Morphological characterization of the antenna of Torymus sinensis (Hymenoptera: Torymidae) and a comparison within the superfamily Chalcidoidea.

The parasitoid Torymus sinensis (Hymenoptera: Torymidae) has been successfully used in Italy since 2005 for biological control of the invasive cynipid Dryocosmus kuriphilus (Hymenoptera: Cynipidae), highly destructive for the economically relevant Castanea sativa (Fagales: Fagaceae). In order to investigate the morphological aspects related to sensorial behavior, a fine morphology study of the antennae and their sensilla was conducted by scanning electron microscopy on both sexes of T. sinensis. The antennae, composed of a scape, a pedicel and a flagellum with ten flagellomeres, had chaetic sensilla of six subtypes, placoid sensilla of three subtypes, trichoid sensilla, sensilla with a roundish grooved tip, and coeloconic sensilla. The chaetic sensilla of the first three subtypes were found in the scape and in the pedicel, and those of the last three subtypes, together with trichoid, roundish grooved tip and coeloconic sensilla, were found only on flagellomeres. Sexual dimorphism was detected in the morphology of the proper pedicel and the flagellum, and in the presence and distribution of the sensilla and their subtypes. The morphological aspects of the antenna of T. sinensis and of its sensilla were compared with those found in the family Torymidae and in other families of the extremely diverse superfamily Chalcidoidea.

An Overview of Ovarian Calyx Fluid Proteins of Toxoneuron nigriceps (Viereck) (Hymenoptera: Braconidae): An Integrated Transcriptomic and Proteomic Approach.

The larval stages of the tobacco budworm, Heliothis virescens (Fabricius) (Lepidoptera: Noctuidae), are parasitized by the endophagous parasitoid wasp, Toxoneuron nigriceps (Viereck) (Hymenoptera: Braconidae). During the injections of eggs, this parasitoid wasp also injects into the host body the secretion of the venom gland and the calyx fluid, which contains a polydnavirus (T. nigriceps BracoVirus: TnBV) and the Ovarian calyx fluid Proteins (OPs). The effects of the OPs on the host immune system have recently been described. In particular, it has been demonstrated that the OPs cause hemocytes to undergo a number of changes, such as cellular oxidative stress, actin cytoskeleton modifications, vacuolization, and the inhibition of hemocyte encapsulation capacity, which results in both a loss of hemocyte functionality and cell death. In this study, by using a combined transcriptomic and proteomic analysis, the main components of T. nigriceps ovarian calyx fluid proteins were identified and their possible role in the parasitic syndrome was discussed. This study provides useful information to support the analysis of the function of ovarian calyx fluid proteins, to better understand T. nigriceps parasitization success and for a more thorough understanding of the components of ovarian calyx fluid proteins and their potential function in combination with other parasitoid factors.

Lucilia sericata (Diptera: Calliphoridae) as Agent of Myiasis in a Goose in Italy and a Review of Myiasis by This Species in Birds.

Myiasis is a type of parasitosis by larvae of Diptera that may affect vertebrates, including wild and domestic birds. Traumatic myiasis was discovered in a domestic goose, Anser anser domesticus L. (Anseriformes: Anatidae), in June 2020 in a rural area of the region Calabria (Southern Italy). The myiasis was caused by Lucilia sericata (Meigen) (Diptera: Calliphoridae). In Italy, this was the first case of myiasis by L. sericata ever described in a bird. It was also the first case of myiasis detected in a goose in Italy. The description of the case is integrated by a discussion on nonhematophagous dipteran larvae causing myiasis in birds and by an updated and detailed review of literature cases of myiasis by L. sericata in birds reported worldwide, useful for monitoring and management of dipteran species of medical and veterinary interest.

A Historical Collection of Termites in Ferrara: Recovery, Cataloguing and Geographical Analyses.

