppression or replacement of vector populations without hampering environments or causing any public health risk. These methods require the mass rearing of the target species in large facilities sized to produce millions of sterile males, as already well established for a number of insects of agricultural importance. Assessing the performance of released males in Sterile Insect Technique (SIT) control programs is of the utmost importance for the success of the operation. Besides the negative effects of mass rearing and sterilization, the handling of sterilized insects and shipment to distant areas may also negatively impact the quality of sterilized males. The aim of the current study was to design and executive quality control (QC) tests for sterilized Aedes albopictus (Asian tiger mosquito) males delivered by air shipment from the mass production facility located in Italy to Greece and Montenegro field release sites. Mass reared mosquito strains were based on biological materials received from Italy, Greece and Montenegro. Tests conducted at the mass rearing facility before transportation revealed a rather high residual female contamination following mechanical sex separation (approximately 1.5% females, regardless of the mosquito strain). Irradiated males of all three mosquito strains induced high levels of sterility to females. Shipment lasting approximately 24 h resulted in approximately 15% mortality, while when shipment lasted nearly two days this increased to almost 40%. The flight ability of sterilized males following one day transportation time was satisfactory (over 60%). The response of sterile males to food and water starvation was comparable and slightly lower than that of wild non-transported males. Longevity of sterile males was shorter than that of wild counterparts and it seems it was not affected by mating to wild females. Both mating propensity and mating competitiveness for wild virgin females was higher for the wild, control males compared to the sterile, transported ones. Overall, the performance of sterile male Ae. albopictus delivered from the mass rearing facility of Italy to Greece in approximately 24 h was satisfactory. Transportation lasting two days or longer incurred detrimental effects on males, which called into question the outcome of the SIT release programs. In conclusion, our results demonstrate the need of quality control procedures, especially when sterile male production facilities are not near to the releasing point. Transportation could be a serious drawback for the implementation of Sterile Insect Releases and, consequently, it is important to establish an efficient and fast transportation of sterilized males in advance

Integrated Deadenylase Genetic Association Network and Transcriptome Analysis in Thoracic Carcinomas

The poly(A) tail at the 3&prime; end of mRNAs determines their stability, translational efficiency, and fate. The shortening of the poly(A) tail, and its efficient removal, triggers the degradation of mRNAs, thus, regulating gene expression. The process is catalyzed by a family of enzymes, known as deadenylases. As the dysregulation of gene expression is a hallmark of cancer, understanding the role of deadenylases has gained additional interest. Herein, the genetic association network shows that CNOT6 and CNOT7 are the most prevalent and most interconnected nodes in the equilibrated diagram. Subsequent silencing and transcriptomic analysis identifies transcripts possibly regulated by specific deadenylases. Furthermore, several gene ontologies are enriched by common deregulated genes. Given the potential concerted action and overlapping functions of deadenylases, we examined the effect of silencing a deadenylase on the remaining ones. Our results suggest that specific deadenylases target unique subsets of mRNAs, whilst at the same time, multiple deadenylases may affect the same mRNAs with overlapping functions

Proteomic Analysis of Human Angiogenin Interactions Reveals Cytoplasmic PCNA as a Putative Binding Partner

Human Angiogenin (hAng) is a member of the ribonuclease A superfamily and a potent inducer of neovascularization. Protein interactions of hAng in the nucleus and cytoplasm of the human umbilical vein cell line EA.hy926 have been investigated by mass spectroscopy. Data are available via ProteomeXchange with identifiers PXD006583 and PXD006584. The first gel-free analysis of hAng immunoprecipitates revealed many statistically significant potential hAng-interacting proteins involved in crucial biological pathways. Surprisingly, proliferating cell nuclear antigen (PCNA), was found to be immunoprecipitated with hAng only in the cytoplasm. The hAng–PCNA interaction and colocalization in the specific cellular compartment was validated with immunoprecipitation, immunoblotting, and immunocytochemistry. The results revealed that PCNA is predominantly localized in the cytoplasm, while hAng is distributed both in the nucleus and in the cytoplasm. hAng and PCNA colocalize in the cytoplasm, suggesting that they may interact in this compartment