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Abstract Background Pyrethroid resistance is one of the major threats for effectiveness of insecticide-treated bed nets (ITNs) in malaria vector control. Genotyping of mutations in the voltage gated sodium channel (VGSC) gene is widely used to easily assess the evolution and spread of pyrethroid target-site resistance among malaria vectors. L1014F and L1014S substitutions are the most common and best characterized VGSC mutations in major African malaria vector species of the Anopheles gambiae complex. Recently, an additional substitution involved in pyrethroid resistance, i.e. V402L, has been detected in Anopheles coluzzii from West Africa lacking any other resistance alleles at locus 1014. The evolution of target-site resistance mutations L1014F/S and V402L was monitored in An. coluzzii and Anopheles arabiensis specimens from a Burkina Faso village over a 10-year range after the massive ITN scale-up started in 2010. Methods Anopheles coluzzii (N = 300) and An. arabiensis (N = 362) specimens collected both indoors and outdoors by different methods (pyrethrum spray catch, sticky resting box and human landing collections) in 2011, 2015 and 2020 at Goden village were genotyped by TaqMan assays and sequencing for the three target site resistance mutations; allele frequencies were statistically investigated over the years. Results A divergent trend in resistant allele frequencies was observed in the two species: 1014F decreased in An. coluzzii (from 0.76 to 0.52) but increased in An. arabiensis (from 0.18 to 0.70); 1014S occurred only in An. arabiensis and slightly decreased over time (from 0.33 to 0.23); 402L increased in An. coluzzii (from 0.15 to 0.48) and was found for the first time in one An. arabiensis specimen. In 2020 the co-occurrence of different resistance alleles reached 43% in An. coluzzii (alleles 410L and 1014F) and 32% in An. arabiensis (alleles 1014F and 1014S). Conclusions Overall, an increasing level of target-site resistance was observed among the populations with only 1% of the two malaria vector species being wild type at both loci, 1014 and 402, in 2020. This, together with the co-occurrence of different mutations in the same specimens, calls for future investigations on the possible synergism between resistance alleles and their phenotype to implement local tailored intervention strategies.

Stingless bees (Hymenoptera, Meliponini) are a large and diverse group including 59 extant groups, representing the main pollinators of Amazon forests. Among those, Trigona is one of the largest endemic genera of Neotropical Meliponini. In this work, we updated the molecular phylogeny of Trigona proposed by Rasmussen and Camargo (2008), including data from 59 new specimens collected in 2020 in the forests of northern Peru, through a multigene phylogenetic approach combining sequences from four gene fragments (16S, ArgK, EF-1a, opsin). Our results confirmed the monophyly of Trigona and of all proposed subgenera, except Aphaneura. In addition, most Trigona species-groups resulted monophyletic but the ‘spinipes’ and ‘pallens’ groups appeared paraphyletic and polyphyletic, respectively. Moreover, the cohesion of the “fulviventris” species group was hindered by the inclusion of T. williana (previously included in the “pallens” group) within this clade. Finally, we provided further evidence for a subdivision into two (geographically) distinct clades within T. guianae in northern Peruvian Amazon, which highlighted the importance of Neotropical biogeographical barriers in Meliponini divergence and evolution. Finally, to avoid misidentifications of Trigona specimens, the need for a robust taxonomic revision based on a cladistic approach of the whole genus is discussed.