Termites are an insect group relevant for recycling of organic matter, but they are also biodeteriogenic and may cause serious damages to wooden structures (including historical buildings and ancient libraries) in anthropogenic environments. The collection of Italian and foreign termites gathered over the years by Antonio Springhetti, Professor of Zoology at the University of Ferrara (Ferrara, Italy) and internationally renowned entomologist, contains over 44,000 specimens, collected by Springhetti during his field campaigns or donated by other entomologists from all over the world. The collection is currently preserved at the Department of Life Sciences and Biotechnology of the University of Ferrara. Unfortunately, all documents, publications and notes concerning the Springhetti Collection were lost; thus, in 2020, the collection was completely re-catalogued within the University Museum System and analyzed in detail. The collection contains specimens dating back to 1878 and represents not only a valuable scientific tool for studies on these ecologically relevant insects that may cause damages to historical buildings, ancient books and artworks but also an important cultural asset for the University Museum System.

Analysis of Insect Pests in an 18th-Century Historical Pharmacy: A Case Study in Ferrara.

Historical pharmacies are valuable artistic, cultural, and scientific assets. In Italy, 482 historical pharmacies have been catalogued thus far, among which is the Navarra-Bragliani Pharmacy in Ferrara, whose wood furniture dates back to the 18th century. A study on insects causing biodeterioration was carried out for the first time on this valuable asset in order to monitor and preserve it. Insect samples were collected during surveys and examined by stereomicroscopy and scanning electron microscopy, using taxonomical keys for identification. The results revealed the presence of three species of Coleoptera-the first one specialised to attack wood materials, the second one polyphagous, and the third one feeding on materials of animal origin. The data obtained in this investigation may be useful for the preservation of the precious furnishing of the historical Navarra-Bragliani Pharmacy for performing similar studies on other ancient pharmacies aimed to protect these valuable cultural and scientific assets.

Hydrogen peroxide toxicity on auditory cells: An in vitro study.

In recent decades, interest has increased in the role of reactive oxygen species (ROS) in health and disease. The ROS are key causative factors in several hearing loss pathologies including ototoxicity, noise trauma, cochlear ageing and ischemic injury. In order to investigate ROS effects on inner ear cells and counteract them, we developed an in vitro model of oxidative stress by exposing the inner ear cell line OC-k3 to hydrogen peroxide (H2O2) at concentrations able to affect in vivo cellular components but allowing cell survival. The treatment with high concentrations (20 and 30 µM) resulted in reduction of cell viability, activation of apoptosis/necrosis and alteration of morphology, cell cycle progression and antioxidant defences. The ROS effects in inner ear cells are difficult to assess in vivo. Organocultures may provide preservation of tissue architecture but involve ethical issues and can be used only for a limited time. An in vitro model that could be commercially available and easy to handle is necessary to investigate inner ear oxidative stress and the ways to counteract it. The OC-k3 line is a suitable in vitro model to study ROS effects on inner ear cells because the observed cell alterations and damages were similar to those reported in studies investigating ROS effects of ototoxic drugs, noise trauma and cochlear ageing.

An integrated transcriptomic and proteomic approach to identify the main Torymus sinensis venom components.

During oviposition, ectoparasitoid wasps not only inject their eggs but also a complex mixture of proteins and peptides (venom) in order to regulate the host physiology to benefit their progeny. Although several endoparasitoid venom proteins have been identified, little is known about the components of ectoparasitoid venom. To characterize the protein composition of Torymus sinensis Kamijo (Hymenoptera: Torymidae) venom, we used an integrated transcriptomic and proteomic approach and identified 143 venom proteins. Moreover, focusing on venom gland transcriptome, we selected additional 52 transcripts encoding putative venom proteins. As in other parasitoid venoms, hydrolases, including proteases, phosphatases, esterases, and nucleases, constitute the most abundant families in T. sinensis venom, followed by protease inhibitors. These proteins are potentially involved in the complex parasitic syndrome, with different effects on the immune system, physiological processes and development of the host, and contribute to provide nutrients to the parasitoid progeny. Although additional in vivo studies are needed, initial findings offer important information about venom factors and their putative host effects, which are essential to ensure the success of parasitism.

Cutaneous myiasis in cats and dogs: Cases, predisposing conditions and risk factors.

