Identification and Quantification of the Most Abondant Hemocytes in the Pine Processionary Caterpillar; ThaumetopoeaPityocampa (Notodontidae)

Abstract Thaumetopoeapityocampa is an important pine pest in the Mediterranean basin and central Europe. The insecte is a butterfly, whose larvae are called pine processionary caterpillars. To understand the resistance mechanism of the insect in order to proceed of its control and to diminuate their effect in nature, it was necessary to study of the immune system and reactions of the larva in different stages. The aim of our work is to identify the hemocytes formula of the caterpillar during the larval stages L2, L3, and L4, as well as the quantification of the different cells during each stage. After extraction of the hemolymph by centrifugation, the cells were placed in culture medium and then incubated. Microscopic observation has shown that prohemocytes population appear early in hemolymph, they differentiate into plasmatocytes and granulocytes during the advanced stages. However, the quantification process (THC), carried out in a Malassez counting chamber has shown that granulocytes are the most abundant cell population in the hemolymph of the insect larvae. To investigate the role of hemocytes in immune responses, we have co-incubate T. pityocampa cells with bacteria, entomopathogenic nematodes, and synthetic beads. Both humoral and cellular encapsulation processes have been observed early in larval stages, all hemocytes seem to be involved in the formation of nodules and capsules against bacteria and microbeads. At the opposite, entomopathogenic nematodes (Steinernemafeltiae) were not recognized and encapsulated, but their presence can strongly damage host hemocytes

Comparison of the immune response of Galleria mellonella and Rhynchophorus ferrugineus after infection with the entomopathogenic nematode Steinernema carpocapsae

The red palm weevil, Rhynchophorus ferrugineus (Olivier 1790) (Coleoptera: Curculionidae), a pest of palm trees, and Galleria mellonella L. (Lepidoptera: Pyralidae), a pest of beehives, were used as models to deepen our understanding of the host-entomopathogenic nematode relationship. The effects of the entomopathogenic nematode Steinernema carpocapsae (Nematoda: Steinernematidae) on the immune activity of R. ferrugineus and G. mellonella larvae were compared. The ability to complete the life cycle of S. carpocapsae in the two insects was also evaluated. Although S. carpocapsae is effective in the biological control of the red weevil, it is not able to complete its cycle within the host, whereas the same nematode successfully reproduces in G. mellonella. On the basis of previous laboratory data and a systematic literature review, the nematode-induced effects on the immune response of the Coleoptera and the Lepidoptera were compared. In particular, the immunocompetent haemocyte population (number and cell types), the activity of the prophenoloxidase- phenoloxidase system (proPO) and finally the inhibition and proliferation of the Gram-negative symbiont bacterium Xenorhabdus nematophila (Enterobacteraceae) were analysed

Preliminary toxicity Evaluation of a porphyrin photosensitizer in an alternative preclinical Model

In photodynamic therapy (PDT), a photosensitizer (PS) excited with a specific wavelength, and in the presence of oxygen, gives rise to photochemical reactions that lead to cell damage. Over the past few years, larval stages of the G. mellonella moth have proven to be an excellent alternative animal model for in vivo toxicity testing of novel compounds and virulence testing. In this article, we report a series of preliminary studies on G. mellonella larvae to evaluate the photoinduced stress response by a porphyrin (PS) (TPPOH). The tests performed evaluated PS toxicity on larvae and cytotoxicity on hemocytes, both in dark conditions and following PDT. Cellular uptake was also evaluated by fluorescence and flow cytometry. The results obtained demonstrate how the administration of PS and subsequent irradiation of larvae affects not only larvae survival rate, but also immune system cells. It was also possible to verify PS’s uptake and uptake kinetics in hemocytes, observing a maximum peak at 8 h. Given the results obtained in these preliminary tests, G. mellonella appears to be a promising model for preclinical PS tests

Distributed Storage for the Provision of Ancillary Services to the Main Grid: Project PRESTO

AbstractThis paper presents the three-year PRESTO research project (2013-2015). PRESTO is a self-funded project developed by the Department of Energy of Politecnico di Milano in cooperation with FIAMM Storage, Elvi Energy and MCM Energy Lab (an Italian spin-off). Within the project, experimental tests and numerical simulations were performed in order to evaluate the effectiveness of an Energy Storage System (ESS) in the provision of ancillary services to the main grid. This paper focuses specifically on the experimental and numerical analyses carried out in the project to develop an innovative control law for the primary frequency regulation, able to maximize the performances of the regulating service and effectively manage the ESS state of charge

