Luca Mazzon

Translocation of Neonicotinoid Insecticides From Coated Seeds to Seedling Guttation Drops: A Novel Way of Intoxication for Bees

ABSTRACT The death of honey bees, Apis mellifera L., and the consequent colony collapse disorder causes major losses in agriculture and plant pollination worldwide. The phenomenon showed increasing rates in the past years, although its causes are still awaiting a clear answer. Although neonicotinoid systemic insecticides used for seed coating of agricultural crops were suspected as possible reason, studies so far have not shown the existence of unquestionable sources capable of delivering directly intoxicating doses in the fields. Guttation is a natural plant phenomenon causing the excretion of xylem fluid at leaf margins. Here, we show that leaf guttation drops of all the corn plants germinated from neonicotinoid-coated seeds contained amounts of insecticide constantly higher than 10 mg/1, with maxima up to 100 mg/1 for thiamethoxam and clothianidin, and up to 200 mg/1 for imidacloprid. The concentration of neonicotinoids in guttation drops can be near those of active ingredients commonly applied in field sprays for pest control, or even higher. When bees consume guttation drops, collected from plants grown from neonicotinoid-coated seeds, they encounter death within few minutes.

Fatal powdering of bees in flight with particulates of neonicotinoids seed coating and humidity implication

Losses of honeybees have been reported in Italy concurrent with the sowing of corn coated with neonicotinoids using a pneumatic drilling machine. Being unconvinced that solid particles containing systemic insecticide, falling on the vegetation surrounding the sown area, could poison bees foraging on contaminated nectar and pollen, the effect of direct aerial powdering was tested on foragers in free flight near the drilling machine. Bees were conditioned to visit a dispenser of sugar solution whilst a drilling machine was sowing corn along the flight path. Samples of bees were captured on the dispenser, caged and held in the laboratory. Chemical analysis showed some hundred nanograms of insecticide per bee. Nevertheless, caged bees, previously contaminated in flight, died only if kept in conditions of high humidity. After the sowing, an increase in bee mortality in front of the hives was also observed. Spring bee losses, which corresponded with the sowing of corn‐coated seed, seemed to be related to the casual encountering of drilling machine during foraging flight across the ploughed fields.

First report in italy of the exotic mosquito species Aedes (Finlaya) koreicus, a potential vector of arboviruses and filariae

BackgroundIn the Veneto region (north-eastern Italy) an entomological surveillance system has been implemented since the introduction of the Asian tiger mosquito (Aedes albopictus) in 1991. During the routine monitoring activity in a tiger mosquito-free area, an unexpected mosquito was noticed, which clearly did not belong to the recorded Italian fauna.FindingsAt the end of May 2011, twelve larvae and pupae were collected in a small village in Belluno province (Veneto region) from a single manhole. Ten adults reared in the laboratory were morphologically and genetically identified as Aedes (Finlaya) koreicus (Edwards, 1917), a species native to Southeast Asia. The subsequent investigations carried out in the following months in the same village provided evidence that this species had become established locally. Entomological and epidemiological investigations are currently ongoing in the surrounding area, to verify the eventual extension of the species outside the village and to trace back the route of entry into Italy.ConclusionsThis is the first report in Italy of the introduction of the exotic mosquito Ae. koreicus. This species has been shown experimentally to be competent in the transmission of the Japanese encephalitis virus and of the dog heartworm Dirofilaria immitis and is considered a potential vector of other arboviruses. Thus, the establishment of this species may increase the current risk or pose new potential threats, for human and animal health. This finding considerably complicates the entomological monitoring of the Asian tiger mosquito Ae. albopictus in Italy and stresses the importance of implementing the entomological surveillance for the early detection of and the rapid response against invasive mosquito species.

Aerial powdering of bees inside mobile cages and the extent of neonicotinoid cloud surrounding corn drillers

Sudden losses of bees have been observed in spring during maize sowing. The death of bees has been correlated with the use of neonicotinoid‐coated seed and the toxic particulates emitted by pneumatic drilling machines. The contamination of foragers in flight over the ploughed fields has been hypothesized. The airborne contamination has been proven, both with bees inside fixed cages around the field and in free flight near the driller. A new trial involving mobile cages has been established and consists of making rapid passes with single bees inside cages fixed to an aluminium bar. The bar was moved by two operators at different distances from the working drilling machine. A single pass was shown as sufficient to kill all the bees exposed to exhaust air on the emission side of the drill, when bees were subsequently held in high relative humidity. The extent of toxic cloud around driller was evaluated at the height of 0.5, 1.8 and 3.5 m and proved to be about 20 m in diameter, with an ellipsoidal shape. The shape may be influenced by working speed of the drill and environmental parameters, and is easily shown by adding talc powder to the seed in the machine hopper. A new driller equipment was evaluated consisting of two tubes inclined towards the soil that direct the exhaust air towards the ground. The survival rate of the bees was not substantially increased using the modified drill and was lower than 50%. Chemical analyses show up to 4000 ng of insecticide in single bees with an average content around 300 ng. Similar quantities were observed at increased distances from the modified or unmodified drillers. This new evaluation of bee mortality in the field is an innovative biological test to verify the hypothetical efficiency (or not) of driller modifications.