Two cases of cutaneous myiasis diagnosed in 2018 in Emilia-Romagna region (northern Italy) were reported. The first one, described in a domestic cat Felis silvestris catus L. (Carnivora: Felidae) and caused by Calliphora vicina Robineau-Desvoidy (Diptera: Calliphoridae), was the first one of this type ever reported in Italy in cats. The second one was described in a domestic dog Canis lupus familiaris L. (Carnivora: Canidae) and caused by Lucilia sericata (Meigen) (Diptera: Calliphoridae) and was unusual because it occurred in absence of lesions. An extensive literature search on cutaneous myiasis in these two domestic animal species was performed in order to draw attention to predisposing conditions and risk factors.

Immune Response After Cochlear Implantation.

A cochlear implant (CI) is an electronic device that enables hearing recovery in patients with severe to profound hearing loss. Although CIs are a successful treatment for profound hearing impairment, their effectivity may be improved by reducing damages associated with insertion of electrodes in the cochlea, thus preserving residual hearing ability. Inner ear trauma leads to inflammatory reactions altering cochlear homeostasis and reducing post-operative audiological performances and electroacoustic stimulation. Strategies to preserve residual hearing ability led to the development of medicated devices to minimize CI-induced cochlear injury. Dexamethasone-eluting electrodes recently showed positive outcomes. In previous studies by our research group, intratympanic release of dexamethasone for 14 days was able to preserve residual hearing from CI insertion trauma in a Guinea pig model. Long-term effects of dexamethasone-eluting electrodes were therefore evaluated in the same animal model. Seven Guinea pigs were bilaterally implanted with medicated rods and four were implanted with non-eluting ones. Hearing threshold audiograms were acquired prior to implantation and up to 60 days by recording compound action potentials. For each sample, we examined the amount of bone and fibrous connective tissue grown within the scala tympani in the basal turn of the cochlea, the cochleostomy healing, the neuronal density, and the correlation between electrophysiological parameters and histological results. Detection of tumor necrosis factor alpha, interleukin-6, and foreign body giant cells showed that long-term electrode implantation was not associated with an ongoing inflammation. Growth of bone and fibrous connective tissue around rods induced by CI was reduced in the scala tympani by dexamethasone release. For cochleostomy sealing, dexamethasone-treated animals showed less bone tissue growth than negative. Dexamethasone did not affect cell density in the spiral ganglion. Overall, these results support the use of dexamethasone as anti-inflammatory additive for eluting electrodes able to protect the cochlea from CI insertion trauma.

Empirical "integrated disease management" in Ferrara during the Italian plague (1629-1631).

Plague, a highly infective disease caused by Yersinia pestis (Proteobacteria: Enterobacteriales), ravaged Europe from 1347 over the course of more than 450 years. During the Italian Plague (1629-1631), the disease was rampaging in the entire Northern Italy down to Tuscany, but the city of Ferrara was relatively spared, in spite that the economic activities were maintained with highly affected cities, such as Milan, through the relevant salt commerce. The aim of the study is to evaluate the hygiene rules that were effective in preventing the spread of the plague in Ferrara in 1630, by examining historical documents and reports. According to these documents, a kind of empirical "integrated disease management" was carried out, using remedies including compounds with bactericidal, anti-parasite and repellent activity, and by technical strategies including avoidance of possible plague carriers. The anti-plague remedies and technical strategies used in ancient Ferrara are critically analysed using a multidisciplinary approach (pharmaceutic, medical, epidemiologic and entomological) and compared to current prevention protocols.

Evaluation of larvicidal activity of esters of 4-mercapto-2-butenoic acid against Aedes albopictus (Diptera: Culicidae).