Abstract Background Meloidae (blister beetles) are known to synthetize cantharidin (CA), a toxic and defensive terpene mainly stored in male accessory glands (MAG) and emitted outward through reflex-bleeding. Recent progresses in understanding CA biosynthesis and production organ(s) in Meloidae have been made, but the way in which self-protection is achieved from the hazardous accumulation and release of CA in blister beetles has been experimentally neglected. To provide hints on this pending question, a comparative de novo assembly transcriptomic approach was performed by targeting two tissues where CA is largely accumulated and regularly circulates in Meloidae: the male reproductive tract (MRT) and the haemolymph. Differential gene expression profiles in these tissues were examined in two blister beetle species, Lydus trimaculatus (Fabricius, 1775) (tribe Lyttini) and Mylabris variabilis (Pallas, 1781) (tribe Mylabrini). Upregulated transcripts were compared between the two species to identify conserved genes possibly involved in CA detoxification and transport. Results Based on our results, we hypothesize that, to avoid auto-intoxication, ABC, MFS or other solute transporters might sequester purported glycosylated CA precursors into MAG, and lipocalins could bind CA and mitigate its reactivity when released into the haemolymph during the autohaemorrhaging response. We also found an over-representation in haemolymph of protein-domains related to coagulation and integument repairing mechanisms that likely reflects the need to limit fluid loss during reflex-bleeding. Conclusions The de novo assembled transcriptomes of L. trimaculatus and M. variabilis here provided represent valuable genetic resources to further explore the mechanisms employed to cope with toxicity of CA in blister beetle tissues. These, if revealed, might help conceiving safe and effective drug-delivery approaches to enhance the use of CA in medicine.

Stingless bees (Hymenoptera: Meliponini) are corbiculate and eusocial bees, including over 500 species distributed in the tropical and sub-tropical regions of the world. They establish perennial colonies using a large variety of nesting sites, as well as colonies of ants, wasps and termites. Compared to termitophily, myrmecophily is an uncommon feature and little studied aspect among Meliponini. In Peru there are few records relating to these types of associations, and in general little is known about the architecture of the nests of bees and ants that live together. For the first time, the nest architecture of Trigona cfr. cilipes in association with Dolichoderus quadridenticulatus is described from the Peruvian Amazon. The record of another association, Paratrigona sp. - Camponotus sp. is also reported. As the nests are arranged, the types of building materials used and some behavioral defense observations of ants and bees are also provided.

Abstract Background Meligethes are pollen-beetles associated with flowers of Rosaceae as larvae. This genus currently consists of 63 known species in two subgenera, Meligethes and Odonthogethes, predominantly occurring in the eastern Palaearctic. We analyzed 74 morphological and ecological characters (169 states) of all species, as well as of 11 outgroup species from 7 Meligethinae genera (including Brassicogethes), to investigate their phylogeny. We also conducted a parallel molecular analysis on 9 Meligethes, 9 Odonthogethes, 3 Brassicogethes and 2 Meligethinus species based on DNA sequence data from mitochondrial (COI, 16S) and nuclear (CAD) genes. Results Morphological phylogenetic reconstructions supported the monophyly of the whole genus and clades corresponding to purported subgenera Meligethes s.str. and Odonthogethes. Main species-groups were mostly confirmed, however some unresolved polytomies remained. Molecular data placed members of Brassicogethes (including 42 mostly W Palearctic species associated with Brassicaceae) as sister to Odonthogethes, with this clade being sister to Meligethes s.str. This phylogenetic scenario suggests that monophyletic Meligethes s.str., Odonthogethes and Brassicogethes should be regarded alternatively as three subgenera of a monophyletic Meligethes, or three genera in a monophyletic genus-complex, with mutually monophyletic Brassicogethes and Odonthogethes. Molecular analyses estimated the origin of this lineage at ca. 14–15 Mya from a common stem including Meligethinus. Conclusions We hypothesize that the ancestor of Meligethes specialized on Rosaceae in the Middle Miocene (likely in Langhian Age) and subsequently radiated during Late Miocene and Plio-Pleistocene maintaining a trophic niche on this plant family. This radiation was primarily due to geographic isolation in E Asiatic mountain systems. Combined evidence from morphology, ancestral state parsimony reconstruction of host-plant associations and molecular evidence suggested that Rosoideae (Rosa spp.) represented the ancestral hosts of Meligethes s.str., followed by an independent shift of ancestral Odonthogethes (ca. 9–15 Mya) on Rubus (Rosoideae) and members of Rosaceae Spiraeoideae. Other ancestral Odonthogethes probably shifted again on the unrelated plant family Brassicaceae (maybe 8–14 Mya in S China), allowing a rapid westward radiation of the Brassicogethes clade.