Antispila oinophylla new species (Lepidoptera, Heliozelidae), a new North American grapevine leafminer invading Italian vineyards: taxonomy, DNA barcodes and life cycle

Abstract A grapevine leafminer Antispila oinophylla van Nieukerken & Wagner, sp. n., is described both from eastern North America (type locality: Georgia) and as a new important invader in North Italian vineyards (Trentino and Veneto Region) since 2006. The species is closely related to, and previously confused with Antispila ampelopsifoliella Chambers, 1874, a species feeding on Virginia creeper Parthenocissus quinquefolia (L.) Planchon., and both are placed in an informal Antispila ampelopsifoliella group. Wing pattern, genitalia, and DNA barcode data all confirm the conspecificity of native North American populations and Italian populations. COI barcodes differ by only 0–1.23%, indicating that the Italian populations are recently established from eastern North America. The new species feeds on various wild Vitis species in North America, on cultivated Vitis vinifera L. in Italy, and also on Parthenocissus quinquefolia in Italy. North American Antispila feeding on Parthenocissus include at least two other species, one of which is Antispila ampelopsifoliella. Morphology and biology of the new species are contrasted with those of North American Antispila Hübner, 1825 species and European Holocacista rivillei (Stainton, 1855). The source population of the introduction is unknown, but cases with larvae or pupae, attached to imported plants, are a likely possibility. DNA barcodes of the three European grapevine leafminers and those of all examined Heliozelidae are highly diagnostic. North American Vitaceae-feeding Antispila form two species complexes and include several as yet unnamed taxa. The identity of three out of the four previously described North American Vitaceae-feeding species cannot be unequivocally determined without further revision, but these are held to be different from Antispila oinophylla. In Italy the biology of Antispila oinophylla was studied in a vineyard in the Trento Province (Trentino-Alto Adige Region) in 2008 and 2009. Mature larvae overwinter inside their cases, fixed to vine trunks or training stakes. The first generation flies in June. An additional generation occurs from mid-August onwards. The impact of the pest in this vineyard was significant with more than 90% of leaves infested in mid-summer. Since the initial discovery in 2006, the pest spread to several additional Italian provinces, in 2010 the incidence of infestation was locally high in commercial vineyards. Preliminary phylogenetic analyses suggest that Antispila is paraphyletic, and that the Antispila ampelopsifoliella group is related to Coptodisca Walsingham, 1895, Holocacista Walsingham & Durrant, 1909 and Antispilina Hering, 1941, all of which possess reduced wing venation. Vitaceae may be the ancestral hostplant family for modern Heliozelidae.

Correlation of bois noir disease with nettle and vector abundance in northern Italy vineyards

The phytoplasmas associated with bois noir (BN) tuf-type a are transmitted to grapevines by means of Hyalesthes obsoletus Signoret using Urtica dioica L. as the inoculum source. In the period 2003–2008, a research was carried out in six vineyards of northern Italy where a large amount of nettles in the ditches surrounding vineyards was detected. The aim was to establish the nettle control effects on the presence of H. obsoletus and new symptomatic grapevines. PCR and RFLP analyses showed the presence of the BN tuf-type a phytoplasmas in symptomatic grapevines of all vineyards and in the vector. Weed control strategies along ditches were always associated with decreases in nettle coverage and H. obsoletus numbers. A reduction in the percentage of new symptomatic grapevines occurred in all vineyards except for one vineyard where the least reduction in nettle and H. obsoletus was observed. The decrease in H. obsoletus population was significantly correlated with a decrease in nettles along ditches, and the reduction in H. obsoletus population was correlated with a decrease in incidence of new symptomatic grapevines compared with the incidence before the application of weed control measures. Nettle weeding can reduce the incidence of BN tuf-type a, but weed control costs and impacts need to be considered.

