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.

Effects of Acibenzolar-S-methyl on the Probing Behaviour and Mortality of Cacopsylla pyri on Pear Plants.

European pear psylla, Cacopsylla pyri, is one of the worst pests of pear in Europe. We investigated whether acibenzolar-S-methyl (ASM) application on pear plants might affect the behaviour in C. pyri. The elicitor was applied on pear potted plants, and after 48 h, we confirmed the ASM-mediated induction of several Pathogenesis-Related protein (PR) coding genes. At the same time, an in-depth analysis was performed on the probing behaviour of adults and nymphs of C. pyri on ASM-treated pear plants by the EPG-DC system, as well as the assessment of young nymphs' survival 7 days after the ASM application. The elicitor application weakly interfered with C. pyri nymphs probing behaviour and survival, while it did not affect adult stages. These data confirm previous observations obtained on C. pyricola and suggest that the elicitor does not represent a viable tool in the control of pear psylla species, especially if used alone, but it might be used in integrated management strategies focused on other plant pathogens such as Erwinia amylovora.

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.

DNA barcoding of the horsefly fauna (Diptera, Tabanidae) of Croatia with notes on the morphology and taxonomy of selected species from Chrysopsinae and Tabaninae.

In the Croatian fauna, horseflies (Tabanidae) are represented by 78 species belonging to two subfamilies, five tribes, and 10 genera. Identification of these species is based on morphological characteristics. In this study, 43 species of horseflies were analyzed. The highest number of species (19) belongs to the genus Tabanus, followed by the genera Hybomitra with seven species, Haematopota with six species, Chrysops with four species, Atylotus and Philipomyia with two species each, and the genera Silvius, Dasyrhamphis, and Heptatoma with one species each. The standard DNA barcoding region of the mitochondrial cytochrome c oxidase gene, subunit I (COI), was sequenced and compared to the Barcode of Life Database (BOLD). Our analyses confirmed our morphological identifications and added 16 new Barcode Index Numbers (BINs) for Tabanidae to BOLD. Potential problems in the systematics and taxonomy of this family are highlighted.

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.

First report of the presence of Necrodes littoralis (L.) (Coleoptera: Silphidae) on a human corpse in Italy.

The colonization of a human body by Necrodes littoralis (L.) (Coleoptera: Silphidae) is reported for the first time in Italy. This species is both necrophagous and predator of necrophagous fauna. The body colonized by the coleopteran was found indoors, in an advanced decomposition stage, in a suburban area of Cosenza (Calabria, Southern Italy) in November. Insects (adults, puparia and larvae) were collected on and around the body. Puparia and larvae were raised in the laboratory until the adult stage for morphological identification, which was carried out through taxonomical keys. Besides N. littoralis, also the presence of Calliphora vicina Robineau-Desvoidy, Chrysomya albiceps (Wiedemann) (Diptera: Calliphoridae), Hydrotaea dentipes (Fabricius) (Diptera: Muscidae), and Creophilus maxillosus (L.) (Coleoptera: Staphylinidae) was detected. Necrodes littoralis is a species of forensic interest because it may colonize human and vertebrate corpses and has been reported elsewhere in Europe.

Investigations on Arthropods Associated with Decay Stages of Buried Animals in Italy.

Burial could be used by criminals to conceal the bodies of victims, interfering with the succession of sarcosaprophagous fauna and with the evaluation of post-mortem interval. In Italy, no experimental investigation on arthropods associated with buried remains has been conducted to date. A first experimental study on arthropods associated with buried carcasses was carried out in a rural area of Arcavacata di Rende (Cosenza), Southern Italy, from November 2017 to May 2018. Six pig carcasses (Susscrofa Linnaeus) were used, five of which were buried in 60-cm deep pits, leaving about 25-cm of soil above each carcass, and one was left above ground. One of the buried carcasses was periodically exhumed to evaluate the effects of disturbance on decay processes and on arthropod fauna. The other four carcasses were exhumed only once, respectively after 43, 82, 133, and 171 days. As expected, the decay rate was different among carcasses. Differences in taxa and colonization of arthropod fauna were also detected in the above ground and periodically exhumed carcasses. In carcasses exhumed only once, no arthropod colonization was detected. The results showed that a burial at about 25 cm depth could be sufficient to prevent colonization by sarcosaprophagous taxa and these data could be relevant in forensic cases involving buried corpses.

