Thomas Bawin

Reintroduction of the invasive mosquito species Aedes albopictus in Belgium in July 2013.

Since its first report in 2000, the invasive mosquito Aedes albopictus was not found any more during the different entomological inspections performed at its place of introduction in Belgium between 2001 and 2012. In July 2013, one adult male was captured at the same site (a platform of imported used tires located in Vrasene, Oost-Vlaanderen Province), during a monitoring using CO2-baited trap. This finding suggests the reintroduction of the species in Belgium via the used tire trade.

Could alternative solanaceous hosts act as refuges for the tomato leafminer, Tuta absoluta?

The tomato leafminer, Tuta absoluta (Lepidoptera: Gelechiidae), is a widespread devastating pest reported to develop on economically important solanaceous plants. The characterization of its effective host range could help to understand and prevent the dispersion behavior of the insect in the environment. In this study, the ability of T. absoluta to locate and develop on wild (Solanum nigrum, Atropa belladonna, Datura stramonium) and cultivated (Solanum tuberosum) solanaceous plant species under laboratory conditions was assessed. Dual-choice behavioral assays performed in flying tunnels (S. tuberosum vs. another plant) revealed that adult distribution and female oviposition did not differ between Solanum species, which were preferred to the other tested plants. The volatile molecules released by each tested plant species provide some explanations in the observed behavioral discrimination: S. nigrum and S. tuberosum volatile profiles were similar and were presenting quantitative and qualitative differences with the other tested solanaceous plants. To determine whether the host plant choice was adaptive or not, we have finally conducted fitness assays, by rearing T. absoluta larvae on each plant species and have shown that Solanum species allowed higher larval survivability and lower development time (from egg to adult emergency) compared to the other plants. We conclude that Solanum species are suitable host plants for T. absoluta, but other solanaceous plant species could be opportunistically colonized with fewer incidences.

A fungal biofilm reactor based on metal structured packing improves the quality of a Gla::GFP fusion protein produced by Aspergillus oryzae

Fungal biofilm is known to promote the excretion of secondary metabolites in accordance with solid-state-related physiological mechanisms. This work is based on the comparative analysis of classical submerged fermentation with a fungal biofilm reactor for the production of a Gla::green fluorescent protein (GFP) fusion protein by Aspergillus oryzae. The biofilm reactor comprises a metal structured packing allowing the attachment of the fungal biomass. Since the production of the target protein is under the control of the promoter glaB, specifically induced in solid-state fermentation, the biofilm mode of culture is expected to enhance the global productivity. Although production of the target protein was enhanced by using the biofilm mode of culture, we also found that fusion protein production is also significant when the submerged mode of culture is used. This result is related to high shear stress leading to biomass autolysis and leakage of intracellular fusion protein into the extracellular medium. Moreover, 2-D gel electrophoresis highlights the preservation of fusion protein integrity produced in biofilm conditions. Two fungal biofilm reactor designs were then investigated further, i.e. with full immersion of the packing or with medium recirculation on the packing, and the scale-up potentialities were evaluated. In this context, it has been shown that full immersion of the metal packing in the liquid medium during cultivation allows for a uniform colonization of the packing by the fungal biomass and leads to a better quality of the fusion protein.

Infestation Level Influences Oviposition Site Selection in the Tomato Leafminer Tuta absoluta (Lepidoptera: Gelechiidae)

The tomato leafminer, Tuta absoluta (Lepidoptera: Gelechiidae), is a devastating pest that develops principally on solanaceous plants throughout South and Central America and Europe. In this study, we tested the influence of three levels of T. absoluta infestations on the attraction and oviposition preference of adult T. absoluta. Three infestation levels (i.e., non-infested plants, plants infested with 10 T. absoluta larvae, and plants infested with 20 T. absoluta larvae) were presented by pairs in a flying tunnel to groups of T. absoluta adults. We found no differences in terms of adult attraction for either level of infestations. However, female oviposition choice is influenced by larvae density on tomato plants. We discuss the underlying mechanisms and propose recommendations for further research.