Immune Response of Drosophila suzukii Larvae to Infection with the Nematobacterial Complex Steinernema carpocapsae-Xenorhabdus nematophila

Entomopathogenic nematodes have been proposed as biological agents for the control of Drosophila suzukii, an invasive pest of small-stone and soft-skinned fruits. Larvae of the fly are susceptible to Steinernema carpocapsae infection but the reaction of immune defenses of the host are unknown. To determine the immune response, larvae were infected with S. carpocapsae and Xenorhabdus nematophila to evaluate the effector mechanisms of both humoral and cellular processes. The symbiont bacteria presented an inhibitory effect on the phenoloxidase cascade with a low level of melanization. Besides, X. nematophila activated the synthesis of putative antimicrobial peptides on the hemolymph of infected larvae. However, those peptides presented a lower antimicrobial activity compared to hemolymph from larvae infected with non-symbiont bacteria. Xenorhabdus nematophila avoided also the phagocytosis response of hemocytes. During in vitro and in vivo assays, S. carpocapsae was not encapsulated by cells, unless the cuticle was damaged with a lipase-treatment. Hemocyte counts confirmed differentiation of lamellocytes in the early phase of infection despite the unrecognition of the nematodes. Both X. nematophila and S. carpocapsae avoided the cellular defenses of D. suzukii larvae and depressed the humoral response. These results confirmed the potential of entomopathogenic nematodes to control D. suzukii

Monitoring and Management of Inland Waters: Insights from the Most Inhabited Italian Region

Monitoring of freshwaters allows the detection of the impacts of multiple anthropic uses and activities on aquatic ecosystems, and an eco-sustainable management of natural resources could limit these impacts. In this work, we highlighted two main issues affecting inland waters, referring to findings from the most inhabited Italian region (Lombardy, approximately 10 M inhabitants): the first issue is lake pollution by old generation pesticides, the second is river development for hydropower. In both cases, some management strategies reducing the anthropic impacts on freshwaters were discussed: organic farming and biocontrol as an alternative to diffuse pollution by agrochemicals; environmental flows and controlled sediment flushing operations to limit the hydropower impact on rivers. Although the two mentioned issues were discussed separately in this paper, the management of water resources should be carried out in a comprehensive way, accounting for the multiple impacts affecting freshwater ecosystems, including those related to the climate changes

Entomopathogenic potential of bacteria associated with soil-borne nematodes and insect immune responses to their infection

Soil-borne nematodes establish close associations with several bacterial species. Whether they confer benefits to their hosts has been investigated in only a few nematode-bacteria systems. Their ecological function, therefore, remains poorly understood. In this study, we isolated several bacterial species from rhabditid nematodes, molecularly identified them, evaluated their entomopathogenic potential on Galleria mellonella larvae, and measured immune responses of G. mellonella larvae to their infection. Bacteria were isolated from Acrobeloides sp., A. bodenheimeri, Heterorhabditis bacteriophora, Oscheius tipulae, and Pristionchus maupasi nematodes. They were identified as Acinetobacter sp., Alcaligenes sp., Bacillus cereus, Enterobacter sp., Kaistia sp., Lysinibacillus fusiformis, Morganella morganii subsp. morganii, Klebsiella quasipneumoniae subsp. quasipneumoniae, and Pseudomonas aeruginosa. All bacterial strains were found to be highly entomopathogenic as they killed at least 53.33% G. mellonella larvae within 72h post-infection, at a dose of 106 CFU/larvae. Among them, Lysinibacillus fusiformis, Enterobacter sp., Acinetobacter sp., and K. quasipneumoniae subsp. quasipneumoniae were the most entomopathogenic bacteria. Insects strongly responded to bacterial infection. However, their responses were apparently little effective to counteract bacterial infection. Our study, therefore, shows that bacteria associated with soil-borne nematodes have entomopathogenic capacities. From an applied perspective, our study motivates more research to determine the potential of these bacterial strains as biocontrol agents in environmentally friendly and sustainable agriculture

Acinetobacter nematophilus sp. nov., Alcaligenes nematophilus sp. nov., Enterobacter nematophilus sp. nov., and Kaistia nematophila sp. nov., Isolated from Soil-Borne Nematodes and Proposal for the Elevation of Alcaligenes faecalis subsp. faecalis, Alcaligenes faecalis subsp. parafaecalis, and Alcaligenes faecalis subsp. phenolicus to the Species Level.