Characterization of Halyomorpha halys TAR1 reveals its involvement in (E)-2-decenal pheromone perception.

In insects, tyramine receptor 1 (TAR1) has been shown to control several physiological functions, including olfaction. We investigated the molecular and functional profile of the Halyomorpha halys type 1 tyramine receptor gene (HhTAR1) and its role in olfactory functions of this pest. Molecular and pharmacological analyses confirmed that the HhTAR1 gene codes for a true TAR1. RT-qPCR analysis revealed that HhTAR1 is expressed mostly in adult brain and antennae as well as in early development stages (eggs, 1st and 2nd instar nymphs). In particular, among the antennomeres that compose a typical H. halys antenna, HhTAR1 was more expressed in flagellomeres. Scanning electron microscopy investigation revealed the type and distribution of sensilla on adult H. halys antennae: both flagellomeres appear rich in trichoid and grooved sensilla, known to be associated with olfactory functions. Through an RNAi approach, topically delivered HhTAR1 dsRNA induced a 50% downregulation in gene expression after 24 h in H. halys 2nd instar nymphs. An innovative behavioural assay revealed that HhTAR1 RNAi-silenced 2nd instar nymphs were less susceptible to the alarm pheromone component (E)-2 decenal as compared with controls. These results provide critical information concerning the role of TAR1 in olfaction regulation, especially alarm pheromone reception, in H. halys. Furthermore, considering the emerging role of TAR1 as target of biopesticides, this work opens the way for further investigation on innovative methods for controlling H. halys.

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.

Wound myiasis in a wild boar by Lucilia caesar (Diptera: Calliphoridae): First case and current status of animal myiasis by this species.

The first case of myiasis caused by Lucilia caesar (L.) (Diptera: Calliphoridae) in a wild boar, Sus scrofa L. (Artiodactyla: Suidae) is described. The myiasis occurred in October 2019 in Southern Italy and the identification of the agent was based on adult male morphology. The wild boar had a wound on its right side, near the neck, which was largely infested by larvae. The ecology, distribution and current literature status about cases of animal myiasis by this species is also included.

Egg data validation in quantitative monitoring of Aedes albopictus in Emilia-Romagna region, Italy.

The monitoring of Aedes urban species is usually conducted by ovitraps, which can provide a good estimate of population density at low cost and relatively easy management. When the monitoring network is managed by many operators, as in the case of the Emilia-Romagna regional plan, it becomes necessary to develop a quality control methodology capable of highlighting the possible data incoherency and ensuring that the monitoring system provides reliable data.This paper presents the procedure applied in the egg counting phase as developed in Emilia-Romagna in order to check the data quality and validate the data before being included in the database.Several steps have been identified and protocols developed to serve quality control such as training of technicians and intralaboratory quality check, daily random double counting of Masonite paddles and interlaboratory periodic comparison.The qualitative test showed that all operators were able to distinguish Aedes albopictus eggs from other mosquito species eggs. The quantitative test showed significant differences between the 11 operators, with a relative error resulting in the range of 0.1-25.8%.The daily internal double counting of randomly extracted Masonite slides was performed by the coordinator showing a relative error in the range 2.47-2.63% without statistical differences, confirming a good alignment of the operators with the coordinator throughout the monitoring period.The interlaboratory comparison trial produced an average relative error of 7.20% showing a sufficient alignment between the three laboratories.Moreover a new time-saving methods in egg counting was developed and tested in real condition.

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.

Urogenital myiasis caused by Lucilia sericata (Diptera: Calliphoridae) in a domestic rabbit in Italy.

The report describes a case of urogenital myiasis in a domestic rabbit Oryctolagus cuniculus L. (Lagomorpha: Leporidae) caused by Lucilia sericata (Meigen; Diptera: Calliphoridae) in region Emilia-Romagna (Northern Italy). The case, occurring in June 2018, is the first one involving L. sericata as an agent of myiasis in a domestic rabbit in Italy. Species identification was based on morphological investigations of males through identification keys. The rabbit developed the urogenital myiasis as a consequence of chronic enteritis causing an accumulation of faeces in the perianal and perineal region.

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.

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.