Betraying its presence: Identification of the chemical signal released by Tuta absoluta-infested tomato plants that guide generalist predators toward their prey

Plants modify their volatile chemical signature under pest infestation, which might directly or indirectly improve their defence against threats. These chemical signals have potential in integrated pest management strategies. Macrolophus pygmaeus (Rambur) [Heteroptera: Miridae] is a generalist predatory insect widely used to control the tomato leaf miner, Tuta absoluta (Meyrick) [Lepidoptera: Gelechiidae]. Based on a previous study demonstrating the ability of this mirid species to discriminate non-infested versus T. absoluta-infested tomato plants, our objective was to identify plant volatile chemicals (herbivore-induced plant volatiles—HIPVs) guiding the behaviour of such a generalist predator towards its prey. First, we used coupled gas chromatography-mass spectrometry-electroantennographic detection analysis to screen for active antenna components from the volatile blend released by T. absoluta-infested tomato plants. Dose responses associated with each isolated HIPV were also performed using an electroantennograph. Subsequently, behavioural assays were conducted in a double-choice olfactometer to analyse and identify the behaviourally active chemicals eliciting olfactory responses. Twenty-one total compounds induced antennal responses and six of the 21 evoked positive attractions in M. pygmaeus: (E)hex-2-enal, 2-carene, α-pinene, β-phellandrene, hexanal, and linalool. A synthetic blend of active HIPVs induced olfactory responses as well as attraction in the bioassays. Our results provided evidence the generalist mirid predator M. pygmaeus uses chemical cues from infested tomato plants to identify plants infested by prey. We discussed how these results can be used to improve existing biological approaches to control the tomato leaf miner, T. absoluta.

Molecular detection of six (endo-) symbiotic bacteria in Belgian mosquitoes: first step towards the selection of appropriate paratransgenesis candidates

Actually, the use of symbiotic bacteria is one of alternative solution to avoid vector resistance to pesticides. In Belgium, among 31 identified mosquito species, 10 were considered as potential vectors. Given to introduction risks of arbovirosis, the purpose of this study was to investigate the presence of symbiosis bacteria in potential mosquito vectors. Eleven species caught from 12 sites in Belgium were used: Culex pipiens s.l., Culex torrentium, Culex hortensis, Anopheles claviger, Anopheles maculipennis s.l., Anopheles plumbeus, Culiseta annulata, Ochlerotatus geniculatus, Ochlerotatus dorsalis, Aedes albopictus, and Coquillettidia richiardii. Six genera of symbiotic bacteria were screened: Wolbachia sp., Comamonas sp, Delftia sp., Pseudomonas sp., Acinetobacter sp., and Asaia sp. A total of 173 mosquito individuals (144 larvae and 29 adults) were used for the polymerase chain reaction screening. Wolbachia was not found in any Anopheles species nor Cx. torrentium. A total absence of Comamonas and Delftia was observed in all species. Acinetobacter, Pseudomonas, and Asaia were found in most of species with a high prevalence for Pseudomonas. These results were discussed to develop potential strategy and exploit the variable occurrence of symbiotic bacteria to focus on them to propose biological ways of mosquito control.

Diversity and ecology survey of mosquitoes potential vectors in Belgian equestrian farms: A threat prevention of mosquito-borne equine arboviruses

Emergence of West Nile Virus was recently recorded in several European countries, which can lead to severe health problems in horse populations. Europe is also at risk of introduction of mosquito-borne equine alphavirus from Americas. Prevention of these arboviruses requires a clear understanding of transmission cycles, especially their vectors. To characterize mosquito fauna, their ecology and identify potential vectors of equine arboviruses in Belgium, entomological surveys of six equestrian farms located in the Wolloon Region were conducted during 2011-2012. The harvest of mosquitoes was based on larval sampling (272 samples from 111 breeding sites) and monthly adults trapping (CO2-baited traps, Mosquito Magnet Liberty Plus). Among 51,493 larvae and 319 adult mosquitoes collected, morphological identification showed the presence of 11 species: Anopheles claviger (Meigen), An. maculipennis s.l. (Meigen), An. plumbeus (Stephens), Culex hortensis (Ficalbi), Cx. territans (Walker), Cx. pipiens s.l. L., Cx. torrentium (Martini), Coquillettidia richiardii (Ficalbi), Culiseta annulata (Schrank), Aedes cantans (Meigen), Ae. geniculatus (Olivier). Molecular identification of Cx. pipiens species complex allowed the detection of three molecular forms, Pipiens (92.3%), Molestus (4.6%) and Hybrid (3.1%). Larvae of Cx. pipiens sl and Cx. torrentium were omnipresent and the most abundant species. Water troughs, ponds and slurry (liquid manure) were the most favorable breeding sites of mosquito larvae. Based upon behavior and ecology of the identified mosquito species, Studied Belgian equestrian farms seem to provide a suitable environment and breeding sites for the proliferation of potential vectors of arboviruses and those being a real nuisance problem for horses and neighboring inhabitants.