Aedes albopictus (Skuse) (Diptera: Culicidae), an aggressive and annoying vector of several arbovirus including Chikungunya and Zika, is a serious health problem worldwide. Control of this mosquito is difficult because of high adaptability, egg resistance to dehydration and ability to exploit many man-made microhabitats. The most effective strategy appears the control of larval population. Based on previous data showing a larvicidal effect of plant extracts containing sulfhydryl and isothiocyanate compounds, we evaluated by bioassays the toxicity of three synthetic esters of 4-mercapto-2-butenoic acid on larvae of A. albopictus in comparison to cypermethrin. Among the compounds tested, the most effective was n-octyl 4-mercapto-2-butenoate, about 5 times more effective than ethyl 4-mercaptobut-2-enoate and about 20 times more effective than menthyl 4-mercaptobut-2-enoate. We advance the hypothesis that the larvicidal properties of n-octyl 4-mercapto-2-butenoate are due to its hydrophobic alkyl chain, longer than that of the other two compounds. This chain confers to the molecule the ability to spread on water surface and interfere with larval respiration. The larvicidal activity of n-octyl 4-mercapto-2-butenoate against A. albopictus appears interesting and may be developed after toxicological evaluation on vertebrates and humans, and environmental toxicity tests in compliance with WHO and ECDC rules.

Myiasis in domestic cats: a global review.

Myiasis is an infestation caused by larvae of Diptera in humans and other vertebrates. In domestic cats, Felis silvestris catus L. (Carnivora: Felidae), four dipteran families have been reported as agents of obligatory and facultative myiasis: Oestridae, Calliphoridae, Sarcophagidae and Muscidae. Among agents of obligatory myiasis, the most frequent genus is Cuterebra Clark (Oestridae) and the most frequent species is Cochliomyia hominivorax (Coquerel) (Calliphoridae). Among the agents of facultative myiasis, the most frequent species is Lucilia sericata (Meigen) (Calliphoridae). A survey of myiasis in cats reported in literature shows that the cases are distributed worldwide and linked to the geographical range of the dipteran species. Factors favouring the occurrence of myiasis in cats are prowling in infested areas, poor hygiene conditions due to diseases and/or neglect, and wounds inflicted during territorial or reproductive competition. The aim of the review is to provide an extended survey of literature on myiasis in cats, as general information and possible development of guidelines for veterinarians, entomologists and other researchers interested in the field.

Geographical origin of Vitis vinifera cv. Cannonau established by the index of bioaccumulation and translocation coefficients.

Geochemical fingerprints in grape require an identification of major and trace elements that show correlations between concentrations in soil and in plant tissues: these correlations are relevant to discriminate grapes according to geographical origin. The Vitis vinifera cultivar Cannonau is used to produce the renowned Italian controlled designation of origin (DOC) wine "Cannonau" from Sardinia. Two Cannonau vineyards located in Sardinia Region were studied to establish the relationship between geochemical features of vineyard soil and chemical composition of leaves and grape berries. Major and trace elements were determined by X-ray fluorescence and inductively coupled plasma-mass spectrometry in soil, leaf, and grape berry samples. The index of bioaccumulation and the translocation coefficients were also calculated for all elements. Data from the two study areas were compared by a non-parametric test and multivariate statistics (principal component analysis). The results showed a specific assimilation of these elements in leaf and grape berry from vine in two different soils. Moreover, geochemical characterization and statistical analysis enabled to discriminate the cultivar "Cannonau" according to geographical origin. The results showed that the elements that could establish a reliable correlation between the soil vineyard and leaves and grape berries from the two study areas were B, Sr, and Zr. These elements may therefore be used as geochemical fingerprints to identify the geographic origin of V. vinifera cv. Cannonau in the two study areas.

Discrimination of geographical origin of hop (Humulus lupulus L.) using geochemical elements combined with statistical analysis.

Beer is a popular alcoholic beverage worldwide, traditionally made from water, barley and hop (Humulus lupulus L.) strobili. The strobili contain lupulin glands whose components (mostly bitter acids and polyphenols) confer unique and locally different flavours to beer types. It is therefore relevant for brewers and consumers to precisely know the geographical origin of hop plants used for high-quality beer. Hop plants belonging to the variety Hallertau Perle, grown in two locations, Cavalese and Imèr, of the Trentino Region (Italy) were analysed to establish a direct relationship between the chemical elements detected in soil and in plant parts. Chemical elements were determined by X-ray fluorescence and inductively coupled plasma mass spectrometry in soil, leaf and strobili samples from Cavalese and Imèr. The data from the two areas were compared by a nonparametric test (Mann-Whitney) and multivariate statistics (principal component analysis and partial least squares discriminant analysis). The geochemical characterization and the statistical analyses showed different concentrations of major and trace elements in soil and plant parts from the two areas. A reliable correlation could be established between some elements in soil and strobili samples, that is Nb, Fe, Rb and Zr for Cavalese and Mg, Ni, Zn and Zr for Imèr. These elements could therefore be used as geochemical fingerprints to identify the geographical origin of strobili from the two study areas, an approach useful to verify the origin of hop plants for the production of high-quality beer.