Abstract Background Neutrophils are one of the key players in the human innate immune system (HIIS). In the event of an insult where the body is exposed to inflammation triggering moieties (ITMs), neutrophils are mobilized towards the site of insult and antagonize the inflammation. If the inflammation is cleared, neutrophils go into a programmed death called apoptosis. However, if the insult is intense or persistent, neutrophils take on a violent death pathway called necrosis, which involves the rupture of their cytoplasmic content into the surrounding tissue that causes local tissue damage, thus further aggravating inflammation. This seemingly paradoxical phenomenon fuels the inflammatory process by triggering the recruitment of additional neutrophils to the site of inflammation, aimed to contribute to the complete neutralization of severe inflammation. This delicate balance between the cost and benefit of the neutrophils’ choice of death pathway has been optimized during the evolution of the innate immune system. The goal of our work is to understand how the tradeoff between the cost and benefit of the different death pathways of neutrophils, in response to various levels of insults, has been optimized over evolutionary time by using the concepts of evolutionary game theory. Results We show that by using evolutionary game theory, we are able to formulate a game that predicts the percentage of necrosis and apoptosis when exposed to various levels of insults. Conclusion By adopting an evolutionary perspective, we identify the driving mechanisms leading to the delicate balance between apoptosis and necrosis in neutrophils’ cell death in response to different insults. Using our simple model, we verify that indeed, the global cost of remaining ITMs is the driving mechanism that reproduces the percentage of necrosis and apoptosis observed in data and neutrophils need sufficient information of the overall inflammation to be able to pick a death pathway that presumably increases the survival of the organism.

Stingless bees (Hymenoptera: Meliponini) are among the most important pollinators of tropical forests. Peru is considered a hotspot of biodiversity of Meliponini, but many areas of this country (e.g., Peruvian Amazon) remain unexplored. We aimed to produce a first inventory of stingless bee species dwelling in humid and seasonally dry forests of northern Peru by combining traditional (morphologically-based) taxonomy and DNA barcoding. Specimens were collected in 2020 at five sites located in San Martin and Piura regions. We identified 12 genera of Meliponini. Among those, Trigona and Plebeia were the most abundant (45.9% and 12.8% respectively), whereas Nannotrigona and Scaura were the least represented ones (2.3%). We assigned a reliable species identification to about 30% of specimens (Trigona amazonensis, T. muzoensis, T. williana, Partamona testacea, Scaura tenuis, Tetragona goettei, and Tetragonisca angustula). Yet, more than a half of the specimens received a provisional identification (e.g., Geotrigona cf. fulvohirta, T. cf. amalthea, T. cf. fuscipennis, T. cf. hypogea, Melipona cf. cramptoni, Partamona cf. epiphytophila, Ptilotrigona cf. perenae, Scaura cf. latitarsis, Tetragona cf. clavipes, Trigonisca cf. atomaria). We also highlighted an extensive polyphyly that affected a number of currently recognized species (e.g., T. fulviventris, T. guianae, Plebeia franki, P. frontalis, M. eburnea, M. illota), whose members were split into various clades. Finally, 16% of individuals failed to be identified at the species level (Trigona sp. 1, T. sp. 2, Nannotrigona sp., Partamona sp., Scaptotrigona sp. 1, S. sp. 2, Trigonisca sp. 1, and Trigonisca sp. 2). We discuss our findings according to the current faunistic and biogeographic knowledge of Meliponini in Peru and the Neotropical region. We also remark on the importance of conducting a taxonomic revision of stingless bees and improving both their morphology-based identification keys and BOLD repository. Finally, we claim that integrative taxonomy shall be strongly implemented to truly assess the biodiversity of Neotropical stingless bees, allowing conserving these important pollinators and the associated traditional meliponiculture in an effective manner.