Structure and distribution of the sensilla on the antennae of Tuta absoluta (Lepidoptera: Gelechiidae)

The tomato leafminer, Tuta absoluta (Lepidoptera: Gelechiidae), is a widespread devastating pest that develops on tomato and other economically important solanaceous crops. Current semiochemically-based management strategies still fail to significantly reduce damages and need to be improved. Here we describe under scanning and transmission electron microscopy the structure and distribution of the sensilla that are displayed on adult antennae. These were similar in size between males (3424.4±135.3μm) and females (3292.1±111.5μm), being segmented into a scape, a pedicel, and a distal filiform flagellum. Eight morphological sensilla types were observed on both sexes: Böhms bristles, sensilla squamiformia, sensilla trichodea, sensilla basiconica (two subtypes), sensilla chaetica, sensilla coeloconica, sensilla auricillica, and sensilla styloconica. The main sexual dimorphism was related to the higher abundance of sensilla trichodea in males, twice as abundant as in females. The putative functional significance of the different sensilla types regarding the insect ecology is discussed based on the available literature. This work provides descriptions of the antennae and related sensory structures. We expect these results to help develop further electrophysiological investigations aiming to a better understanding of T. absoluta olfaction.

Production of two entomopathogenic Aspergillus species and insecticidal activity against the mosquito Culex quinquefasciatus compared to Metarhizium anisopliae

ABSTRACT The spore productivity and insecticidal activity of two opportunistic insect pathogenic Aspergillus species (namely: Aspergillus clavatus Desmazieres and Aspergillus flavus Link (Ascomycota: Eurotiales, Trichocomaceae)) were compared to Metarhizium anisopliae sensu lato (Metchnikoff) Sorokin (Ascomycota: Hypocreales, Clavicipitaceae) for mosquito (Diptera: Culicidae) control. The production of aerial spores on wheat bran and white rice was investigated in solid-, semi-solid-, and liquid-state media supplemented with a nutritive solution. Wheat bran-based media increased the spore yield in solid-state from three to sevenfold: A. clavatus produced 48.4 ± 5.2 and 15.7 ± 1.6 × 108 spores/g, A. flavus produced 22.3 ± 4.1 and 3.1 ± 2.5 × 108 spores/g, and M. anisopliae produced 39.6 ± 6.5 and 13.1 ± 2.6 × 108 spores/g of wheat bran or white rice, respectively. A. clavatus, A. flavus and M. anisopliae spores harvested from wheat bran-based solid-state media showed lethal concentrations (LC50) of 1.1, 1.8, and 1.3 × 108 spores/ml against Culex quinquefasciatus Say larvae in 72 h. Because A. clavatus and M. anisopliae displayed similar features when cultured under these conditions, our results suggest that insect pathogenic Aspergillus species may be as productive and virulent against mosquito larvae as a well-recognised entomopathogenic fungus.

Oviposition deterrent activity of basil plants and their essentials oils against Tuta absoluta (Lepidoptera: Gelechiidae)

The leafminer Tuta absoluta Meyrick (Lepidoptera: Gelechiidae) is one of the most important pests of tomato, reducing crop yields by up to 100% in greenhouses and fields, in several countries globally. Because synthetic insecticides lead to resistance and have adverse effects on natural enemies and the health of producers, alternative control methods are needed. In this study, we assessed the oviposition-deterring effect of basil plants, Ocimum gratissimum L. and O. basilicum L. (Lamiaceae), using dual-choice behavioural assays performed in flight tunnels. We found that both plants significantly reduced T. absoluta oviposition behaviour on a tomato plant located nearby. To evaluate the potential effect of basil volatile organic compounds, we formulated essential oils of both plant species in paraffin oil, and observed a similar oviposition-deterring effect. Gas chromatography analyses detected 18 constituents in these essential oils which the major constituents included thymol (33.3%), p-cymene (20.4%), γ-terpinene (16.9%), myrcene (3.9%) in O. gratissimum and estragol (73.8%), linalool (8.6%), β-elemene (2.9%) and E-β-ocimene (2.6%) in O. basilicum. Twenty and 33 compounds were identified of the volatiles collected on O. gratissimum and O. basilicum plants, respectively. The main components include the following: p-cymene (33.5%), γ-terpinene (23.6%), α-terpinene (7.2%), α-thujene (6.7%) and E-α-bergamotene (38.9%) in O. gratissimum, and methyl eugenol (26.1%), E-β-ocimene (17.7%), and linalool (9.4%) in O. basilicum. Four compounds (α-pinene, β-pinene, Myrcene, Limonene) were common in essential oils and plants. Our results suggest the valuable potential of basil and associated essential oils as a component of integrated management strategies against the tomato leafminer.