Ultrastructural investigations on spermatogenesis and spermatozoan morphology in the endangered Adriatic sturgeon, Acipenser naccarii (Chondrostei, Acipenseriformes).

Spermatogenesis was investigated in the Adriatic sturgeon, Acipenser naccarii, by light and electron microscopy. The testis of the unrestricted type had a germinal compartment composed of lobules containing germ cells and Sertoli cells, and separated by a basal lamina from the interstitial compartment, in which Leydig and myoid cells were detected for the first time in Acipenseridae. Spermatogenesis occurred in spermatocysts produced when Sertoli cells became associated with type A spermatogonia of subsequent generations, which produced a clone of synchronized aligned spermatogonia. In primary spermatocytes at zygo-pachytene stage, the large spherical nucleus contained synaptonemal complexes. The smaller secondary spermatocytes were ovoid with a central round nucleus and scarce cytoplasm. Spermatids were interconnected by cytoplasmic bridges until early spermiogenesis. Chromatin initially condensed as long, twisted, and nonhomogeneous fibers and finally as a compact structure made of thick filaments. Early spermatids showed the flagellum, the primordia of centriole complex and of "implantation fossa," followed by the acrosomal vesicle formed by Golgi complexes and a fibrous body associated to centriole complex. The spermatozoan head had 10 postero-lateral projections and a trapezoidal nucleus, a cylindrical midpiece with six to eight mitochondria, the centriole complex, and a "9 + 2" flagellum with a pair of lateral fins. Three helical endonuclear canals crossed the nucleus from the acrosome base to the implantation fossa; their spiralization and that of chromatin fibers suggest a spiral twisting of the nucleus during spermiogenesis. The Sertoli cells performed phagocytosis of degenerating spermatids and spermatozoa. Significant interindividual differences were detected in most morphological parameters of spermatozoa. Data on spermatogenesis in A. naccarii and morphometric measurements on mature spermatozoa provide information about the reproductive biology of the species useful not only for phylogenetic studies but also for evaluation of sperm quality for artificial reproduction projects and restocking of this and other critically endangered sturgeon species.

Ultrastructural characterization of sensilla and microtrichia on the antenna of female Haematopota pandazisi (Diptera: Tabanidae).

The haematophagous females of the cleg fly Haematopota pandazisi (Kröber) (Diptera: Tabanidae) are a common pest in areas inhabited by wild and domestic ungulates in southern Europe, North Africa and Anatolia. A morphological investigation by scanning electron microscopy (SEM) was carried out for the first time on the antennae of females of H. pandazisi, with special attention to the type and distribution of sensilla and microtrichia. The typical brachyceran antenna is divided into three regions: the scape, the pedicel and the flagellum, which is the longest of the three and is composed of four flagellomeres. The scape and pedicel are characterized by only one type of microtrichium and chaetic sensillum, whereas five types of microtrichia and sensilla were identified on the flagellum and classified according to shape and distribution. The sensilla are of the chaetic, clavate, basiconic, trichoid and coeloconic types; the latter with either a basiconic or grooved peg inside. The results obtained in this study were compared to those found in other species in the family Tabanidae and other Diptera, with special attention to haematophagous species.

Ecdysteroidogenesis and development in Heliothis virescens (Lepidoptera: Noctuidae): Focus on PTTH-stimulated pathways.