Protease inhibitors are widely studied since the unrestricted activity of proteases can cause extensive organ lesions. In particular, elastase activity is involved in the pathophysiology of acute lung injury, for example during SARS-CoV-2 infection, while serine proteases and thrombin-like proteases are involved in the development and/or pathology of the nervous system. Natural protease inhibitors have the advantage to be reversible and with few side effects and thus are increasingly considered as new drugs. Kunitz-type protease inhibitors (KTPIs), reported in the venom of various organisms, such as wasps, spiders, scorpions, and snakes, have been studied for their potent anticoagulant activity and widespread protease inhibitor activity. Putative KTPI anticoagulants have been identified in transcriptomic resources obtained for two blister beetle species, Lydus trimaculatus and Mylabris variabilis. The KTPIs of L. trimaculatus and M. variabilis were characterized by combined transcriptomic and bioinformatics methodologies. The full-length mRNA sequences were divided on the base of the sequence of the active sites of the putative proteins. In silico protein structure analyses of each group of translational products show the biochemical features of the active sites and the potential protease targets. Validation of these genes is the first step for considering these molecules as new drugs for use in medicine.

Members of the family Meloidae are known to produce cantharidin, a highly toxic monoterpene found in their hemolymph and exuded as droplets capable of deterring many predators. As a nuptial gift, males transfer large amounts of cantharidin to females via a spermatophore, which is formed by specific accessory glands containing high concentrations of this terpene. Using light, electron and ion beam microscopy, the ultrastructural features of the three pairs of male accessory glands as well as the glandular part of the vasa deferentia were comparatively investigated in seven species of blister beetles belonging to five different tribes and two subfamilies. All gland pairs examined share common features such as mesodermal derivation, the presence of muscle sheath, a developed rough endoplasmic reticulum, abundant mitochondria, secretory vesicles, and microvillated apical membranes. Within the same species, glands exhibit distinctive features, suggesting that each pair is responsible for the formation of a specific substance. The vasa deferentia, while showing many similarities within the family, often exhibit features unique to each of the individual species investigated, whereas the accessory glands of the first and second pairs display the highest degree of ultrastructural variability. A comparison across the species shows an interesting constancy limited to ultrastructural features in the third pair of accessory glands. The similarities and differences among the species are discussed in the light of the available literature and in relation to the potential role that blister beetles’ male accessory glands could play in the storage and management of cantharidin.

Cerambyx cerdo is a longhorn beetle widely distributed in southern and central Europe. This saproxylic beetle is generally associated with oak forests where there are mature or partially dead and sun-exposed trees. Its populations are currently threatened by forest practices such as the removal of partially dead trees and the decline in the number of old oak trees situated in open or semi-open landscapes. Thus, C. cerdo has been included in Annexes II and IV of the Habitats Directive. The present paper is part of a special issue on monitoring of saproxylic beetles which are protected in Europe, based on the research carried out during the LIFE-MIPP project, with a revision of the current knowledge on systematics, ecology and conservation of C. cerdo. The main aim of the present paper is to test different monitoring methods in order to develop a quick and reproducible protocol for the conservation of this species. The methods tested were: artificial sap attracting the adults, baited traps, VES (visual encounter survey) and collecting remains of predation along transects. Based on these results, a detailed monitoring method for C. cerdo using baited trap is proposed in this paper, together with a discussion on its constraints, spatial validity and possible interferences. In order to assess the conservation status of populations of C. cerdo in Europe and to compare populations over time, a method for the calculation of a reference value, based on the monitoring method, is provided.

One aim of the MIPP Project (http://www.lifemipp.eu) was to develop non-invasive monitoring methods for selected saproxylic beetles. In this paper, a method is proposed for monitoring the larvae of Osmoderma eremita in their natural habitat (i.e. hollow trees), using a conservation detection dog (CDD). Wood mould sampling (WMS), the standard method to detect hermit beetles and other saproxylic insects inside tree hollows, is time-consuming and exposes the target species and the whole saproxylic communities to some risks. In contrast, CDDs pose no risk to the species living inside trees while, at the same time, offer a powerful tool for surveying the insects. In this paper, the methods applied to train the dog are presented, together with the results for accuracy (the overall proportion of correct indications), sensitivity (the proportion of correct positive indications) and specificity (the proportion of correct negative indications) obtained once the CDD had been fully trained. Results are presented for nitrocellulose filters with the odour of the target species, for larvae living inside hollow trees, for frass and for the remains of adults. A comparison of the efficiency between CDD and WMS showed that employing the dog was much less time-consuming than WMS. The literature on training CDDs for nature conservation tasks, with particular reference to cases involving Coleoptera, was also reviewed.

