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Morphostructural investigations have been carried out on Apis mellifera workers treated with single controlled acute exposures to a biopesticide containing Bacillus thuringiensis (Bt), to detect midgut changes until 96 h. Our findings show concentration-dependence of these changes, reflecting in different degrees on both mortality and behaviour. In particular, some midgut changes are also found 96 h after treatment. Our results show that the tested product does not affect survival at presumable environmental concentrations, so confirming the lesser toxicity of Bt-based biopesticides compared to other pesticides. However, in the light of the discovered long-term changes, we discuss the opportunity of taking into account possible chronic exposures to Bt-based products on A. mellifera. honeybees / Bacillus thuringiensis var. aizawai /kurstaki / bioassay / mortality / morphostructural changes Electronic supplementary material The online version of this article (

BackgroundPioneering research on “Mediterranean Kala-Azar” carried out by Adler and Theodor early in the past century (~1930s) had identified Catania city (Sicily) as a major focus of the disease nowadays known as zoonotic visceral leishmaniasis (VL). Despite the fact that disease in both humans and dogs has continued to be highly prevalent in the Catania province up to the present times, research on Leishmania vectors in this urban focus dates back to that distant period. This study aimed to evaluate the persistence and current composition of the sand fly fauna in urban environments of Catania in recent years, 2006 and 2013.MethodsIn 2006 fifty-one suitable collecting sites were identified within 44 sub-units of a grid drawn to include the urban Catania area. In 2013 the survey was restricted to four of the most productive and representative sites resulting from the 2006 survey. In both periods 3 collections per month were performed using standard sticky traps set for 3 days in wall holes/cavities along public roads, from the end of April through December.Results43/51 sites (84.3%) were found positive for sand flies. The 2006 collections accounted for a total of 4341 specimens including six species. Among competent Leishmania vector species, P. perniciosus was the most prevalent (36.5%) being identified in all sand fly-positive sites, with significant abundance in those of the old city centre. Other species of interest were P. sergenti (2.5%) and P. neglectus (1.5%). The 2013 survey produced 1130 sand flies, of which 39.5% were P. perniciosus, 1.6% P. sergenti and 0.7% P. neglectus. A search for Leishmania DNA in a small sample of 72 P. perniciosus females revealed 11% infection prevalence.ConclusionsOur findings from an old urban focus of leishmaniasis demonstrate that phlebotomine sand flies have adapted fairly well to the drastic environmental changes that have occurred in cities of the Western world in the past century and still represent a potential risk for Leishmania transmission.

Comparing the type series of Ramazzottius thulini with specimens of Ramazzottius oberhaeuseri, from the locus typicus, we confirmed there are morphological and metric differences between these two species. This evidence convinced us that R. thulini should be considered a bona species. A new species, Ramazzottius libycus sp. nov., is also described. Ramazzottius libycus sp. nov. differs from R. oberhaeuseri in having more slender internal claws I-III and anterior claws IV, and other metric differences relative to all claws. The new species differs from R. thulini in details of the cuticular ornamentation, stylet supports inserted on the buccal tube in a slightly more caudal position, internal claws slightly different in shape, and some metric differences relative to all claws. It appears R. thulini and R. libycus sp. nov. are sibling species of R. oberhaeuseri and therefore the authors expect that there are other cryptic species included under the R. oberhaeuseri name still awaiting to be recognized and described.

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