Post-embryonic development and molting in insects are regulated by endocrine changes, including prothoracicotropic hormone (PTTH)-stimulated ecdysone secretion by the prothoracic glands (PGs). In Lepidoptera, two pathways are potentially involved in PTTH-stimulated ecdysteroidogenesis, mitogen-activated protein kinase (MAPK) and phosphoinositide 3-kinase/protein kinase B/target of rapamycin (PI3K/Akt/TOR). We investigated the potential roles of both these pathways in Heliothis virescens ecdysteroidogenesis. We identified putative proteins belonging to MAPK and PI3K/Akt/TOR signaling cascades, using transcriptomic analyses of PGs from last (fifth) instar larvae. Using western blots, we measured the phosphorylation of 4E-BP and S6K proteins, the main targets of TOR, following the in vitro exposure of PGs to brain extract containing PTTH (hereafter referred to as PTTH) and/or the inhibitors of MAPK (U0126), PI3K (LY294002) or TOR (rapamycin). Next, we measured ecdysone production, under the same experimental conditions, by enzyme immunoassay (EIA). We found that in Heliothis virescens last instar larvae, both pathways modulated PTTH-stimulated ecdysteroidogenesis. Finally, we analyzed the post-embryonic development of third and fourth instar larvae fed on diet supplemented with rapamycin, in order to better understand the role of the TOR pathway in larval growth. When rapamycin was added to the diet of larvae, the onset of molting was delayed, the growth rate was reduced and abnormally small larvae/pupae with high mortality rates resulted. In larvae fed on diet supplemented with rapamycin, the growth of PGs was suppressed, and ecdysone production and secretion were inhibited. Overall, the in vivo and in vitro results demonstrated that, similarly to Bombyx mori, MAPK and PI3K/Akt/TOR pathways are involved in PTTH signaling-stimulated ecdysteroidogenesis, and indicated the important role of TOR protein in H. virescens systemic growth.

Chemical elements as fingerprints of geographical origin in cultivars of Vitis vinifera L. raised on the same SO4 rootstock.

The uptake of major and trace elements in grapevine (Vitis vinifera L.) can be influenced by soil, climate, geographic origin, and rootstock type. Rootstocks were mainly selected to resist phylloxera and for specific tolerance to lime, mineral uptake, drought, and salinity. The relationship among concentrations of major, trace, and rare earth elements was studied in soil and leaves from two Italian grapevine cultivars, "Cabernet Sauvignon" and "Corvina," employed to produce renowned controlled designation of origin (DOC) wines. The cultivars were raised on the same rootstock SO4 in two different areas of the Veneto Region (Northern Italy). The elements were studied by X-ray fluorescence and inductively coupled plasma mass spectrometry, and data were elaborated by non-parametric tests and multivariate linear discrimination analysis. The related index of bioaccumulation was calculated to define the specific assimilation of the elements from soil to leaves. A statistically significant correspondence between soil and leaf samples was observed for Mg, Sm, V, and Zr. The results allowed to discriminate soil and leaf samples of the two cultivars according to geographical provenance, possibly providing geochemical markers (fingerprints) useful against fraudulent use of DOC wine labels.

Saharan dust particles in snow samples of Alps and Apennines during an exceptional event of transboundary air pollution.

Southern European countries are often affected in summer by transboundary air pollution from Saharan dust. However, very few studies deal with Saharan dust pollution at high altitudes in winter. In Italy, the exceptional event occurred on February 19, 2014, colored in red the entire mountain range (Alps and Apennines) and allowed to characterize the particulate matter deposited on snow from a morphological and chemical point of view. Snow samples were collected after this event in four areas in the Alps and one in the Apennines. The particulate matter of the melted snow samples was analyzed by scanning electron microscopy with energy dispersive X-ray spectrometry (SEM-EDS) and by inductively coupled plasma mass spectrometry (ICP-MS). These analyses confirmed the presence of Saharan dust particle components in all areas with similar percentages, supported also by the positive correlations between Mg-Ca, Al-Ca, Al-Mg, and Al-K in all samples.

Correction: Coenzyme Q10 plus Multivitamin Treatment Prevents Cisplatin Ototoxicity in Rats.

[This corrects the article DOI: 10.1371/journal.pone.0162106.].