Four bacterial strains, A-IN1T, A-TC2T, E-TC7T, and K-TC2T, isolated from soil-borne nematodes of the species Oscheius tipulae and Acrobeloides bodenheimeri, were found to represent new species of the genera Acinetobacter, Alcaligenes, Enterobacter, and Kaistia, respectively. In this study, we described these new species using a polyphasic taxonomic approach that included wholegenome and whole-proteome phylogenomic reconstructions, core genome sequence comparisons, and phenotypic characterization. Phylogenomic reconstructions using whole-genome and wholeproteome sequences show that A-IN1T is closely related to Acinetobacter guillouiae DSM 590T and to Acinetobacter bereziniae LMG 1003T. The dDDH values between A-IN1T and these latest strains are 25.1 and 39.6%, respectively, which are below the 70% divergence threshold for prokaryotic species delineation. A-TC2T is closely related to Alcaligenes faecalis subsp. faecalis DSM 30030T and to Alcaligenes faecalis subsp. phenolicus DSM 16503T. The dDDH values between A-TC2T and these latest strains are 47.0 and 66.3%, respectively. In addition, the dDDH values between Alcaligenes faecalis subsp. faecalis DSM 30030T, Alcaligenes faecalis subsp. phenolicus DSM 16503T, and Alcaligenes faecalis subsp. parafaecalis are always lower than 70%, demonstrating that the three strains represent species within the genus Alcaligenes rather than subspecies within Alcaligenes faecalis. E-TC7T is closely related to Enterobacter kobei DSM 13645T, Enterobacter chuandaensis 090028T, and to Enterobacter bugandensis STN0717-56T. The dDDH values between E-TC7T and these strains are 43.5, 42.9, and 63.7%, respectively. K-TC2T is closely related to Kaistia terrae DSM 21341T and to Kaistia defluvii JCM 18034T. The dDDH values between these strains are 29.2 and 30.7%, respectively. Several biochemical tests allow to differentiate the type strains of the newly described species from the type strains of their more closely related species. Based on the results of this polyphasic taxonomic approach, the following new species are proposed: Acinetobacter nematophilus sp. nov. with A-IN1T (=CCM 9231T =CCOS 2018T) as the type strain, Alcaligenes nematophilus sp. nov. with A-TC2T (=CCM 9230T =CCOS 2017T) as the type strain, Enterobacter nematophilus sp. nov. with E-TC7T (=CCM 9232T =CCOS 2020T) as the type strain, and Kaistia nematophila sp. nov. with K-TC2T (=CCM 9239T =CCOS 2022T) as the type strain. In addition, we propose the elevation of Alcaligenes faecalis subsp. faecalis, Alcaligenes faecalis subsp. parafaecalis, and Alcaligenes faecalis subsp. phenolicus to the species level. Therefore, we propose the creation of Alcaligenes parafaecalis sp. nov. with DSM 13975T as the type strain, and Alcaligenes phenolicus sp. nov. with DSM 16503T as the type strain. Our study contributes to a better understanding of the biodiversity and phylogenetic relationships of bacteria associated with soil-borne nematodes

Measurement of the top quark forward-backward production asymmetry and the anomalous chromoelectric and chromomagnetic moments in pp collisions at √s = 13 TeV

Abstract The parton-level top quark (t) forward-backward asymmetry and the anomalous chromoelectric (d̂ t) and chromomagnetic (μ̂ t) moments have been measured using LHC pp collisions at a center-of-mass energy of 13 TeV, collected in the CMS detector in a data sample corresponding to an integrated luminosity of 35.9 fb−1. The linearized variable AFB(1) is used to approximate the asymmetry. Candidate t t ¯ events decaying to a muon or electron and jets in final states with low and high Lorentz boosts are selected and reconstructed using a fit of the kinematic distributions of the decay products to those expected for t t ¯ final states. The values found for the parameters are AFB(1)=0.048−0.087+0.095(stat)−0.029+0.020(syst),μ̂t=−0.024−0.009+0.013(stat)−0.011+0.016(syst), and a limit is placed on the magnitude of | d̂ t| < 0.03 at 95% confidence level. [Figure not available: see fulltext.