Rosalia alpina (Linnaeus, 1758) is a large longhorn beetle (Coleoptera: Cerambycidae) which is protected by the Habitats Directive and which typically inhabits beech forests characterised by the presence of mature, dead (or moribund) and sun-exposed trees. A revision of the current knowledge on systematics, ecology and conservation of R. alpina is reported. The research was carried out as part of the LIFE MIPP project which aims to find a standard monitoring method for saproxylic beetles protected in Europe. For monitoring this species, different methods were tested and compared in two areas of the Apennines, utilising wild trees, logs and tripods (artificially built with beech woods), all potentially suitable for the reproduction of the species. Even if all methods succeeded in the survey of the target species, these results showed that the use of wild trees outperformed other methods. Indeed, the use of wild trees allowed more adults to be observed and required less intensive labour. However, monitoring the rosalia longicorn on wild trees has the main disadvantage that they can hardly be considered “standard sampling units”, as each tree may be differently attractive to adults. Our results demonstrated that the most important factors influencing the attraction of single trunks were wood volume, sun-exposure and decay stage. Based on the results obtained during the project LIFE MIPP, as well as on a literature review, a standard monitoring method for R. alpina was developed.

Polymerase chain reaction (PCR)-based genotyping of mutations in the voltage-sensitive sodium channel (vssc) associated with resistance to pyrethroid insecticides is widely used and represents a potential early warning and monitoring system for insecticide resistance arising in mosquito populations, which are vectors of different human pathogens. In the secondary vector Aedes albopictus—an Asian species that has invaded and colonized the whole world, including temperate regions—sequencing of domain II of the vssc gene is still needed to detect the V1016G mutation associated with pyrethroid resistance. In this study we developed and tested a novel allele-specific PCR (AS-PCR) assay to genotype the V1016G mutation in this species and applied it to the analysis of wild populations from Italy. The results confirm the high accuracy of the novel AS-PCR and highlight frequencies of the V1016G allele as >5% in most sampling sites, with peaks of 20–45% in coastal touristic sites where pyrethroid treatments are extensively implemented, mostly for mosquito nuisance reduction. The high frequency of this mutation observed in Italian Ae. albopictus populations should serve as a warning bell, advocating for increased monitoring and management of a phenomenon which risks neutralizing the only weapon today available to counteract (risks of) arbovirus outbreaks.

Alkaline phosphatase (AP) is an enzyme that exhibits anti-inflammatory effects by dephosphorylating inflammation triggering moieties (ITMs) like bacterial lipopolysaccharides and extracellular nucleotides. AP administration aims to prevent and treat peri- and post-surgical ischemia reperfusion injury in cardiothoracic surgery patients. Recent studies reported that intravenous bolus administration and continuous infusion of AP in patients undergoing coronary artery bypass grafting with cardiac valve surgery induce an increased release of liver-type “tissue non-specific alkaline phosphatase” (TNAP) into the bloodstream. The release of liver-type TNAP into circulation could be the body's way of strengthening its defense against a massive ischemic insult. However, the underlying mechanism behind the induction of TNAP is still unclear. To obtain a deeper insight into the role of AP during surgery, we developed a mathematical model of systemic inflammation that clarifies the relation between supplemented AP and TNAP and describes a plausible induction mechanism of TNAP in patients undergoing cardiothoracic surgery. The model was validated against clinical data from patients treated with bovine Intestinal AP (bIAP treatment) or without AP (placebo treatment), in addition to standard care procedures. We performed additional in-silico experiments adding a secondary source of ITMs after surgery, as observed in some patients with complications, and predicted the response to different AP treatment regimens. Our results show a strong protective effect of supplemented AP for patients with complications. The model provides evidence of the existence of an induction mechanism of liver-type tissue non-specific alkaline phosphatase, triggered by the supplementation of AP in patients undergoing cardiac surgery. To the best of our knowledge this is the first time that a quantitative and validated numerical model of systemic inflammation under clinical treatment conditions is presented.