A unique midgut-associated bacterial community hosted by the cave beetle Cansiliella servadeii (Coleoptera: Leptodirini) reveals parallel phylogenetic divergences from universal gut-specific ancestors

BackgroundCansiliella servadeii (Coleoptera) is an endemic troglobite living in deep carbonate caves in North-Eastern Italy. The beetle constantly moves and browses in its preferred habitat (consisting in flowing water and moonmilk, a soft speleothem colonized by microorganisms) self-preens to convey material from elytra, legs, and antennae towards the mouth. We investigated its inner and outer microbiota using microscopy and DNA-based approaches.ResultsAbundant microbial cell masses were observed on the external appendages. Cansiliella’s midgut is fully colonized by live microbes and culture-independent analyses yielded nearly 30 different 16S phylotypes that have no overlap with the community composition of the moonmilk. Many of the lineages, dominated by Gram positive groups, share very low similarity to database sequences. However for most cases, notwithstanding their very limited relatedness with existing records, phylotypes could be assigned to bacterial clades that had been retrieved from insect or other animals’ digestive traits.ConclusionsResults suggest a history of remote separation from a common ancestor that harboured a set of gut-specific bacteria whose functions are supposedly critical for host physiology. The phylogenetic and coevolutionary implications of the parallel occurrences of these prokaryotic guilds appear to apply throughout a broad spectrum of animal diversity. Their persistence and conservation underlies a possibly critical role of precise bacterial assemblages in animal-bacteria interactions.

First record of the grape leafhopper Erythroneura vulnerata Fitch (Hom., Cicadellidae) in Europe

Abstract:  The leafhopper Erythroneura vulnerata Fitch is native to North America, where it infests wild and cultivated grapes. In July 2004, E. vulnerata was recorded for the first time on Vitis vinifera L. (cv. Cabernet Sauvignon) in north‐eastern Italy (Veneto region). This record is assumed to be the first in Europe. Preliminary observations on the pest distribution, seasonal abundance and the extent of symptoms in north‐eastern Italy are reported.

Green fluorescent protein (GFP)-labeling of enterobacteria associated with fruit flies (Diptera: Tephritidae) and persistence in their natural host Rhagoletis completa Cresson.

Fruit flies (Diptera: Tephritidae) are a highly successful, widespread group of insects that cause economic damage in agriculture. Data available so far on the composition of the bacterial community associated with their digestive tract indicate that members of Enterobacteriaceae are the species most often isolated. Bacteria naturally occurring in insect guts may be engineered and used to study the spatial and functional interactions of microbes within the insect system and offer one route to meet the demand for novel insect pest management strategies. With this aim we introduced by conjugation the gfp gene carried by the suicide plasmid pTn5gfpmut1 into Klebsiella oxytoca and Raoultella (formerly Klebsiella ) spp. strains isolated from the oesophageal bulb of the fruit flies Ceratitis capitata (Wiedemann) and Rhagoletis completa Cresson, respectively. The GFP-encoding gene was stably maintained in two tested transgenic strains, both originally isolated from R. completa. In one case, GFP-labeled bacterial cells were used to feed larvae and adults of the original host. Genetically modified bacteria were able to colonize the gut of larvae and persisted through all larval instars to pupal stage.

Seasonal and Daily Activity Patterns of Mosquito (Diptera: Culicidae) Vectors of Pathogens in Northeastern Italy

ABSTRACT The seasonal and daily activity of mosquito vectors of pathogens affecting animals and humans were studied in northeastern Italy at a site within the Po River Delta Park. A CDC-CO2 trap and a gravid trap were operated at 2-h intervals for 24 h every 15 d from May to October 2010. Overall, 5,788 mosquitoes comprising six species were collected, namely Culex pipiens L. (75.1% of total), Aedes caspius (Pallas) (15.2%), Aedes vexans (Meigen) (6.9%), Anopheles maculipennis s.l. Meigen (2.6%), Culiseta annulata (Schrank) (0.2%), and Culex modestus Ficalbi (<0.1%). The relative abundance of these species increased from May until the beginning of July and then decreased, disappearing at the beginning of October. The diel host-seeking patterns and oviposition site-seeking patterns were species specific and were differentially affected by the ecological variables recorded at the day and hour of mosquito collection or two weeks before collection. Knowledge of the seasonal and daily host-seeking patterns of mosquitoes highlights the time periods of the day and the seasons of potential exposure for animals and humans to mosquito-borne pathogens, therefore delineating the best time for the application of preventive measures. Furthermore, knowledge of the oviposition site-seeking activity of the mosquitoes optimizes the capture of gravid females, thereby enhancing the likelihood of detecting pathogens.