It is still unclear under which conditions temporary combined antiretroviral therapy (cART) results in a prolonged remission after interruption. Clinical trials have contradicting reposts about the effect of cART during primary HIV infection on the disease progression. Here we propose that the apparent contradiction is due the presence of a window of opportunity for cART treatment observed in the in silico studies. We study non-linear correlations in the HIV dynamics over time using information theory. This approach requires a large dataset of CD4+ T lymphocytes and viral load concentrations over time. Since it is unfeasible to collect the required amount of data in clinical trials we use C-ImmSim, a clinically validated in silico model of the HIV infection, to simulate the HIV infection and temporary cART in 500 virtual patients for a period of 6 years post infection in time steps of 8 hours. We validate the results of our model with two published clinical trials of temporary cART in acute infection and analyse the impact of cART on the immune response. Our quantitative analysis predicts a "window of opportunity" of about ten months after the acute phase during which a temporary cART has significantly longer-lasting beneficial effects on the immune system as compared to treatment during the chronic phase. This window may help to explain the controversial outcomes of clinical trials that differ by the starting time and duration of the short-term course cART and provides a critical insight to develop appropriate protocols for future clinical trials.

In this paper we present additional information on taxonomy, ecological preferences and conservation biology of the rare and poorly known Sicilian endemic Ochthebius biltoni Jäch & Delgado, 2017. It represents one of the most outstanding endemic elements of the Sicilian and Italian water beetle fauna.

Carpophilus zeaphilus Dobson, 1969 (Coleoptera, Nitidulidae, Carpophilinae) is an Afrotropical species that has become widespread in Sub-Saharan Africa, the Arabian Peninsula, and southern Mediterranean areas in recent years. The species was first recorded from Europe in Portugal and Spain nearly thirty years ago, and it was later intercepted in Sicily near Trapani in 1991. A few specimens of this species were collected in April, 2015 in a sparsely forested area near Rome, which suggests a recent acclimatization into peninsular Italy. Specimens were taken on flowering trees of Prunus spinosa L. (Rosaceae), an unusual occurrence for most introduced species of Carpophilinae that are normally associated with rotten fruit and other decomposing vegetal matter.

Results of a molecular analysis on the European hermit beetles (the Osmoderma eremita species-complex), recently published in a companion paper, are shortly dis- cussed and commented. These results indicate a clear-cut distinction between two clades. The first one includes the W-European O. eremita Scopoli, 1763, and the two Italian endemic taxa O. italicum Sparacio, 2000 and O. cristinae Sparacio, 1994, from southern peninsular Italy and Sicily, respectively. The second one includes the widespread E-European O. barnabita Motschulsky, 1845 (nom. resurr.), and the southern Balcanic O. lassallei Baraud & Tauzin, 1991 from Greece and European Turkey. Within the two clades, molecular data well support a specific rank for O. lassallei and O. barn­abita on one side, and of O. eremita and O. cristinae on the other side, while the taxonomic position of O. italicum, more closely related with O. eremita, is still uncertain, waiting for analysis of additional specimens of this very rare taxon. Current geographical distribution, interspecific genetic diversification, and relatively low levels of intraspecific genetic divergence in O. eremita sensu stricto, are hypothesized to be the result of multiple speciation events (mainly occurred in refugial forest areas of the Italian and Balkan peninsulas and Sicily before and during the Pleistocene glacial peaks), followed by fast post-glacial northward and westward expansion of some species. The need of further genetic data on the rare and threatened hermit beetle species and the importance of more detailed information on their distribution ranges are emphasized and discussed, in order to plan conservation strategies in the near future. An updated worldwide checklist of the species of the genus Osmoderma is finally presented.

"Far-West" Africa is known to be a secondary contact zone between the two major malaria vectors Anopheles coluzzii and A. gambiae. We investigated gene-flow and potentially adaptive introgression between these species along a west-to-east transect in Guinea Bissau, the putative core of this hybrid zone. To evaluate the extent and direction of gene flow, we genotyped site 702 in Intron-1 of the para Voltage-Gated Sodium Channel gene, a species-diagnostic nucleotide position throughout most of A. coluzzii and A. gambiae sympatric range. We also analyzed polymorphism in the thioester-binding domain (TED) of the innate immunity-linked thioester-containing protein 1 (TEP1) to investigate whether elevated hybridization might facilitate the exchange of variants linked to adaptive immunity and Plasmodium refractoriness. Our results confirm asymmetric introgression of genetic material from A. coluzzii to A. gambiae and disruption of linkage between the centromeric "genomic islands" of inter-specific divergence. We report that A. gambiae from the Guinean hybrid zone possesses an introgressed TEP1 resistant allelic class, found exclusively in A. coluzzii elsewhere and apparently swept to fixation in West Africa (i.e. Mali and Burkina Faso). However, no detectable fixation of this allele was found in Guinea Bissau, which may suggest that ecological pressures driving segregation between the two species in larval habitats in this region may be different from those experienced in northern and more arid parts of the species' range. Finally, our results also suggest a genetic subdivision between coastal and inland A. gambiae Guinean populations and provide clues on the importance of ecological factors in intra-specific differentiation processes.

The LIFE project “MIPP” - Monitoring of Insects with Public Participation (11 NAT/IT/000252) is focused on selected insect species (five Coleoptera, three Lepidoptera, one Orthoptera), all included in the annexes II and IV of the Habitats Directive (HD) 92/43/EEC. One important aim is a citizen science initiative where every person may become a citizen scientist and collect faunistic data on the above species throughout Italy. Another objective of the project MIPP is the development of standard methods for monitoring the conserva- tion status of the five target beetle species. One innovative method employed is a sniffer-dog (“Osmodog”), trained to find the rare and endangered hermit beetle, Osmoderma eremita, which lives in veteran, hollow trees. The dog detects the strong smell of mature peach produced by adult males and an odor produced by the larvae. Another objective of the project MIPP is the dissemination of topics such as HD, Natura 2000, importance of dead-wood, Life projects, insect monitoring and conservation.

Continuous antiretroviral therapy is currently the most effective way to treat HIV infection. Unstructured interruptions are quite common due to side effects and toxicity, among others, and cannot be prevented. Several attempts to structure these interruptions failed due to an increased morbidity compared to continuous treatment. The cause of this failure is poorly understood and often attributed to drug resistance. Here we show that structured treatment interruptions would fail regardless of the emergence of drug resistance. Our computational model of the HIV infection dynamics in lymphoid tissue inside lymph nodes, demonstrates that HIV reservoirs and evasion from immune surveillance themselves are sufficient to cause the failure of structured interruptions. We validate our model with data from a clinical trial and show that it is possible to optimize the schedule of interruptions to perform as well as the continuous treatment in the absence of drug resistance. Our methodology enables studying the problem of treatment optimization without having impact on human beings. We anticipate that it is feasible to steer new clinical trials using computational models.

The main Afrotropical malaria vector, Anopheles gambiae sensu stricto, is undergoing a process of sympatric ecological diversification leading to at least two incipient species (the M and S molecular forms) showing heterogeneous levels of divergence across the genome. The physically unlinked centromeric regions on all three chromosomes of these closely related taxa contain fixed nucleotide differences which have been found in nearly complete linkage disequilibrium in geographic areas of no or low M-S hybridization. Assays diagnostic for SNP and structural differences between M and S forms in the three centromeric regions were applied in samples from the western extreme of their range of sympatry, the only area where high frequencies of putative M/S hybrids have been reported. The results reveal a level of admixture not observed in the rest of the range. In particular, we found: i) heterozygous genotypes at each marker, although at frequencies lower than expected under panmixia; ii) virtually all possible genotypic combinations between markers on different chromosomes, although genetic association was nevertheless detected; iii) discordant M and S genotypes at two X-linked markers near the centromere, suggestive of introgression and inter-locus recombination. These results could be indicative either of a secondary contact zone between M and S, or of the maintenance of ancestral polymorphisms. This issue and the perspectives opened by these results in the study of the M and S incipient speciation process are discussed.